Isopentenyl xanthone compound as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medicines, and discloses an isopentenyl xanthone compound as well as a preparation method and application thereof. The isopentenyl xanthone compound is any one of compounds shown in a formula 1 to a formula 9, is separated and extracted from secondary metabolites of Aspergillus astroides, has excellent anti-tumor activity and anti-inflammatory activity, can be applied to preparation of anti-tumor drugs and anti-inflammatory drugs, and can be used for preparing anti-tumor drugs and anti-inflammatory drugs. The method has great practical value.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an isopentenylxanthone compound and a preparation method and application thereof. Background Art

[0002] Fungi are an important source of bioactive compounds. Due to their short life cycle and strong adaptability to external media, they can produce biologically significant and diverse secondary metabolites. Aspergillus is a typical filamentous fungus, and its secondary metabolites include alkaloids, terpenes, anthraquinones, xanthones and other types. Aspergillus asteroides belongs to the genus Aspergillus. At present, relevant research on its secondary metabolites is still lacking. The types currently reported are mainly various types of xanthone compounds. The secondary metabolites of this species need to be further explored.

[0003] Xanthone is a class of oxygenated heterocyclic molecules containing dibenzo-γ-pyrrolone. Its derivatives show a variety of biological activities. Compounds with a xanthone skeleton often have good therapeutic application potential. The most abundant subclass obtained from plant and fungal sources is prenylated xanthone. Therefore, it is of great significance to conduct a systematic study on Aspergillus stellatus and search for active prenylated xanthone compounds from its secondary metabolites. Summary of the invention

[0004] The purpose of the present invention is to overcome the problems that the current types of isopentenylxanthone compounds are relatively few, and the activity of current anti-tumor and anti-inflammatory drugs needs to be further improved, and to provide an isopentenylxanthone compound and its preparation method and application. The isopentenylxanthone compound of the present invention is separated and extracted from the secondary metabolites of Aspergillus asteroides, and has excellent anti-tumor and anti-inflammatory activities, can be used to prepare anti-tumor drugs and anti-inflammatory drugs, and has great practical value.

[0005] In order to achieve the above object, the present invention provides an isopentenylxanthone compound, wherein the isopentenylxanthone compound is any one of the compounds shown in Formula 1 to Formula 9;

[0006]

[0007] The second aspect of the present invention provides a method for preparing the isopentenylxanthone compound, the method comprising the following steps:

[0008] (1) inoculating the hyphae of Aspergillus asteroides on a potato dextrose agar medium, followed by constant temperature culture to obtain a seed culture medium;

[0009] (2) cutting the seed culture medium containing mycelium into pieces and inoculating them into sterilized rice culture medium for fermentation culture;

[0010] (3) extracting the fermentation product obtained in step (2) with ethanol, and then concentrating under reduced pressure to obtain a total extract;

[0011] (4) suspending the total extract with water, then extracting with ethyl acetate, and then concentrating under reduced pressure to obtain ethyl acetate extract;

[0012] (5) subjecting the ethyl acetate extract to normal phase column chromatography to obtain five components Fr.1 to Fr.5;

[0013] (6) Component Fr.3 is subjected to reverse phase column chromatography to obtain 10 components Fr.3.1 to Fr.3.10;

[0014] (7) Component Fr.3.4 is purified by normal phase column chromatography and then by high performance liquid chromatography to obtain the compound represented by formula 6;

[0015] (8) Component Fr.3.7 is subjected to normal phase column chromatography to obtain seven components Fr.3.7.1 to Fr.3.7.7;

[0016] (9) purifying component Fr.3.7.3 by high performance liquid chromatography to obtain the compound represented by formula 1;

[0017] (10) purifying component Fr.3.7.4 by high performance liquid chromatography to obtain the compound represented by Formula 2 and the compound represented by Formula 3;

[0018] (11) purifying component Fr.3.7.5 by high performance liquid chromatography to obtain the compound represented by Formula 4 and the compound represented by Formula 5;

[0019] (12) Component Fr.3.8 was subjected to normal phase column chromatography to obtain 9 components Fr.3.8.1 to Fr.3.8.9;

[0020] (13) Separating component Fr.3.8.6 by gel column chromatography to obtain three components Fr.3.8.6.1 to Fr.3.8.6.3;

[0021] (14) Component Fr.3.8.6.2 is purified by high performance liquid chromatography to obtain the compound of Formula 7, the compound of Formula 8 and the compound of Formula 9.

