Application of dicaffeoylglucose in treatment and / or prevention of ischemic heart diseases

Abstract

The invention discloses application of dicaffeoylglucose in treatment and / or prevention of ischemic heart diseases, belongs to the technical field of biological medicines, and provides application of dicaffeoylglucose or pharmaceutically acceptable salt or ester thereof in preparation of medicines for treatment and / or prevention of ischemic heart diseases. A medicine for treating and / or preventing ischemic heart diseases comprises the dicaffeoylglucose or the pharmaceutically acceptable salt or ester of the dicaffeoylglucose. The invention discloses application of dicaffeoylglucose in preparation of a medicine for promoting heart injury repair and regeneration. A medicine for promoting heart injury repair and regeneration comprises dicaffeoylglucose. A zebra fish heart injury high-throughput screening model is utilized to find that the natural compound dicaffeoylglucose extracted from the root of pseudolargetrifoliolious bugbane has a good effect of promoting heart injury repair, so that a new way is opened up for treating myocardial infarction and other ischemic heart diseases, and the pain of a patient is relieved; the social and economic burden of cardiovascular diseases is reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of dicaffeoyl glucose in treating and / or preventing ischemic heart disease. Background Art

[0002] Myocardial infarction is a cardiovascular disease with a high morbidity and mortality rate today. Ischemic cardiomyopathy caused by myocardial infarction has seriously affected human health and increased the socioeconomic burden. Acute myocardial ischemia causes irreversible death of myocardial cells. After the death of adult myocardial cells in mammals, the damaged site cannot be repaired by new myocardial cells, but is replaced by collagen-rich scars. However, the scars lacking elasticity cannot restore the original function of the heart, leading to compensatory hypertrophy of the remaining myocardial cells, which in turn causes secondary loss of myocardial cells and ultimately leads to heart failure. Although currently commonly used medical drugs, interventional treatments and other measures can increase local blood supply after myocardial infarction, they cannot fundamentally repair the necrotic myocardial tissue and heart function. Therefore, how to promote cardiac repair and regeneration is an important scientific issue that needs to be urgently addressed in the field of cardiovascular research.

[0003] The zebrafish model has unique advantages in cardiac repair and regeneration research. Its cardiac structure and function are similar to those of humans to a certain extent, and the embryos are transparent, making it easy to observe the repair process after cardiac injury, and the effects of compounds can be evaluated quickly and efficiently.

[0004] Experiments have confirmed that regulating the proliferation of endogenous cardiomyocytes through various strategies is an important way to promote cardiac repair and regeneration and improve cardiac function after myocardial infarction. The strategy of inducing endogenous cardiomyocyte proliferation through chemical small molecule drugs is still in its infancy. There are many kinds of plants with different forms in nature, which have a large number of natural product molecules with complex and diverse structures. These natural product molecules can not only participate in important physiological activities of plants as endogenous small molecules, but also are an important source of many innovative drug molecules. From quinine to artemisinin, natural products have long played an important role in the treatment of many serious diseases. There is no doubt that the development of natural small molecule drugs that promote cardiac repair and regeneration and the exploration of their mechanism of action will provide new targets and therapeutic strategies for the treatment of myocardial infarction and ischemic diseases.

[0005] Dicaffeoyl glucose (3,4-di-O-(E)-caffeoyl-α / β-D-glucopyranoside, E7) is a new natural compound extracted from the root of Aruncus sylvester. The compound is high in Aruncus sylvester and is easy to separate and purify. Aruncus sylvester Kostel.ex Maxim is a perennial herb of the genus Aruncus in the family Rosaceae. It is one of the seven medicines of Shaanxi. It is commonly used in folk medicine to treat muscle and bone pain due to fatigue. It has the effects of relaxing muscles, activating collaterals, relieving pain and anti-inflammatory. Various extracts of this plant show strong free radical scavenging activity and have certain effects in antioxidant, anti-inflammatory and antibacterial aspects. However, at present, there are no relevant reports on the use of dicaffeoyl glucose in the treatment of cardiovascular diseases, especially in the treatment of myocardial infarction or injury. Summary of the invention

[0006] In order to solve the above technical problems, the present invention proposes the use of dicaffeoyl glucose in the treatment and / or prevention of ischemic heart disease. By using a zebrafish heart injury high-throughput screening model, it is found that the natural compound dicaffeoyl glucose extracted from the root of Pseudo-Cimicifuga has a good effect in promoting the repair of heart damage, which opens up a new way for the treatment of myocardial infarction and other ischemic heart diseases, alleviates the pain of patients, and reduces the socioeconomic burden of cardiovascular diseases.

