CN108546247B

CN108546247B - Application of alkaloid compound in preparation of anti-obesity drugs

Info

Publication number: CN108546247B
Application number: CN201810595611.3A
Authority: CN
Inventors: 刘晓萌; 吴长景; 杨明生; 崔香; 毛建航; 李凤
Original assignee: Zhoukou Normal University
Current assignee: Zhoukou Normal University
Priority date: 2018-06-11
Filing date: 2018-06-11
Publication date: 2020-11-24
Anticipated expiration: 2038-06-11
Also published as: CN108546247A

Abstract

The invention relates to an application of an alkaloid compound in preparation of anti-obesity drugs. The alkaloid compound has a structure shown in a formula I, or pharmaceutically acceptable salt thereof; the application of the structural analogue of the compound in the preparation of anti-obesity drugs singly or in the form of a composition. Extracting Aspergillus terreus from seaAspergillus terreusML-44 is subjected to fermentation culture to obtain a fermentation product containing the compound of the formula I, and the fermentation product is separated and purified to obtain the compound of the formula I. Wherein, the Aspergillus terreus ML-44 has a preservation number of CGMCC No.15664, a preservation date of 2018, 04 and 17 days, and the preservation place is China general microbiological culture Collection center (CGMCC). The invention discovers that the compound and the structural analogue thereof have anti-obesity activity for the first time, and provides a lead compound for developing novel anti-obesity medicaments.

Description

Application of alkaloid compound in preparation of anti-obesity drugs

Technical Field

The invention relates to an application of an alkaloid compound in preparation of anti-obesity drugs.

Background

With The development of socioeconomic, The incidence of obesity in our country is increasing year by year, and The number of patients is increasing from 240 ten thousand in 1975 to 9000 ten thousand in 2014, which accounts for 14.3% of The worldwide obesity population (NCD Risk patient organization, Trends in adolt body-mass index in 200 countries from 1975 to 2014: a porous analysis of 1698 porous-based diabetes patients with 19.2 million partitionings, The Lancet, 2016, 387, 1377-. Obesity is closely related to The development of a series of chronic diseases such as type II diabetes, cardiovascular and cerebrovascular diseases, cancer, etc. (Smith and Minson, Obesity and adipokines: efficiencies on sympathic overview. J Physiol, 2012, 590, 1787-. The rapid development of obesity, including childhood obesity, not only imposes a huge economic burden on society, but also necessarily endangers the long-term sustainable development of socioeconomic performance in China (T2 DM) in Germany: devilpment and cost-of-illneness analysis. Gesundheitswesen, 2006, 68, 600-. Therefore, obesity prevention and treatment has become a great concern in the field of medical research.

The method for treating obesity mainly comprises reducing intake of energy and food, increasing energy consumption, inhibiting differentiation and proliferation of fat cells, inhibiting fat production, increasing decomposition and oxidation of fat, etc. In the aspect of drug therapy, scholars at home and abroad are around the pathogenesis of obesity, and develop a series of drugs aiming at different targets, such as lipid-lowering drugs, inhibition of fat absorption (orlistat) in intestinal tracts, promotion of fat metabolism and the like; or the adrenergic medicine is directly acted on the central nervous system to play the roles of suppressing appetite, reducing food intake and reducing weight. However, the medicaments generally have the prominent problems of obvious side effect, low cure rate, obvious rebound after stopping administration and the like, and have limited clinical application value. Therefore, the screening and finding of more ideal anti-obesity drugs have important practical significance.

