Polyketone compounds diamorphone C and D, and preparation method and application thereof

Abstract

The invention discloses polyketone compounds diaporthone C and D as well as a preparation method and application of the polyketone compounds diaporthone C and D. According to the present invention, the compound diapothrone C and the compound diapothrone D are prepared and separated from the fermentation culture of the marine fungus diapothrone FS722, and the compound diapothrone C and the compound diapothrone D are prepared and obtained through the separation of the fermentation culture of the marine fungus diapothrone FS722; experiments prove that the IC50 value range of the compounds diapothrone C and D to glioma cells SF-268, breast cancer cells MCF-7, liver cancer cells HepG-2 and non-small cell lung cancer cells A549 is 1.50-23.26 [mu] M, and the compounds diapothrone C and D show remarkable anti-tumor activity and can be used for preparing anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical biotechnology, and in particular relates to polyketide compounds diaporthrone C and D, and a preparation method and application thereof. Background Art

[0002] The incidence and mortality rates of malignant tumors are rising annually, posing an increasingly severe challenge to human health and lifespan. A comprehensive assessment of commonly used clinical cancer treatments reveals that chemotherapy continues to play an irreplaceable role and remains a crucial component of combined treatment approaches. Therefore, the search for new, highly effective, and low-side-effect anti-tumor drugs, and the development of more effective chemotherapy agents for cancer prevention and treatment, is a crucial focus for researchers worldwide.

[0003] Marine fungi have long held a prominent position in the research of marine natural products, owing to their significant advantages, including short cultivation cycles, sustainable development, and ease of industrialization. Their secondary metabolites are structurally diverse and highly active, and, due to their amenability to transcriptional and metabolic regulation and ecological sustainability, have become a research hotspot in natural product chemistry, pharmacology, synthetic chemistry, and synthetic biology. Therefore, marine fungal secondary metabolites hold broad application prospects in the development of anti-tumor drugs. Summary of the Invention

[0004] The first object of the present invention is to provide polyketide compounds diaporthrone C and D with anti-tumor activity, whose structures are shown in formula (I):

[0005]

[0006]

[0007] A second object of the present invention is to provide a method for preparing polyketide compounds diaporthrone C and D. The diaporthrone C and D compounds are isolated and prepared from a fermentation culture of the marine fungus Diaporthe hongkongensis FS722, and specifically comprise the following steps:

[0008] (1) A solid fermentation culture of the marine fungus Diaporthe hongkongensis FS722 was prepared, and the solid fermentation culture was extracted with ethyl acetate. The extraction was repeated three times, and the ethyl acetate extract was concentrated to obtain a brown-black paste-like crude extract.

[0009] (2) The crude extract was subjected to silica gel column chromatography using petroleum ether-ethyl acetate in a volume ratio of 80:20, 70:30, 60:40, 50:50, 0:100 and methanol 100% as eluents, gradient elution, thin layer chromatography (TLC) monitoring and analysis of the eluted fractions, and similar spots were merged according to the color and relative shift value (Rf) after the color development of the dot plate map, and finally 5 components (Fr.1 to Fr.5) were obtained. Component Fr.3 with Rf = 0.3-0.7 was obtained by eluting with petroleum ether-ethyl acetate in a volume ratio of 60:40 and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 5:1 v / v;

