CN111139188B - Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs - Google Patents
Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs Download PDFInfo
- Publication number
- CN111139188B CN111139188B CN202010017838.7A CN202010017838A CN111139188B CN 111139188 B CN111139188 B CN 111139188B CN 202010017838 A CN202010017838 A CN 202010017838A CN 111139188 B CN111139188 B CN 111139188B
- Authority
- CN
- China
- Prior art keywords
- derivative
- derivatives
- heteroterpene
- preparation
- ethyl acetate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/10—Spiro-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J73/00—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms
- C07J73/001—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom
- C07J73/003—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom by oxygen as hetero atom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/181—Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P33/00—Preparation of steroids
- C12P33/20—Preparation of steroids containing heterocyclic rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a novel skeleton heteroterpene derivative derived from marine fungi and application thereof in preparing anti-inflammatory drugs. The invention obtains a strain of marine fungus Aspergillus terreus GZU-311 by separation and screening, the strain is preserved in Guangdong province microorganism strain preservation center 12 and 17 days 2019, and the preservation number is GDMCC No: 60789. further separating and identifying the heteroterpene derivatives from the fermentation culture products of the fungi, wherein the derivatives have stronger NO production resistance activity, the anti-inflammatory activity of the derivatives is remarkably improved by about 2 times compared with that of the positive control indomethacin, and the derivatives have great safety factors; therefore, the diterpenoid derivative provided by the invention has good clinical application prospect in the preparation of anti-inflammatory drugs, especially in the preparation of drugs for inhibiting the generation of NO induced by LPS.
Description
Technical Field
The invention belongs to the technical field of medicines. More particularly, relates to a novel skeleton heteroterpene derivative derived from marine fungi and application thereof in preparing anti-inflammatory drugs.
Background
Inflammation is a defensive response that occurs when an organism is subjected to an external antigenic stimulus. The macrophage is an important immune cell in the organism, and has the functions of regulating and controlling inflammatory factors, resisting infection, resisting tumor and regulating immunity. Macrophages and their inflammatory mediators released are now thought to be involved in the development and progression of a variety of inflammatory diseases, such as atherosclerosis, rheumatoid arthritis, inflammatory bowel disease, sepsis, gouty arthritis, and the like.
At present, there are two main types of anti-inflammatory drugs commonly used in clinic: non-steroidal anti-inflammatory drugs and steroidal anti-inflammatory drugs. Although both anti-inflammatory drugs have certain clinical anti-inflammatory effects, a series of adverse reactions, side effects and tolerance, such as gastric mucosa injury, liver injury, kidney injury and the like, are generated after long-term use of the drugs in large quantities, and great burden and injury are brought to patients. Therefore, there is a need to find new anti-inflammatory drugs to replace the existing non-steroidal anti-inflammatory drugs and steroidal anti-inflammatory drugs to solve the tolerance of patients to the existing anti-inflammatory drugs and the adverse reactions of the existing anti-inflammatory drugs to the patients.
The secondary metabolite of the marine fungus is an important component of a natural product, has the characteristics of sustainability, environmental friendliness, abundant and various metabolites and the like, and is always an important source for natural drug screening; meanwhile, secondary metabolites of marine fungi have wide physiological activities such as antibacterial, antitumor, immunoregulatory, anti-inflammatory, enzyme inhibition and the like. At present, the search for new drug source molecules from marine fungi has become a hot spot of international and domestic research. In the previous research work, the applicant obtains a marine fungus Diaporthe sp.GZU-1021 by separation and screening from sea crab trunks, and obtains a naphthoquinone compound by fermentation production of the marine fungus, wherein the naphthoquinone compound has good anti-inflammatory activity, can obviously inhibit NO production induced by LPS, and shows that the marine fungus has potential in the aspect of fermentation production of the compound with anti-inflammatory activity. Therefore, more marine fungi are separated and screened to ferment and produce more compounds with anti-inflammatory activity, and the method has important significance for treating inflammatory diseases.
