Application of SMOX in preparation of medicine for treating hepatocyte inflammation

Abstract

The invention belongs to the technical field of biological medicine, and discloses application of SMOX in preparation of a medicine for treating hepatocyte inflammation. The expression of SMOX in AML12 cells is stimulated by LPS (Lipopolysaccharide). Inflammation response induced by LPS in AML12 cells can be inhibited through inhibition of SMOX. LPS can induce beta-catenin to be transferred from cytoplasm to a cell nucleus, and down-regulation or inhibition of SMOX can partially reverse the process. An SMOX inhibitor MDL72527 or SMOX knockout mouse is used for intervention in vivo, so that liver function impairment can be remarkably improved, liver inflammation can be reduced, beta-catenin nuclear translocation in liver tissues can be reduced, and carbon tetrachloride-induced mouse liver fibrosis can be relieved. The invention provides a new treatment strategy for hepatitis and hepatic fibrosis, and provides a new thought for inhibiting early liver cancer.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of SMOX in the preparation of drugs for treating hepatocellular inflammation. Background Technology

[0002] The liver plays a vital role in the body's life activities. Through biosynthesis, biotransformation, and detoxification, it participates not only in the metabolism of substances such as proteins, lipids, and carbohydrates, but also in the metabolism of drugs, alcohol, and toxins. At the same time, the liver is also an organ frequently attacked by various pathogenic factors or diseases; abnormal metabolism, drugs, and microorganisms can all cause liver damage.

[0003] It has been confirmed that most liver injuries are accompanied by inflammatory responses, and liver fibrosis, cirrhosis, liver failure, and cancer are the most common forms of liver disease progression and outcomes. Therefore, for liver diseases of different etiologies, it is necessary to fully understand their association with inflammation and to carry out targeted treatment to slow disease progression and improve quality of life.

[0004] Liver fibrosis is the common pathological basis for the progression of various chronic liver diseases to cirrhosis. Its essence lies in the imbalance between the synthesis and degradation of extracellular matrix in the liver, leading to the excessive deposition of extracellular matrix, especially collagen. The continued development of liver fibrosis results in dysfunction and failure of the corresponding organs, seriously endangering life and health. Slowing down, preventing, or even reversing liver fibrosis is of great significance for reducing the occurrence of serious complications, improving patients' quality of life, and increasing survival rates. Therefore, the development of anti-liver fibrosis drugs has significant application value.

[0005] Spermine oxidase (SMOX) is a FAD-dependent enzyme that specifically oxidizes spermine (SPM) to produce spermidine (SPD), hydrogen peroxide (H2O2), and the aldehyde 3-aminopropionaldehyde (3-AP). It plays a dominant role in the catabolism of polyamines in mammals. In addition to its traditional metabolic functions, SMOX, as a metabolic enzyme, also regulates cell proliferation, survival, apoptosis, stress responses, and tumor microenvironment remodeling through post-translational autoacetylation.

[0006] Existing technology: Application of nonsteroidal anti-inflammatory drugs (NSAIDs) to treat hepatocellular inflammation

[0007] Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used medications for treating hepatocellular inflammation. These drugs reduce the production of prostaglandins by inhibiting cyclooxygenase (COX), thereby alleviating the inflammatory response.

[0008] Problems with existing technology

[0009] 1. Side effects: Although NSAIDs can effectively relieve inflammation and pain in the short term, long-term use can cause a range of side effects, such as gastrointestinal damage, kidney damage, and cardiovascular events.

[0010] 2. Limited therapeutic efficacy: NSAIDs primarily reduce inflammation by inhibiting prostaglandin production, but this does not address the underlying cause of hepatocellular inflammation. For some severe or chronic hepatitis cases, the therapeutic effect of NSAIDs may be limited.

[0011] 3. Irreversible nuclear translocation of β-catenin: The mechanism of action of NSAIDs is not related to the regulation of β-catenin, so they cannot reverse the nuclear translocation of β-catenin, which limits their application in the treatment of some diseases associated with β-catenin nuclear translocation (such as liver cancer). Summary of the Invention

[0012] In view of the problems existing in the prior art, the present invention provides the application of SMOX in the preparation of drugs for treating hepatocellular inflammation.

[0013] This invention is achieved through the application of SMOX in the preparation of drugs for treating hepatocellular inflammation.