Screening of lipid having regulation function from natureThe generated and metabolized micromolecular substance is one of the hot spots of the research in the Obesity field at present, Natural Products provide rich chemical diversity, and can provide lead compounds for developing new Natural weight-reducing drugs (Fu et al, Natural Products with Anti-Obesity Effects and Difference Mechanisms of action, J age Food Chem, 2016, 64, 9571-. The secondary metabolite of the microorganism is an important source of new drugs and lead structures thereof, and the research of screening and obtaining natural products with related activity for regulating lipid metabolism from the microorganism is rarely reported. Screening by taking pluripotent mesenchymal stem cells C3H10T1/2 as a cell model, finding that the fermentation products of three marine-derived fungi have the activity of inhibiting the lipogenic differentiation of C3H10T1/2, wherein Aspergillus terreus ML-44 (Aspergillus terreusML-44) has a lower minimum effective concentration. Further activity-tracking isolation an alkaloid compound methyl 3,4,5-trimethoxy-2- (2- (nicotinamido) benzamido) benzoate (formula I) was isolated and identified from a large number of ML-44 fermentation products. The compounds of the type reported at present have five in total, and are formed by connecting two molecules of anthranilic acid or its derivative and one molecule of 3-picolinic acid (nicotinic acid) through amido bond or further performing intramolecular ring formation (Arai et al, Metabolic products of the family of chemical industries, Inc., etc.)Aspregillus terreus. VI. Metabolites of the strain IFO 8835. (3). The isolation and chemical structures of colorless metabolites. Chem Pharm Bull, 1981, 29, 1005–1012.; Wang et al., Three new compounds from Aspergillus terreus PT06-2 grown in a high salt medium, Mar Drugs, 2011, 9, 1368–1378; He et al., Asperterrestide A, a Cytotoxic Cyclic Tetrapeptide from the Marine-Derived Fungus Aspergillus terreus SCSGAF0162. J Nat Prod, 2013, 76, 1182–1186; Chen et al., Structurally diverse secondary metabolites from a deep-sea-derived fungus Penicillium chrysogenumSCSIO 41001 and the biological evaluation, Fitoterapia, 2017, 117: 71-78). Few reports are available on the biological activity of the compounds, especially in the regulation of lipidsThe biological activity in the aspects of metabolism and obesity resistance is not reported.

Disclosure of Invention

The invention aims to provide application of an alkaloid compound in preparation of anti-obesity drugs. The alkaloid compound has obvious effects of inhibiting adipocyte differentiation in vitro and resisting obesity in vivo, namely the compound has good anti-obesity function.

The structure of the alkaloid compound provided by the invention is shown as formula I:

I

the present invention also provides structural analogs of compounds of formula I: use of terramide a, terramide B, terramide C, terramide D, or a pharmaceutically acceptable salt thereof, alone or in combination, for the manufacture of an anti-obesity medicament.

The invention also relates to Aspergillus terreus (A)Aspergillus terreus) ML-44, its preserving number is CGMCC number 15664, preserving date is 2018, 17.04.2018, preserving unit is China general microbiological culture Collection center (CGMCC), the address is No. 3 of Xilu No.1 of Beijing city facing Yang district, institute of microbiology of China academy of sciences, zip code 100101. The strain is separated from a sample of the digestive tract of pacific oyster, and is identified as aspergillus terreus (a), (b) and (c) through taxonomic studyAspergillus terreus）。

The preparation method of the compound of formula I provided by the invention comprises the following steps:

mixing the above-mentioned Aspergillus terreus (A), (B), (C), (Aspergillus terreus) ML-44 is subjected to fermentation culture to obtain a fermentation product containing the compound shown in the formula I, and the fermentation product is separated and purified to obtain the compound. Other Aspergillus producers capable of producing compounds of formula I may also be used for the fermentation culture.

The separation and purification includes conventional methods for separation and purification of natural products, such as extraction, column chromatography, high performance liquid preparation, etc., which are well known to those skilled in the art.

The invention provides a specific preparation method of a compound shown in the formula I, which comprises the following steps:

1) mixing the above-mentioned Aspergillus terreus (A), (B), (C), (Aspergillus terreus) ML-44 performing rice solid fermentation culture to obtain a fermentation product;

2) ultrasonically extracting the obtained fermented product with 50-90% (v/v) acetone aqueous solution, filtering, concentrating the filtrate under reduced pressure until the filtrate does not contain acetone, and extracting the residual suspension with ethyl acetate to obtain ethyl acetate extract of the fermented product;

3) concentrating the ethyl acetate extract obtained in the step 2) under reduced pressure to dryness to obtain an ethyl acetate extract;

4) sequentially separating the ethyl acetate total extract by silica gel column chromatography (petroleum ether ‒ dichloromethane ‒ methanol gradient elution), Sephadex LH-20 column chromatography (methanol elution) and ODS column chromatography (water → 100% methanol gradient elution) to obtain column chromatography components containing the compound;

5) the column chromatography fraction containing the compound was separated by HPLC.

Wherein, the acetone aqueous solution in the step 2) is 70-90% (v/v), preferably 75-85% (v/v), and more preferably 80% (v/v).