Disclosure of Invention
The invention aims to overcome the defects and defects that the existing anti-inflammatory drugs have side effects, adverse reactions and tolerance to patients, and provides a novel skeleton heteroterpene derivative derived from marine fungi and application thereof in preparing anti-inflammatory drugs.
The invention aims to provide an Aspergillus terreus GZU-311 strain of marine fungus.
The invention also aims to provide the application of the marine fungus Aspergillus terreus GZU-311 in the fermentation production of the diterpenoid derivatives.
The invention also aims to provide a hetero-terpenoid derivative.
The invention also aims to provide a preparation method of the diterpenoid derivative.
The invention also aims to provide application of the diterpenoid derivatives in preparation of anti-inflammatory drugs.
The above purpose of the invention is realized by the following technical scheme:
the invention firstly provides a strain of marine fungus Aspergillus terreus GZU-311, which has been deposited in Guangdong province microorganism culture collection center in 2019, 12 and 17 days, and the deposit number is GDMCC No: 60789, the preservation address is Guangzhou institute for microorganisms of Guangdong province, No. 59 building, No. 5 building, of Michelia Tokoro, Michelia Tourette, Guangzhou, City.
The application of the marine fungus Aspergillus terreus GZU-311 in the fermentation production of the diterpenoid derivatives also belongs to the protection scope of the invention.
The invention also provides a hetero-terpenoid derivative, wherein the structural formula of the derivative is shown as the formula (I), the formula (II) or the formula (III):
the invention also provides a preparation method of the diterpenoid derivative, which is prepared by fermenting and producing the Aspergillus terreus GZU-311 marine fungus.
Preferably, the preparation method of the diterpenoid derivative comprises the following steps:
s1, inoculating Aspergillus terreus GZU-311 to a liquid culture medium, and culturing to obtain a seed solution;
s2, inoculating the seed liquid obtained in the step S1 into a solid rice culture medium, and standing for 30-60 days at 28-35 ℃ to obtain a fermentation culture product;
s3, extracting the fermentation culture product obtained in the step S2 with methanol for 2-5 times, concentrating an extracting solution, and extracting the obtained concentrated extract with ethyl acetate to obtain an ethyl acetate crude extract;
s4, separating the ethyl acetate crude extract obtained in the step S3 by using normal phase silica gel chromatography, performing gradient elution by using petroleum ether/ethyl acetate, collecting 10-50% petroleum ether/ethyl acetate part, and performing separation and purification by using silica gel, gel and C-18 reverse phase column chromatography separation technology to obtain the diterpenoid derivative.
Preferably, the formula of the slant culture medium in step S1 is: by mass ratio, 0.3 percent of glucose, 0.1 percent of yeast extract, 0.1 to 0.5 percent of peptone, 1.5 to 2.5 percent of agar, 1.5 to 4 percent of sodium chloride and the balance of water.
More preferably, the formula of the slant culture medium in step S1 is: by mass ratio, glucose 0.3%, yeast extract 0.1%, peptone 0.5%, agar 2.5%, sodium chloride 3%, and water in balance.
Preferably, the temperature for the cultivation in step S1 is 28-35 ℃.
More preferably, the temperature of the cultivation in step S1 is 30 ℃.
Preferably, the culturing time in the step S1 is 4-10 d.
More preferably, the incubation time of step S1 is 6 d.
Preferably, the formulation of the solid rice culture medium in step S2 is: rice, by mass ratio: 1, sea water: 1 to 2.
More preferably, the formulation of the solid rice culture medium in step S2 is: rice, by mass ratio: 1, sea water: 1.
preferably, the temperature of the standing in step S2 is 35 ℃.
Preferably, the standing time in step S2 is 60 d.
Preferably, the number of methanol extractions in step S3 is 3.