[0010] Fr.3 was separated into nine subfractions, Fr.3.1 to Fr.3.9, by gradient elution with a reverse-phase column (MeOH / H₂O, 30:70 → 100:0, v / v). Fraction Fr.3.2, obtained by eluting with a 30:70 MeOH / H₂O volume ratio and developing with n-hexane:ethyl acetate = 1:1 v / v, was collected and developed by thin-layer chromatography (TLC) with n-hexane:ethyl acetate = 1:1 v / v, yielding an Rf of 0.5-0.6. Fr.3.2 was separated into four fractions, Fr.3.2.1 to Fr.3.2.4, by normal-phase column (CH₂Cl₂ / MeOH, 200:1 → 50:1, v / v). Fraction Fr.3.2.1, obtained by eluting with a 100:1 CH₂Cl₂ / MeOH volume ratio and developing with n-hexane:ethyl acetate = 1:1 v / v, was collected and developed by TLC with an Rf of 0.5-0.6. Fr.3.2.1 was then separated by Sephadex LH-20 column (methanol elution) to remove the pigment, and then diaporthrone D (t R=35.1 min). Elution with MeOH / H₂O (volume ratio: 60:40) was collected and developed by thin-layer TLC (n-hexane:ethyl acetate = 10:1 v / v) to afford fraction Fr.3.8 with an Rf of 0.7-0.8. Elution of Fr.3.8 with a normal phase column (CH₂Cl₂ / MeOH, 500:1 → 60:1, v / v) afforded six fractions, Fr.3.8.1 to Fr.3.8.6. Elution with CH₂Cl₂ / MeOH (200:1, v / v) afforded fraction Fr.3.8.5 with an Rf of 0.7-0.8. Fr.3.8.5 was further separated by a normal phase column (CH2Cl2 / MeOH, 500:1→60:1, v / v) to obtain four fractions Fr.3.8.5.1 to Fr.3.8.5.4. The fractions eluted with CH2Cl2 / MeOH, 100:1, v / v were collected and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 8:1 v / v to obtain Fr.3.8.5.4 with Rf = 0.7-0.8. Fr.3.8.5.4 was separated by semi-preparative HPLC to obtain diaporthrone C (t R =17.5min).

[0011] Furthermore, components Fr.3.2.1 and Fr.3.8.5.4 were separated and purified by HPLC as follows: component Fr.3.2.1 was subjected to semi-preparative HPLC using a YMCpack ODS-A / AQ column, the mobile phase was acetonitrile / water in a volume ratio of 19:81, the flow rate was 2 mL / min, and the elution fractions with a retention time of 35.1 min were collected to obtain compound diaporthrone D; component Fr.3.8.5.4 was subjected to semi-preparative HPLC using a YMCpack ODS-A / AQ column, the mobile phase was acetonitrile / water in a volume ratio of 67:33, the flow rate was 2 mL / min, and the elution fractions with a retention time of 17.5 min were collected to obtain compound diaporthrone C.

[0012] The preparation of the solid fermentation culture of the marine fungus Diaporthe hongkongensis FS722 specifically comprises the following steps: picking FS722 mycelium and inoculating it into a potato glucose liquid culture medium, culturing it at 28°C and 120r / min for 5 days to obtain a seed solution, then inoculating the seed solution into a rice culture medium at an inoculum rate of 0.1mL / g, and culturing it at 28°C for 30 days to obtain a solid fermentation culture of FS722, wherein the potato glucose liquid culture medium is prepared by the following method per liter: boiling 200g of potatoes with 500mL of pure water for 20 minutes, filtering to obtain potato juice, and then adding 20g of glucose, 3g of KH2PO4, 1.5g of MgSO4, and 100mg of vitamin B1. mg, made up to 1000 mL with water and sterilized; the rice culture medium is prepared by the following method: 480 g of rice is mixed with 600 mL of a crude sea salt solution with a mass volume ratio of 0.5 mg / mL and sterilized.

[0013] A third object of the present invention is to provide the use of the polyketide compounds diaporthrone C and / or diaporthrone D, or pharmaceutically acceptable salts thereof, in the preparation of antitumor drugs, preferably drugs for glioma, breast cancer, liver cancer, or non-small cell lung cancer.

[0014] The present invention found through experiments that the IC values of compounds diaporthrone C and D on glioma cells SF-268, breast cancer cells MCF-7, liver cancer cells HepG-2, and non-small cell lung cancer cells A549 were 50 The values ranged from 1.50 to 23.26 μM. The positive control adriamycin had an IC value of 1.50 to 23.26 μM for the above four tumor cell lines. 50 The values were 1.44, 1.25, 1.12 and 1.39 μM respectively. This result indicates that the compounds diaporthrone C and D of the present invention have very significant anti-tumor activity.