[0007] To achieve the above object, the present invention provides the use of dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof in the preparation of a drug for treating and / or preventing ischemic heart disease.

[0008] Preferably, the structural formula of the dicaffeoyl glucose is as shown in Formula I:

[0009]

[0010] Preferably, the ischemic heart disease is acute myocardial infarction caused by insufficient blood supply, myocardial hypoxia or excessive myocardial oxygen consumption due to various reasons.

[0011] The present invention provides a medicine for treating and / or preventing ischemic heart disease, and the medicine comprises dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof.

[0012] Preferably, the drug further comprises one or more pharmaceutically acceptable carriers; the carriers are conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers or lubricants in the pharmaceutical field.

[0013] Preferably, the drug can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, or mucosal tissue by injection, spraying, nasal drops, eye drops, penetration, absorption, or physical or chemical mediation.

[0014] The present invention also provides the use of the dicaffeoyl glucose or its pharmaceutically acceptable salt or ester in the preparation of a drug for promoting cardiac injury repair and regeneration.

[0015] Preferably, the dicaffeoyl glucose can promote the repair and regeneration of zebrafish heart damage.

[0016] The present invention also provides the use of the dicaffeoyl glucose in preparing a drug for promoting heart damage repair and regeneration.

[0017] The present invention also provides a medicine for promoting the repair and regeneration of cardiac damage, and the medicine comprises the dicaffeoyl glucose.

[0018] Compared with the prior art, the present invention has the following advantages and technical effects:

[0019] The present invention focuses on myocardial infarction, a cardiovascular disease with high morbidity and mortality. By using a zebrafish heart injury high-throughput screening model, it is found that the natural compound dicaffeoylglucose extracted from the root of Pseudo-Cimicifuga has a good effect in promoting heart injury repair. The network pharmacology target analysis technology is used to lock in the tyrosine kinase Src as its core target. This key discovery provides extremely important clues and direction points for explaining the molecular regulatory network of dicaffeoylglucose in the process of heart injury repair.

[0020] The present invention provides the medicinal value of dicaffeoyl glucose in treating and / or preventing ischemic diseases and a new target for the treatment of myocardial infarction. The invention is committed to developing a new cardiovascular drug preparation with dicaffeoyl glucose as the main active ingredient, thereby opening up a new way for the treatment of myocardial infarction and other ischemic heart diseases, alleviating the pain of patients and reducing the socioeconomic burden of cardiovascular diseases.

[0021] The present invention proves through experiments that dicaffeoyl glucose can promote the repair and regeneration of zebrafish heart after damage. Therefore, dicaffeoyl glucose can be used in the preparation of drugs for promoting heart repair and regeneration and drugs for treating or preventing heart disease and cardiovascular disease and other related fields, providing a new drug and treatment idea for treating or preventing heart damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0023] Figure 1 Figure 2 shows that dicaffeoyl glucose promotes zebrafish heart repair and regeneration, where A is a schematic diagram for screening active natural products that promote heart repair and regeneration after heart injury, B is the proportion of heart repair and regeneration after treatment with different active natural products, * indicates that there is a significant difference between different experimental groups and the injury group, C is the proportion of heart repair and regeneration after treatment with different concentrations of dicaffeoyl glucose, * indicates that there is a significant difference between different experimental groups and the injury group, D is a representative heart phenotype of the sham injury group, with a scale of 100 μm, E is a representative heart phenotype of the injury control group, with a scale of 100 μm, and F is a representative heart phenotype of the dicaffeoyl glucose treatment group, with a scale of 100 μm;

[0024] Figure 2 The treatment of dicaffeoylglucose promotes the recovery of ventricular volume after zebrafish heart injury, where A is a representative image of 3D volume reconstruction of the heart in the sham injury group, with a scale of 20 μm, B is a representative image of 3D volume reconstruction of the heart in the injury control group, with a scale of 20 μm, C is a representative image of 3D volume reconstruction of the heart in the dicaffeoylglucose treatment group, with a scale of 20 μm, and D is the heart volume statistics of the sham injury group, injury control group, and dicaffeoylglucose treatment group. * indicates significant differences between different treatment groups;

[0025] Figure 3 This is the target analysis of dicaffeoylglucose, where A is the target related to dicaffeoylglucose and cardiac repair and regeneration, and B is the dicaffeoylglucose-cardiac repair and regeneration target action network;

[0026] Figure 4 This is the docking analysis of dicaffeoylglucose and target molecules, where A is the binding energy of dicaffeoylglucose and SRC, MMP9 and HIF1α, and B is the docking pattern diagram of dicaffeoylglucose and SRC. DETAILED DESCRIPTION

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] The experimental methods used in the following examples are all conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified. The zebrafish used in the following examples are zebrafish fry that were born and hatched for three days. The zebrafish used in the following examples are zebrafish fry of the strain Tg (cmlc2: eCFP-NTR). The MTZ used in the following examples to induce zebrafish heart damage is produced by Tianjin Heowns Biochemical Technology Company with the product number 443-48-1 and the trade name Metronidazole.