[0022] Preferably, the constant temperature culture conditions include: a temperature of 20-30° C. and a time of 5-12 days.

[0023] Preferably, the fermentation culture conditions include: a temperature of 20-30° C. and a time of 40-50 days.

[0024] Preferably, in step (5), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether-ethyl acetate-methanol in a volume ratio of 10:1:0-10:10:0-10:10:1.

[0025] Preferably, in step (6), the eluent of the reverse phase column chromatography is a methanol-water mixed solution with a volume ratio of 20:80-80:20.

[0026] Preferably, in step (7), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:1.

[0027] Preferably, in step (8), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:1.

[0028] Preferably, in step (12), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 10:1 to 0:1.

[0029] The third aspect of the present invention provides a use of the above-mentioned isopentenylxanthone compounds in the preparation of anti-tumor drugs.

[0030] A fourth aspect of the present invention provides a use of the above-mentioned isopentenylxanthone compounds in the preparation of anti-inflammatory drugs.

[0031] In the present invention, 9 isopentenyl shantones (compounds shown in formulas 1 to 9) are obtained by separating and purifying the ethyl acetate extract of the fungus Aspergillus asteroides. In addition, the specific structures of the 9 isopentenyl shantones are determined as shown in formulas 1 to 9 by spectral analysis techniques such as nuclear magnetic resonance spectroscopy. The isopentenyl shantones of the present invention are further evaluated for anti-tumor activity and anti-inflammatory activity, proving that the isopentenyl shantones of the present invention all have certain anti-tumor activity and anti-inflammatory activity, wherein the compound shown in formula 4, the compound shown in formula 8 and the compound shown in formula 9 show obvious anti-tumor activity, and the compound shown in formula 9 has obvious anti-tumor effect. In addition, the compound shown in formula 1, the compound shown in formula 4, the compound shown in formula 7 and the compound shown in formula 8 have significant anti-inflammatory activity. DETAILED DESCRIPTION

[0032] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0033] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0034] The isopentenylxanthone compound of the present invention is any one of the compounds shown in Formula 1 to Formula 9;

[0035]

[0036] In the present invention, the compound shown in formula 1 is named compound 1, the compound shown in formula 2 is named compound 2, the compound shown in formula 3 is named compound 3, the compound shown in formula 4 is named compound 4, the compound shown in formula 5 is named compound 5, the compound shown in formula 6 is named compound 6, the compound shown in formula 7 is named compound 7, the compound shown in formula 8 is named compound 8, and the compound shown in formula 9 is named compound 9.

[0037] In the present invention, compounds 1 to 9 are extracted from secondary metabolites (fermentation products) of the fungus Aspergillus asteroides.

[0038] The present invention further provides a method for preparing the isopentenylxanthone compound, which comprises the following steps:

[0039] (1) inoculating the hyphae of Aspergillus asteroides on a potato dextrose agar medium, followed by constant temperature culture to obtain a seed culture medium;

[0040] (2) cutting the seed culture medium containing mycelium into pieces and inoculating them into sterilized rice culture medium for fermentation culture;

[0041] (3) extracting the fermentation product obtained in step (2) with ethanol, and then concentrating under reduced pressure to obtain a total extract;

[0042] (4) suspending the total extract with water, then extracting with ethyl acetate, and then concentrating under reduced pressure to obtain ethyl acetate extract;

[0043] (5) subjecting the ethyl acetate extract to normal phase column chromatography to obtain five components Fr.1 to Fr.5;

[0044] (6) Component Fr.3 is subjected to reverse phase column chromatography to obtain 10 components Fr.3.1 to Fr.3.10;

[0045] (7) Component Fr.3.4 is purified by normal phase column chromatography and then by high performance liquid chromatography to obtain the compound represented by formula 6;

[0046] (8) Component Fr.3.7 is subjected to normal phase column chromatography to obtain seven components Fr.3.7.1 to Fr.3.7.7;