[0015] A fourth object of the present invention is to provide an anti-tumor drug comprising as an active ingredient at least one of the polyketide compounds diaporthrone C and D, or a pharmaceutically acceptable salt thereof. Preferably, the anti-tumor drug is an anti-glioma, breast cancer, liver cancer, or non-small cell lung cancer drug.

[0016] A fifth object of the present invention is to provide the use of the marine fungus Diaporthe hongkongensis FS722 in the preparation of the polyketide compounds diaporthrone C and / or diaporthrone D.

[0017] Compared with the prior art, the advantages of the present invention are:

[0018] The present invention separates and prepares compounds diaporthrone C and D from the marine fungus Diaporthe hongkongensis FS722. These compounds have significant anti-tumor activity and can be used to prepare anti-tumor drugs. They provide candidate compounds for the research and development of new anti-tumor drugs and provide a scientific basis for the development and utilization of natural active substances derived from marine microorganisms.

[0019] The marine fungus Diaporthe hongkongensis FS722 of the present invention was disclosed in the non-patent literature (webpage) on February 14, 2025, and its URL is: https: / / www.ncbi.nlm.nih.gov / nuccore / PV082656.1 / . The applicant also holds the strain and guarantees that it will be provided to the public within 20 years from the date of application. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Compound 1 (diaporthrone C) 1 H NMR spectrum;

[0021] Figure 2 Compound 1 (diaporthrone C) 13 C NMR spectrum;

[0022] Figure 3 is the COSY spectrum of compound 1 (diaporthrone C);

[0023] Figure 4 is the HSQC spectrum of compound 1 (diaporthrone C);

[0024] Figure 5 is the HMBC spectrum of compound 1 (diaporthrone C);

[0025] Figure 6 is the NOESY spectrum of compound 1 (diaporthrone C);

[0026] Figure 7 is the HR-ESIMS spectrum of compound 1 (diaporthrone C);

[0027] Figure 8 The structural formula of compound 1 (diaporthrone C) and 1 H- 1 H COSY and key HMBC signals;

[0028] Figure 9 It is NOE related to compound 1 (diaporthrone C);

[0029] Figure 10 is the calculated and measured ECD curve of compound 1 (diaporthrone C);

[0030] Figure 11 is compound 2 (diaporthrone D) 1 H NMR spectrum;

[0031] Figure 12 is compound 2 (diaporthrone D) 13 C NMR spectrum;

[0032] Figure 13 is the COSY spectrum of compound 2 (diaporthrone D);

[0033] Figure 14 is the HSQC spectrum of compound 2 (diaporthrone D);

[0034] Figure 15 is the HMBC spectrum of compound 2 (diaporthrone D);

[0035] Figure 16 is the NOESY spectrum of compound 2 (diaporthrone D);

[0036] Figure 17 is the HR-ESIMS spectrum of compound 2 (diaporthrone D);

[0037] Figure 18 The structural formula of compound 2 (diaporthrone D) and 1 H- 1 H COSY and key HMBC signals;

[0038] Figure 19 It is NOE related to compound 2 (diaporthrone D);

[0039] Figure 20 These are the calculated and measured ECD curves of compound 2 (diaporthrone D). DETAILED DESCRIPTION

[0040] The following examples are provided to further illustrate the present invention, but are not intended to limit the present invention.

[0041] Example 1

[0042] 1. Isolation, purification and identification of the marine fungus Diaporthe hongkongensis FS722

[0043] The strain was collected by our research group in June 2017 from seafloor sediment at a depth of 1428 m in the South China Sea, Indian Ocean, at the coordinates 110°59'04″E, 18°00'47″N. It was identified as Diaporthe hongkongensis by ITS sequencing and has the NCBI gene accession number PV082656. It is deposited in the Medicinal Fungi Research Group, Institute of Microbiology, Guangdong Academy of Sciences.

