Application of epihevetide in preparation of medicine for treating heart failure
By reversing heart failure using epiheveadride, the problems of toxic and inadequate supply of existing heart failure treatment drugs are solved, and new drugs with low toxic side effects and stronger therapeutic activity are provided to meet the needs of heart failure treatment.
Patent Information
- Application Number
- CN202411502656.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-25
- Publication Date
- 2025-01-10
AI Technical Summary
Existing heart failure treatment drugs have toxic side effects and cannot meet clinical needs, especially in end-stage patients, where the supply of heart transplant organs is limited and cannot meet the needs.
Epiheveadride is used as a treatment drug for heart failure. Epiheveadride and Pharmacy are used to reverse the problem of decreased heart rate, ejection fraction, cardiac output and shortening of the ventricular short axis, as well as the increase in end-diastolic ventricular volume, end-systolic ventricular volume and SV-BA distance. The combination of acceptable carriers or adjuvants on the above combinations form different dosage forms to improve therapeutic effect.
epiheveadride significantly reverses verapamil-induced heart failure, has low toxic side effects, and provides a new candidate as a heart failure drug, with broad market prospects and better therapeutic activity than digoxin.
Smart Images
Figure CN119280221A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to application of epiheveadride in preparing a heart failure therapeutic drug, belonging to the technical field of biomedicine. Background Art
[0002] The human heart consists of two atria and two ventricles, which provide oxygen and nutrition to the human body through systemic and pulmonary circulation. According to epidemiological surveys, cardiovascular disease has become the "number one killer" of human health. Among them, heart failure is a progressive cardiovascular disease, and its penetrance has shown a significant upward trend with age. Heart failure is characterized by the gradual deterioration of heart function, leading to arrhythmias and edema, and ultimately death. About 50% of patients diagnosed with heart failure may die within 5 years. Most heart failures are chronic processes, usually caused by long-term hypertension or cardiovascular disease. Acute heart failure may occur due to sudden deterioration of chronic heart failure, infection or chemotherapy. Commonly used drugs for the treatment of heart failure include hydrochlorothiazide, captopril, digoxin, metoprolol, etc., but these drugs have certain toxic side effects. Hydrochlorothiazide has a probability of causing patients to develop hyperuricemia, etc. Captopril must be used when heart function is asymptomatic to be effective, and digoxin will have poisoning symptoms such as arrhythmias when used. Once metoprolol is discontinued, clinical symptoms may worsen. Heart transplantation is the only effective treatment for patients with end-stage heart failure, but the supply of heart organs is limited and cannot meet the huge clinical demand.
[0003] Model organisms are important experimental subjects in the field of life science and medical research, and play a very important role in human disease research and drug development. Zebrafish is one of the commonly used model organisms. Compared with model organisms such as mice and rats, zebrafish have unique advantages such as small size, in vitro fertilization, in vitro development, strong fertility, rapid development, and full body transparency in early development. In addition, zebrafish are highly homologous to humans at the genome level. The heart of zebrafish consists of a single atrium and a single ventricle, and its development process is highly similar to that of mammals. About 5 hours after fertilization, the heart begins to develop; about 22 hours, the heart begins to contract; about 28 hours, the atrium and ventricle are formed; about 48 hours, the valve is formed, and the heart matures. Because the zebrafish is transparent in the early stage of development, its physiological phenomena such as heart beating and contraction can be clearly observed. In summary, zebrafish has excellent application potential in the fields of studying human heart disease, developing related therapeutic drugs, or screening and evaluating cardiotoxic substances.
[0004] Epiheveadride is an oxidized derivative of the nonsteroidal compound dihydroepiheveadride. Studies have found that epiheveadride has certain antibacterial activity and has excellent medicinal potential. However, research on epiheveadride in the heart is still very limited, especially as an anti-heart failure drug, which has not yet been reported. Summary of the invention
[0005] In view of the deficiencies in the prior art, the present invention provides an application of epiheveadride in preparing a drug for treating heart failure.
[0006] The technical solution of the present invention is as follows:
[0007] Use of epiheveadride in the preparation of a drug for treating heart failure, wherein the chemical structure of the epiheveadride is as shown in Formula I:
[0008]
[0009] Preferably according to the present invention, the epiheveadride can reverse the decrease in heart rate, ejection fraction, cardiac output and ventricular short-axis shortening rate and the increase in end-diastolic ventricular volume, end-systolic ventricular volume and SV-BA distance caused by heart failure.
