A long-chain alkenone compound, a preparation method and application thereof
The long-chain enone compounds extracted from Trichoderma harzianum ZN-4 have solved the cardiotoxicity problem caused by anthracycline drugs, providing a low-toxicity and highly effective cardioprotective drug suitable for the recovery and prevention of myocardial cell damage.
Patent Information
- Application Number
- CN202510194674.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-21
- Publication Date
- 2026-03-03
- Estimated Expiration
- 2045-02-21
AI Technical Summary
Existing anthracycline drugs can damage cardiomyocytes and cause cardiotoxicity during cancer treatment. Traditional drugs such as dextromethorphan also have adverse reactions. There is a need to find low-toxicity and highly effective cardiomyocyte protective drugs.
Novel compounds (R)-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5E-en-2-one and harzianopyrone were extracted from the fermentation products of Trichoderma harzianum fungus ZN-4 and obtained through a multi-step separation and purification method. These compounds possess cardioprotective activity and can be used to prepare cardioprotective drugs.
These compounds can effectively restore cardiomyocyte damage caused by doxorubicin, have low cytotoxicity, and are suitable for use in the preparation of cardioprotective drugs and preventive health foods, reducing the cardiotoxicity of anthracycline drugs.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of biotechnology, specifically to a method for obtaining *Trichoderma harzianum* fungus (… Trichoderma harzianum Long-chain ketone compounds extracted from ZN-4 and their applications. Background Technology
[0002] Anthracyclines (ANTs) are chemotherapy drugs used to treat cancer, with doxorubicin, epirubicin, and daunorubicin being the main representative drugs. Anthracyclines are commonly used to treat various cancers, including solid tumors, breast cancer, soft tissue sarcomas, lymphoma, and leukemia, and have shown good efficacy. However, during treatment, they can easily damage the patient's cardiomyocytes, leading to serious cardiovascular diseases and posing a serious threat to the patient's life, with harm no less than that caused by cancer itself. ANT-induced cardiotoxicity (AIC) often manifests as left ventricular diastolic dysfunction, gradually developing into irreversible congestive cardiomyopathy, chronic heart failure, and even death.
[0003] Current research on drugs for the prevention and treatment of AIC (Acute Cardiomyopathy) mainly focuses on traditional drugs such as dextrorazoxane and beta-blockers. Dextrorazoxane, also known as dextrorazoxane, is a chemotherapy adjuvant and has a significant effect on reducing the cardiotoxicity caused by this class of drugs. It is often used in combination with anthracycline chemotherapy drugs in clinical practice to reduce the cardiotoxicity caused by these anthracyclines. However, dextrorazoxane also has some adverse reactions, such as affecting changes in platelets and neutrophils, and high-dose use can easily cause bone marrow suppression (Qian Jun et al. Prevention and treatment strategies of anthracycline cardiotoxicity with traditional Chinese and Western medicine. World Science and Technology - Modernization of Traditional Chinese Medicine, 2024, 26(7):1706-1714.). Therefore, it is necessary to find new cardioprotective drugs with low toxicity and high efficacy.
[0004] Compared to terrestrial microorganisms, marine microorganisms are able to tolerate the extreme conditions unique to the ocean, such as high salinity, high pressure, low oxygen, and low light. This unique living environment leads to diversity in species, genetic composition, and ecological functions among marine microorganisms. The special characteristics of the marine environment, coupled with advancements in marine microbial resource acquisition technologies, have brought unprecedented opportunities for the research of natural pharmaceutical compounds derived from marine microorganisms. Marine fungi are a rich source of bioactive secondary metabolites. Research on marine bioactive substances has revealed many compounds derived from marine organisms exhibiting diverse activities, such as anticancer, antidiabetic, antiviral, anticoagulant, and anti-inflammatory pharmacological activities. Discovering natural products with specific structural types using marine fungi as raw materials is of great significance for the development of marine drugs.
