Application of dicaffeoylglucose in treatment and / or prevention of ischemic heart diseases
By screening caffeoyl glucose from the root of *Cimicifuga foetida* using a zebrafish model and targeting tyrosine kinase Src as the key target, the study promoted cardiac damage repair, solved the regeneration problem after myocardial infarction, and achieved cardiac function recovery.
Patent Information
- Application Number
- CN202511886782.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Priority Date
- 2024-12-19
- Filing Date
- 2025-12-15
- Publication Date
- 2026-01-27
AI Technical Summary
Current technologies cannot effectively promote cardiac repair and regeneration after myocardial infarction, leading to heart failure, and there is a lack of effective treatment methods.
Using a high-throughput screening model for cardiac injury in zebrafish, we discovered dicaffeoyl glucose, a natural compound extracted from the root of *Cimicifuga foetida*. Through network pharmacology target analysis, we identified tyrosine kinase Src as the key target, which promotes cardiac injury repair.
Disaccharyl glucose significantly improves the repair and regeneration rate of zebrafish heart after injury, providing a new treatment approach for myocardial infarction and ischemic heart disease, reducing patient suffering and socioeconomic burden.
Smart Images
Figure CN121401276A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of dicaffeoyl glucose in the treatment and / or prevention of ischemic heart disease. Background Technology
[0002] Myocardial infarction (MI) is a cardiovascular disease with high morbidity and mortality rates. Ischemic cardiomyopathy caused by MI seriously affects human health and increases the socioeconomic burden. Acute myocardial ischemia causes irreversible death of cardiomyocytes. After the death of cardiomyocytes in adult mammals, the damaged area cannot be repaired by new cardiomyocytes, but is replaced by collagen-rich scar tissue. However, the inelastic scar tissue cannot restore the original function of the heart, leading to compensatory hypertrophy of the remaining cardiomyocytes, which in turn causes secondary loss of cardiomyocytes, ultimately leading to heart failure. Although commonly used medical drugs and interventional treatments can increase local blood supply after myocardial infarction, they cannot fundamentally repair the necrotic myocardial tissue and cardiac function. Therefore, how to promote cardiac repair and regeneration is an important scientific problem that urgently needs to be solved in the field of cardiovascular research.
[0003] Zebrafish models have unique advantages in cardiac repair and regeneration research. Their heart structure and function are similar to those of humans to some extent, and the transparent nature of the embryos makes it easy to observe the repair process after cardiac injury, enabling rapid and efficient evaluation of the effects of compounds.
[0004] Experiments have confirmed that regulating endogenous cardiomyocyte proliferation through multiple strategies is an important pathway to promote cardiac repair and regeneration and improve cardiac function after myocardial infarction. The strategy of inducing endogenous cardiomyocyte proliferation through small molecule chemical drugs is still in its early stages. The diverse and morphologically varied plants in nature possess a large number of structurally complex and diverse natural product molecules. These natural product molecules can participate in important physiological activities of plants as endogenous small molecules, and are also important sources of many innovative drug molecules. From quinine to artemisinin, natural products have long played an important role in the treatment of various serious diseases. Undoubtedly, developing natural small molecule drugs that promote cardiac repair and regeneration and exploring their mechanisms of action will provide new targets and treatment strategies for myocardial infarction and ischemic diseases.
[0005] 3,4-di-O-(E)-caffeoyl-α / β-D-glucopyranoside (E7) is a novel natural compound extracted from the root of *Aruncus sylvester Kostel*. This compound is abundant in *Aruncus sylvester Kostel* and is easily isolated and purified. *Aruncus sylvester Kostel* is a perennial herb belonging to the genus *Aruncus* in the family Rosaceae. It is one of the seven traditional Chinese medicines of Shaanxi Province and is commonly used in folk medicine to treat muscle and bone pain caused by overexertion, possessing the effects of relaxing muscles and tendons, relieving pain, and reducing inflammation. Various extracts of this plant exhibit strong free radical scavenging activity and show certain effects in antioxidation, anti-inflammation, and antibacterial activity. However, currently, there are no reports on the application of 3,4-di-O-(E)-caffeoyl-α / β-D-glucopyranoside in the treatment of cardiovascular diseases, especially in the treatment of myocardial infarction or injury. Summary of the Invention
[0006] To address the aforementioned technical problems, this invention proposes the application of dicaffeoyl glucose in the treatment and / or prevention of ischemic heart disease. Using a zebrafish high-throughput screening model for cardiac injury, it was found that dicaffeoyl glucose, a natural compound extracted from the root of *Cimicifuga foetida*, has a good effect on promoting the repair of cardiac damage. This opens up new avenues for the treatment of myocardial infarction and other ischemic heart diseases, alleviates patient suffering, and reduces the socioeconomic burden of cardiovascular diseases.