[0047] (9) purifying component Fr.3.7.3 by high performance liquid chromatography to obtain the compound represented by formula 1;

[0048] (10) purifying component Fr.3.7.4 by high performance liquid chromatography to obtain the compound represented by Formula 2 and the compound represented by Formula 3;

[0049] (11) purifying component Fr.3.7.5 by high performance liquid chromatography to obtain the compound represented by Formula 4 and the compound represented by Formula 5;

[0050] (12) Component Fr.3.8 was subjected to normal phase column chromatography to obtain 9 components Fr.3.8.1 to Fr.3.8.9;

[0051] (13) Separating component Fr.3.8.6 by gel column chromatography to obtain three components Fr.3.8.6.1 to Fr.3.8.6.3;

[0052] (14) Component Fr.3.8.6.2 is purified by high performance liquid chromatography to obtain the compound of Formula 7, the compound of Formula 8 and the compound of Formula 9.

[0053] In the present invention, the Aspergillus asteroides used was purchased from the strain bank of China General Microbiological Culture Collection Center (CGMCC), and the purchased strain serial number is CGMCC 3.6292.

[0054] In a preferred embodiment, in step (1), the conditions of the constant temperature culture include: a temperature of 20-30°C and a time of 5-12 days. Specifically, the temperature of the constant temperature culture is 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C; and the time of the constant temperature culture is 5 days, 6 days, 7 days, 8 days, 10 days, 11 days or 12 days.

[0055] In a specific embodiment, the specific process of step (1) includes: using an inoculation loop to inoculate the hyphae of Aspergillus asteroides on a potato dextrose agar medium, then culturing at a constant temperature, and after three consecutive generations of replication, culturing the Aspergillus asteroides strain in large quantities to obtain a seed culture medium.

[0056] In the present invention, in the rice culture medium, the solid-liquid ratio of rice to water is 180-220 g: 200 mL. Specifically, the solid-liquid ratio of rice to water can be 180 g: 200 mL, 190 g: 200 mL, 200 g: 200 mL, 210 g: 200 mL or 220 g: 200 mL.

[0057] In a preferred embodiment, in step (2), the sterilization process includes: sterilizing the rice culture medium in a high pressure sterilizer for 20-40 minutes. Specifically, the sterilization time can be 20 minutes, 30 minutes or 40 minutes.

[0058] In a preferred embodiment, in step (2), the fermentation culture conditions include: a temperature of 20-30°C and a time of 40-50 days. Specifically, the fermentation culture temperature may be 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C or 30°C; the fermentation culture time may be 40 days, 41 days, 42 days, 43 days, 44 days, 45 days, 46 days, 47 days, 48 ​​days, 49 days or 50 days.

[0059] In the present invention, in step (3), the fermentation product obtained in step (2) refers to a material comprising a culture medium and mycelium obtained after fermentation culture.

[0060] In a preferred embodiment, in step (3), the number of ethanol extractions is 8-15 times, more preferably 9-12 times. Specifically, the number of ethanol extractions can be 8 times, 9 times, 10 times, 11 times, 12 times, 13 times, 14 times or 15 times.

[0061] In a specific embodiment, in step (3), the solvent ethanol is recovered by concentration under reduced pressure to obtain a total extract.

[0062] In a preferred embodiment, in step (4), the number of extractions with ethyl acetate is 8-13 times, more preferably 9-12 times. Specifically, the number of extractions with ethyl acetate can be 8 times, 9 times, 10 times, 11 times, 12 times or 13 times.

[0063] In a specific embodiment, in step (4), the solvent ethyl acetate is recovered by concentration under reduced pressure to obtain a total extract.

[0064] In a preferred embodiment, in step (5), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether-ethyl acetate-methanol in a volume ratio of 10:1:0-10:10:0-10:10:1, and this eluent is used for gradient elution to obtain five components Fr.1 to Fr.5.

[0065] In a preferred embodiment, in step (6), the eluent of the reverse phase column chromatography is a methanol-water mixed solution with a volume ratio of 20:80-80:20, and this eluent is used for gradient elution to obtain 10 components Fr.3.1 to Fr.3.10.