Preferably, the petroleum ether/ethyl acetate in step S4 refers to a mixture of petroleum ether/ethyl acetate in volume ratio.
Creative research researches show that the diterpenoid derivatives can obviously inhibit NO generation induced by LPS, have good anti-inflammatory activity and can be used for preparing anti-inflammatory drugs. Therefore, the application of the diterpenoid derivatives in the preparation of anti-inflammatory drugs also falls within the protection scope of the invention.
Preferably, the application is the application in the preparation of medicines for inhibiting NO production induced by LPS.
The invention has the following beneficial effects:
the invention provides a novel skeleton heteroterpene derivative derived from marine fungi and application thereof in preparing anti-inflammatory drugs. The invention separates and screens from the body of the Onchidium struma to obtain a strain of marine fungus Aspergillus terreus GZU-311, and separates and identifies the heteroterpene derivative from the fermentation culture product of the fungus, the derivative has stronger activity of resisting NO generation, the anti-inflammatory activity of the derivative is obviously improved by about 2 times compared with that of positive control indomethacin, and the derivative is in IC50Under the concentration, the compound does not show cytotoxic activity and has a great safety factor. Therefore, the diterpenoid derivative provided by the invention has good clinical application prospect in the preparation of anti-inflammatory drugs, especially in the preparation of drugs for inhibiting the generation of NO induced by LPS.
Detailed Description
The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
EXAMPLE 1 obtaining of the Marine fungus Aspergillus terreus GZU-311
1. Experimental methods
The marine fungus was isolated from the trunk of Tugassum maritima, Xuwen county, Zhanjiang province, Guangdong, reference (Nature protocols,2010,5, 480-: extracting DNA of the marine fungus by adopting a DNeasy Plant Mini Kit according to the steps of the instruction, amplifying an ITS region by taking the DNA as a template, sequencing to obtain an ITS-rRNA gene sequence of the marine fungus, and identifying through a sequence result.
2. Results of the experiment
The ITS-rRNA gene sequence of marine fungi is shown as SEQ ID NO.1, similarity retrieval is carried out through a BLAST database, and through comparison, the similarity of the sequence and the Aspergillus terreus fungal sequence reaches over 99 percent; thus, the marine fungus was identified as belonging to the marine fungus Aspergillus terreus, named: the marine fungus Aspergillus terreus GZU-311 is deposited in Guangdong province microorganism culture collection center 12 and 17 months in 2019, and the deposit number is GDMCC No: 60789, the preservation address is Guangzhou institute for microorganisms of Guangdong province, No. 59 building, No. 5 building, of Michelia Tokoro, Michelia Tourette, Guangzhou, City.
Example 2 isolation and identification of Heteroterpenoid derivatives
In this example, the specific experimental method and experimental results of the production of the triterpene derivative by fermenting Aspergillus terreus GZU-311 are as follows:
1. isolation of derivatives
The preparation method of the derivative comprises the following steps:
s1, inoculating the marine fungus Aspergillus terreus GZU-311 obtained by screening in the embodiment 1 into a planar culture medium, then inoculating into a liquid culture medium, and culturing for 6 days at 30 ℃ to obtain a seed solution;
s2, inoculating the seed liquid obtained in the step S1 into a solid rice culture medium, and standing for 60 days at 35 ℃ to obtain a fermentation culture product;
s3, extracting the fermentation culture product obtained in the step S2 for 3 times by using methanol, concentrating an extracting solution, and extracting the obtained concentrated extract by using ethyl acetate to obtain an ethyl acetate crude extract;
s4, separating the ethyl acetate crude extract obtained in the step S3 by using normal phase silica gel chromatography, performing gradient elution by using petroleum ether/ethyl acetate (the volume ratio of the petroleum ether to the ethyl acetate is respectively 10:90, 20:80, 30:70, 40:60, 50:50, 60:40 and 70:30), collecting 10-50% of petroleum ether/ethyl acetate part, and performing separation and purification by using silica gel, gel and C-18 reverse phase column chromatography separation technology to obtain the derivatives 1, 2 and 3;
wherein, the formula of the slant culture medium is as follows: by mass ratio, 0.3% of glucose, 0.1% of yeast extract, 0.5% of peptone, 2.5% of agar, 3% of sodium chloride and the balance of water;
the formula of the solid rice culture medium is as follows: rice, by mass ratio: 1, sea water: 1.
