Application of DDR1 agonist in preparation of medicine for treating acute myocardial infarction
By targeting the DDR1 signaling pathway with entologliflozin and activating epicardial epithelial cells, the problem of difficulty in repairing ventricular structure and function after myocardial infarction in existing technologies is solved, and significant tissue repair and functional recovery effects are achieved.
Patent Information
- Application Number
- CN202511136750.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-14
- Publication Date
- 2025-10-17
AI Technical Summary
Existing methods for treating acute myocardial infarction are difficult to fundamentally repair damaged myocardial tissue and reconstruct normal ventricular structure and function. Traditional drugs mostly focus on inhibiting neuro-endocrine activity or reducing afterload, which makes it difficult to effectively inhibit ventricular remodeling.
Using iprotigozine as a DDR1 agonist, it targets the DDR1 signaling pathway in epicardial epithelial cells, activates multiple downstream signaling pathways, promotes cell migration, regulates matrix remodeling, inhibits excessive fibrosis, and improves myocardial repair and cardiac function.
Etogliflozin significantly improves cardiac function after acute myocardial infarction, inhibits pathological ventricular remodeling, promotes myocardial repair, enhances epicardial activity and angiogenesis, and provides a new treatment strategy to improve prognosis after myocardial infarction.
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Figure CN120789086A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The application belongs to the technical field of biological medicine, and particularly relates to application of a DDR1 agonist in preparation of a drug for treating acute myocardial infarction. BACKGROUND
[0002] Myocardial infarction is one of the main causes of heart failure and death, and its core pathological change is that a large area of myocardial cells is necrotic due to acute interruption of coronary blood flow. The repair process after myocardial infarction is accompanied by ventricular remodeling, that is, abnormal changes in the geometry and structure of the ventricle, including ventricular cavity dilation, wall thinning and fibrous hyperplasia. Although ventricular remodeling is helpful to maintain cardiac output in the early stage, long-term remodeling will lead to reduced ventricular compliance and impaired systolic function, and eventually develop into heart failure or even death. At present, the main clinical treatment is to delay the process of ventricular remodeling through drugs (such as beta blockers, ACE inhibitors) and interventional / surgical means, but the existing treatment means mainly focus on inhibiting neuroendocrine activity or reducing afterload, and it is difficult to fundamentally repair damaged myocardial tissue and rebuild normal ventricular structure and function. Therefore, developing a new treatment strategy that can promote myocardial regeneration and effectively inhibit ventricular remodeling has become an important research direction in the field of cardiovascular diseases.
[0003] In recent years, studies have shown that epicardium is not only a mechanical protective layer of the heart, but also an important regulatory center for myocardial injury repair. During the reactivation process of epicardial epithelial cells after myocardial infarction, epicardial-derived cells can be generated through epithelial-mesenchymal transition (EMT), and these cells can migrate to the infarct border zone and differentiate into fibroblasts, smooth muscle cells and perivascular cells, participating in neovascularization and matrix remodeling. In addition, epicardial epithelial cells also secrete a variety of paracrine factors such as FGF, VEGF, IGF, etc., promoting myocardial cell survival, inhibiting apoptosis, and improving microvascular perfusion. Studies have shown that activating and regulating the reactivation process of epicardial epithelial cells is expected to enhance myocardial repair, inhibit fibrosis and excessive remodeling, and promote ventricular function recovery after myocardial infarction. Therefore, targeting epicardial epithelial cells and their signaling pathways provides a new treatment strategy for improving ventricular remodeling and heart function recovery after myocardial infarction.
[0004] DDR1 (Discoidin Domain Receptor 1) is a receptor tyrosine kinase mainly expressed in epicardial cells in the heart. The main ligand of DDR1 is the extracellular matrix component, especially collagen, which enables it to sense and respond to structural changes in the matrix. During tissue injury and repair, DDR1 activates multiple downstream signaling pathways, including MAPK, PI3K / AKT and NF-κB, through binding to collagen, thereby regulating cell proliferation, migration, differentiation and survival. Studies have shown that DDR1 can promote the migration and closure of wound surface epithelial cells and inhibit excessive fibrosis by regulating the matrix remodeling process, maintaining the stability of tissue structure and function. In addition, DDR1 has been found to be associated with the recruitment and activation of inflammatory cells in chronic injury and fibrotic diseases, suggesting its dual role in tissue repair and pathological remodeling. Therefore, targeting DDR1 signaling pathway is considered a potential therapeutic strategy to regulate the balance of tissue repair and inhibit fibrosis.