[0044] 2. Solid-state fermentation of Diaporthe hongkongensis FS722

[0045] The activated deep-sea fungus FS722 mycelium was inoculated into potato glucose liquid medium (each liter of culture medium was prepared by the following method: 200 g of potatoes were boiled in 500 mL of pure water for 20 minutes, the potato juice was filtered, 20 g of glucose, 3 g of KH2PO4, 1.5 g of MgSO4, and 10 mg of vitamin B1 were added, the volume was made up to 1000 mL with water, and sterilized), and cultured at 28°C and 120 r / min for 5 days to prepare a seed solution. The seed solution was then inoculated into rice medium at an inoculum rate of 0.1 mL / g of rice medium (the medium was prepared by mixing 480 g of rice with 600 mL of a crude sea salt solution with a mass volume ratio of 0.5 mg / mL, autoclaving at 121°C for 20 minutes, and cooling), and cultured at 28°C for 30 days to obtain a solid fermentation culture of FS722.

[0046] 3. Preparation of compounds diaporthrone C and diaporthrone D

[0047] (1) Ethyl acetate was added to the solid fermentation culture of FS722, and the mixture was soaked and extracted for 24 hours. The extraction was repeated three times, and the extract was concentrated to obtain an extract (40 g).

[0048] (2) The crude extract (extract) obtained in step (1) was subjected to silica gel column chromatography, using petroleum ether-ethyl acetate in a volume ratio of 80:20, 70:30, 60:40, 50:50, 0:100 and methanol 100% as eluents, gradient elution, thin layer chromatography (TLC) monitoring and analysis of the eluted fractions, and merging similar spots based on the color and relative shift value (Rf) after the dot plate diagram was developed, and finally 5 components (Fr.1 to Fr.5) were obtained. Component Fr.3 obtained by elution with petroleum ether-ethyl acetate in a volume ratio of 60:40 and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 5:1 v / v was obtained with Rf = 0.3-0.7;

[0049] Fr.3 (7.7 g) was gradient eluted with a reverse phase column (MeOH / H2O, 30:70→100:0, v / v) and divided into nine subfractions Fr.3.1 to Fr.3.9.

[0050] Elution with MeOH / H₂O (volume ratio: 30:70) was performed on TLC with n-hexane:ethyl acetate (v / v) to yield Fraction Fr.3.2 (Rf = 0.5-0.6). Elution with MeOH / H₂O (volume ratio: 60:40) was performed on TLC with n-hexane:ethyl acetate (v / v) to yield Fraction Fr.3.8 (Rf = 0.7-0.8).

[0051] Fr.3.2 was separated into four fractions, Fr.3.2.1 to Fr.3.2.4, using a normal phase column (CH2Cl2 / MeOH, 200:1 → 50:1, v / v). Elution with CH2Cl2 / MeOH (volume ratio: 100:1) was collected and developed by thin-layer chromatography (TLC) with n-hexane:ethyl acetate (volume ratio: 1:1) to afford Fraction Fr.3.2.1 with an Rf of 0.5-0.6. Fraction Fr.3.2.1 was purified by a Sephadex LH-20 column (methanol elution) to remove the pigment. The product was then separated by semi-preparative HPLC using a YMCpack ODS-A / AQ column with a mobile phase of acetonitrile / water (volume ratio: 19:81) at a flow rate of 2 mL / min. The fraction with a retention time of 35.1 min was collected to yield 4.6 mg of diaporthrone D.

