Use of meroterpenoid compound i in preparation of heart failure prevention drug

Abstract

A use of a meroterpenoid compound I in the preparation of a heart failure prevention drug. By using a meroterpenoid compound I isolated and purified from the marine fungus Aspergillus terreus GZU-311 to treat mouse heart failure models induced by doxorubicin, left anterior descending coronary artery ligation, or angiotensin II, it was found that the meroterpenoid compound I can improve cardiac function in mice, reduce cardiomyocyte injury, inhibit myocardial connective tissue thickening, improve disordered arrangement of cardiomyocytes, and reduce inflammatory cell infiltration, thereby having a heart failure prevention effect. Accordingly, the use of the meroterpenoid compound I in the preparation of a heart failure prevention drug is provided. The drug source molecules that can be used for treating heart failure are enriched, which is helpful for the development of heart failure prevention drugs and clinical treatment using same.

Description

Application of heteroterpenoid compound I in the preparation of drugs for treating heart failure Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of heteroterpenoid compound I in the preparation of drugs for treating heart failure. Background Technology

[0002] Heart failure, or HF for short, is a pathophysiological state and clinical syndrome in which the heart's pumping function is reduced, making it unable to adequately pump blood returning from the veins to meet the metabolic needs of the body's tissues. It is characterized by high readmission rates, high disability rates, and high mortality rates.

[0003] Currently, commonly used anti-heart failure drugs in clinical practice mainly include renin-angiotensin system inhibitors, anti-aldosterone drugs, diuretics, beta-blockers, and cardiac glycosides. While these drugs all have certain anti-heart failure effects in clinical practice, long-term use can lead to drug tolerance and a series of side effects and adverse reactions, such as orthostatic hypotension, depression, hyperlipidemia, hypoglycemia, peripheral circulatory disturbances, fatigue, bronchospasm, inhibition of myocardial contractility, and conduction block. Therefore, there is an urgent need to find new drug molecules to replace existing clinical drugs in order to address patient tolerance to existing anti-heart failure drugs and the adverse reactions they cause.

[0004] Secondary metabolites of marine fungi are an important component of natural products, characterized by sustainability, environmental friendliness, rich diversity of metabolites, and minimal side effects, making them a crucial source for natural drug screening. Furthermore, these secondary metabolites possess a wide range of physiological activities, including antibacterial, antitumor, antifibrotic, anti-Alzheimer's disease, anti-heart failure, and enzyme inhibition, injecting new impetus into modern drug innovation and the development of innovative drugs. Currently, the search for new drug molecules from marine fungi has become a research hotspot both internationally and domestically.

[0005] While the development of cardiovascular disease is closely linked to inflammation and myocardial fibrosis, not every compound with anti-inflammatory or anti-fibrotic effects can be used to treat heart failure. For example, drugs prepared from thiazolidinediones can inhibit renal inflammation, suppress renal cell fibrosis, and improve renal microcirculation (DOI:10.3969 / j.issn.1672-6170.2009.01.044.). However, these drugs may cause water and sodium retention and fluid overload, inducing or worsening heart failure and increasing the risk of death from heart failure or cardiovascular disease. Although marine fungi have abundant secondary metabolites, obtaining compounds with anti-heart failure effects from them is not easy. Summary of the Invention

[0006] In view of the above-mentioned problems in the prior art, the present invention provides a heteroterpenoid compound that can be used to prepare drugs for treating heart failure, namely, heteroterpenoid compound I of the present invention.

[0007] The first objective of this invention is to provide the use of heteroterpenoid compound I in the preparation of drugs for treating heart failure.

[0008] A second objective of this invention is to provide a drug for treating heart failure.

[0009] The above-mentioned objective of this invention is achieved through the following technical solution:

[0010] This invention utilizes heteroterpenoid compound I, isolated from a fermentation culture of the marine fungus *Aspergillus terreus* GZU-311, to treat mouse models of heart failure induced by doxorubicin, ligation of the left anterior descending coronary artery, or angiotensin II. The results show that heteroterpenoid compound I exhibits anti-heart failure effects. Therefore, this invention seeks protection for the use of said heteroterpenoid compound I in the preparation of anti-heart failure drugs.

[0011] Specifically, the structural formula of the heteroterpene compound I is shown in formula (I):

[0012] Formula (I).

[0013] Specifically, the heteroterpene compound I can improve cardiac function.