[0005] Ertugliflozin is a SGLT-2 selective inhibitor widely used in the treatment of type 2 diabetes and has shown cardiovascular protective effects in heart failure patients, reducing the risk of hospitalization and improving prognosis. There is no related report on the application of Ertugliflozin as a DDR1 agonist in the preparation of drugs for treating cardiac repair after acute myocardial infarction. SUMMARY
[0006] In view of this, the purpose of the present application is to provide the application of DDR1 agonist in the preparation of drugs for treating acute myocardial infarction. Using virtual drug screening and molecular docking technology, the potential binding mode of multiple clinically approved drugs and DDR1 receptor is systematically analyzed. It is found that the SGLT-2 selective inhibitor Ertugliflozin shows good binding affinity and stable docking conformation with the active site of DDR1, suggesting its potential to agonize DDR1 receptor. Further, by agonizing DDR1 signaling pathway with Ertugliflozin, cell migration is promoted, matrix remodeling is regulated and excessive fibrosis is inhibited in epicardial cells, achieving inhibition of pathological ventricular remodeling, improvement of myocardial repair and maintenance of cardiac function after acute myocardial infarction. This provides a theoretical basis for the new use of old drugs and a new idea for the development of treatment strategies for preventing and treating ventricular remodeling after acute myocardial infarction.
[0007] To achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:
[0008] In a first aspect of the present application, a SGLT-2 selective inhibitor, Ertugliflozin, is provided for use in the preparation of a drug targeting DDR1.
[0009] In a second aspect of the present application, a drug composition targeting DDR1 is provided, which comprises a therapeutically effective amount of SGLT-2 selective inhibitor, and takes the SGLT-2 selective inhibitor, i.e. etogliptin, as an active ingredient.
[0010] Preferably, the concentration of etogliptin in the drug composition targeting DDR1 is 1-1000 μM.
[0011] Preferably, the drug composition targeting DDR1 further comprises a pharmaceutically acceptable carrier, and can be prepared into an injection, an emulsion, a tablet, a powder, a granule, an ointment, a liposome or an oral liquid.
[0012] In a third aspect of the present application, a DDR1 agonist is provided, which comprises a therapeutically effective amount of SGLT-2 selective inhibitor, and takes the SGLT-2 selective inhibitor, i.e. etogliptin, as an active ingredient.
[0013] Preferably, the concentration of etogliptin in the DDR1 agonist is 1-1000 μM.
[0014] Preferably, the DDR1 agonist further comprises a pharmaceutically acceptable carrier, and can be prepared into an injection, an emulsion, a tablet, a powder, a granule, an ointment, a liposome or an oral liquid.
[0015] In a fourth aspect of the present application, the drug composition targeting DDR1 or the DDR1 agonist is used in the preparation of a drug for treating acute myocardial infarction.
[0016] Preferably, the drug targets DDR1, and is used for improving ventricular remodeling and tissue repair after acute myocardial infarction, in particular, for inhibiting pathological ventricular remodeling, improving myocardial repair and maintaining cardiac function after acute myocardial infarction.
[0017] Compared with the prior art, the present application has the following beneficial effects:
[0018] 1. DDR1 receptor agonist etogliptin is obtained based on molecular docking and virtual drug screening, and the activation of DDR1 signaling pathway promotes the reconstruction of epicardium-myocardial repair axis, thereby improving the prognosis after myocardial infarction, and verifying the mechanism and therapeutic potential of DDR1 agonist in acute myocardial infarction from the perspective of cell signal regulation. By establishing a mouse coronary ligation model, combining with etogliptin intervention, and using immunofluorescence staining, histological staining, qPCR, Western blot and other technical means, the influence of etogliptin on epicardial activity, angiogenesis and inflammatory response is comprehensively evaluated. Preliminary results show that the etogliptin treatment group shows more significant cardiac function retention, epicardial WT1 +Cell activation and enhanced angiogenesis suggest that it can promote heart injury repair by activating the DDR1 pathway, and has potential clinical transformation value.
[0019] 2. Compared with traditional cardiovascular drugs, the DDR1 agonist Ertugliflozin has the advantages of strong targeting and clear mechanism, which can directly activate the DDR1 signal pathway related to epicardium, regulate the inflammatory response and tissue remodeling process after myocardial infarction from the source, and achieve more targeted tissue repair effect.