2. identification of derivatives
The following experimental data were obtained by performing structural test analysis on derivatives 1, 2 and 3:
derivative 1 has the formula: c28H32O10High resolution mass spectrometry (HRESI-MS): 527.1902[ M-H]-(cald for C28H31O10 527.1917)；
Derivative 2 has the formula: c28H32O10High resolution mass spectrometry (HRESI-MS): 527.1914[ M-H]-(cald for C28H31O10 527.1917)；
Derivative 3 has the formula: c28H34O10High resolution mass spectrometry (HRESI-MS): 529.2056[ M-H]-(cald for C28H31O10 529.2073)。
Nuclear Magnetic Resonance (NMR) data for derivatives 1, 2 and 3 are shown in table 1, respectively.
TABLE 1 NMR data (CDCl) for derivatives 1, 2 and 33，400MHz/100MHz,ppm)
The structural formulas of the derivatives 1, 2 and 3 are respectively shown as a formula (I), a formula (I I) or a formula (II):
example 3 anti-inflammatory cell screening model and cytotoxicity screening study of Heteroterpenoid derivatives
When exogenous antigen Lipopolysaccharide (LPS) stimulates macrophages, a series of immune reactions are generated, and inflammatory factors NO are released2 -，NO2 -The level may indirectly reflect the degree of inflammation. The Griess reagent is a reagent which can react with NO2 -Azo dye compound with pigment generated by interaction can judge NO by detecting the absorbance of the dye by a microplate reader2 -And thereby using this model to screen for small molecule inhibitors with anti-inflammatory activity. Thus, the present example uses the following experiments to study the anti-inflammatory effects of the triterpene derivatives. The specific experimental methods and experimental results are as follows:
1. anti-inflammatory cell screening model
1) Experimental methods
(1) Culture and treatment of cells
RAW 264.7 cells were cultured in DMEM medium (penicillin 60U/mL, streptomycin 100. mu.g/mL) containing 10% serum and placed in 5% CO2The temperature was set at 37 ℃ in the incubator of (1), and conventional maintenance culture and passage were carried out.
(2) Compound intervention
After the cells had stabilized after one generation of culture, they were plated in 96-well plates at a concentration of 2X 104cells/well, adherent for 24 hours; respectively dissolving the mother liquor of the derivatives 1, 2 and 3 obtained in the example 2 in DMSO, and diluting the mother liquor with a DMEM culture medium to ensure that the concentration of the DMSO is not higher than 0.1%; respectively acting derivatives 1, 2 and 3 (2-50 μ M, final concentration) with different concentration gradients on a 96-well plate (200 μ L, LPS:1 μ g/mL), and setting a positive control group (indomethacin); the NO inhibitory effect was determined using a Griess kit, i.e., 100. mu.L of cell supernatant mixed with an equal volume of Griess reagent (0.2% naphtylenediamine dihydrate and 2% sulfonilamide in 5% H)3PO4) (ii) a The absorbance was measured at 540nm with a microplate reader, according to the addition of derivative 1 and a positive controlJudging the inhibition effect of the compound; sodium nitrite was used to make a standard curve, and the experiment was tested in parallel three times; the formula for calculating the inhibition rate is as follows:
inhibition rate (%) ([ NO ]2 -]LPS-[NO2 -]LPS+sample)/([NO2 -]LPS-[NO2 -]untreated)×100。
2) Results of the experiment
The results of the anti-inflammatory activity test of derivatives 1, 2 and 3 are shown in Table 2, and it can be seen that derivatives 1, 2 and 3 all showed strong anti-NO production activity, IC thereof5017.8. mu.M, 14.1. mu.M, 13.4. mu.M, respectively, and IC of indomethacin as a positive control5024.0. mu.M; the anti-inflammatory activity of the derivatives 1, 2 and 3 is shown to be remarkably improved by about 2 times compared with that of the positive control indometacin.