[0010] A heart failure therapeutic drug comprising an effective amount of epiheveadride.
[0011] Preferably according to the present invention, the heart failure therapeutic drug contains one or more pharmaceutically acceptable carriers or adjuvants.
[0012] More preferably, the auxiliary agent is at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption enhancer, a surfactant or a lubricant.
[0013] Preferably according to the present invention, the heart failure therapeutic drug can reverse the decrease in heart rate, ejection fraction, cardiac output and ventricular short-axis shortening rate and the increase in end-diastolic ventricular volume, end-systolic ventricular volume and SV-BA distance caused by heart failure.
[0014] Preferably according to the present invention, the drug for treating heart failure is in the form of tablets, pills, capsules, oral liquid or injection.
[0015] The beneficial effects of the present invention are as follows:
[0016] The present invention first discovered that epiheveadride has the effect of treating heart failure. The present invention used zebrafish as experimental animals for detection and found that epiheveadride can effectively reverse heart failure induced by verapamil, a non-dihydropyridine calcium channel blocker, and has a significant effect of treating heart failure in the body with little toxic side effects. The present invention can provide candidates for anti-heart failure drugs and has broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 Evaluate photos for safety;
[0018] In the figure: 0 represents the blank control group, 10 represents the group treated with 10 μM epiheveadride concentration, 20 represents the group treated with 20 μM epiheveadride concentration, and 40 represents the group treated with 40 μM epiheveadride concentration.
[0019] Figure 2 These are photos of the heart after treatment with different concentrations of epiheveadride;
[0020] In the figure: 0 represents the blank control group, verapamil represents the modeling group, 10 represents the 10 μM epiheveadride treatment group, 20 represents the 20 μM epiheveadride treatment group, and 40 represents the 40 μM epiheveadride treatment group.
[0021] Figure 3 The bar graph shows the statistical data of cardiac function indexes of zebrafish treated with different concentrations of epiheveadride;
[0022] In the figure: Ctl represents the blank control group, verapamil represents the modeling group, 10 represents the 10μM epiheveadride treatment group, 20 represents the 20μM epiheveadride treatment group, and 40 represents the 40μM epiheveadride treatment group. ### represents P<0.0002 compared with the blank control group; ## represents P<0.0021 compared with the blank control group; # represents P<0.0332 compared with the blank control group. *** represents P<0.0002 compared with verapamil; ** represents P<0.0021 compared with verapamil; * represents P<0.0332 compared with verapamil.
[0023] Figure 4 This is a photo of the heart after treatment with 10 μg / mL digoxin and 40 μM epiheveadride;
[0024] In the figure: 0 represents the blank control group, verapamil represents the modeling group, Y represents the 10 μg / mL digoxin-positive drug group, and 40 represents the 40 μM epiheveadride treatment group.
[0025] Figure 5 This is a statistical bar graph of cardiac function indicators in zebrafish treated with 10μg / mL digoxin and 40μM epiheveadride;
[0026] In the figure: Ctl represents the blank control group, verapamil represents the modeling group, Y represents the 10μg / mL digoxin positive drug group, and 40 represents the 40μM epiheveadride treatment group. ### represents P<0.0002 compared with the blank control group; ## represents P<0.0021 compared with the blank control group; # represents P<0.0332 compared with the blank control group. *** represents P<0.0002 compared with verapamil; ** represents P<0.0021 compared with verapamil; * represents P<0.0332 compared with verapamil. DETAILED DESCRIPTION
[0027] The technical solution of the present invention is further described below in conjunction with the embodiments and the accompanying drawings of the specification, but the protection scope of the present invention is not limited thereto.
[0028] The contents without specific conditions in the examples were carried out under conventional conditions; the reagents or instruments used without indicating the manufacturer were all common commercially available products.
[0029] Epiheveadride can be prepared according to existing methods, such as referring to the method provided in the document "A Pair of New SpirocyclicAlkaloid Enantiomers with TrxR Inhibitory Activities Were Isolated from Marine-Derived Aspergillus ruber TX-M4-1" (WANG Cong, WANG Yufei, etc., 2023).
[0030] Example 1
[0031] Safety evaluation of epiheveadride
[0032] 1. Zebrafish sample pretreatment:
[0033] Healthy and sexually mature AB strain zebrafish were placed in a mating tank at a ratio of 1:1 between male and female, with a partition placed in the middle, and placed in a dark environment. The partition was removed before the lights were turned on the next day, and light stimulation was used to stimulate ovulation. The adult fish were removed half an hour after ovulation, and the ovulation time was controlled within half an hour to reduce the difference in development time between embryos. Fertilized eggs were collected, disinfected and cleaned; then the fertilized eggs were moved to clean zebrafish embryo culture water, and 0.2ppm methylene blue was added, and cultured at 28℃ for 12h.