[0066] In a preferred embodiment, in step (7), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:1, and the eluent is used for elution, and then the obtained eluent is purified by high performance liquid chromatography to obtain the compound shown in Formula 6,

[0067] In a specific embodiment, in step (7), the mobile phase of the HPLC is an acetonitrile-water mixed solution with a volume ratio of 70:30.

[0068] In a preferred embodiment, in step (8), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:1.

[0069] In a specific embodiment, in step (9), the mobile phase of the HPLC is an acetonitrile-water mixed solution with a volume ratio of 90:10.

[0070] In a specific embodiment, in step (10), the mobile phase of the HPLC is an acetonitrile-water mixed solution with a volume ratio of 75:25.

[0071] In a specific embodiment, in step (11), the mobile phase of the HPLC is an acetonitrile-water mixed solution with a volume ratio of 80:20.

[0072] In a preferred embodiment, in step (12), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 10:1 to 0:1.

[0073] In the method of the present invention, in step (13), the eluent of the gel column chromatography is a methanol-dichloromethane mixed solution with a volume ratio of 50:50.

[0074] In a specific embodiment, in step (14), the eluate obtained from the gel column chromatography in step (13) is purified by high performance liquid chromatography, and an acetonitrile-water mixed solution with a volume ratio of 74:26 is used as the mobile phase to purify the compound shown in Formula 7, the compound shown in Formula 8 and the compound shown in Formula 9.

[0075] The present invention further provides an application of the isopentenylxanthone compound in the preparation of anti-tumor drugs.

[0076] The present invention provides an application of the isopentenylxanthone compound in the preparation of an anti-liver cancer drug, an anti-colon cancer drug, an anti-breast cancer drug or an anti-lung cancer drug.

[0077] By evaluating the anti-tumor activity of the isopentenylxanthone compounds of the present invention, it can be seen that the isopentenylxanthone compounds of the present invention exhibit certain anti-tumor activity, and compound 4 shown in formula 4, compound 8 shown in formula 8, and compound 9 shown in formula 9 exhibit significant anti-tumor activity, and compound 9 shown in formula 9 has the highest anti-tumor activity. Specifically, compound 4 shown in formula 4 has significant activity against liver cancer cells, compound 8 shown in formula 8 has significant activity against liver cancer and colon cancer, and compound 9 shown in formula 9 has significant activity against liver cancer cells, colon cancer cells, breast cancer cells, colon epithelial cells, and lung cancer cells.

[0078] The present invention can also provide a use of the isopentenylxanthone compound in the preparation of an anti-inflammatory drug. By evaluating the anti-inflammatory activity of the isopentenylxanthone compounds of the present invention, it can be seen that the isopentenylxanthone compounds of the present invention all have certain anti-inflammatory activity, among which compound 1 shown in formula 1, compound 4 shown in formula 4, compound 7 shown in formula 7, and compound 8 shown in formula 8 have significant anti-inflammatory activity.

[0079] The present invention will be described in detail below through examples, but the protection scope of the present invention is not limited thereto.

[0080] The strain Aspergillus stellatus used in the following examples is stored in Room 414, Building 1, School of Pharmacy, Huazhong University of Science and Technology, numbered 414JZ-112. The strain was purchased from the strain bank of China General Microbiological Culture Collection Center (CGMCC), and the purchase serial number is CGMCC 3.6292. It was identified as Aspergillus stellatus by ITS gene sequencing. ITS sequence number of the strain:

[0081] .

[0082] Example 1

[0083] (1) reviving the Aspergillus stellate strain in a sterile operating table, inoculating the mycelium of the Aspergillus stellate strain on a potato dextrose agar (PDA) medium with an inoculation loop, culturing in an incubator at a temperature of 25° C. for 7 days, and after three consecutive generations, culturing the strain in large quantities to prepare a seed culture medium;

[0084] (2) 200 g of rice and 200 mL of tap water were poured into 1000 mL conical flasks, and 250 flasks were prepared in total. The flasks were sterilized in an autoclave for 30 min to obtain a rice culture medium; the seed culture medium containing mycelium was cut into square pieces, inoculated into the rice culture medium cooled to room temperature (25° C.), and fermented and cultured at a constant temperature of 25° C. for 45 days (the total fermentation amount was 50 kg, calculated by the weight of rice);