Table 2 anti-inflammatory activity test results for derivatives 1, 2 and 3
2. Cytotoxicity screening
1) Experimental methods
(1) Principle of
The chemical name of MTT is 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyl tetrazole bromide salt determined by an MTT method, and the commercial name is as follows: thiazole blue. The detection principle is as follows: the live and dead cells differ in that the presence of succinate dehydrogenase in mitochondria in live cells reduces exogenous MTT to the water-insoluble blue crystalline formazan. Dimethyl sulfoxide (DMSO) can solubilize the formazan deposited within the cells, whose absorbance, measured at 490 or 540nm wavelength, can indirectly reflect the number of cell survivors, depending on the intervening and positive control groups to which the compound is added.
(2) Cytotoxicity screening assay
RAW 264.7 cells were cultured in DMEM medium (penicillin 60U/mL, streptomycin 100. mu.g/mL) containing 10% serum and placed in 5% CO2In the incubator, is provided withThe temperature is kept at 37 ℃;
after the cells had stabilized after one generation of culture, they were plated in 96-well plates at a concentration of 2X 104cells/well, adherent for 24 hours; respectively dissolving the mother liquor of the derivatives 1, 2 and 3 obtained in the example 2 in DMSO, and diluting the mother liquor with a DMEM culture medium to ensure that the concentration of the DMSO is not higher than 0.1%; derivatives 1, 2 and 3 (2-50. mu.M, final concentration) with different concentration gradients were applied to a 96-well plate (200. mu.L, LPS:1 g/mL);
after 24 hours of culture, adding 20 mu L of MTT solution (5mg/mL, prepared by PBS) into each hole, continuing to incubate for 4 hours, terminating the culture, carefully sucking the supernatant, adding 150 mu L of DMSO into each hole, and shaking for 10min to fully dissolve crystals; the 490nm wavelength absorption was detected by a microplate reader and the results recorded.
The inhibition rate was calculated by the following formula: the inhibition ratio (%) [ (a 0-a)/a 0] × 100%, where a0 is the absorbance OD value of the blank and a is the absorbance OD value of the sample.
5 concentrations of derivative 1, 2 and 3 samples were measured, and a dose-inhibition curve was plotted to obtain IC50 values; each sample was assayed in triplicate and the results expressed as mean ± standard deviation.
2) Results of the experiment
The results of cytotoxicity screening of derivatives 1, 2 and 3 are shown in Table 2, and it can be seen that derivatives 1, 2 and 3 are in IC50Under the concentration, the compound does not show cytotoxic activity and has a great safety factor.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Sequence listing
<110> Guangzhou college of traditional Chinese medicine (Guangzhou institute of traditional Chinese medicine)
<120> novel skeleton heteroterpene derivative derived from marine fungi and application thereof in preparation of anti-inflammatory drugs
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 614
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
ccttccgtag gtgaacctgc ggaaggatca ttaccgagtg cgggtcttta tggcccaacc 60
tcccacccgt gactattgta ccttgttgct tcggcgggcc cgccagcgtt tgctggccgc 120
cggggggcga ctcgcccccg ggcccgtgcc cgccggagac cccaacatga accctgttct 180
gaaagcttgc agtctgagtg tgattctttg caatcagtta aaactttcaa caatggatct 240
cttggttccg gcatcgatga agaacgcagc gaaatgcgat aactaatgtg aattgcagaa 300
ttcagtgaat catcgagtct ttgaacgcac attgcgcccc ctggtattcc ggggggcatg 360
cctgtccgag cgtcattgct gccctcaagc ccggcttgtg tgttgggccc tcgtcccccg 420
gctcccgggg gacgggcccg aaaggcagcg gcggcaccgc gtccggtcct cgagcgtatg 480
gggcttcgtc ttccgctccg taggcccggc cggcgcccgc cgacgcattt atttgcaact 540
tgtttttttt ccaggttgac ctcggatcag gtagggatac ccgctgaact taagcatatc 600
aataagcgga ggaa 614
Claims (9)
1. Marine fungusAspergillus terreusGZU-311, wherein the strain has been deposited at 17.12.2019 in the Guangdong province culture Collection with the following deposition numbers GDMCC No: 60789.