[0014] Specifically, the improvement in cardiac function refers to the ability of the heteroterpene compound I to increase cardiac ejection fraction and fractional shortening.

[0015] Specifically, the heteroterpene compound I can reduce systolic and diastolic blood pressure.

[0016] Specifically, the heteroterpene compound I can reduce myocardial cell damage.

[0017] Specifically, the heteroterpene compound I can inhibit the thickening of myocardial connective tissue.

[0018] Specifically, the heteroterpene compound I can improve the disordered arrangement of cardiomyocytes.

[0019] Specifically, the heteroterpene compound I can reduce inflammatory cell infiltration.

[0020] Specifically, the heart failure is chronic heart failure or ischemic heart failure.

[0021] Specifically, the heart failure is doxorubicin- or angiotensin II-induced heart failure.

[0022] Specifically, the heteroterpene compound I was isolated and purified from the fermentation culture of the marine fungus Aspergillus terreus GZU-311.

[0023] Specifically, the culture medium used for fermentation is a solid corn culture medium prepared from corn and seawater.

[0024] Specifically, the preparation method of the heteroterpene compound I includes the following steps:

[0025] S1. Preparation of seed culture of marine fungus Aspergillus terreus GZU-311;

[0026] S2. The seed culture prepared in S1 is inoculated into a solid corn culture medium and cultured to obtain a fermentation culture;

[0027] S3. Extract the fermentation culture obtained from S2 with methanol 2 to 5 times, concentrate the extract, and extract the concentrated extract with ethyl acetate to obtain crude ethyl acetate extract;

[0028] S4. After separating the crude ethyl acetate extract obtained in S3 by normal-phase silica gel chromatography, it was sequentially eluted with petroleum ether / ethyl acetate mixed solutions with volume fractions of 10%, 20%, 30%, 40%, 50%, 60%, and 70% respectively. The 40% petroleum ether / ethyl acetate eluent fraction was collected and then separated and purified by silica gel, gel chromatography, and C-18 reversed-phase column chromatography to obtain the heteroterpene compound I.

[0029] Specifically, the marine fungus Aspergillus terreus strain GZU-311 described in S1 was deposited at the Guangdong Provincial Center for Microbial Culture Collection on December 17, 2019, with accession number GDMCC No: 60789.

[0030] Specifically, the seed culture preparation method described in S1 is as follows: the marine fungus Aspergillus terreus GZU-311 is inoculated into a slant culture medium and then inoculated into a liquid culture medium to obtain the seed culture; the culture temperature is 28 ℃~35 ℃ and the culture time is 4~10 days.

[0031] More specifically, the culture temperature was 30 ℃ and the culture time was 6 days.

[0032] Specifically, S2 involves inoculating the seed culture prepared in S1 into a solid corn culture medium and allowing it to be cultured statically to obtain a fermentation culture; the culture temperature is 28 ℃~35 ℃, and the culture time is 30~60 days.

[0033] More specifically, the culture temperature was 30 ℃ and the culture time was 60 days.

[0034] Specifically, the formula of the slant culture medium described in S1 is as follows: by mass ratio, glucose 0.3%, yeast extract 0.1%, peptone 0.1% to 0.5%, agar 1.5% to 2.5%, sodium chloride 1.5% to 4%, and water to make up to 100%.

[0035] More specifically, the formulation of the slant culture medium described in S1 is as follows: by mass ratio, glucose 0.3%, yeast extract 0.1%, peptone 0.5%, agar 2.5%, sodium chloride 3%, and water to make up to 100%.

[0036] Specifically, the formula for the solid corn culture medium described in S2 is: corn: seawater = 1:1~2 by mass ratio.

[0037] More specifically, the formula for the solid corn culture medium described in S2 is: corn: seawater = 1:1 by mass.

[0038] Specifically, the methanol extraction in S3 is performed three times.

[0039] This invention provides a drug for treating heart failure, which contains the heteroterpenoid compound I described in this invention or a pharmaceutically acceptable salt thereof.

[0040] Specifically, the drug also includes a heteroterpene compound I or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients.

[0041] Specifically, the heteroterpene compound I can be administered alone or in the form of a pharmaceutical composition. The pharmaceutical composition can be formulated into various suitable pharmaceutical preparations depending on the route of administration.