[0005] Trichoderma ( Trichoderma Trichoderma is a ubiquitous genus of filamentous fungi, attracting widespread attention as a biocontrol agent or plant pathogen. The genus contains a variety of chemical components, such as polyketides, peptides, and diketopiperazines, which possess broad biological activities, including antibacterial, antioxidant, anticancer, and antiviral properties, making it an important producer of compounds with potential biotechnological applications. In previous research, our group isolated a strain of *Trichoderma harzianum* from marine sediments. Trichoderma harzianum ZN-4 (C. Zhou, et al . Harzianolides BG: UndescribedButenolides isolated from the fungus Trichoderma harzianum ZN-4. Fitoterapia (2024, 176, 106039) It is of great significance to analyze the secondary metabolites of this fungus and discover natural active substances with medicinal value. Summary of the Invention
[0006] The purpose of this invention is to obtain Trichoderma harzianum ( Trichoderma harzianum A natural active substance with cardiomyocyte protective activity was extracted from the fermentation products of )
[0007] To achieve the above objectives, the present invention adopts the following technical solution:
[0008] This invention is derived from Trichoderma harzianum ( Trichoderma harzianum Two new compounds were isolated from the fermentation products of ZN-4. After structural identification, one of the compounds had the molecular formula C0. 14 H 18 O4, with the structural formula shown in formula (Ⅰ), is named ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one; another compound has the molecular formula C 13 H 18 O4, with the structural formula shown in formula (II), is named harzianopyrone;
[0009] (I); (II).
[0010] The Trichoderma harzianum ( Trichoderma harzianum ZN-4 is a publicly available biomaterial; see reference (C. Zhou, et al. Harzianolides BG: Undescribed Butenolides isolated from the fungus Trichoderma harzianum ZN-4. Fitoterapia , 2024,176,106039).
[0011] This invention provides a method for obtaining Trichoderma harzianum ( Trichoderma harzianum The method for separating and extracting the above-mentioned new compounds from the fermentation products of ZN-4 is described, but the preparation method of the above-mentioned compounds in this invention is not limited to this.
[0012] A method for extracting the long-chain enone compound from fermentation products includes the following steps:
[0013] (1) Trichoderma harzianum ( Trichoderma harzianum ZN-4 was activated and then inoculated into PDB medium for fermentation.
[0014] (2) After the fermentation culture was completed, the fermentation broth was extracted with ethyl acetate to obtain the extract;
[0015] (3) After concentrating the extract, normal-phase silica gel column chromatography was performed. Gradient elution was carried out with dichloromethane / methanol mixtures with volume ratios of 100:1, 50:1, 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9, and 0:10. The fraction eluted with dichloromethane / methanol mixture at a volume ratio of 9:1 was collected. The fraction was then subjected to reverse-phase silica gel column chromatography. Gradient elution was carried out with methanol / water mixtures with volume ratios of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1. Each gradient elution was performed 3 times, and the collected fractions were numbered sequentially from 1 to 27. Fractions numbered 13 to 18 were combined and then separated by high-performance liquid chromatography to obtain the long-chain ketone compound.
[0016] In step (1), Trichoderma harzianum ( Trichoderma harzianum ZN-4 was fermented and cultured.
[0017] Trichoderma harzianum ( Trichoderma harzianum ZN-4 is a fungus that can be fermented using conventional PDB medium, which includes the following ingredients: potatoes, glucose and water, with 200 g of potatoes and 20 g of glucose added per 1 L of water.
[0018] The fermentation culture conditions are static culture at 25-32℃ for 15-40 days. The static culture method refers to culture without shaking flasks.
[0019] Preferably, the fermentation culture temperature is 28-30°C. More preferably, the culture is carried out at 28°C for 21 days, under which the yield of the long-chain enone compound is the highest.
[0020] In step (2), ethyl acetate is used to extract the active ingredients in the fermentation broth to obtain an extract.
[0021] In step (3), the extract is separated by normal-phase silica gel column chromatography, and the resulting fraction is further separated by reversed-phase silica gel column chromatography and high-performance liquid chromatography. Through multi-step separation and purification, long-chain ketone compounds with high purity can be obtained.
[0022] Preferably, the high-performance liquid chromatography separation method includes: using a methanol / water mixture with a volume ratio of 55:45 as an eluent, a peak with a retention time of 19 minutes is a compound with the structural formula shown in formula (I). R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one, with a retention time of 22 minutes, is the compound harzianopyrone with the structural formula shown in formula (II).