[0034] The composition of embryo culture water is as follows:
[0035] CaCl2 0.4mM, NaCl 5mM, MgSO4 0.16mM, KCl 0.17mM, prepared with deionized water.
[0036] 2. epiheveadride treatment:
[0037] The zebrafish larvae described in step 1 were randomly divided into a blank control group and a group treated with different concentrations of epiheveadride, with 3 parallel wells in each group and 15 fish in each well. The following steps were performed: the embryo culture water in the 12-well plate was removed, 2 mL of embryo culture water was added to the control group, 2 mL of embryo culture water containing 10 μM epiheveadride was added to the epiheveadride treatment group (low-dose group), 2 mL of embryo culture water containing 20 μM epiheveadride was added to the epiheveadride treatment group (medium-dose group), and 2 mL of embryo culture water containing 40 μM epiheveadride was added to the epiheveadride treatment group (high-dose group), and the cells were cultured in a 28°C constant temperature incubator for 72 h.
[0038] 3. Epiheveadride safety test:
[0039] The four groups of zebrafish were placed in a tricaine solution with a mass concentration of 0.02% to anesthetize them. Images of 24hpf, 48hpf, and 72hpf zebrafish were collected under a microscope, and the hatching rate and mortality rate were calculated.
[0040] The experimental results are as follows Figure 1 As shown in the figure, compared with the blank control group, the zebrafish treated with different concentrations of epiheveadride developed normally at 24hdf, without death or deformity; all hatched at 48hdf, without death or deformity; and no death or deformity at 72hdf. This result shows that when the concentration of epiheveadride is below 40μM, it is non-toxic to zebrafish embryos and larvae.
[0041] Example 2
[0042] Therapeutic effects of different concentrations of epiheveadride on heart failure in zebrafish
[0043] 1. Zebrafish sample pretreatment:
[0044] Healthy and sexually mature cardiac fluorescent transgenic zebrafish Tg (cmlc2: EGFP) were placed in a mating tank at a ratio of 1:1 between male and female, with a partition placed in the middle, and placed in a dark environment. The partition was removed before the lights were turned on the next day, and light stimulation was used to stimulate ovulation. The adult fish were removed half an hour after ovulation, and the ovulation time was controlled within half an hour to reduce the difference in development time between embryos. Fertilized eggs were collected, disinfected and cleaned; then the fertilized eggs were moved into clean zebrafish embryo culture water, and 0.2ppm methylene blue was added, and the light was controlled and cultured for 48 hours at 28°C and a 14h light / 10h dark cycle.
[0045] 2. epiheveadride treatment:
[0046] The zebrafish larvae described in step 1 were randomly divided into a blank control group, a model group, and a group treated with different concentrations of epiheveadride, with 3 parallel wells in each group and 10 zebrafish in each well. The larvae were treated by the following steps: (1) The embryo culture water in the 24-well plate was removed, and 2 mL of embryo culture water was added to the control group and the model group, 2 mL of embryo culture water containing 10 μM epiheveadride was added to the low-dose epiheveadride treatment group, 2 mL of embryo culture water containing 20 μM epiheveadride was added to the medium-dose epiheveadride treatment group, and 2 mL of embryo culture water containing 40 μM epiheveadride was added to the high-dose epiheveadride treatment group. (1) The cells were incubated in a 28°C incubator with embryo culture water containing epiheveadride for 4 h. (2) The embryo culture water in the 24-well plate was then removed, and 2 mL of embryo culture water was added to the control group, and 2 mL of embryo culture water containing 200 μM verapamil was added to the modeling group and the groups treated with different concentrations of epiheveadride, and the cells were cultured in a 28°C incubator for 30 min.
[0047] 3. Zebrafish heart function index detection:
[0048] The zebrafish in each group were placed in a tricaine solution with a mass concentration of 0.02% to anesthetize them. Heart images showing green fluorescence were collected under a fluorescence microscope. According to the collected images, the image-pro plus 6.0 software was used to calculate the heart rate, end-diastolic ventricular volume, end-systolic ventricular volume, ejection fraction, cardiac output, ventricular short axis shortening rate, and SV-BA distance of each group of zebrafish. GraphPad Prism 8.0.2 software was used for data processing.