[0085] (3) extracting the fermentation product obtained in step (2) with anhydrous ethanol for 10 times, and then concentrating under reduced pressure to recover the ethanol to obtain a brown-black total extract;

[0086] (4) suspending the total extract obtained in step (3) with 10 L of water, then extracting 10 times with ethyl acetate having an equal volume to the water, and concentrating under reduced pressure to recover the solvent to obtain an ethyl acetate extract;

[0087] (5) The obtained ethyl acetate extract was subjected to normal phase column chromatography, and a mixed solution of petroleum ether-ethyl acetate-methanol with a volume ratio of 10:1:0-10:10:0-10:10:1 was used as the mobile phase for gradient elution to obtain 5 components (Fr.1 to Fr.5);

[0088] (6) eluting the component Fr.3 by reverse phase column chromatography using a methanol-water mixed solution with a volume ratio of 20:80-80:20 as an eluent to obtain 10 components (Fr.3.1 to Fr.3.10);

[0089] (7) Component Fr.3.4 was eluted by normal phase column chromatography using a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:1, and the obtained eluate was then purified by high performance liquid chromatography using a mixed solution of acetonitrile and water in a volume ratio of 70:30 as the mobile phase to obtain compound 6 represented by formula 6;

[0090] (8) Component Fr.3.7 was eluted by normal phase column chromatography using a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1 to 0:1 to obtain 7 components (Fr.3.7.1 to Fr.3.7.7);

[0091] (9) Component Fr.3.7.3 was purified by high performance liquid chromatography using a 90:10 by volume acetonitrile-water mixed solution as the mobile phase to obtain compound 1 represented by formula 1;

[0092] (10) Component Fr.3.7.4 was purified by high performance liquid chromatography using a mixed solution of acetonitrile and water in a volume ratio of 75:25 as the mobile phase to obtain compound 2 represented by formula 2 and compound 3 represented by formula 3;

[0093] (11) Component Fr.3.7.5 was purified by high performance liquid chromatography using an acetonitrile-water mixed solution with a volume ratio of 80:20 as the mobile phase to obtain compound 4 represented by formula 4 and compound 5 represented by formula 5;

[0094] (12) Component Fr.3.8 was subjected to normal phase column chromatography and eluted with petroleum ether-ethyl acetate in a volume ratio of 10:1-0:1 to obtain 9 components (Fr.3.8.1 to Fr.3.8.9);

[0095] (13) Component Fr.3.8.6 was separated by gel column chromatography and eluted with a methanol-dichloromethane mixed solution with a volume ratio of 50:50 to obtain three components (Fr.3.8.6.1 to Fr.3.8.6.3);

[0096] (14) Component Fr.3.8.6.2 was purified by high performance liquid chromatography using a mixed solution of acetonitrile and water in a volume ratio of 90:10 as the mobile phase to obtain compound 7 of formula 7, compound 8 of formula 8 and compound 9 of formula 9.

[0097] Test Case

[0098] Test Example 1

[0099] The structures of the compounds represented by Formula 1 to Formula 9 were identified.

[0100] Compounds 1 to 9 shown in Formulas 1 to 9 were subjected to high resolution mass spectrometry (HRESIMS), ultraviolet spectroscopy (UV), infrared spectroscopy (IR), optical rotation spectroscopy (ORD), nuclear magnetic resonance spectroscopy ( 1 H NMR and 13 C NMR), circular dichroism (ECD) tests, and circular dichroism (ECD) calculation, molybdenum salt reaction, Mosher reaction method, and comprehensive analysis of the data to determine the structures of compounds 1 to 9, the results are as follows:

[0101] Compound 1: yellow powder, UV(MeCN)λ max (logε)=247(4.20),271(4.15),292(4.07),382(3.63)nm; ECD(MeCN)λ max (Δε)=206(+4.40),221(+2.82),234(-0.73),252(+1.57),276(-0.48); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 1; 13 The data of CNMR (CDCl3, 150MHz) are shown in Table 2; HRESIMS m / z 431.1108 [M+Na] + (The calculated value is C 23 H 20 O7Na,431.1107);