[0023] This invention demonstrates that the above method can be used to extract *Trichoderma harzianum* (…). Trichoderma harzianum The long-chain enone compounds isolated from ZN-4 fermentation cultures exhibit good cardioprotective activity. Therefore, this invention provides the application of these long-chain enone compounds in the preparation of cardioprotective drugs. This invention uses the compound's anti-Namalwa lymphoma cell activity as its cytotoxicity data, and the results show that it exerts cardioprotective effects while exhibiting low toxicity to human cells, demonstrating biocompatibility.
[0024] Furthermore, the cardioprotective drug is a drug for preventing or treating cardiotoxicity caused by anthracyclines.
[0025] Furthermore, the anthracycline drug is doxorubicin. Studies have shown that the above-mentioned long-chain enone compounds can effectively restore cardiomyocyte damage caused by doxorubicin.
[0026] Furthermore, the manifestations of cardiotoxicity include decreased cardiomyocyte viability.
[0027] The present invention also provides an antitumor pharmaceutical composition comprising a first formulation formed of doxorubicin and a pharmaceutically acceptable carrier, and a second formulation formed of a long-chain enone compound with a structural formula as shown in formula (I) or (II) and a pharmaceutically acceptable carrier. The second formulation is used to mitigate the cardiotoxicity caused by the first formulation during antitumor therapy.
[0028] The beneficial effects of this invention are as follows:
[0029] (1) This invention extracts and isolates two compounds with novel structures from the fermentation culture of marine fungi. The method is simple to operate, has a high extraction yield and high product purity, and is suitable for large-scale production.
[0030] (2) An in vitro experiment to restore doxorubicin-induced H9c2 cardiomyocyte damage demonstrated that the invention provides ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E The compounds -en-2-one and harzianopyrone exhibit good cardioprotective activity. Further cytotoxicity assays demonstrate that the compounds provided by this invention... R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one and harzianopyrone compounds exhibit low cytotoxicity and show promising potential for development in the preparation of cardiovascular protective drugs and preventive health foods. Attached Figure Description
[0031] Figure 1 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one 1 H NMR data (in DMSO- d 6,600 MHz).
[0032] Figure 2 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one 13 C NMR data (in DMSO- d 6,150 MHz).
[0033] Figure 3 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one 1 H- 1H-COSY data (in DMSO- d 6).
[0034] Figure 4 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E HSQC data for -en-2-one (in DMSO- d 6).
[0035] Figure 5 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E HMBC data for -en-2-one (in DMSO- d 6).
[0036] Figure 6 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E Electronic circular dichroism data for -en-2-one (in CH3OH).
[0037] Figure 7 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E The structure of -en-2-one.
[0038] Figure 8 For the compound harzianopyrone 1 H NMR data (in DMSO- d 6,600 MHz).
[0039] Figure 9 For the compound harzianopyrone 13 C NMR data (in DMSO- d 6,150 MHz).
[0040] Figure 10 For the compound harzianopyrone 1 H- 1 H-COSY data (in DMSO- d 6).
[0041] Figure 11 HSQC data for the compound harzianopyrone (in DMSO- d 6).
[0042] Figure 12 HMBC data for the compound harzianopyrone (in DMSO- d 6).
[0043] Figure 13 Electronic circular dichroism chromatogram data for the compound harzianopyrone (in CH3OH).
[0044] Figure 14 Here is the structural formula for the compound harzianopyrone.
[0045] Figure 15 For compounds ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E Analysis of the protective activity of -en-2-one on cardiomyocytes.
[0046] Figure 16 Analysis of the cardioprotective activity of the compound harzianopyrone. Detailed Implementation
[0047] The present invention will be further described below with reference to specific embodiments. These embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and essence of the invention are within the scope of the invention.
[0048] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified.
[0049] Trichoderma harzianum used in the following examples ( Trichoderma harzianum ZN-4 was isolated from marine sediments in previous studies by our research group; see reference (C. Zhou, et al . Harzianolides BG:Undescribed Butenolides isolated from the fungus Trichoderma harzianum ZN-4. Fitoterapia , 2024,176,106039).
[0050] Example 1: Trichoderma harzianum (Trichoderma harzianum Fermentation culture of ZN-4
[0051] Trichoderma harzianum ( Trichoderma harzianum ZN-4 was activated and inoculated into PDB medium, and then statically fermented at 28°C for 21 days.