[0014] Application of SMOX inhibitors in the preparation of drugs for treating hepatocellular inflammation.

[0015] Furthermore, SMOX inhibition or knockout can effectively reduce LPS-induced AML12 hepatocyte inflammatory response.

[0016] Furthermore, downregulation or inhibition of SMOX partially reverses the translocation of β-catenin from the cytoplasm to the nucleus.

[0017] Furthermore, in vivo intervention with the SMOX inhibitor MDL72527 or SMOX knockout mice reduced intrahepatic inflammation, decreased nuclear translocation of β-catenin in liver tissue, and alleviated carbon tetrachloride-induced liver fibrosis in mice.

[0018] Another object of the present invention is to provide a method for determining the use of SMOX in the preparation of a drug for treating hepatocellular inflammation, specifically comprising:

[0019] (1) Using small interfering RNA or the SMOX inhibitor MDL72527, we verified the effect of SMOX inhibition on LPS-induced inflammatory response in the mouse hepatocyte line AML12.

[0020] (2) Western blotting and immunofluorescence were used to determine whether LPS could induce β-catenin to enter the cell nucleus and whether this process could be reversed by interfering with SMOX expression or by using SMOX inhibitors;

[0021] (3) The above hypothesis was verified using SMOX inhibitor MDL72527 and SMOX knockout mice.

[0022] Furthermore, the drug is formulated into a liquid or solid dosage form.

[0023] Furthermore, the drug can be formulated as an injection, oral liquid, granules, powder, tablet, capsule, or external patch.

[0024] Based on the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solution to be protected by this invention are as follows:

[0025] First, the etiology of hepatitis and liver fibrosis: This invention initially reveals the role of SMOX in hepatocellular inflammation and fibrosis.

[0026] Lack of therapeutic targets: There is a lack of effective therapeutic targets for the treatment of hepatitis and liver fibrosis. This invention provides a potential new target, namely SMOX.

[0027] Regulation of the β-catenin pathway: This invention explores how SMOX can influence the nuclear translocation of β-catenin, a signal transduction pathway that plays a key role in many biological processes, including cell proliferation and inflammation.

[0028] Second, it inhibits the inflammatory response: SMOX inhibition or knockout can effectively reduce LPS-induced AML12 hepatocyte inflammatory response.

[0029] Nuclear translocation reversal: Downregulation or inhibition of SMOX partially reverses the translocation of β-catenin from the cytoplasm to the nucleus.

[0030] In vivo validation: In vivo experiments using SMOX inhibitor MDL72527 or SMOX knockout mice confirmed the effects of SMOX, further demonstrating its potential in the treatment of hepatitis and liver fibrosis.

[0031] Treatment strategy: This invention provides a new potential treatment strategy for hepatitis and liver fibrosis, and helps prevent or inhibit early-stage liver cancer.

[0032] Third, significant technological advancements in the application of SMOX in the treatment of hepatocellular inflammation include:

[0033] 1. A novel drug target—SMOX—was proposed and validated.

[0034] Through experiments with SMOX inhibitors or SMOX knockout mice, SMOX was proposed and validated as a potential drug target for treating hepatocellular inflammation.

[0035] 2. Revealing the association between SMOX and β-catenin nuclear translocation.

[0036] This provides researchers with new understanding and insights into the nuclear translocation of SMOX and β-catenin, opening up new directions for the research and treatment of liver diseases.

[0037] 3. Innovation in interdisciplinary research

[0038] By utilizing knowledge from multiple disciplines such as molecular biology, pharmacology, and genetic engineering, comprehensive research has been conducted, promoting the cross-integration and innovation of science and technology.

[0039] 4. A potential new strategy for treating liver fibrosis is proposed.

[0040] An intervention experiment using SMOX inhibitors or SMOX knockout mice in a liver fibrosis model demonstrated a potential new strategy for treating liver fibrosis.

[0041] 5. It provides a new direction for drug development.

[0042] This study validates the potential of SMOX inhibitors (such as MDL72527) in vivo, providing prospects and directions for subsequent drug development and clinical trials.