[0020] 3. Ertugliflozin, as a small molecule compound, has good oral bioavailability, stable pharmacokinetic characteristics and clear safety, and is suitable for long-term chronic medication. It shows significant therapeutic potential in regulating epicardial activity and promoting angiogenesis, and provides a new treatment strategy and transformation prospect for precise intervention of acute myocardial infarction and other serious cardiovascular events. BRIEF DESCRIPTION OF DRAWINGS
[0021] Figure 1 Results of experiments in the examples for identifying Ertugliflozin as a potential agonist of DDR1 based on molecular docking and virtual drug screening.
[0022] Figure 2 Results of experiments in the examples for Ertugliflozin promoting the migration and remodeling of epicardial epithelial cells by enhancing the activity of DDR1 signal pathway.
[0023] Figure 3 Results of experiments in the examples for verifying that Ertugliflozin can alleviate the cardiac function damage of acute myocardial infarction mice, including improving ejection fraction (EF), fractional shortening (FS), left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV).
[0024] Figure 4 Results of experiments in the examples for Ertugliflozin promoting angiogenesis and epicardial epithelial cell migration in the infarct border zone. DETAILED DESCRIPTION
[0025] In order to more fully understand the technical solutions, objectives and advantages of the present application, the following will further describe the technical effects of the present application in detail and completely in combination with the drawings and specific embodiments. Obviously, the described embodiments are only some of the embodiments of the present application, not all. It should be noted that for those skilled in the art, other embodiments obtained without departing from the concept of the present application all belong to the protection scope of the present application.
[0026] The following examples are used to verify that Ertugliflozin can be used as a DDR1 agonist to prepare a drug for treating cardiac repair after acute myocardial infarction, especially for inhibiting pathological ventricular remodeling, improving myocardial repair, and maintaining cardiac function after acute myocardial infarction.
[0027] 1. Virtual drug screening and molecular docking: Based on the three-dimensional crystal structure information of DDR1 receptor, the receptor protein model is first constructed and optimized on the AutoDockVina platform, the active site is defined and the grid parameters are set, and the clinical drug molecule library is subjected to high-throughput virtual screening. Through the scoring of binding energy and the screening of binding sites, candidate molecules with potential binding ability to the active pocket of DDR1 are screened out. Subsequently, Discovery Studio software is used for further molecular docking simulation of these candidate molecules, including conformation search, binding mode optimization and binding energy fine calculation, analysis of the spatial complementarity, hydrogen bond, hydrophobic interaction and key residue interaction of the molecules in the active pocket. Finally, PyMOL is used for three-dimensional structure visualization of protein-ligand complex to verify the rationality and stability of the docking conformation, and to screen out candidate molecules with optimal binding mode and high affinity.
[0028] 2. Animal grouping: Male C57BL / 6 WT mice (8-10 weeks old, about 25 g) were purchased from Shanghai Slac Laboratory Animal Center of Chinese Academy of Sciences, and all mice were randomly divided into four groups after adaptive feeding for one week:
[0029] ① Sham control group (Sham+Vehicle group), open chest without coronary artery ligation, and intraperitoneal injection of the same volume of solvent after operation;
[0030] ② Sham drug group (Sham+Ert group), also open chest without coronary artery ligation, and intraperitoneal injection of Ertugliflozin solution after operation;
[0031] ③ Myocardial infarction model control group (MI+Vehicle group), intraperitoneal injection of the same volume of solvent after left coronary artery ligation modeling;
[0032] ④ Myocardial infarction model drug group (MI+Ert group), intraperitoneal injection of Ertugliflozin solution after left coronary artery ligation modeling.
[0033] All animal experiment schemes strictly follow the "Regulations of the People's Republic of China on the Management of Experimental Animals" and the ethical standards for experimental animals, and have been approved by the Animal Experiment Ethics Committee of Zhongshan Hospital, Fudan University.
[0034] 3. Epicardial cell culture: Epicardial cell line was purchased from Hangzhou Meisen Cell Technology Co., Ltd. and maintained under the recommended culture conditions by the supplier. Cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum (FBS), 1% penicillin-streptomycin at 37°C in a 5% CO2 humidified incubator, and routinely passaged at 80%-90% confluence.