[0052] The PDB culture medium formula is: potatoes, glucose, and water, with 200 g of potatoes and 20 g of glucose added per 1 L of water. It is then autoclaved at 121℃ for 20 min.
[0053] Example 2: Preparation of long-chain enone compounds
[0054] Trichoderma harzianum ( Trichoderma harzianum After fermentation, ZN-4 was extracted and concentrated with ethyl acetate, and then separated by normal-phase silica gel column chromatography (200-300 mesh, 1 kg; silica gel column size L 50 mm, diameter 12 mm). Gradient elution was performed with dichloromethane / methanol mixtures with volume ratios of 100:1, 50:1, 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9, and 0:10. The fraction eluted from the dichloromethane / methanol mixture with a volume ratio of 9:1 was collected.
[0055] The fraction was subjected to reversed-phase silica gel column chromatography with methanol / water (1:9-9:1) as the eluent. Gradient elution was performed sequentially with methanol / water mixtures of volume ratios of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1, with 1.5 L eluted per gradient. Fractions were collected in 0.5 L increments using an automated collector, resulting in 27 fractions, numbered 1 to 27. Based on the similarity of their components, the fractions were combined to form seven subfractions, named 5-1 to 5-7. Specifically, 5-1: 1-3; 5-2: 4; 5-3: 5-10; 5-4: 11-12; 5-5: 13-18; 5-6: 19-24; 5-7: 25-27.
[0056] The fractions were then separated by high performance liquid chromatography. The peak with a retention time of 19 minutes for subfraction 5-5 was designated as compound 1, and the peak with a retention time of 22 minutes was designated as compound 2, using a methanol / water mixture with a volume ratio of 55:45 as the eluent.
[0057] Example 3: Structural Identification of Long-Chain Enone Compounds
[0058] The purity of the obtained compounds was determined by HPLC. The structures of samples with a purity greater than 98% were identified by mass spectrometry and nuclear magnetic resonance (NMR). NMR was measured using a JEOL 600 MHz spectrometer with TMS as an internal standard. High-resolution mass spectrometry was performed using an AB Sciex 5500 Q-TRAP.
[0059] According to the one-dimensional NMR of compound 1 (see Table 1), Figure 1 - 2 According to the results of mass spectrometry analysis, the molecular formula of compound 1 is C13C12C ... 14 H 18 O4. Results of analysis by two-dimensional NMR and electron circular dichroism spectroscopy (see...) Figure 3 - 6 The structure of the compound was confirmed, and by consulting databases and literature, compound 1 was identified as a new compound. R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one, structure as follows Figure 7 As shown.
[0060] Table 1. NMR data of compound 1
[0061] Position <![CDATA[ δ C (ppm), type]]> <![CDATA[ δ H (ppm) mult. ( J in Hz)]]> 1 104.8, CH 6.26, s 2 156.7, C - 3 110.1, C - 4 156.7, C - 5 104.8, CH 6.26, s 6 137.5, C - 7 79.5, CH 4.76, s 8 210.2, C - 9 <![CDATA[36.7, CH2]]> 2.40, m 10 <![CDATA[26.5, CH2]]> 1.99, q (7.5) 11 130.5, CH 5.23, dt (15.6, 5.2) 12 125.4, CH <![CDATA[5.25, m a ]]> 13 <![CDATA[18.1, CH3]]> 1.49, d (4.7) 14 <![CDATA[8.9, CH3]]> 1.85, s
[0062] According to the one-dimensional NMR of compound 2 (see Table 2), Figure 8 - 9 According to the results of mass spectrometry analysis, the molecular formula of compound 2 is C2. 13 H 18 O4. Results of analysis by two-dimensional NMR and electron circular dichroism spectroscopy (see...) Figure 10 - 13 The structure of the compound was confirmed. By consulting databases and literature, compound 2 was identified as a new compound, harzianopyrone, with the structure shown below. Figure 14 As shown.