The invention provides an extract of aspergillus terreus fermentation product, which contains the compound shown in the formula I and structural analogues thereof. Specifically, the extract is an ethyl acetate extract, an ethyl acetate total extract or a chromatography component of the aspergillus terreus fermentation product. The extract can be prepared by referring to the corresponding steps of the above preparation method of the compound of the present invention. Specifically, the extract can be prepared by extraction, column chromatography and high performance liquid chromatography.

The invention provides application of an extract of an aspergillus terreus fermented product in preparing an anti-obesity drug.

The invention provides application of the aspergillus terreus in preparing a compound or a fermentation product extract of a formula I.

In summary, the present invention provides a medicament comprising the structure of formula I or a pharmaceutically acceptable salt thereof, the medicament comprising a pharmaceutically acceptable adjuvant; the medicine is injection, tablet, pill, capsule, suspension or emulsion, etc.

The term "pharmaceutically acceptable salt" in the present invention may be a pharmaceutically acceptable inorganic or organic salt. The compounds of formula I of the present invention having basic groups can form pharmaceutically acceptable salts with inorganic acids, such as sulfate, hydrochloride, hydrobromide, phosphate; pharmaceutically acceptable salts can also be formed with organic acids such as acetates, oxalates, citrates, gluconates, succinates, tartrates, p-toluenesulfonates, methanesulfonates, benzoates, lactates, maleates, and the like.

The compound of the formula I and the structural analogs thereof can be combined with various pharmaceutically acceptable carriers, excipients or auxiliary materials to prepare anti-obesity medicaments for treating obesity and related metabolic syndromes.

The compounds of the present invention may be administered alone or in the form of a pharmaceutical composition. The route of administration may be oral, parenteral or topical. The pharmaceutical composition can be formulated into various suitable dosage forms according to the administration route.

Pharmaceutical compositions of the compounds of the present invention may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, buccal, topical, parenteral, e.g. subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or via an external reservoir. Among them, oral, intraperitoneal or intravenous administration is preferable.

When administered orally, the compounds of the present invention may be formulated in any orally acceptable dosage form, including but not limited to tablets, capsules, aqueous solutions or suspensions. Among these, carriers for tablets generally include lactose and corn starch, and additionally, lubricating agents such as magnesium stearate may be added. Diluents used in capsule formulations generally include lactose and dried corn starch. Aqueous suspension formulations are generally prepared by mixing the active ingredient with suitable emulsifying and suspending agents. Optionally, some sweetener, aromatic or colorant may be added into the above oral preparation.

When applied topically to the skin, the compounds of the present invention may be formulated in a suitable ointment, lotion, or cream formulation wherein the active ingredient is suspended or dissolved in one or more carriers. Carriers that may be used in ointment formulations include, but are not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; carriers that can be used in lotions or creams include, but are not limited to: mineral oil, sorbitan monostearate, tween 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The compounds of the present invention may also be administered in the form of sterile injectable preparations, including sterile injectable aqueous or oleaginous suspensions or solutions. Among the carriers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, the sterilized fixed oil may also be employed as a solvent or suspending medium, such as a monoglyceride or diglyceride.

It is further noted that the dosage and method of administration of the compounds of the present invention will depend upon a variety of factors including the age, weight, sex, physical condition, nutritional status, the activity level of the compound, time of administration, metabolic rate, severity of the condition, and the subjective judgment of the treating physician. The preferred dosage is between 0.01-100 mg/kg body weight/day.

The invention provides a preliminary cell activity test of a compound shown in a formula I, wherein the compound shown in the formula I can inhibit mouse embryonic mesenchymal C3H10T1/2 cells from differentiating into fat cells and obviously inhibit fat accumulation in the cells; the in-vivo activity pre-experimental result shows that the composition can obviously inhibit the occurrence of male drosophila obesity induced by high-fat diet, and the weight of the drosophila is maintained at a lower level. Therefore, the compound has good anti-obesity function, and provides a lead compound for developing novel anti-obesity medicaments.

Drawings

FIG. 1 Compound I significantly inhibited adipogenic differentiation of C3H10T1/2 cells a) blank control, b) Compound I (20. mu.M).

Figure 2 compound I significantly inhibited high fat-induced male drosophila obesity.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by manufacturers, and are all conventional products available on the market.