[0049] The experimental results are as follows Figure 2 , 3 As shown in the figure, compared with the blank control group, the heart function of zebrafish in the modeling group decreased, including: decreased heart rate, increased ventricular volume at the end of diastole and systole, decreased ejection fraction, cardiac output and ventricular short axis shortening rate, increased distance between venous sinus (SV) and arterial bulb (BA) (SV-BA), etc., and the above differences were extremely statistically significant, indicating that verapamil induced heart failure in zebrafish. Compared with the modeling group, the heart function of zebrafish in each dose group of epiheveadride was significantly restored, including: increased heart rate, reduced ventricular volume at the end of diastole and systole, increased ejection fraction, cardiac output and ventricular short axis shortening rate, and decreased SV-BA distance. This shows that epiheveadride effectively reversed the heart failure of zebrafish caused by verapamil, and the reversal effect was proportional to the concentration of epiheveadride.
[0050] Example 3
[0051] Comparison of the therapeutic effects of digoxin and epiheveadride on heart failure in zebrafish
[0052] 1. The pretreatment of zebrafish samples was the same as in Example 2.
[0053] 2. epiheveadride treatment:
[0054] The zebrafish larvae described in step 1 were randomly divided into a blank control group, a modeling group, a digoxin-positive drug group, and an epiheveadride-treated group, with 3 parallel wells in each group and 10 fish in each well. The following steps were performed: (1) The embryo culture water in the 12-well plate was removed, and 2 mL of embryo culture water was added to the control group and the modeling group, 2 mL of embryo culture water containing 10 μg / mL digoxin was added to the digoxin-positive drug group, and 2 mL of embryo culture water containing 40 μM verapamil was added to the epiheveadride-treated group, and the cells were cultured in a 28°C constant temperature incubator for 4 h; (2) The embryo culture water in the 12-well plate was removed, 2 mL of embryo culture water was added to the control group, and 2 mL of embryo culture water containing 200 μM verapamil was added to the modeling group, the digoxin-positive drug group, and the epiheveadride-treated group, and the cells were cultured in a 28°C constant temperature incubator for 30 min.
[0055] 3. The detection of zebrafish cardiac function indexes is the same as in Example 2.
[0056] The experimental results are as follows Figure 4 , 5As shown in the figure, compared with the blank control group, the heart rate of zebrafish in the modeling group was significantly reduced, and the ventricular volume at the end of diastole and systole and the SV-BA distance were significantly increased. This shows that verapamil caused heart failure in zebrafish and the modeling was successful. Compared with the modeling group, both digoxin and epiheveadride effectively improved the cardiac function of zebrafish, and their heart rate, ventricular volume at the end of diastole and systole and SV-BA distance were restored. Statistical tests showed that epiheveadride had a better effect than digoxin in restoring the heart rate, ventricular volume at the end of diastole and systole and SV-BA distance of zebrafish.
[0057] The present invention discovers for the first time that epiheveadride has a therapeutic effect on heart failure, and compared with digoxin, a known drug for the treatment of heart failure, epiheveadride has a stronger activity for the treatment of heart failure. The present invention can provide a candidate for a drug for the treatment of heart failure, and has a broad market prospect.
Claims
1. Use of epiheveadride in the preparation of a drug for treating heart failure, wherein the chemical structure of epiheveadride is shown in Formula I:
2. The use according to claim 1, characterized in that: The epiheveadride can reverse the decrease in heart rate, ejection fraction, cardiac output and ventricular short-axis shortening rate and the increase in end-diastolic ventricular volume, end-systolic ventricular volume and SV-BA distance caused by heart failure.
3. A drug for treating heart failure, characterized in that: The medicament includes an effective amount of epiheveadride.
4. The heart failure therapeutic drug according to claim 3, characterized in that: The medicine contains one or more pharmaceutically acceptable carriers or adjuvants.
5. The heart failure therapeutic drug according to claim 4, characterized in that: The auxiliary agent is at least one of a sustained-release agent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant or a lubricant.
6. The heart failure therapeutic drug according to claim 3, characterized in that: The drug can reverse the decrease in heart rate, ejection fraction, cardiac output and ventricular short axis shortening rate and the increase in end-diastolic ventricular volume, end-systolic ventricular volume and SV-BA distance caused by heart failure.
7. The heart failure therapeutic drug according to claim 3, characterized in that: The drug dosage form is tablet, pill, capsule, oral solution or injection.