[0102] Compound 2: yellow powder, UV(MeCN)λ max(logε)=260(4.24),275(4.36),297(3.82),393(3.64)nm; ECD(MeCN)λ max (Δε)=207(+9.50),226(+1.66),258(-1.32),286(+1.38),334(-0.64); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 1; 13 The data of CNMR (CDCl3, 150MHz) are shown in Table 2; HRESIMS m / z 405.1315 [M+Na] + (The calculated value is C 22 H 22 O6Na,405.1314);

[0103] Compound 3: yellow powder, UV(MeCN)λ max (logε)=241(4.18),274(4.24),296(3.77),391(3.57)nm; ECD(MeCN)λ max (Δε)=207(+7.88),229(+1.80),257(-0.23),280(+1.06),333(-0.48); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 1; 13 The data of CNMR (CDCl3, 150MHz) are shown in Table 2; HRESIMS m / z 461.1566 [M+Na] + (The calculated value is C 25 H 26 O7Na,461.1576);

[0104] Compound 4: yellow powder, UV(MeCN)λ max (logε)=241(4.38),273(4.49),387(3.77)nm; ECD(MeCN)λ max (Δε)=221(+3.71),248(-2.57),269(-4.50),296(+0.79),333(-1.22); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 1; 13 The data of C NMR (CDCl3, 150 MHz) are shown in Table 2; HRESIMS m / z 519.1995 [M+Na] + (The calculated value is C28 H 32 O8Na,519.1995);

[0105] Compound 5: yellow powder, UV(MeCN)λ max (logε)=241(4.37),273(4.50),387(3.77)nm; ECD(MeCN)λ max (Δε)=219(+4.95),245(-2.78),260(-1.32),270(-4.38),294(+1.04),335(-1.09); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 3; 13 The data of CNMR (CDCl3, 150MHz) are shown in Table 2; HRESIMS m / z 505.1832 [M+Na] + (The calculated value is C 27 H 30 O8Na,505.1838);

[0106] Compound 6: yellow powder, UV(MeCN)λ max (logε)=240(4.08),272(4.18),383(3.47)nm; ECD(MeCN)λ max (Δε)=221(+1.52),250(-1.44),269(-1.77),332(-0.70); 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 3; 13 The data of C NMR (CDCl3, 150 MHz) are shown in Table 2; HRESIMS m / z 535.1940 [M+Na] + (The calculated value is C 28 H 32 O9Na,535.1944);

[0107] Compound 7: yellow powder, UV(MeCN)λ max (logε)=241(4.33),275(4.45),393(3.73)nm; 1 The data of H NMR (C5D5N, 600 MHz) are shown in Table 3; 13 The data of C NMR (C5D5N, 150 MHz) are shown in Table 2; HRESIMS m / z 535.2301 [M+Na] + (The calculated value is C29 H 36 O8Na,535.2308);

[0108] Compound 8: yellow powder, UV(MeCN)λ max (logε)=241(4.42),275(4.54),393(3.51)nm; 1 The data of H NMR (C5D5N, 400 MHz) are shown in Table 3; 13 The data of C NMR (C5D5N, 100 MHz) are shown in Table 2; HRESIMS m / z 535.2301 [M+Na] + (The calculated value is C 29 H 36 O8Na,535.2308);

[0109] Compound 9: yellow powder, UV(MeCN)λ max (logε)=241(4.40),275(4.51),393(3.79)nm; 1 The data of H NMR (CDCl3, 600 MHz) are shown in Table 3; 13 The data of C NMR (CDCl3, 150 MHz) are shown in Table 2; HRESIMS m / z 535.2305 [M+Na] + (The calculated value is C 29 H 36 O8Na,535.2308).

[0110] Table 1 Compound 1 to Compound 4 in deuterated chloroform solvent at 600 MHz 1 H NMR data

[0111]

[0112]

[0113] Table 2 Compounds 1 to 9 13 C NMR data

[0114]

[0115] Note: a 150MHz data in deuterated chloroform solvent; b 100 MHz data in deuterated pyridine solvent; c 150 MHz data in deuterated pyridine solvent.