In the following examples, the compound I, i.e. the compound of formula I, is a fungus of the invention from a marine originAspergillus terreusML-44 fermentation product ethyl acetate extract, and the anti-obesity function is found for the first time.

The arabic numerals indicate the corresponding index bits.

I

In the structural study of the following examples, ESI-MS was measured using an LCQ fly LC MS (Thermo Scientific, USA) and NMR spectra were measured using a superconducting NMR spectrometer (500 MHz, model III 500, Avance, Bruke, Switzerland)¹H-NMR，125 MHz ¹³C-NMR).

Example 1: microbial fermentation culture and preparation of compounds

1. Fermentation culture and extraction treatment of fermented product

1) Production strain

The producer of the present example for the fermentative production of Compound I is Aspergillus terreus isolated from the digestive tract of Pacific oyster (A. pacificus) ((II. RTM.))Aspergillus terreus) The ML-44 strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC number 15664.

2) Fermentation culture

Culturing at 4 deg.C according to conventional methodTaking out Aspergillus in refrigeratorAspergillus terreus) ML-44 tube slant, scrape appropriate amount of spores with inoculating loop under aseptic condition, streak-inoculate on newly prepared 5 PDA solid media (composition: glucose 2%, agar 2%, NaCl 1.5%, prepared from 20% potato water decoction), activating and culturing for 5 days in an incubator at 28 ℃, respectively washing with 10ml of sterile water to obtain spores, and combining and dispersing all the spores in 100 ml of sterile water to obtain spore suspension. The spore suspension was inoculated into 24 1000 ml Erlenmeyer flasks each containing a rice solid medium (prepared by adding 150 ml of natural seawater to 80 g of commercially available rice per flask, standing overnight, and sterilizing at 121 ℃ for 30 min), and allowed to stand at room temperature for 35 days.

2. Extraction treatment and preparation of ethyl acetate extract

Soaking and suspending the fermented product with 80% acetone aqueous solution (500 ml per bottle), ultrasonic treating for 1h, leaching at room temperature for 24h, filtering with 4 layers of gauze, and repeating the operation for 3 times. Mixing filtrates, concentrating under reduced pressure until acetone is removed, extracting the residual water layer about 3L with equal volume of ethyl acetate for 3 times to obtain fermented product ethyl acetate extract, and concentrating under reduced pressure to dry to obtain 66.2 g ethyl acetate extract.

3. Column chromatography separation of ethyl acetate extract and preparation of column chromatography component containing target compounds I-V

Dissolving 66.2 g of the ethyl acetate extract with 200ml of dichloromethane-methanol (v/v 1: 1), filtering insoluble substances, adding 90 g of chromatographic silica gel (200-mesh and 300-mesh) for adsorption and sample mixing, drying by distillation under reduced pressure, adding the mixture to a silica glass pressure-reduced column (120 g of silica gel and 4.8 multiplied by 18.0 cm of a column bed), and performing gradient elution chromatography by using a petroleum ether-dichloromethane-methanol solvent system to obtain a plurality of fractions. According to the thin-layer analysis result of silica gel, merging corresponding fractions, and obtaining 8 components according to the elution polarity sequence: fr-1 (12.4 g, petroleum ether), Fr-2 (6.3 g, petroleum ether-dichloromethane 1:1 elution), Fr-3 (4.5 g, petroleum ether-dichloromethane 1:2 elution), Fr-4 (11.6 g, dichloromethane elution), Fr-5 (15.8 g, dichloromethane-methanol 99:1 elution), Fr-6 (6.2 g, dichloromethane-methanol 95:5 elution), Fr-7 (1.4 g, dichloromethane 9:1 elution), Fr-8 (1.9 g, dichloromethane-methanol 7:3 elution).

Dissolving Fr-4 (11.6 g) with about 50 ml methanol, loading on Sephadex LH-20 column, eluting with methanol, and collecting 18 fractions (each 30 ml); according to the HPLC analysis, the fractions ML-4 gel chromatography fractions 1-18 were combined into 8 fractions: Fr-4-1-Fr-4-8. After the fraction Fr-4-5 (about 1.8 g) was dissolved in methanol and filtered through a 0.45 μm filter, it was separated by a QuikSep medium-low pressure chromatography system (Huideyi, Beijing, China) with 80% methanol (flow rate 10 ml/min, room temperature) on a glass column (bed 1.8 cm. times.30 cm) pre-packed with ODS. Taking 7 components according to an ultraviolet detection result: ML-4-5-1-ML-4-5-7. Wherein Fr-4-5-2 contains the target compound I.