[0042] Optionally, the pharmaceutical preparation is an oral preparation, an injectable preparation, a topical transdermal preparation, or a sustained-release preparation.

[0043] Specifically, the oral formulation can be formulated in any orally acceptable form, including but not limited to tablets, capsules, aqueous solutions, or aqueous suspensions. The carriers used in the tablets generally include lactose and corn starch, and lubricants such as magnesium stearate may also be added. The diluents used in the capsule formulations generally include lactose and dried corn starch. The aqueous suspensions typically involve mixing the active ingredient with suitable emulsifiers and suspending agents.

[0044] Optionally, sweeteners, flavorings, or colorings may also be added to the above-mentioned oral formulations.

[0045] Specifically, the injectable formulation includes liquid injections, powders for injection, or tablets for injection. The injectable formulation can be administered in a sterile injectable form, including sterile water or oil suspensions or sterile injectable solutions. Optional carriers and solvents used in the injectable formulation include water, Ringer's solution, isotonic sodium chloride solution, and sterile non-volatile oils such as monoglycerides and diglycerides.

[0046] Specifically, the topical transdermal preparations include sprays, gels, emulsions, ointments, lotions, creams, or gel patches. The carriers of the ointments include, but are not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsified wax, and water. The carriers of the lotions or creams include, but are not limited to: mineral oil, sorbitan monostearate, Tween 60, hexadecyl ester wax, hexadecene aromatic alcohol, 2-octyldodecyl alcohol, benzyl alcohol, and water.

[0047] Specifically, the pharmaceutically acceptable excipients include one or more of the following: drug carriers, surfactants, buffers, disintegrants, binders, fillers, lubricants, excipients, solubilizers, flavorings, and colorings.

[0048] Specifically, the pharmaceutical preparation can be administered in any of the following ways: orally, by spray inhalation, rectal administration, nasal administration, topical administration, and enteral administration; wherein, enteral administration includes subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, and / or intracranial injection or infusion, or administration via an external implantation device.

[0049] Specifically, the drug formulation is preferably administered orally, intraperitoneally, or intravenously.

[0050] It should also be noted that the dosage and method of use of the heteroterpenoid compound I described in this invention depend on many factors, including the patient's age, weight, sex, natural health condition, nutritional status, the activity intensity of the compound, the time of administration, the metabolic rate, the severity of the disease, and the subjective judgment of the treating physician.

[0051] The present invention has the following beneficial effects:

[0052] This invention utilizes a heteroterpene compound I, isolated and purified from the marine fungus *Aspergillus terreus* GZU-311, to treat a mouse heart failure model induced by doxorubicin, ligation of the left anterior descending coronary artery, or angiotensin II. The results show that heteroterpene compound I can improve cardiac function in mice, reduce cardiomyocyte damage, inhibit myocardial connective tissue thickening, improve cardiomyocyte disorganization, and reduce inflammatory cell infiltration. Therefore, this invention provides the application of heteroterpene compound I in the preparation of anti-heart failure drugs. This invention enriches the pool of drug molecules that can be used to treat heart failure, contributing to the development of anti-heart failure drugs and their clinical application, and has promising clinical prospects. Attached Figure Description

[0053] Figure 1 shows the echocardiograms of the doxorubicin-induced mouse heart failure model and the mouse heart failure model treated with the aforementioned heteroterpene compound I.

[0054] Figure 2 shows the cardiac function test results of the mouse heart failure model induced by doxorubicin and the mouse heart failure model treated with the aforementioned heteroterpene compound I; A in the figure is the calculated ejection fraction (EF), and B in the figure is the calculated fractional shortening (FS); # in the figure indicates P < 0.05; This indicates that P < 0.05; This indicates that P < 0.01; This means P < 0.001.

[0055] Figure 3 shows HE staining images of heart sections from a mouse heart failure model induced by doxorubicin and a mouse heart failure model treated with the aforementioned heteroterpene compound I. The scale bar is 100 μm.

[0056] Figure 4 shows Masson staining images of heart sections from a mouse heart failure model induced by doxorubicin and a mouse heart failure model treated with the aforementioned heteroterpene compound I. The scale bar is 50 μm.

[0057] Figure 5 shows the echocardiograms of a mouse heart failure model induced by ligation of the left anterior descending coronary artery and the mouse heart failure model treated with the aforementioned heteroterpene compound I.