[0043] 6. Provides new disease models and research platforms

[0044] SMOX knockout mice have become a new model for studying hepatocellular inflammation and liver fibrosis, providing a new platform for basic research on the disease and drug screening. Attached Figure Description

[0045] To more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the embodiments of the present invention will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0046] Figure 1 This is a flowchart illustrating the method for using SMOX in preparing a drug for treating hepatocellular inflammation, as provided in this embodiment of the invention. Detailed Implementation

[0047] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0048] This invention addresses the following problems and deficiencies in the prior art, achieving significant technological advancements:

[0049] Limitations of hepatitis treatment: Existing hepatitis treatments may not be effective in controlling the inflammatory response, especially in cases of chronic liver disease and liver fibrosis.

[0050] Incomplete understanding of inflammatory pathways: The incomplete understanding of the molecular mechanisms and regulatory pathways of hepatocellular inflammation limits the development of new drugs and innovative treatment strategies.

[0051] Side effects of existing treatments: Some existing treatments may be accompanied by significant side effects or are insufficient to target specific pathological pathways.

[0052] To address the problems existing in the prior art, this invention identifies the key role of SMOX in hepatocyte inflammation, particularly in LPS (lipopolysaccharide)-induced AML12 hepatocyte inflammatory responses.

[0053] Application of SMOX inhibitors: SMOX inhibitors, such as MDL72527, are used to reduce intrahepatic inflammation and alleviate liver fibrosis.

[0054] Nuclear metastasis of β-catenin: It was found that downregulation or inhibition of SMOX can partially reverse the metastasis of β-catenin from the cytoplasm to the nucleus, which is an important target for the treatment of hepatitis.

[0055] Diversity of drug formulations: This invention proposes to formulate SMOX inhibitors into different forms of drugs, such as injections, oral solutions, granules, tablets, etc., to meet the needs of different patients.

[0056] Experimental validation: The role of SMOX in the treatment of hepatitis was validated by small interfering RNA, Western blotting, immunofluorescence, and the use of SMOX inhibitors / knockout mice.

[0057] The technical effects and significant technological advancements brought about by this invention in solving the problems of the prior art are as follows:

[0058] Novel therapeutic target: Identifying SMOX as a novel target for the treatment of hepatitis cell inflammation provides a new approach to hepatitis treatment.

[0059] Reduce liver inflammation and liver fibrosis: The use of SMOX inhibitors reduces liver inflammation and helps alleviate liver fibrosis.

[0060] Reducing nuclear metastasis of β-catenin: By influencing the nuclear metastasis of β-catenin, SMOX inhibitors offer a novel mechanism for the treatment of hepatitis.

[0061] Diversity of drug formulations: The most suitable drug formulation can be selected according to the different needs and treatment conditions of patients.

[0062] Experimental verification: Through various experimental methods, this invention verified the effectiveness of SMOX inhibitors in treating hepatocellular inflammation.

[0063] In view of the problems existing in the prior art, the present invention provides the application of SMOX in the preparation of drugs for treating hepatocellular inflammation.

[0064] The application of SMOX provided in this invention in the preparation of a drug for treating hepatocellular inflammation.

[0065] Application of SMOX inhibitors in the preparation of drugs for treating hepatocellular inflammation.

[0066] Furthermore, SMOX inhibition or knockout can effectively reduce LPS-induced AML12 hepatocyte inflammatory response.

[0067] Furthermore, downregulation or inhibition of SMOX partially reverses the translocation of β-catenin from the cytoplasm to the nucleus.

[0068] Furthermore, in vivo intervention with the SMOX inhibitor MDL72527 or SMOX knockout mice reduced intrahepatic inflammation, decreased nuclear translocation of β-catenin in liver tissue, and alleviated carbon tetrachloride-induced liver fibrosis in mice.

[0069] Another object of the present invention is to provide a method for determining the use of SMOX in the preparation of a drug for treating hepatocellular inflammation, specifically comprising:

[0070] (1) Using small interfering RNA or the SMOX inhibitor MDL72527, we verified the effect of SMOX inhibition on LPS-induced inflammatory response in the mouse hepatocyte line AML12.

[0071] (2) Western blotting and immunofluorescence were used to determine whether LPS could induce β-catenin to enter the cell nucleus and whether this process could be reversed by interfering with SMOX expression or by using SMOX inhibitors;

[0072] (3) The above hypothesis was verified using SMOX inhibitor MDL72527 and SMOX knockout mice.

[0073] Furthermore, the drug is formulated into a liquid or solid dosage form.