[0035] 4. Model preparation: Mouse myocardial infarction model was prepared by left coronary artery ligation. The experimental animals were operated under sterile conditions, and after anesthesia, tracheal intubation was performed and connected to a small animal respirator to maintain ventilation, and the depth of anesthesia was confirmed by toe pinch reflex. The mouse was taken in a supine position, and after routine disinfection of the left chest, the skin and intercostal muscles were incised, the 3-4 intercostal space was separated and exposed, and the left chest was exposed to expose the heart. A non-traumatic 6-0 silk thread was wrapped and ligated about 1-2 mm below the origin of the left coronary artery, and the myocardium of the apex and left ventricular anterior wall immediately appeared pale or discolored to confirm successful ischemia. The sham operation group was only threaded without ligation. After ligation, a small amount of sterile normal saline was injected into the chest to exhaust, and the chest wall and skin were sutured layer by layer, the tracheal intubation was removed, and the mouse was placed on a warming pad for recovery. The respiratory and activity status were closely monitored, and analgesic drugs and antibiotics were given after operation to prevent infection, and the mental state, body weight change and wound healing of the mouse were observed and recorded daily.
[0036] 5. Main observation indexes:
[0037] 5.1 Heart ultrasound detection of mouse left heart structure and function. High-resolution small animal heart ultrasound imaging system (Vevo 3100, VisualSonics Company) was used to detect the mouse heart noninvasively, and the left ventricular structure and function were evaluated in real time. During the experiment, the mouse was placed on a thermostatic platform and anesthetized, a high-frequency probe was used to obtain short-axis or long-axis two-dimensional images and M-mode images of the heart, left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were measured, and EF, FS, LVEDV, LVESV and other indexes were calculated, the left heart structure and pumping function of the mouse were evaluated, and quantitative basis was provided for cardiovascular disease model research.
[0038] 5.2 Heart histological staining. In heart histological analysis, hematoxylin-eosin (H&E) staining was first used to evaluate the morphology of mouse heart paraffin sections, and histological changes such as myocardial structure, cell arrangement and fibrosis were observed. Subsequently, immunofluorescence staining was performed, and CD31 + α-SMA + antibodies were used for double-labeling detection to identify mature vascular structures, and WT1 + α-SMA +perivascular epicardial cells to explore the histological features of cardiac angiogenesis and epicardial activation.
[0039] 5.3 Western blot analysis. Western blot was used to analyze the expression of DDR1 signaling pathway in epicardial cells under drug stimulation. Total proteins of treated cells were extracted and separated by SDS-PAGE electrophoresis, then transferred to PVDF membrane. Specific primary antibodies against DDR1 and its downstream signaling molecules (such as NF-KB, p-NF-KB, MMP9, ADGRA2, etc.) and HRP-labeled secondary antibodies were used for incubation in turn. Protein bands were detected by chemiluminescence method to evaluate the influence of drug treatment on the activation level of DDR1 pathway, and to reveal its molecular mechanism in the response regulation of epicardial cells.
[0040] In the following examples, WT mice were purchased from Shanghai Slac Laboratory Animal Center of Chinese Academy of Sciences, immunofluorescence antibodies were purchased from BD Company of the United States, Western blot antibodies were purchased from Abeam Company of the United States, and Ertugliflozin was purchased from MCE Company of the United States.
[0041] Example 1: Ertugliflozin is identified as a potential agonist of DDR1 by structure-based virtual screening
[0042] Based on the three-dimensional structure of DDR1 receptor, molecular simulation and virtual drug screening strategy were used for structure-based drug design to find effective agonists of DDR1 signaling pathway. Through homology modeling and molecular docking technology, the binding mode of candidate small molecules and DDR1 active site was simulated, and based on the scoring of binding energy, molecular dynamics simulation and conformational stability evaluation, the interaction stability and agonistic potential of the receptor were systematically analyzed. The screening results showed that small molecule compound Ertugliflozin could stably bind to multiple key amino acid sites of DDR1, showing high binding affinity and good conformational stability, as shown in Figure 1
[0043] Example 2: Ertugliflozin promotes epicardial cell migration and activates DDR1 signaling pathway
[0044] By in vitro cell scratch test, it was found that after treatment under hypoxic conditions, Ertugliflozin significantly promoted the migration ability of epicardial epithelial cells, showing that the closing speed of the scratch area was significantly accelerated. Further, Western blot was used to detect the expression of DDR1 signal pathway related proteins, and the results showed that Ertugliflozin could significantly enhance the phosphorylation level of DDR1 receptor under hypoxic conditions, and up-regulate the expression of its downstream p-NF-κB, MMP9 and ADGRA2, suggesting that it could enhance the migration phenotype of epicardial epithelial cells by activating DDR1 signal pathway in a low oxygen environment. The above experimental results show that Ertugliflozin can effectively regulate cell migration related pathways in a simulated myocardial infarction hypoxic microenvironment, and has a potential heart repair promoting effect, such as Figure 2 as shown in the following.