[0058] Figure 6 shows the cardiac function test results of a mouse heart failure model induced by ligation of the left anterior descending coronary artery and the mouse heart failure model treated with the aforementioned heteroterpene compound I; A in the figure is the calculated ejection fraction (EF), and B in the figure is the calculated fractional shortening (FS); ### in the figure indicates P < 0.001; This means P < 0.001.

[0059] Figure 7 shows HE staining images of heart sections from a mouse heart failure model induced by ligation of the left anterior descending coronary artery and a mouse heart failure model treated with the aforementioned heteroterpene compound I. The scale bar is 50 μm.

[0060] Figure 8 shows the diastolic and systolic blood pressure of a mouse model of angiotensin II-induced heart failure and mice treated with the aforementioned heteroterpene compound I.

[0061] Figure 9 shows echocardiograms of a mouse heart failure model induced by angiotensin II and a mouse heart failure model treated with the aforementioned heteroterpene compound I.

[0062] Figure 10 shows the cardiac function test results of the angiotensin II-induced heart failure model in mice and the mouse heart failure model treated with the aforementioned heteroterpene compound I; in the figure This indicates that P < 0.05; This means P < 0.0001. Embodiments of the present invention

[0063] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0064] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0065] Example 1: Obtaining Heteroterpenoid Compound I from Marine Fungi

[0066] The heteroterpene compound I described in this invention was isolated and purified from the fermentation culture of the marine fungus Aspergillus terreus GZU-311. The marine fungus Aspergillus terreus GZU-311 was deposited on December 17, 2019, at the Guangdong Provincial Microbial Culture Collection Center (GDMCC No.: 60789), located at the Guangdong Institute of Microbiology, 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Relevant isolation and identification information has been published in patent application CN111139188A.

[0067] The isolation, purification, and identification process of the heteroterpene compound I is as follows:

[0068] 1. Separation of compounds

[0069] The method for separating compounds includes the following steps:

[0070] (1) The marine fungus Aspergillus terreus GZU-311 was inoculated into a slant culture medium (the formula of the slant culture medium is: by mass ratio, glucose 0.3%, yeast extract 0.1%, peptone 0.5%, agar 2.5%, sodium chloride 3%, water to make up to 100%), and then inoculated into a liquid culture medium (the formula is the same as that of the slant culture medium, except that it does not contain agar), and cultured at 30 ℃ for 6 days to obtain the seed culture;

[0071] (2) The seed culture was inoculated into solid corn culture medium (the formula of solid corn culture medium is: corn: seawater = 1:1 by mass ratio), and allowed to stand at 30 ℃ for 60 days to obtain fermentation culture (fermentation stock solution).

[0072] (3) The fermentation culture was extracted with methanol three times, the extract was concentrated into a paste, and the concentrated extract was extracted with ethyl acetate to obtain crude ethyl acetate extract;

[0073] (4) After separating the crude ethyl acetate extract by normal phase silica gel chromatography, it was eluted sequentially with petroleum ether / ethyl acetate mixed solutions with volume fractions of 10%, 20%, 30%, 40%, 50%, 60%, and 70% respectively. The 40% petroleum ether / ethyl acetate eluent was collected and then separated and purified sequentially by silica gel, gel and C-18 reversed phase column chromatography. The purified compounds were identified.

[0074] 2. Identification of compounds

[0075] The structures of the compounds obtained from the above separation and purification were analyzed using high-resolution mass spectrometry and nuclear magnetic resonance (NMR), and the following experimental data were obtained:

[0076] The molecular formula of the compound is: C 26 H 32 O9, High-resolution mass spectrometry (HRESI-MS): 487.1959 [MH] - (cald for C) 26 H 31 O9487.19626).

[0077] The structural formula of the compound is shown below (Formula (I)):

[0078] Formula (I).

[0079] The NMR data of the compounds are shown in Table 1.

[0080] Table 1 NMR data (CDCl3, 400 MHz / 100 MHz, ppm)

[0081]

[0082] Upon identification, the purified compound was determined to be a heteroterpene compound, which was named Heterpene Compound I.

[0083] Example 2: The role of heteroterpenoid compound I in the treatment of doxorubicin-induced heart failure

[0084] 1. Experimental Methods

[0085] Eight-week-old male C57BL / 6 mice were randomly divided into six groups (n=8 per group): control group, model group, low-dose group of heteroterpenoid compound I (I (10 mg / kg)), medium-dose group of heteroterpenoid compound I (I (20 mg / kg)), high-dose group of heteroterpenoid compound I (I (40 mg / kg)) and positive control group of satrocubitril / valsartan (XST).