[0052] Fr.3.8 was eluted with a normal phase column (CH2Cl2 / MeOH, 500:1→60:1, v / v) to obtain 6 fractions Fr.3.8.1~Fr.3.8.6. The fractions eluted with (CH2Cl2 / MeOH, 200:1, v / v) were collected and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 10:1 v / v to obtain component Fr.3.8.5 with Rf = 0.7-0.8. Fr.3.8.5 was further fractionated using a normal phase column (CH2Cl2 / MeOH, 500:1→60:1, v / v) to obtain four fractions, Fr.3.8.5.1 to Fr.3.8.5.4. The fraction eluted with CH2Cl2 / MeOH, 100:1, v / v, was collected and developed by thin-layer chromatography (TLC) with n-hexane:ethyl acetate = 8:1 v / v to obtain fraction Fr.3.8.5.4 with an Rf = 0.7-0.8. Fraction Fr.3.8.5.4 was subjected to semi-preparative HPLC using a YMCpack ODS-A / AQ column with a mobile phase of acetonitrile / water (67:33, v / v) at a flow rate of 2 mL / min. The fraction with a retention time of 17.5 min was collected to obtain 2.5 mg of compound diaporthrone C.

[0053] 4. Structural identification of compounds diaporthrone C and D

[0054] 1 H-NMR, 13 C-NMR and HMBC NMR spectra were measured using a Bruker Advance-500 NMR spectrometer with tetramethylsilane (TMS) as the internal standard; ESI-MS data were measured using a VG Autospec-3000 mass spectrometer; and UV spectra were measured using a Shimadzu UV-2600 spectrophotometer. The structure was identified as follows:

[0055] like Figure 1-20 As shown, Figure 1 Compound 1 (diaporthrone C) 1 H NMR spectrum; Figure 2 is compound 1 13 CNMR spectrum; Figure 3 is the COSY spectrum of compound 1; Figure 4 is the HSQC spectrum of compound 1; Figure 5 is the HMBC spectrum of compound 1; Figure 6 is the NOESY spectrum of compound 1; Figure 7 is the HR-ESIMS spectrum of compound 1; Figure 8 is the structural formula of compound 1 and 1 H- 1 H COSY and key HMBC signals; Figure 9It is NOE related to compound 1; Figure 10 This is the calculated and measured ECD curve of compound 1. Figure 11 It is the compound 2 (diaporthrone D) 1 H NMR spectrum; Figure 12 is compound 2 13 C NMR spectrum; Figure 13 is the COSY spectrum of compound 2; Figure 14 is the HSQC spectrum of compound 2; Figure 15 is the HMBC spectrum of compound 2; Figure 16 is the NOESY spectrum of compound 2; Figure 17 is the HR-ESIMS spectrum of compound 2. Figure 18 is the structural formula of compound 2 and 1 H- 1 H COSY and key HMBC signals; Figure 19 It is NOE related to compound 2; Figure 20 This is the ECD calculated and measured curve of compound 2.

[0056] New compound 1 (diaporthrone C), white powder, easily soluble in MeOH, HR-ESI-MS data of compound 1 (m / z 299.1859 [M+H] + , the calculated value is 299.1853) and its molecular formula can be determined to be C 16 H 26 O5, the unsaturation value is 4. 1 H NMR spectrum (Table 1) showed that one methyl group δ H 0.89 (H3-16), 6 methylene protons δ H 1.31-1.42 (9H, m), 1.37 (1H, m, H-11a), 1.51 (1H, m, H-11b), 1.76 (1H, m, H-10b), 5 oxymethyl signals δ H 3.63 (1H, dd, H-9), 3.70 (1H, dd, H-9), 3.94 (1H, dd, H-8), 4.52 (1H, dt, H-4), 4.15 (1H, dd, H-5), 2 olefin proton signals δ H 5.96 (1H, dd, H-7) and 6.87 (1H, dd, H-7). 13 C NMR and HSQC spectra showed the presence of 16 carbon resonance signals, including one carbonyl carbon (δ C202.6), 1 methyl group, 6 methylene groups, and 8 methine groups (5 oxygen-linked carbon atoms and 1 olefinic carbon). The functional groups listed above (carbonyl group, double bond) account for a total of 2 degrees of unsaturation. Combined with the remaining 2 degrees of unsaturation, it is speculated that compound 1 should have a bicyclic structure.