[0007] To achieve the above objectives, the present invention provides the use of dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof in the preparation of medicaments for the treatment and / or prevention of ischemic heart disease.
[0008] Preferably, the structural formula of the dicaffeoyl glucose is shown in Formula I: .
[0009] Preferably, the ischemic heart disease is acute myocardial infarction caused by multiple factors such as insufficient blood supply, myocardial hypoxia, or excessive myocardial oxygen consumption due to various causes.
[0010] The present invention also provides the use of the dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof in the preparation of a drug that promotes the repair and regeneration of cardiac damage.
[0011] Preferably, the dicaffeoyl glucose can promote the repair and regeneration of zebrafish heart damage, and the concentration of dicaffeoyl glucose in the drug is 100~500μM.
[0012] The present invention also provides the application of the dicaffeoyl glucose in the preparation of a drug that promotes the repair and regeneration of cardiac damage, wherein the concentration of dicaffeoyl glucose in the drug is 100~500μM.
[0013] Compared with the prior art, the present invention has the following advantages and technical effects: This invention focuses on myocardial infarction, a cardiovascular disease with high incidence and mortality. Using a zebrafish high-throughput screening model for cardiac injury, it was discovered that dicaffeoyl glucose, a natural compound extracted from the root of *Cimicifuga foetida*, has a good effect on promoting cardiac injury repair. Using network pharmacology target analysis technology, tyrosine kinase Src was identified as its core target. This key discovery provides extremely important clues and directions for elucidating the molecular regulatory network of dicaffeoyl glucose in the cardiac injury repair process.
[0014] This invention discloses the medicinal value of dicaffeoyl glucose in the treatment and / or prevention of ischemic diseases and a new therapeutic target for myocardial infarction. The aim is to develop novel cardiovascular drug formulations with dicaffeoyl glucose as the main active ingredient, thereby opening new avenues for the treatment of myocardial infarction and other ischemic heart diseases, alleviating patient suffering, and reducing the socioeconomic burden of cardiovascular disease.
[0015] This invention demonstrates through experiments that dicaffeoyl glucose can promote the repair and regeneration of zebrafish hearts after damage. Therefore, dicaffeoyl glucose can be used in the preparation of drugs that promote cardiac repair and regeneration, as well as in the treatment or prevention of heart disease and cardiovascular diseases, providing a new drug and treatment approach for the treatment or prevention of heart damage. Attached Figure Description
[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments 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.
[0017] Figure 1 The diagram shows how dicaffeoyl glucose promotes the regeneration of zebrafish cardiomyocytes. In the diagram, A is a schematic diagram of screening active natural products that promote the repair and regeneration of the heart after injury; B is the proportion of heart repair and regeneration after treatment with different active natural products; * indicates a significant difference between different experimental groups and the injury group; and C is the proportion of heart repair and regeneration after treatment with different concentrations of dicaffeoyl glucose; * indicates a significant difference between different experimental groups and the injury group. Figure 2 The target analysis of dicaffeoyl glucose is as follows: A represents the target related to dicaffeoyl glucose and cardiac repair and regeneration, and B represents the network of action of dicaffeoyl glucose and cardiac repair and regeneration targets. Figure 3The docking analysis of dicaffeoyl glucose with target molecules is shown in Figure A, where A represents the binding energy of dicaffeoyl glucose with SRC, MMP9 and HIF1α, and B represents the docking mode diagram of dicaffeoyl glucose with SRC. Figure 4 A volcano plot of transcription sequencing data; Figure 5 A statistical chart of GO function categories; Figure 6 Bubble chart for GO enrichment analysis; Figure 7 Chord plot for GO enrichment analysis; Figure 8 A statistical chart of KEGG pathway classification; Figure 9 This is a graph showing the enrichment of the KEGG pathway. Detailed Implementation
[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0019] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.
[0020] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.
[0021] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.
[0022] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.