[0086] The daily oral gavage dosages were as follows: triterpenoid compound I (dissolved in 10% DMSO + physiological saline) was administered at doses of 10, 20, and 40 mg / kg; positive control drug XST was administered at 40 mg / kg; and the control and model groups received an equal volume of blank solvent (10% DMSO + physiological saline). Simultaneously, doxorubicin was administered subcutaneously once weekly at a single dose of 5 mg / kg, while the control group received an equal volume of physiological saline. Specific oral gavage dosages were determined based on the mice's body weight; for example, "10 mg / kg" means 10 mg for a mouse weighing 1 kg.

[0087] On day 29 after gavage administration, the cardiac structure and function of mice in each group were evaluated using the VINNO 6 small animal high-resolution micro-ultrasound imaging system, and the ejection fraction (EF) and fractional shortening (FS) were calculated. Subsequently, the mice were dissected, and embedded sections of the heart were obtained for HE staining and Masson staining.

[0088] 2. Data Processing

[0089] The experimental results were statistically analyzed using GraphPad Prism 8.0 software. The results are expressed as Mean±SEM, and one-way ANOVA was used for inter-group comparisons.

[0090] 3. Experimental Results

[0091] (1) Evaluation of cardiac function

[0092] Figure 1 shows the echocardiograms of the doxorubicin-induced mouse heart failure model and the mouse heart failure model treated with the aforementioned heteroterpene compound I; Figure 2 shows the cardiac function test results of the doxorubicin-induced mouse heart failure model and the mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figures 1 and 2, compared to the control group, the model group mice showed significantly decreased cardiac ejection fraction (EF) and fractional shortening (FS), and significantly increased left ventricular end-systolic and end-diastolic volumes, indicating a significant decrease in doxorubicin-induced cardiac function. In contrast, mice treated with heteroterpene compound I showed significantly increased cardiac ejection fraction and fractional shortening, and decreased left ventricular end-systolic and end-diastolic volumes, which significantly improved doxorubicin-induced cardiac function in mice.

[0093] (2) HE staining of cardiac sections

[0094] Figure 3 shows HE staining images of heart sections from a mouse heart failure model induced by doxorubicin and a mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figure 3, compared to the control group, the model group mice showed significant cardiomyocyte damage, while heart sections from mice treated with 40 mg / kg heteroterpene compound I showed little or no cardiomyocyte damage, with cells arranged regularly and densely. This indicates that the aforementioned heteroterpene compound I can reduce cardiomyocyte damage and has a certain protective effect on cardiac tissue.

[0095] (3) Masson staining of heart sections

[0096] Figure 4 shows Masson staining images of heart sections from a mouse heart failure model induced by doxorubicin and a mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figure 4, the control group showed only a small area of ​​connective tissue thickening, while the model group showed a significant and large area of ​​connective tissue thickening. Compared with the model group, treatment with heteroterpene compound I at a dose of 40 mg / kg significantly reduced doxorubicin-induced connective tissue thickening.

[0097] Example 3: Effect of heteroterpenoid compound I on ischemic heart failure induced by ligation of the left anterior descending coronary artery.

[0098] 1. Experimental Methods

[0099] (1) Experimental materials

[0100] Eight-week-old male C57BL / 6 mice were purchased from Guangdong Zhiyuan Biomedical Technology Co., Ltd., and were fed standard feed with free access to water and food.

[0101] (2) Experimental modeling

[0102] The model was established by in situ ligation of the left anterior descending coronary artery (LAD) via open-chest surgery. Mice were first anesthetized by intraperitoneal injection of 0.25% sodium pentobarbital, and a ventilator cannula was inserted. Ventilation parameters were: tidal volume 11cc, respiratory ratio 1:2, respiratory rate 144 breaths / min. The wound was covered with saline-soaked gauze. A 1 cm incision was made in the intercostal space between the 3rd and 4th ribs on the left side of the sternum. Two retractors were used to open the heart, exposing it. The pericardium was opened, and a standard 6-0 silk suture and a round needle were used to insert the needle at the bifurcation of the left coronary artery and the posterior descending artery, approximately 3-4 mm below the left atrial appendage, to a depth of about 1 mm and a width of about 2 mm. After insertion, the suture was tightened, and the electrocardiogram was immediately observed. The chest was closed upon the appearance of ST segment elevation or a wide QRS complex. Four weeks post-surgery, small animal ultrasound was performed to assess cardiac function. Successful modeling was defined as an ejection fraction (EF) of less than 45%.