[0057] Table 1 NMR data of compound 1 (deuterated chloroform)

[0058]

[0059] The planar structure of compound 1 was finally determined by a series of two-dimensional spectroscopic analyses. The relative configuration of compound 1 was determined by HH coupling constants and NOE correlations ( Figure 9 ). According to J 3 H-4 / H-5 (8.2Hz), J 3 H-5 / H-6 (11.0Hz) and J 3 H-6 / H-7 The large coupling constant (10.2 Hz) indicates that H-4, H-5, H-6, and H-7 must be located in vertical bonds, so that H-5 / H-7 are in the same direction and H-4 / H-6 are in opposite directions. Assuming that H-7 is in β orientation, according to H-7 / H2-10b (δ H 4.15 / δ H 1.76) and H-8 / H2-10a(δ H 3.70 / δ H 1.42) correlation peak, indicating that H-5 / H-7 / H-8 / H-10 are in the same β direction. H 2.72 / δ H 3.94) are all correlated, so H-4 / H-6 / H-9 have the same orientation, which is in the α orientation. By comparing the experimental and ECD spectra ( Figure 10 ) was used to determine the absolute configuration of compound 1 as 4S, 5S, 6S, 7S, 8S, 9R.

[0060] The new compound 2, named diaporthrone D, is a yellow oil, soluble in MeOH. According to the HR-ESI-MS data of compound 2 (m / z 325.1665[MH] - , the calculated value is 325.1667) is determined as C 16 H 18 O4, unsaturation value is 5.

[0061] The H-C spectrum data of compound 2 (Table 2) are very similar to those of compound 1, but there are two more double bond signals δ H6.15(1H,dd,H-2),δ H 6.93 (1H, dd, H-3), δ C 129.5(C-2), δ C 150.1(C-3). Further analysis showed that from H-3(δ H 6.93) to C-1(δ C 186.6) and C-5(δ C 76.1), from H-2(δ H 6.15) to C-4(δ C 70.2) and C-6(δ C 134.6), indicating that the additional double bond is located between C-2 and C-3 and satisfies one additional degree of unsaturation.

[0062] Table 2 NMR data of compound 2 (deuterated methanol)

[0063]

[0064] The relative configuration of compound 2 was determined by NOESY spectrum analysis. H 4.60 / δ H 3.60) has a significant NOE correlation, indicating that H-4 and H-9 are coplanar and assumed to be α-oriented. H 4.01 / δ H 1.56) between H-8 and H-9, indicating that H-8 and H-9 are not facing. In addition, because there is no NOESY correlation between H-5 and H-9, it is speculated that H-5 should be β-oriented. The absolute configuration of compound 2 was further determined by quantum chemical ECD calculation. The calculated ECD spectrum completely matches the experimental ECD spectrum of compound 2 ( Figure 20 ). Therefore, the absolute configuration of compound 2 was finally determined to be 4S, 5S, 8S, 9R.

[0065] The target compound 1 separated by the above method was named as compound diaporthrone C, and the target compound 2 was named as compound diaporthrone D. Their structural formulas are shown in formula (I):

[0066]

[0067] Example 2

[0068] The antitumor activity of compounds diaporthrone C and D was tested using the SRB method (Skehan P, Storeng R, Dominic S. New colorimetric cytotoxicity assay for anticancer-drug screening [J]. J Natl Cance Inst, 1990, 82: 1107-1112.).

[0069] 1. Assay Reagents: Dissolve diaporthrone C and D, compounds prepared herein, in dimethyl sulfoxide (DMSO) to obtain a 10 mM stock solution. The stock solution was then diluted in RPMI-1640 medium to various concentrations: 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.125 μM, 1.525 μM, and 0.78 μM. A positive control was an aqueous solution of doxorubicin.