[0023] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the materials and reagents used in the following examples are commercially available. The zebrafish used in the following examples are three-day-old zebrafish juveniles. The zebrafish used in the following examples are Tg(cmlc2:eCFP-NTR) strain juveniles. The MTZ used in the following examples to induce cardiac injury in zebrafish is Metronidazole, manufactured by Tianjin Heowns Biochemical Technology Co., Ltd., with product number 443-48-1 and trade name.
[0024] The materials used in this invention are sourced as follows: The E3 culture medium is formulated with deionized water as solvent, 5 mM NaCl, 0.17 mM KCl, 0.4 mM CaCl2, and 0.33 mM MgSO4; all raw materials used in the preparation were purchased from Sinopharm Group Co., Ltd.; DMSO was purchased from Beijing Solarbio Co., Ltd., catalog number D8370, trade name dimethyl sulfoxide.
[0025] Example (1) Establishing a zebrafish heart chemogenetic damage model Transgenic strain Tg(cmlc2:eCFP-NTR) was mated with wild-type adult zebrafish, and zebrafish embryos (>200) were collected and incubated in E3 medium at 28.5℃ for 48 hours (E3 medium was changed daily). The zebrafish embryos were examined under a stereomicroscope, and embryos with positive CFP in the heart were selected for subsequent cardiac injury experiments. CFP-positive embryos developed to day 3 were placed in 12-well plates, with more than 20 embryos per well. The normal control group was treated with DMSO, while the experimental groups were treated with 10 mM Mtz for 2 hours. On day 5 of embryonic development, the pericardial edema rate of each group was counted using a stereomicroscope. The treatment conditions for the zebrafish cardiac injury model were determined to be 10 mM Mtz treatment for 2 hours on CFP-positive embryos developed to day 3.
[0026] (2) Screening for active natural products that promote the repair and regeneration of the heart after injury. The transgenic strain Tg(cmlc2:eCFP-NTR) was mated with wild-type adult zebrafish, and zebrafish embryos (>200) were collected and cultured in E3 medium. These embryos were then incubated in a 28.5℃ incubator for 48 hours (with the E3 medium changed daily). The zebrafish embryos were examined under a stereomicroscope, and embryos with positive CFP in the heart were selected for subsequent cardiac injury experiments. Figure 1As shown in Figure A, when the embryos developed to day 3, CFP-positive embryos were placed in a 12-well plate and treated with 10 μM Mtz for 2 hours. Then, different natural products (screening concentration of 100 μM, freshly prepared) were added to each well and treated for 4 hours. After treatment, the embryos were transferred to E3 culture medium. On day 4 of embryo development, the pericardial edema and other conditions of the zebrafish heart were observed and statistically analyzed.
[0027] (3) Determine the optimal concentration of dicaffeoyl glucose to promote repair and regeneration in zebrafish. Transgenic strain Tg(cmlc2:eCFP-NTR) was mated with wild-type adult zebrafish, and zebrafish embryos (>200) were collected and incubated in E3 medium at 28.5℃ for 48 hours (E3 medium was changed daily). The zebrafish embryos were examined under a stereofluorescence microscope, and embryos with positive CFP in their hearts were selected for subsequent cardiac injury experiments. On day 3 of embryonic development, CFP-positive embryos were divided into three groups: a sham injury group, an injury control group, and an injury experimental group. The concentrations of dicaffeoyl glucose in the injury experimental group were 100 μM, 200 μM, and 500 μM, respectively. The proportion of cardiac repair and regeneration in zebrafish embryos was assessed on days 4 and 5 of embryonic development to determine the optimal concentration of dicaffeoyl glucose for promoting cardiac repair and regeneration in zebrafish.
[0028] (4) Examine the effect of dicaffeoyl glucose on promoting the repair and regeneration of zebrafish heart. On day 3 of zebrafish embryonic development, samples were divided into a sham-damage group, a damage control group, and a dicaffeoyl glucose treatment group, and treated accordingly. On day 4 of embryonic development, the proportion of zebrafish heart repair and regeneration in each group was statistically analyzed using a stereomicroscope. Subsequently, the samples were embedded in low-melting-point agarose and three-dimensional imaging was performed using a two-photon light-sheet fluorescence microscope to detect heart volume.