[0116] Table 3 Compounds 5 to 91 H NMR data

[0117]

[0118]

[0119] Note: a 150MHz data in deuterated chloroform solvent; b 100 MHz data in deuterated pyridine solvent; c 150 MHz data in deuterated pyridine solvent.

[0120] Test Example 2

[0121] The anti-tumor activities of Compounds 1 to 9 represented by Formulae 1 to 9 were evaluated.

[0122] Test method: The CCK-8 method was used to screen the activity of five cell types: HepG2 (liver cancer cells), RKO (colon cancer cells), MCF-7 (breast cancer cells), NCM460 (colon epithelial cells), and A549 (lung cancer cells).

[0123] The above five types of cells were seeded in a 96-well plate at a density of 5000 cells / well. After incubation for 24 hours, different concentrations of compounds 1 to 9 (the concentrations of compound 1 were 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM and 1.25 μM, respectively, and the concentrations of compounds 2 to 9 were the same as those of compound 1) were used to treat the five types of cells after incubation for 72 hours. Then 10 μL of WST-8 solution was added to each well, and incubated for another 4 hours. The absorbance at 450 nm of each well was measured using an ELISA reader. Each cell line was measured at least three times. IBM SPSS Statistics 25 was used to calculate the half maximal inhibitory concentration (IC) value through the dose-response curve. 50 ), and the results are shown in Table 4.

[0124] Table 4

[0125] Compound <![CDATA[IC 50 (HepG2)]]> <![CDATA[IC 50 (RKO)]]> <![CDATA[IC 50 (MCF-7)]]> <![CDATA[IC 50 (NCM460)]]> <h2 style=";text-align:left;direction:ltr"><![CDATA[IC <h2 style=";text-align:left;direction:ltr"> 50 <h2 style=";text-align:left;direction:ltr"> (A549)]]><h2 style=";text-align:left;direction:ltr"> Compound 4 16.65±0.95 >40 >40 >40 >40 Compound 8 18.26±1.80 10.07±1.35 >40 >40 >40 Compound 9 20.81±0.96 8.62±1.18 4.98±0.28 3.35±0.58 5.66±0.86

[0126] According to the data in Table 4, compound 9 has significant inhibitory activity against RKO, MCF-7, NCM460 and A549 cell lines, IC 50 The values ​​were 8.62μM, 4.98μM, 3.35μM and 5.66μM, respectively. For HepG2 cell line, compounds 4, 8 and 9 showed moderate inhibitory activity, with IC 50 The values ​​were 16.65, 18.26 and 20.81 μM, respectively. In addition, compound 8 also had an inhibitory effect on the RKO cell line, with IC 50The results show that compound 4 shown in formula 4, compound 8 shown in formula 8 and compound 9 shown in formula 9 show significant anti-tumor activity, and compound 9 shown in formula 9 has the highest anti-tumor activity.

[0127] Test Example 3

[0128] The anti-inflammatory activities of Compounds 1 to 9 represented by Formulae 1 to 9 were evaluated.

[0129] The anti-inflammatory activity of compounds 1 to 9 was evaluated by inhibiting NO (nitric oxide) production in RAW 264.7 cells (mouse mononuclear macrophage leukemia cells) induced by LPS (lipopolysaccharide) at non-toxic concentrations, simulating inflammatory response.

[0130] Test method: RAW 264.7 cells were cultured at 37°C and 5% carbon dioxide, and 10% fetal bovine serum and penicillium were added to the culture medium. RAW 264.7 cells were seeded in 96-well plates. After pre-incubation for 24 hours, RAW 264.7 cells were treated with compounds 1 to 9 at concentrations of 80 to 2.5 μM (the concentrations of compound 1 were 80 μM, 40 μM, 20 μM, 10 μM, 5 μM and 2.5 μM, respectively, and the concentrations of compounds 2 to 9 were the same as those of compound 1) for 2 hours, and then stimulated with LPS (1 μg / mL) for 12 hours, and the NO content was determined by the Griess method. The cell culture supernatant (50 μL) was mixed with 100 μL of Griess reagent (1% sulfonamide, 0.1% naphthalenediamine dihydrochloride, 2% phosphoric acid) in a 96-well plate at room temperature for 10 min, and the optical density was measured at 510 nm using a microplate reader. The results are shown in Table 5.