[0070] The tumor cell lines used in this experiment were glioma cells SF-268, breast cancer cells MCF-7, liver cancer cells HepG-2, and non-small cell lung cancer cells A549.

[0071] 2. Experimental method: SF-268, MCF-7, HepG-2 and A549 cells in the logarithmic growth phase were digested with trypsin and counted with trypan blue staining. After the cell viability was greater than 95% as determined by trypan blue exclusion test, the cell concentration was adjusted to 3×10 cells / mL with fresh RPMI-1640 medium. 4 Cells were seeded in a 96-well plate at a concentration of 180 μL of cell suspension per well. Three blank wells were set to zero and incubated at 37°C, 5% CO₂ for 24 hours. After cell attachment, 20 μL of a solution of the aforementioned compounds, diaporthrone C and D, was added to each well. 20 μL of RPMI-1640 medium was added to the negative control, and doxorubicin was used as a positive control. After incubation at 37°C, 5% CO₂ for 72 hours, the cells were fixed with 50 μL of a 50% (v / v) solution of cold trichloroacetic acid. After incubation at 4°C for 1 hour, the cells were washed five times with distilled water and air-dried. Then, 100 μL / well of a 4 mg / mL solution of sulforhodamine B (SRB) prepared in a 1% (v / v) glacial acetic acid solution was added. The cells were stained at room temperature for 30 minutes, the supernatant was removed, and the cells were washed five times with a 1% (v / v) solution of glacial acetic acid and air-dried. Finally, 200 μL / well of 10 mmol / mL Tris solution was added, and the absorbance (A) at 570 nm was measured using a microplate reader. The inhibition rate of the drug on cell growth was calculated using the following formula: Cell growth inhibition rate (%) = (1-A 样品组 / A 对照组 )×100%.

[0072] 3. Experimental Results: The cytotoxicity of diaporthrone C and D, compounds prepared in the present invention, against four tumor cell lines is shown in Table 3. These results demonstrate that diaporthrone C and D, compounds prepared in the present invention, possess significant anti-tumor activity. Therefore, the present invention provides candidate compounds for the research and development of new anti-tumor drugs and provides a scientific basis for the development and utilization of natural active substances derived from deep-sea microorganisms.

[0073] Table 3 Inhibitory effects of compounds diaporthrone C and D on cancer cells

[0074]

[0075] The above are merely preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be construed as limiting the present invention, and the scope of protection of the present invention should be determined by the scope defined in the claims. Persons skilled in the art will appreciate that improvements and modifications may be made without departing from the spirit and scope of the present invention, and such improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Any compound represented by formula (I):

2. A method for preparing the compound according to claim 1, characterized in that: It is isolated and prepared from the fermentation culture of the marine fungus Diaporthehongkongensis FS722.

3. The preparation method according to claim 2, characterized in that The specific steps include: (1) preparing a solid fermentation culture of the marine fungus Diaporthe hongkongensis FS722, extracting the solid fermentation culture with ethyl acetate, and concentrating the extract to obtain a crude extract; (2) The crude extract was chromatographed on a silica gel column, and eluted with a gradient of petroleum ether-ethyl acetate in a volume ratio of 80:20, 70:30, 60:40, 50:50, 0:100 and methanol 100%, and the fraction Fr.3 obtained by eluting with a volume ratio of petroleum ether-ethyl acetate of 60:40 was collected and developed by TLC thin layer chromatography with n-hexane: ethyl acetate = 5:1 v / v to obtain Rf = 0.3-0.7; Fr.3 was eluted with a MeOH / H2O system in a volume ratio of 30:70→100:0 on a reverse phase column, and the fractions obtained by eluting with a volume ratio of MeOH / H2O of 30:70 and developed by TLC thin layer chromatography with n-hexane: ethyl acetate = 1:1 v / v to obtain Rf = 0.5-0. .6 component Fr.3.2 and the component Fr.3.8 with Rf=0.7-0.8 obtained by eluting with MeOH / H2O in a volume ratio of 60:40 and developing by TLC thin layer chromatography with n-hexane:ethyl acetate=10:1 v / v; Fr.3.2 was eluted with a normal phase column with a CH2Cl2 / MeOH system in a volume ratio of 200:1→50:1 gradient, and the component Fr.3.2.1 with Rf=0.5-0.6 obtained by eluting with CH2Cl2 / MeOH in a volume ratio of 100:1 and developing by TLC thin layer chromatography with n-hexane:ethyl acetate=1:1 v / v was collected. After the pigment was removed from Fr.3.2.1, diaporthrone was separated by semi-preparative HPLC. D; Fr.3.8 was eluted with a CH2Cl2 / MeOH system in a volume ratio of 500:1→60:1 by normal phase column, and the component eluted with a CH2Cl2 / MeOH volume ratio of 200:1 was collected and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 10:1 v / v to obtain component Fr.3.8.5 with Rf = 0.7-0.