[0103] (3) Experimental grouping and data measurement

[0104] The experimental mice were divided into the following 4 groups (5 mice in each group): blank control group (thoracotomy and suturing, without ligation of coronary arteries, Sham), model group (CHF), model group + heteroterpenoid compound I group (I (20 mg / kg)) and model group + sacubitril / valsartan group (XST).

[0105] The dosages administered via gavage were as follows: the model group plus group I of heteroterpenoids (dissolved in 10% DMSO + physiological saline) was administered at 20 mg / kg; the model group plus sacubitril / valsartan was administered at 26 mg / kg; the blank control group and the model group were given an equal volume of solvent (10% DMSO + physiological saline). The specific gavage dosage was determined based on the mouse's body weight; for example, "20 mg / kg" means that a mouse weighing 1 kg would receive 20 mg of the drug. The medication was administered via gavage once daily for 4 weeks.

[0106] On day 29 after gavage administration, the cardiac structure and function of mice in each group were evaluated using the VINNO 6 small animal high-resolution micro-ultrasound imaging system, and the ejection fraction (EF) and fractional shortening (FS) were calculated. Subsequently, the mice were dissected, and embedded sections of the heart were obtained for HE staining.

[0107] 2. Data Processing

[0108] Experimental data were statistically analyzed using GraphPad Prism 8.0 software. The results are expressed as Mean±SEM, and one-way ANOVA was used for intergroup comparisons.

[0109] 3. Experimental Results

[0110] (1) Evaluation of cardiac function

[0111] Figure 5 shows the echocardiograms of the mouse heart failure model induced by ligation of the left anterior descending coronary artery and the mouse heart failure model treated with the aforementioned heteroterpene compound I. Figure 6 shows the cardiac function test results of the mouse heart failure model induced by ligation of the left anterior descending coronary artery and the mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figure 6, compared with the blank control group, the ejection fraction (EF) and fractional shortening (FS) of the heart in the model group mice were significantly decreased, and the cardiac function of the mice in the model group was significantly reduced. Compared with the model group, treatment with heteroterpene compound I at a dose of 20 mg / kg can significantly improve the EF and FS values ​​of the heart failure model induced by ligation of the left anterior descending coronary artery and improve the cardiac function of mice with heart failure.

[0112] (2) HE staining of cardiac sections

[0113] Figure 7 shows HE staining images of heart sections from a mouse heart failure model induced by ligation of the left anterior descending coronary artery and a mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figure 7, compared with the control group, the model group exhibited disordered cardiomyocyte arrangement and extensive inflammatory cell infiltration. Compared with the model group, treatment with heteroterpene compound I at a dose of 20 mg / kg significantly reduced inflammatory cell infiltration and maintained cardiomyocyte arrangement.

[0114] Example 4: The role of heteroterpenoid compound I in the treatment of angiotensin II-induced heart failure.

[0115] 1. Experimental Methods

[0116] Six male C57BL / 6J mice aged 6–8 weeks were randomly assigned to the control group (n=6) and treated with PBS, while the remaining mice were treated with angiotensin II (Ang II) to establish a chronic heart failure model. Ang II was dissolved in sterile PBS containing 0.01 mol / L acetic acid. Under sterile conditions, the Ang II solution was carefully loaded into an Alzet osmotic pump to avoid air bubble formation. The osmotic pump was immersed in physiological saline and incubated at 37 °C for 48 h to activate its osmotic system (flow rate set at 1.5 μg / min / kg). After gas anesthesia with isoflurane, an incision of about 1 cm was made in the back of the neck of each mouse, and the osmotic pump was implanted subcutaneously. The control group mice were implanted with osmotic pumps containing PBS. Postoperatively, each mouse was injected intraperitoneally with 600 units of penicillin for three consecutive days to prevent infection.