[0033] Sources of materials used in the present invention: The formula of E3 culture medium is deionized water as solvent, 5mM NaCl, 0.17mM KCl, 0.4mM CaCl2, 0.33mM MgSO4; the raw materials used for preparation are purchased from Sinopharm Group Co., Ltd.; DMSO is purchased from Beijing Solebow Company with item number D8370 and trade name dimethyl sulfoxide.

[0034] Example

[0035] (1) Establishment of a zebrafish cardiac chemical genetic injury model

[0036] Transgenic strain Tg(cmlc2:eCFP-NTR) was mated with wild-type adult zebrafish to collect zebrafish embryos (>200) in E3 culture medium and placed in a 28.5℃ constant temperature incubator for 48h (E3 culture medium was changed once a day). Zebrafish embryos were placed under a stereo fluorescence microscope, and embryos with positive cardiac CFP were selected for subsequent cardiac injury experiments. CFP-positive embryos on the third day of development were placed in a 12-well plate, with more than 20 embryos per well. The normal control group was treated with DMSO, and the experimental groups were treated with 2mM, 5mM, and 10mM Mtz for 2h. On the fifth day of embryonic development, the cardiac pericardial edema rate of each group was counted using a stereo microscope, and the treatment conditions of the zebrafish cardiac injury model were determined to be 10mM Mtz treatment for 2h for CFP-positive embryos on the third day of development.

[0037] (2) Screening for active natural products that promote repair and regeneration after cardiac injury

[0038] The transgenic strain Tg(cmlc2:eCFP-NTR) was mated with wild-type adult zebrafish to collect zebrafish embryos (>200) in E3 culture medium and incubated in a 28.5℃ constant temperature incubator for 48h (E3 culture medium was changed once a day). The zebrafish embryos were placed under a stereofluorescence microscope and embryos with positive cardiac CFP were selected for subsequent cardiac injury experiments. Figure 1 As shown in A, when the embryos develop to the third day, CFP-positive embryos are placed in a 12-well plate. After the embryos are treated with 10 Mm Mtz for 2 hours, different natural products (screening concentration is 100 μM, prepared immediately before use) are added to each well for 4 hours and then transferred to E3 culture medium. On the fourth day of embryonic development, the zebrafish cardiac pericardial edema and other conditions are observed and counted.

[0039] (3) Determine the optimal concentration of dicaffeoyl glucose to promote zebrafish repair and regeneration

[0040] The transgenic strain Tg (cmlc2: eCFP-NTR) was mated with wild-type adult zebrafish to collect zebrafish embryos (>200) in E3 culture medium and place them in a 28.5℃ constant temperature incubator for 48h (E3 culture medium was changed once a day). The zebrafish embryos were placed under a stereofluorescence microscope, and embryos with positive cardiac CFP were selected for subsequent cardiac injury experiments. When the embryos developed to the third day, the CFP-positive embryos were divided into three groups, namely, the sham injury group, the injury control group and the injury experimental group. The test concentrations of dicaffeoyl glucose in the injury experimental group were 100μM, 200μM and 500μM, respectively. The proportion of zebrafish embryonic heart repair and regeneration was evaluated on the 4th and 5th days of embryonic development, and the optimal concentration of dicaffeoyl glucose to promote zebrafish heart repair and regeneration was determined.

[0041] (4) Examining the effect of dicaffeoyl glucose on zebrafish cardiac repair and regeneration

[0042] On the third day of zebrafish embryonic development, the samples were divided into a sham injury group, an injury control group, and a dicaffeoyl glucose treatment group, and were treated accordingly. On the fourth day of embryonic development, the proportion of zebrafish heart repair and regeneration in each group was counted using a stereo microscope. 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 the heart volume.