[0045] Example 3: Ertugliflozin improves the deterioration of heart function in acute myocardial infarction mice
[0046] Echocardiography was used to dynamically evaluate the heart function of acute myocardial infarction mice at D0, D4, D7 and D14. The results showed that the Ertugliflozin treatment group had a significant improvement in multiple key heart function parameters compared with the control group, specifically: EF and FS increased, indicating improved left ventricular pumping function, LVEDV and LVESV decreased, further supporting its inhibitory effect on ventricular dilation and remodeling. In addition, LVEDD and LVESD also decreased at the corresponding time points, indicating that the degree of cardiac chamber dilation was effectively controlled. HE staining results further verified the morphological basis of functional improvement, showing that the infarct area was significantly reduced. The above experimental results show that Ertugliflozin can effectively alleviate the deterioration of heart function after acute myocardial infarction, improve ventricular remodeling, and has a good heart protective effect, such as Figure 3 as shown in the following.
[0047] Example 4: Ertugliflozin promotes angiogenesis and epicardial epithelial cell migration in the infarct border zone
[0048] The effect of Ertugliflozin on promoting angiogenesis and epicardial epithelial cell migration after myocardial infarction was evaluated by immunofluorescence staining. The results showed that in the infarct myocardial area, whether in the infarct border zone or in the epicardial area, Ertugliflozin treatment increased the number of CD31 + α-SMA + double positive coated blood vessels, suggesting that it can effectively promote the generation of mature blood vessels. At the same time, in the epicardial area, WT1 and α-SMA double staining results showed that WT1 +Cell significantly increased, and epicardial area of WT1 + α-SMA + The number of double positive cells also increased significantly, indicating that Ertugliflozin can activate epicardial WT1 + cells and promote their transformation to mesenchymal phenotype and migration to myocardium. The above experimental results show that Ertugliflozin can synergistically promote the tissue repair process after myocardial infarction by activating epicardial cells and enhancing angiogenesis.
[0049] In summary, the present application takes DDR1 as a target, and verifies that Ertugliflozin can be used as a DDR1 agonist to prepare a drug for improving ventricular remodeling and tissue repair after acute myocardial infarction, which has a broad application prospect.
[0050] The above is only a preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.
Claims
1. Use of an SGLT-2 selective inhibitor in the preparation of a drug targeting DDR1, wherein the SGLT-2 selective inhibitor is ertugliflozin.
2. A pharmaceutical composition targeting DDR1, characterized in that: The invention comprises a therapeutically effective amount of an SGLT-2 selective inhibitor as an active ingredient, wherein the SGLT-2 selective inhibitor is Ertugliflozin.
3. The pharmaceutical composition targeting DDR1 according to claim 2, characterized in that The concentration of ertogliflozin is 1-1000 μM.
4. The pharmaceutical composition targeting DDR1 according to claim 2, characterized in that The pharmaceutical composition targeting DDR1 further comprises a pharmaceutically acceptable carrier and can be prepared into an injection, emulsion, tablet, powder, granule, ointment, liposome or oral solution.
5. A DDR1 agonist, characterized in that The invention comprises a therapeutically effective amount of an SGLT-2 selective inhibitor as an active ingredient, wherein the SGLT-2 selective inhibitor is Ertugliflozin.
6. The DDR1 agonist according to claim 5, characterized in that The concentration of ertogliflozin is 1-1000 μM.
7. The DDR1 agonist according to claim 5, characterized in that The DDR1 agonist further comprises a pharmaceutically acceptable carrier and can be prepared into an injection, emulsion, tablet, powder, granule, ointment, liposome or oral solution.
8. Use of the pharmaceutical composition targeting DDR1 according to any one of claims 2 to 4, or the DDR1 agonist according to any one of claims 5 to 7, in the preparation of a medicament for treating acute myocardial infarction.
9. The use according to claim 8, characterized in that The drug targets DDR1 and is used to improve ventricular remodeling and tissue repair after acute myocardial infarction.
10. The use according to claim 8, characterized in that The drug targets DDR1 and is used to inhibit pathological ventricular remodeling, improve myocardial repair and maintain cardiac function after acute myocardial infarction.