[0029] (5) Network pharmacology analysis of the target of caffeoyl glucose in promoting cardiac repair and regeneration Network pharmacology, as a bioinformatics tool, is widely used in discovering bioactive components, drug targets, and mechanisms of action. By taking the intersection of 100 targets interacting with dicaffeoyl glucose and 4626 targets related to cardiac repair and regeneration, 70 potential targets for dicaffeoyl glucose to promote cardiac repair and regeneration were obtained (e.g., ...). Figure 2(A). These targets were uploaded to the STRING database with a confidence level of 0.4, and the interaction network between dicaffeoyl glucose and cardiac repair and regeneration targets was analyzed. The CytoNCA plugin was used to calculate the betweenness center number of network nodes; nodes with higher betweenness centers are considered more important and have stronger interactions. Based on the betweenness center number, 70 common targets were ranked, revealing that tyrosine kinase Src, matrix metalloproteinase MMP9, and hypoxia-inducible factor HIF-1α had relatively high interactions with dicaffeoyl glucose, with tyrosine kinase Src exhibiting the strongest interaction with dicaffeoyl glucose (e.g., Src). Figure 2 (B)
[0030] (6) Molecular docking analysis of the binding energy between dicaffeoyl glucose and the target To assess the binding energy (i.e., affinity) between dicaffeoyl glucose and its targets, molecular docking analysis was performed on the top three candidate target proteins, SRC, MMP9, and HIF1α, with dicaffeoyl glucose. The results showed that the binding energies between the three targets and dicaffeoyl glucose were -9.5 kcal / mol, -8.4 kcal / mol, and -7.6 kcal / mol, respectively. Figure 3 (A). Binding energy reflects the likelihood of binding between the receptor and ligand. The lower the binding energy, the higher the affinity between the receptor and ligand, and the more stable the conformation. The results show that SRC has the lowest binding energy, suggesting that SRC can bind efficiently to dicaffeoyl glucose (e.g., ...). Figure 3 (B)
[0031] (7) Transcriptomics research Volcano plot of transcriptome sequencing data (e.g.) Figure 4 The study showed that, compared with the simple injury group, the dicaffeoyl glucose treatment group induced significant changes in gene expression profiles. Specifically, the expression of genes such as the key enzyme in glucose metabolism (PGM2), endoplasmic reticulum shaping protein (REEP2), and RNA exonucleoside component (EXOSC5) was significantly downregulated, while the expression of tRNA processing enzyme (ELAC1) and non-coding RNA transcript (FRA5AS1, si_ch1073-340i21.3) showed a significant upregulation trend. This differential expression pattern suggests that dicaffeoyl glucose may mediate metabolic reprogramming of cardiomyocytes through a non-coding RNA regulatory network: while alleviating the metabolic burden of glycolysis and endoplasmic reticulum stress (PGM2 downregulation, REEP2 downregulation), it enhances protein translation efficiency by optimizing RNA processing and metabolic homeostasis (ELAC1 upregulation, EXOSC5 downregulation), thereby promoting the transformation of damaged cardiomyocytes from an acute stress state to a regenerative and structural remodeling state.
[0032] GO Functionality Classification Statistics Chart (e.g.) Figure 5This study further revealed the distribution characteristics of differentially expressed genes at three levels: biological processes (BP), cellular components (CC), and molecular functions (MF), showing an overall trend of "upregulation of synthetic and metabolic functions." In biological processes (BP), metabolic processes and cellular processes not only had the highest gene enrichment but also a significant proportion of upregulated genes (dark bars), confirming that the cells in the treatment group were in a metabolically active state, undergoing large-scale material and energy exchange to support regeneration. In molecular functions (MF), catalytic activity and binding activity were dominant. High expression of catalytic activity (especially upregulated components) directly corresponds to increased activity of the aforementioned metabolic enzymes (such as glutathione metabolites) and modifying enzymes (such as NuA4 acetyltransferase); while the enrichment of binding activity suggests extensive intracellular protein-protein interactions and the binding of transcription factors to DNA. The enrichment of cellular anatomical entities within the cellular component (CC) further underscores the physical repair of cellular entity structures, such as Z-discs and cytoskeleton remodeling.
[0033] Further GO enrichment analysis (GO enrichment analysis bubble chart as shown) Figure 6 As shown, the GO enrichment analysis chord diagram is as follows: Figure 7 As shown in the figure, differentially expressed genes were significantly enriched in the NuA4 histone acetyltransferase complex, ribonucleoprotein complex, and Z-disc cellular components. This result indicates that dicaffeoyl glucose promotes myocardial repair through a multidimensional molecular mechanism: firstly, it promotes chromatin opening and transcriptional activation through NuA4 complex-mediated histone acetylation modification; secondly, it enhances mRNA processing and transport using the ribonucleoprotein complex, increasing protein synthesis rates; and finally, it promotes the expression and assembly of Z-disc-related proteins, completing sarcomere structural reconstruction. This cascade reaction, from epigenetic regulation to protein translation and then to cytoskeleton remodeling, constitutes the potential molecular mechanism by which this drug promotes myocardial regeneration.