[0063] Table 2. NMR data of compound 2
[0064] Position <![CDATA[ δ C (ppm), type]]> <![CDATA[ δ H (ppm) mult. ( J in Hz)]]> 1 - - 2 165.7, C - 3 97.2, C - 4 165.3, C - 5 101.3, CH 5.97, s 6 161.1, C - 7 <![CDATA[41.9, CH2]]> 2.45, m 2.34, dd (14.3, 8.3) 8 67.7, CH 3.67, m 9 <![CDATA[37.3, CH2]]> 1.37, m 10 <![CDATA[28.8, CH2]]> 2.02, m 1.95, m 11 131.6, CH 5.36, dt (15.3, 4.9) 12 125.0, CH 5.37, m 13 <![CDATA[18.3, CH3]]> 1.56, d (4.9) 14 <![CDATA[8.9, CH3]]> 1.70, s
[0065] Example 4: ( R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E Analysis of the cardioprotective activity of -en-2-one and harzianopyrone compounds
[0066] (1) Solution preparation
[0067] DMEM complete medium: 89% DMEM basal medium + 10% FBS + 1% antibiotics;
[0068] DMEM maintenance medium: 98.8% DMEM basal medium + 1% antibiotics;
[0069] PBS (1×) 1 L: NaCl 8 g + KCl 0.2 g + Na2HPO4·12H2O 3.58 g + KH2PO4 0.24 g, filter through a 0.22 μm membrane, tighten the cap to sterilize, and tighten again after natural cooling;
[0070] Cell cryopreservation solution: 80% DMEM basal medium + 10% FBS + 10% DMSO.
[0071] (2) Cell viability determination
[0072] H9c2 cells were treated with 1.0 μM doxorubicin for 24 h to establish a cell model. The protective effect against doxorubicin-induced cardiomyocyte damage was then determined using this cell model in combination with cell survival assays.
[0073] First, take H9c2 cells in the logarithmic growth phase, 7 × 10⁶ cells per well. 3 Cells were seeded in 96-well plates and cultured for 24 h in an incubator at 37°C and 5% CO2. Cells were divided into three groups: a damage model group, an experimental group, and a positive control group. The model group was treated with 1.0 μM doxorubicin, while the treatment groups were simultaneously given doxorubicin and different concentration gradients of (…). R )-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E -en-2-one or harzianopyrone, the positive control group was given doxorubicin and the positive control drug dextrorazoxane (20 μM), with 3 replicates per group. Cell viability was measured by MTT assay (10 μL of 5 mg / mL MTT solution was added to each well, incubated for 4 h, the supernatant was discarded, 150 μL of DMSO was added, and the OD value was measured at 570 nm) to assess the repair effect of the drug on doxorubicin-induced damage.
[0074] Survival rate (%) = (mean OD of sample group - mean OD of blank group) / (mean OD of negative group - mean OD of blank group) × 100%.
[0075] (3) Experimental results
[0076] Table 3. Survival rate of H9c2 cardiomyocytes damaged by doxorubicin under the action of the compound
[0077]
[0078] For compounds ( R)-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5 E H9C2 cardioprotective experiments were conducted using -en-2-one and harzianopyrone, and the results were as follows: Figure 15 and Figure 16 As shown, both compounds significantly improved cell viability at 5 μM. Furthermore, the compound harzianopyrone showed better efficacy at 10 μM. At certain concentrations, both compounds were able to restore H9c2 cells damaged by doxorubicin, and this effect was not concentration-dependent.
[0079] Example 5: Cytotoxicity analysis of long-chain enone compounds
[0080] (1) Reagent preparation
[0081] 0.4% SRB solution: Weigh 0.8 g SRB, dissolve in 200 mL 1% acetic acid, and store at room temperature.
[0082] 50% TCA solution: Weigh 50g of TCA, add water to make up to 100 mL, and store at 4℃.
[0083] 10 mM Tris-base solution: Weigh 0.6057 g Tris-base, add water to a final volume of 500 mL, pH 10.5, and store at 4 °C.
[0084] (2) Experimental instruments
[0085] CO2 incubator (Thermo), mini shaker (Kylin-Bell Lab instruments), microplate reader (MD, M5 model).