[0043] (5) Network pharmacology analysis of the targets of dicaffeoylglucose in promoting cardiac repair and regeneration

[0044] As a bioinformatics tool, network pharmacology is widely used in discovering bioactive ingredients, drug therapeutic targets and mechanisms of action. The intersection of 100 targets acting on dicaffeoyl glucose and 4626 targets related to cardiac repair and regeneration was taken, and 70 potential targets of dicaffeoyl glucose in promoting cardiac repair and regeneration were obtained (such as Figure 3 A in the middle). These targets were uploaded to the STRING database, and the confidence was set to 0.4. The interaction network between dicaffeoyl glucose and cardiac repair and regeneration targets was analyzed. The CytoNCA plug-in was used to calculate the betweenness center number of the network nodes. The larger the value, the more important the node is in the network and the higher the interaction relationship. The 70 common targets were arranged according to the betweenness center number, and it was found that tyrosine kinase Src, matrix metalloproteinase MMP9, and hypoxia-inducible factor HIF-1α had a high interaction relationship with dicaffeoyl glucose, among which tyrosine kinase Src had the highest interaction relationship with dicaffeoyl glucose (such as Figure 3 (middle B).

[0045] (6) Molecular docking analysis of the binding energy between dicaffeoyl glucose and the target

[0046] In order to evaluate the binding energy (i.e. affinity) between dicaffeoyl glucose and the target, the top three candidate target proteins SRC, MMP9 and HIF1α were subjected to molecular docking analysis with dicaffeoyl glucose, and it was found that the binding energies of the three targets and dicaffeoyl glucose were -9.5 kcal / mol, -8.4 kcal / mol and -7.6 kcal / mol, respectively (e.g. Figure 4 The binding energy reflects the possibility of binding between the receptor and the ligand. The lower the binding energy, the higher the affinity between the receptor and the ligand, and the more stable the conformation. The results show that SRC has the lowest binding energy, suggesting that SRC can bind to dicaffeoyl glucose efficiently (e.g. Figure 4 (middle B).

[0047] (7) Results

[0048] The test results are shown in the figure. Based on the zebrafish heart chemical genetic damage model, natural products that promote heart repair and regeneration were screened. It was found that dicaffeoyl glucose has a good effect on promoting heart repair and regeneration. Compared with the injury control group, the proportion of heart repair and regeneration in the dicaffeoyl glucose treatment group increased to 50% (as shown in Figure 2). Figure 1 Further investigation of the effects of different concentrations of dicaffeoyl glucose on cardiac repair and regeneration revealed that 200 μM dicaffeoyl glucose had the best cardiac repair and regeneration effect, with the proportion of cardiac repair and regeneration increasing to 73% (Fig. Figure 1 Representative cardiac phenotypes of the sham injury group, injury control group, and dicaffeoyl glucose treatment group (Fig. Figure 1 Middle D, Figure 1 ZhongE and Figure 1 F) The experimental results showed that compared with the sham injury group ( Figure 2 Middle A and Figure 2 D), the ventricular volume of the injured control group was significantly decreased (49% ± 0.04) (Fig. Figure 2 Medium B and Figure 2 After dicaffeoyl glucose treatment, the volume of the heart ventricle (86% ± 0.02) was significantly higher than that of the injury control group (49% ± 0.04) (Fig. Figure 2 Middle B, Figure 2 Middle C and Figure 2 Middle D), indicating that dicaffeoyl glucose treatment can significantly improve ventricular volume after cardiac injury and rescue heart failure caused by irreversible damage.

[0049] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof in the preparation of a drug for treating and / or preventing 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 is an acute myocardial infarction caused by insufficient blood supply, myocardial hypoxia or excessive myocardial oxygen consumption due to various reasons.

4. A drug for treating and / or preventing ischemic heart disease, characterized in that: The drug comprises the dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof as claimed in claim 1.

5. The drug according to claim 4, characterized in that: The drug also includes one or more pharmaceutically acceptable carriers; the carriers are conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers or lubricants in the pharmaceutical field.

6. The drug according to claim 4, characterized in that: The drug can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, or mucosal tissue by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation.

7. Use of dicaffeoyl glucose or a pharmaceutically acceptable salt or ester thereof as claimed in claim 1 in the preparation of a drug for promoting cardiac injury repair and regeneration.

8. The use according to claim 7, characterized in that: The dicaffeoyl glucose can promote the repair and regeneration of zebrafish heart damage.

9. Use of dicaffeoyl glucose as claimed in claim 1 in preparing drugs for promoting cardiac injury repair and regeneration.

10. A drug for promoting the repair and regeneration of cardiac damage, characterized in that: The medicine comprises the dicaffeoyl glucose as claimed in claim 1.

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