[0074] Furthermore, the drug can be formulated as an injection, oral liquid, granules, powder, tablet, capsule, or external patch.

[0075] Example 1: Application of SMOX inhibitor MDL72527 in the treatment of hepatocellular inflammation

[0076] 1. Drug Synthesis: First, the SMOX inhibitor MDL72527 needs to be synthesized. Synthetic methods for this compound can be found in existing scientific literature or purchased from commercial suppliers.

[0077] 2. Drug administration: MDL72527 can be administered orally or by injection to laboratory animals (e.g., mice). The dosage and frequency of administration should be determined based on the severity of inflammation and the animal's weight.

[0078] 3. Inflammation assessment: During treatment, the severity of hepatitis and the effectiveness of treatment can be assessed through methods such as blood biochemistry analysis, liver tissue pathology examination, and determination of inflammatory markers.

[0079] 4. Assessment of β-catenin nuclear translocation: The effect of MDL72527 on β-catenin nuclear translocation can be assessed by immunofluorescence staining and confocal microscopy.

[0080] 5. Results Analysis: By comparing the results of the treatment group and the control group, the effectiveness of MDL72527 in treating hepatocellular inflammation can be evaluated.

[0081] Example 2: Application of SMOX knockout mice in the treatment of hepatocellular inflammation

[0082] 1. Production of SMOX knockout mice: SMOX knockout mouse models are created using gene editing technologies such as CRISPR / Cas9.

[0083] 2. Inflammation induction: LPS or carbon tetrachloride were used to induce hepatitis in mice by injection.

[0084] 3. Inflammation assessment: Similar to Example 1, the severity of hepatitis and the effectiveness of treatment are assessed through methods such as blood biochemistry analysis, liver tissue pathology examination, and determination of inflammatory markers.

[0085] 4. Assessment of β-catenin nuclear translocation: Similarly, the effect of SMOX knockout on β-catenin nuclear translocation can be assessed by immunofluorescence staining and confocal microscopy.

[0086] 5. Results Analysis: Finally, the effects of SMOX knockout on hepatocellular inflammation and its therapeutic efficacy were evaluated by analyzing the experimental data.

[0087] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions, and improvements made by those skilled in the art within the scope of the technology disclosed in the present invention, and within the spirit and principles of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. Application of SMOX in the preparation of drugs for treating hepatocellular inflammation.

2. Application of SMOX inhibitors in the preparation of drugs for treating hepatocellular inflammation.

3. The use of SMOX as described in claim 1 in the preparation of a medicament for treating hepatocellular inflammation, characterized in that, SMOX inhibition or knockout can effectively reduce LPS-induced AML12 hepatocyte inflammatory response.

4. The use of SMOX as described in claim 1 in the preparation of a medicament for treating hepatocellular inflammation, characterized in that, Downregulation or inhibition of SMOX partially reverses the translocation of β-catenin from the cytoplasm to the nucleus.

5. The use of SMOX as described in claim 1 in the preparation of a medicament for treating hepatocellular inflammation, characterized in that, In vivo intervention with the SMOX inhibitor MDL72527 or SMOX knockout mice reduced intrahepatic inflammation, reduced nuclear translocation of β-catenin in liver tissue, and alleviated carbon tetrachloride-induced liver fibrosis in mice.

6. A method for determining the use of SMOX in the preparation of a drug for treating hepatocellular inflammation as described in any one of claims 1 to 5, specifically comprising: (1) Using small interfering RNA or the SMOX inhibitor MDL72527, we verified the effect of SMOX inhibition on LPS-induced inflammatory response in the mouse hepatocyte line AML12. (2) Western blotting and immunofluorescence were used to determine whether LPS could induce β-catenin to enter the cell nucleus and whether this process could be reversed by interfering with SMOX expression or by using SMOX inhibitors; (3) The above hypothesis was verified using SMOX inhibitor MDL72527 and SMOX knockout mice.

7. The use of SMOX as described in claim 1 in the preparation of a medicament for treating hepatocellular inflammation, characterized in that, The drug is formulated as a liquid or solid preparation.

8. The use of SMOX as described in claim 1 in the preparation of a medicament for treating hepatocellular inflammation, characterized in that, The drug is formulated as an injection, oral liquid, granules, powder, tablet, capsule, or external patch.

Images