[0086] (3) Method
[0087] Namalwa cells in logarithmic growth phase were selected, and after centrifugation, the cell concentration was adjusted to 2 × 10⁶ cells / mL using RPMI 1640 medium containing 10% fetal bovine serum. 4Cells were seeded at a rate of 190 μL / well in 96-well plates and cultured at 37°C and 5% CO2 for 24 h. 10 μL of sample solution (final concentration: compound 20 µM) was added to each drug treatment well, and 20 µM 5-FU was added to each positive control well. The plates were cultured at 37°C and 5% CO2 for 3 days. The culture medium was discarded, and 100 μL of pre-chilled 50% TCA-fixed cells (4°C) was gently added. The plates were incubated for 5 min, then moved to 4°C and incubated for 1 h. The fixative was discarded, and the cells were washed 5 times with distilled water to remove TCA and air-dried for 1 h. 80 μL of 0.4% SRB solution was added to each well, and the plates were stained at room temperature for 30 min. The staining solution was discarded, and the cells were washed 5 times with 1% acetic acid to remove unbound SRB and air-dried. 150 μL of 10 mM Tris-base (pH=10.5) was added to dissolve the cells, and the plates were shaken on a micro-oscillator for 5 min. M5 microplate reader for OD measurement 510 nm value.
[0088] (4) Calculation of results
[0089] Tumor cell growth inhibition rate (%) = (OD) 对照 -OD 药物 ) / (OD 对照 -OD 空白 ) × 100%.
[0090] (5) Experimental results
[0091] Table 4. Namalwa cell survival rate of the compound
[0092]
[0093] The cytotoxicity of compounds 1 and 2 to Namalwa cell lines was further determined at a concentration of 20 µM. The data showed that the survival rate of Namalwa cell lines was 11.2% for the positive control drug 5-Fu and 5.6% for the positive control drug As2O3. Compounds 1 and 2 showed low cytotoxicity to Namalwa cell lines.
Claims
1. A long-chain enone compound, characterized in that, The structural formulas of the long-chain ketene compounds are shown in any of formulas (I)-(II).
2. The method for preparing the long-chain enone compound according to claim 1, characterized in that, Includes the following steps: (1) Trichoderma harzianum ZN-4 was activated and inoculated into PDB medium for fermentation culture; (2) After the fermentation culture was completed, the fermentation broth was extracted with ethyl acetate to obtain the extract; (3) After concentrating the extract, normal-phase silica gel column chromatography was performed. Gradient elution was carried out with dichloromethane / methanol mixtures with volume ratios of 100:1, 50:1, 9:1, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9, and 0:
10. The fraction eluted with dichloromethane / methanol mixture at a volume ratio of 9:1 was collected. The fraction was then subjected to reverse-phase silica gel column chromatography. Gradient elution was carried out with methanol / water mixtures with volume ratios of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:
1. Each gradient elution was performed 3 times. The collected fractions were numbered 1 to 27. Fractions numbered 13 to 18 were combined and then separated by high-performance liquid chromatography to obtain the long-chain ketone compound.
3. The preparation method according to claim 2, characterized in that, In step (1), the fermentation culture conditions are static culture at 25-32℃ for 15-40 days.
4. The preparation method according to claim 2, characterized in that, In step (3), the high performance liquid chromatography separation method includes: under the eluent of a methanol / water mixture with a volume ratio of 55:45, the peak with a retention time of 19 minutes is the compound (R)-1-(3,5-dihydroxy-4-methylphenyl)-1-hydroxyhept-5E-en-2-one with the structural formula as shown in formula (I), and the peak with a retention time of 22 minutes is the compound harzianopyrone with the structural formula as shown in formula (II).
5. The use of the long-chain enone compound as described in claim 1 in the preparation of cardioprotective drugs.
6. The application as described in claim 5, characterized in that, The cardioprotective drugs mentioned are those used to prevent or treat cardiotoxicity caused by anthracyclines.
7. The application as described in claim 6, characterized in that, The anthracycline drug in question is doxorubicin.
8. The application as described in claim 6, characterized in that, The manifestations of cardiotoxicity include decreased cardiomyocyte viability.
9. An antitumor drug composition, characterized in that, The pharmaceutical composition comprises a first formulation formed of doxorubicin and a pharmaceutically acceptable carrier, and a second formulation formed of a long-chain enone compound with a structure as shown in formula (I) or formula (II) and a pharmaceutically acceptable carrier;