4. HPLC preparation of Compound I

Component Fr-4-5-2 (180 mg) containing compound I was dissolved in 5 ml of methanol, filtered through a 0.45 μm filter, and subjected to HPLC separation using a Gemini C18 preparative column (21.2 mm. times.250 mm) using a Waters 2545 type HPLC system (column temperature 26 ℃, flow rate 10 ml/min with 68% methanol as the mobile phase, 0.5 ml per sample injection, detection wavelength 210 and 254 nm) to give compound I (62 mg,t _R = 19.6 min）。

physicochemical constants and spectral data of Compound I

The compound I is colorless needle-like crystals (MeOH), m.p. 137-139-^oC. Positive ion ESI-MS:m/z 466 [M + H]⁺，488 [M + Na]⁺，931 [2M + H]⁺(ii) a Negative ion ESI-MS:m/z 464 [M - H]^-，490 [M + Cl]^-。¹H NMR（500 MHz，CDCl₃）：12.20 (1H, s, 2-NH), 9.20 (1H, br s, 2ʹʹ-NH), 9.19 (1H, d, J = 1.90 Hz, H-2ʹ), 8.79 (1H, br d, J = 7.85, H-3), 8.67 (1H, dd, J = 4.80, 1.90 Hz, H-4ʹ), 8.21 (1H, dt, J = 7.90, 1.90 Hz, H-6ʹ), 7.87 (1H, dd, J= 7.85, 1.10 Hz, H-6), 7.56 (1H, td, J = 7.85, 1.10 Hz, H-4), 7.33 (1H, dd, J= 7.90, 4.80 Hz, H-5ʹ), 7.24 (1H, s, H-6ʹʹ), 7.19 (1H, br t, J = 7.85 Hz, H-5), 3.94 (3H, s, 5ʹʹ-OCH₃), 3.874 (3H, s, 4ʹʹ-OCH₃), 3.867 (3H, s, 3ʹʹ-OCH₃), 3.78 (3H, s, 7ʹʹ-OCH₃)。¹³C NMR（125 MHz，CDCl₃）：168.2 (C-7ʹʹ), 167.2 (C-7), 164.0 (C-7ʹ), 152.6 (C-4ʹ), 151.5 (C-5ʹʹ), 149.2 (C-4ʹʹ), 148.9 (C-2ʹ), 146.9 (C-3ʹʹ), 140.4 (C-2), 135.2 (C-6ʹ), 133.6 (C-4), 130.6 (C-1ʹ), 127.9 (C-6), 125.7 (C-2ʹʹ), 123.8 (C-5), 123.6 (C-5ʹ), 121.8 (C-3), 120.4 (C-1), 119.1 (C-1ʹʹ), 108.8 (C-6ʹʹ), 61.24 (4ʹʹ-OCH₃), 61.23 (3ʹʹ-OCH₃), 56.5 (5ʹʹ-OCH₃), 52.7 (7ʹʹ-OCH₃). The NMR data were assigned in comparison with literature data.

Example 2: compound I inhibits adipogenic differentiation of C3H10T1/2 cells

1) Growth inhibition of C3H10T1/2 cells by Compound I

The MTT method is adopted to detect the influence of the compound I on the vitality of the C3H10T1/2 cells. Taking C3H10T1/2 cells in logarithmic growth phase, preparing the cell density to be 2 x 10 by using fresh DMEM medium⁵Each/ml cell suspension was inoculated into a 96-well plate at 200. mu.l/well, and after 12 hours of incubation at 37 ℃ 2. mu.l each of the sample solutions was added to each well of the sample group, and 2. mu.l each of the methanol solution was added to each well of the blank control group, and the mixture was treated at 37 ℃ for 48 hours. Mu.l of precooled 5mg/ml MTT solution (prepared by PBS solution) is added into each well, after incubation for 4h at 37 ℃, the mixture is centrifuged at 4 ℃ and 2000 rpm for 10 min, supernatant is absorbed, 150 mu.l DMSO is added into each well, the mixture is placed on a microplate reader to be fully shaken to completely dissolve MTT purple products, and the OD value at 570 nm of each well is measured. In the experiment, three parallel holes are respectively arranged on the sample and the blank control group, the OD average value is taken, and the ratio of IR percent to (OD) is calculated_{Blank space}-OD_{Sample (I)})/OD_{Blank space}X 100% equation, calculate the inhibition rate (IR%) of the sample on C3H10T1/2 cells. In the MTT assay, compound I did not show any cytotoxic or growth inhibitory effect on C3H10T1/2 cells at the action concentration of 100. mu.M, as examined by microscopy.