[0034] KEGG pathway classification statistics chart (e.g.) Figure 8The study showed that dicaffeoyl glucose not only regulates metabolism but also broadly activates signal transduction networks related to environmental information processing. In the "environmental information processing" category, the MAPK signaling pathway, neuroactive ligand-receptor interaction, and cytokine-cytokine receptor interaction showed the highest enrichment of genes. Furthermore, classic regeneration and survival signaling pathways such as FoxO, Wnt, mTOR, and Apelin were also significantly enriched. This indicates that dicaffeoyl glucose exerts its effects by constructing a complex signal regulatory network: the enrichment of MAPK and mTOR pathways suggests that the drug activates upstream commands for cell proliferation and protein synthesis (corresponding to the aforementioned ribosome biosynthesis); the involvement of FoxO and Wnt pathways suggests fine-tuning of the cell cycle and differentiation state; and the presence of Apelin and the calcium signaling pathway is directly associated with improved myocardial contractility and potential regulation of angiogenesis. Furthermore, the enrichment of cardiac muscle contraction and adrenergic signaling in the "Organismal Systems" category further confirms the restoration of the cardiac cell contraction / excitation coupling mechanism in the treatment group from a functional perspective.
[0035] KEGG pathway enrichment analysis further revealed the systemic role of dicaffeoyl glucose in remodeling the metabolic network of cardiomyocytes (e.g. Figure 9 The most significantly enriched lipoic acid metabolism and glutathione metabolism pathways indicate that this therapeutic strategy not only optimizes substrate utilization by regulating key energy metabolism enzymes (such as the aforementioned changes in PGM2), but more importantly, establishes a highly efficient antioxidant defense system. The activation of the metabolic pathways of lipoic acid and glutathione, as key intracellular antioxidants and mitochondrial coenzymes, suggests that intracellular redox homeostasis has been effectively restored, thereby mitigating secondary damage from reactive oxygen species (ROS) to damaged myocardium. Furthermore, the enrichment of amino sugar and nucleotide sugar metabolism suggests that cells are actively synthesizing glycoconjugates through the hexosamine biosynthesis pathway (HBP), providing the necessary substrate basis for extracellular matrix (ECM) remodeling and intercellular signaling communication.
[0036] (8) Results The experimental results are shown in the figure. Based on the zebrafish cardiac chemogenetic injury model, natural products promoting cardiac repair and regeneration were screened, and dicaffeoyl glucose was found to have a good effect on promoting cardiac repair and regeneration. Compared with the injury control group, the proportion of cardiac repair and regeneration in the dicaffeoyl glucose treatment group increased to 50% (e.g., ...). Figure 1 (Middle B). Further investigation into the effects of different concentrations of dicaffeoyl glucose on cardiac repair and regeneration revealed that 200 μM dicaffeoyl glucose treatment resulted in the best cardiac repair and regeneration effect, increasing the proportion of cardiac repair and regeneration to 73% (e.g., Figure 1 (C)
[0037] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.
Claims
1. The use of dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof in the preparation of medicaments for the treatment and / or prevention of ischemic heart disease.
2. The application according to claim 1, characterized in that, The structural formula of the dicaffeoyl glucose is shown in Formula I: 。 3. The application according to claim 1, characterized in that, The ischemic heart disease mentioned above refers to acute myocardial infarction caused by multiple factors such as insufficient blood supply, myocardial hypoxia, or excessive myocardial oxygen consumption due to various causes.
4. The use of the dicaffeoyl glucose as described in claim 1 or a pharmaceutically acceptable salt or ester thereof in the preparation of a drug for promoting the repair and regeneration of cardiac damage.
5. The application according to claim 4, characterized in that, The dicaffeoyl glucose can promote the repair and regeneration of zebrafish heart damage, and the concentration of dicaffeoyl glucose in the drug is 100~500μM.
6. The use of dicaffeoyl glucose as described in claim 1 in the preparation of drugs that promote cardiac injury repair and regeneration, characterized in that, The concentration of dicaffeoyl glucose in the drug is 100~500μM.