[0117] Two weeks after modeling, blood pressure was measured. The successfully modeled mice were divided into 6 groups (n=6 per group): Model group: continued Ang II intervention; Positive drug group (LCZ696): given sacubitril / valsartan sodium tablets at a dose of 30 mg / kg (sacubitril / valsartan dissolved in ddH2O); Low-dose group of heteroterpenoid compound I (I 2.5 mg / kg): given heteroterpenoid compound I at a dose of 2.5 mg / kg; Medium-dose group of heteroterpenoid compound I (I 5 mg / kg): given heteroterpenoid compound I at a dose of 5 g / kg; High-dose group of heteroterpenoid compound I (I 10 mg / kg): given heteroterpenoid compound I at a dose of 10 mg / kg; Control group: received only PBS treatment. The daily oral gavage dosages were as follows: terpene compound I (dissolved in 10% DMSO + physiological saline) was administered at doses of 2.5, 5, and 10 mg / kg, respectively; the positive control drug LCZ696 was administered at 30 mg / kg; the control and model groups received an equal volume of blank solvent (10% DMSO + physiological saline). For 4 weeks, blood pressure was monitored weekly from start to finish. At the end of the experiment, echocardiography and blood pressure were performed, and blood and heart samples were collected from the mice and stored at -80°C. Four weeks after oral gavage administration, the cardiac structure and function of the mice in each group were evaluated using a VINNO 6 small animal high-resolution micro-ultrasound imaging system, and ejection fraction (EF) and fractional shortening (FS) were calculated.

[0118] 2. Data Processing

[0119] The experimental results were statistically analyzed using GraphPad Prism 8.0 software. The results are expressed as Mean±SEM, and one-way ANOVA was used for inter-group comparisons.

[0120] 3. Experimental Results

[0121] (1) Blood pressure assessment

[0122] Figure 8 shows the angiotensin II-induced heart failure model in mice and the diastolic and systolic blood pressure of mice treated with the aforementioned heteroterpene compound I. As shown in Figure 8, compared with the control group, the systolic and diastolic blood pressure of the model group mice were significantly increased; compared with the model group, treatment with heteroterpene compound I significantly reduced the systolic and diastolic blood pressure of the mice.

[0123] (2) Evaluation of cardiac function

[0124] Figure 9 shows the echocardiograms of the angiotensin II-induced heart failure model in mice and the mouse heart failure model treated with the aforementioned heteroterpene compound I; Figure 10 shows the cardiac function test results of the angiotensin II-induced heart failure model in mice and the mouse heart failure model treated with the aforementioned heteroterpene compound I. As shown in Figures 9 and 10, compared with the control group, the ejection fraction (EF) and fractional shortening (FS) of the heart in the model group mice were significantly decreased, indicating a significant reduction in angiotensin II-induced cardiac function. Compared with the model group, the ejection fraction and fractional shortening of the heart in mice treated with heteroterpene compound I were significantly increased, which can significantly improve angiotensin II-induced cardiac function in mice.

[0125] The above results indicate that the heteroterpenoid compound I described in this invention has an anti-heart failure effect and can be used to prepare drugs for treating heart failure, showing good prospects for clinical application.

[0126] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The application of heteroterpenoid compound I in the preparation of drugs for treating heart failure, characterized in that, The structural formula of the heteroterpene compound I is shown in formula (I): Formula (I).

2. The application according to claim 1, characterized in that, The heteroterpene compound I was isolated and purified from the fermentation culture of the marine fungus Aspergillus terreus GZU-311; the culture medium used for fermentation was a medium prepared from corn and seawater.

3. The application according to claim 1, characterized in that, The heteroterpene compound I can improve cardiac function.

4. The application according to claim 3, characterized in that, The heteroterpene compound I can increase cardiac ejection fraction and fractional shortening.

5. The application according to claim 1, characterized in that, The heteroterpene compound I can reduce myocardial cell damage.

6. The application according to claim 1, characterized in that, The heteroterpene compound I can inhibit the thickening of myocardial connective tissue.

7. The application according to claim 1, characterized in that, The heteroterpene compound I can improve the disordered arrangement of cardiomyocytes.

8. The application according to claim 1, characterized in that, The heteroterpene compound I can reduce inflammatory cell infiltration.

9. A drug for treating heart failure, characterized in that, It contains the heteroterpene compound I as described in claim 1 or a pharmaceutically acceptable salt thereof.

10. The drug according to claim 9, characterized in that, It also contains pharmaceutically acceptable excipients.

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