2) Compound I inhibits C3H10T1/2 cell differentiation into adipocytes

C3H10T1/2 cells are inoculated into a 6-well plate and cultured until the cells are fully paved into a 80% dish bottom, and then a fresh induction culture medium (DMEM culture medium +10% fetal calf serum +0.5 mM isobutyl methylxanthine +0.1 mM indomethacin +1.0 mu M dexamethasone +10 mu g/mL insulin +1% streptomycin double-antibody solution) is added for culture for 2 days; changing a differentiation culture medium (DMEM culture medium +10% fetal calf serum +1.0 mu g/mL insulin +1% streptomycin double antibody solution) and continuing to culture for 2 days; the growth medium was changed (DMEM medium +10% fetal bovine serum +1% streptomycin double antibody solution) and the culture was continued for 4 days (fresh medium was changed every 2 days). The treatment group was supplemented with 2. mu.l each of the sample solutions to each of the culture media, and the control group was not subjected to the same procedure. On the 8 th day of culture, oil red O staining and cytomicroscopic examination were performed, respectively. The staining results show that most cells in the blank group successfully differentiated into mature adipocytes, and oil drops in the cells were large and large, while the cells in the treated group had few oil drops (fig. 1), indicating that compound I can very significantly inhibit the adipogenic differentiation of C3H10T1/2 cells.

Example 3: compound I inhibits the development of high-fat-induced male Drosophila obesity

The body weight of the drosophila melanogaster in each group was measured by dividing 3-day-old male drosophila melanogaster into 3 groups, culturing the control group in a minimal medium, administering a high-fat medium (minimal medium supplemented with 15% lard) to the HFD-induced group, administering a high-fat medium supplemented with Compound I (final concentration of 0.1 mg/ml) to the treated group, and continuously treating for 20 days. As can be seen from the results of the test in FIG. 2, the fruit fly weight can be significantly increased by the culture in the high-fat medium, resulting in the formation of obese fruit flies; concurrent treatment with compound I significantly inhibited HFD-induced drosophila obesity.

Although specific embodiments of the invention have been described in detail, it will be appreciated by those skilled in the art that, based upon the overall teachings of the disclosure, various modifications and alternatives to those details could be developed and still be encompassed by the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims

1. An application of an alkaloid compound in preparing anti-obesity drugs is disclosed, wherein the alkaloid compound has a structure shown in a formula I:

I。

2. use according to claim 1, the alkaloid compound being replaced by an analogue of the structure:

。

3. use according to claim 1, characterized in that it is a process for the preparation of the alkaloid compounds, characterized in that it comprises the following steps:

1) mixing A. terreus (A. terreus)Aspergillus terreus) ML-44 performing rice solid fermentation culture to obtain a fermentation product;

2) ultrasonically extracting the obtained fermented product with 50-90% acetone aqueous solution, filtering, concentrating the filtrate under reduced pressure until the filtrate does not contain acetone, and extracting the residual suspension with ethyl acetate to obtain ethyl acetate extract of the fermented product;

4) sequentially carrying out silica gel column chromatography, petroleum ether ‒ dichloromethane ‒ methanol gradient elution, Sephadex LH-20 column chromatography, methanol elution and ODS column chromatography, and water → 100% methanol gradient elution on the ethyl acetate extract, and separating to obtain column chromatography components containing the compound;

5) separating the column chromatography fraction containing said compound by HPLC;

the preservation number of the aspergillus terreus ML-44 is CGMCC No. 15664.

4. Use according to claim 3, characterized in that the concentration of the aqueous acetone solution in step 2) is 80%, v/v.

5. Use according to claim 1, characterized in that the medicament is an injection, a tablet, a pill, a capsule, a suspension or an emulsion.