[0131] Table 5

[0132] Compound <![CDATA[IC 50 (mean±SD)]]> Compound 1 9.39±0.93 Compound 2 42.12±1.93 Compound 3 45.23±0.93 Compound 4 12.61±1.32 Compound 5 49.78±1.56 Compound 6 46.50±0.88 Compound 7 7.30±0.69 Compound 8 5.16±0.17 Compound 9 43.26±1.78

[0133] From the data in Table 5, it can be seen that compounds 1 to 9 all have certain anti-inflammatory activity, among which compounds 1, 4, 7 and 8 can significantly reduce the production of NO (nitric oxide) induced by LPS (lipopolysaccharide), and their IC 50 The values ​​were 9.39 μM, 12.61 μM, 7.30 μM and 5.16 μM, respectively, showing significant anti-inflammatory activity.

[0134] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. An isopentenylxanthone compound, characterized in that: The isopentenylxanthone compound is any one of the compounds shown in Formula 1 to Formula 9; 2. A method for preparing the isopentenylxanthone compound according to claim 1, characterized in that: The preparation method comprises the following steps: (1) inoculating the hyphae of Aspergillus asteroides on a potato dextrose agar medium, followed by constant temperature culture to obtain a seed culture medium; (2) cutting the seed culture medium containing mycelium into pieces and inoculating them into sterilized rice culture medium for fermentation culture; (3) extracting the fermentation product obtained in step (2) with ethanol, and then concentrating under reduced pressure to obtain a total extract; (4) suspending the total extract with water, then extracting with ethyl acetate, and then concentrating under reduced pressure to obtain ethyl acetate extract; (5) subjecting the ethyl acetate extract to normal phase column chromatography to obtain five components Fr.1 to Fr.5; (6) Component Fr.3 is subjected to reverse phase column chromatography to obtain 10 components Fr.3.1 to Fr.3.10; (7) Component Fr.3.4 is purified by normal phase column chromatography and then by high performance liquid chromatography to obtain the compound represented by formula 6; (8) Component Fr.3.7 is subjected to normal phase column chromatography to obtain seven components Fr.3.7.1 to Fr.3.7.7; (9) purifying component Fr.3.7.3 by high performance liquid chromatography to obtain the compound represented by formula 1; (10) purifying component Fr.3.7.4 by high performance liquid chromatography to obtain the compound represented by Formula 2 and the compound represented by Formula 3; (11) purifying component Fr.3.7.5 by high performance liquid chromatography to obtain the compound represented by Formula 4 and the compound represented by Formula 5; (12) Component Fr.3.8 was subjected to normal phase column chromatography to obtain 9 components Fr.3.8.1 to Fr.3.8.9; (13) Separating component Fr.3.8.6 by gel column chromatography to obtain three components Fr.3.8.6.1 to Fr.3.8.6.3; (14) Component Fr.3.8.6.2 is purified by high performance liquid chromatography to obtain the compound of Formula 7, the compound of Formula 8 and the compound of Formula 9.

3. The preparation method according to claim 2, characterized in that: The conditions of the constant temperature culture include: a temperature of 20-30° C. and a time of 5-12 days.

4. The preparation method according to claim 2, characterized in that: The fermentation culture conditions include: a temperature of 20-30° C. and a time of 40-50 days.

5. The preparation method according to claim 2, characterized in that: In step (5), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether-ethyl acetate-methanol in a volume ratio of 10:1:0-10:10:0-10:10:

1.

6. The preparation method according to claim 2, characterized in that: In step (6), the eluent of the reverse phase column chromatography is a methanol-water mixed solution with a volume ratio of 20:80-80:

20.

7. The preparation method according to claim 2, characterized in that: In step (7), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:

1.

8. The preparation method according to claim 2, characterized in that: In step (8), the eluent of the normal phase column chromatography is a mixed solution of petroleum ether and ethyl acetate in a volume ratio of 8:1-0:

1.

9. Use of the isopentenylxanthone compound according to claim 1 in the preparation of anti-tumor drugs.

10. Use of the isopentenylxanthone compound according to claim 1 in the preparation of anti-inflammatory drugs.

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