8. Fr.3.8.5 was eluted with a CH2Cl2 / MeOH system in a volume ratio of 500:1→60:1 by normal phase column, and the component eluted with a CH2Cl2 / MeOH volume ratio of 100:1 was collected and developed by TLC thin layer chromatography with n-hexane:ethyl acetate = 8:1 v / v to obtain component Fr.3.8.5.4 with Rf = 0.7-0.

8. Fr.3.8.5.4 was separated by semi-preparative HPLC to obtain diaporthrone C.

4. The preparation method according to claim 3, characterized in that After the pigment was removed from Fr.3.2.1, diaporthrone D was separated by semi-preparative HPLC. Specifically, a YMCpack ODS-A / AQ column was used, the mobile phase was acetonitrile / water in a volume ratio of 19:81, the flow rate was 2 mL / min, and the elution fraction with a retention time of 35.1 min was collected to obtain the compound diaporthrone D; Fr.3.8.5.4 was separated by semi-preparative HPLC to obtain diaporthrone C. Specifically, a YMCpack ODS-A / AQ column was used, the mobile phase was acetonitrile / water in a volume ratio of 67:33, the flow rate was 2 mL / min, and the elution fraction with a retention time of 17.5 min was collected to obtain the compound diaporthrone C.

5. The preparation method according to claim 3, characterized in that The step (1) of preparing the solid fermentation culture of the marine fungus Diaporthe hongkongensis FS722 specifically comprises the following steps: picking the mycelium of Diaporthe hongkongensis FS722 and inoculating it into a potato glucose liquid culture medium, culturing it at 28°C and 120r / min for 5 days to obtain a seed liquid, and then inoculating the seed liquid into a rice culture medium at an inoculum amount of 0.1mL / g, and culturing it at 28°C for 30 days to obtain a solid fermentation culture; the potato glucose liquid culture medium is prepared by the following method per liter: boiling 200g of potatoes with 500mL of pure water for 20min, filtering to obtain potato juice, and then adding 20g of glucose, 3g of KH2PO4, 1.5g of MgSO4, and 10% of vitamin B1. mg, made up to 1000 mL with water and sterilized; the rice culture medium is prepared by the following method: 480 g of rice is mixed with 600 mL of a crude sea salt solution with a mass volume ratio of 0.5 mg / mL and sterilized.

6. Use of the compound diaporthrone C and / or diaporthrone D according to claim 1, or a pharmaceutically acceptable salt thereof, in the preparation of an antitumor drug.

7. The use according to claim 6, characterized in that The anti-tumor drug is a drug for glioma, breast cancer, liver cancer, or non-small cell lung cancer.

8. An anti-tumor drug, characterized in that: A method comprising as an active ingredient at least one of the compounds diaporthrone C and diaporthrone D according to claim 1, or a pharmaceutically acceptable salt thereof.

9. Use of the marine fungus Diaporthe hongkongensis FS722 in the preparation of the compound diaporthrone C and / or diaporthrone D according to claim 1.

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