2. a heteroterpenoid derivative having the structural formula (I), formula (I I) or formula (II):
3. a process for the preparation of the triterpenoid derivatives according to claim 2, which comprises using the marine fungus according to claim 1Aspergillus terreusGZU-311 is produced by fermentation.
4. The method of claim 3, comprising the steps of:
S1A method for culturing the marine fungus of claim 1Aspergillus terreusGZU-311 inoculating liquid culture medium, and culturing to obtain seed solution;
s2, inoculating the seed liquid obtained in the step S1 into a solid rice culture medium, and standing for 30-60 days at 28-35 ℃ to obtain a fermentation culture product;
s3, extracting the fermentation culture product obtained in the step S2 with methanol for 2-5 times, concentrating an extracting solution, and extracting the obtained concentrated extract with ethyl acetate to obtain an ethyl acetate crude extract;
s4, separating the ethyl acetate crude extract obtained in the step S3 by normal phase silica gel chromatography, performing gradient elution by petroleum ether/ethyl acetate, collecting 10-50% petroleum ether/ethyl acetate part, and performing separation and purification by silica gel, gel and C-18 reverse phase column chromatography separation technology to obtain the triterpenoid derivative.
5. The method according to claim 4, wherein the formula of the slant medium in step S1 is: by mass ratio, 0.3 percent of glucose, 0.1 percent of yeast extract, 0.1 to 0.5 percent of peptone, 1.5 to 2.5 percent of agar, 1.5 to 4 percent of sodium chloride and the balance of water.
6. The method according to claim 4, wherein the temperature of the culture in the step S1 is 28-35 ℃; the culturing time of the step S1 is 4-10 days.
7. The method according to claim 4, wherein the solid rice medium of step S2 is prepared by: rice, by mass ratio: seawater = 1: 1 to 2.
8. Use of the heteroterpenoid derivative of claim 2 for the preparation of an anti-inflammatory agent.
9. The use according to claim 8, in the manufacture of a medicament for inhibiting LPS-induced NO production.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010017838.7A CN111139188B (en) | 2020-01-08 | 2020-01-08 | Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010017838.7A CN111139188B (en) | 2020-01-08 | 2020-01-08 | Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111139188A CN111139188A (en) | 2020-05-12 |
| CN111139188B true CN111139188B (en) | 2021-11-26 |
Family
ID=70523996
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010017838.7A Active CN111139188B (en) | 2020-01-08 | 2020-01-08 | Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111139188B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111154658B (en) * | 2020-01-13 | 2021-11-26 | 广州中医药大学(广州中医药研究院) | Marine fungus, novel skeleton heteroterpene derivative prepared from marine fungus, and preparation method and application of novel skeleton heteroterpene derivative |
| CN111407764B (en) * | 2020-03-27 | 2021-09-10 | 广州中医药大学(广州中医药研究院) | Application of heteroterpene derivative in preparation of medicine for resisting non-alcoholic steatohepatitis and hepatic fibrosis |
| CN112521398B (en) * | 2020-07-30 | 2022-03-15 | 上海交通大学医学院附属仁济医院 | Sponge epiphyte-derived open-loop rearrangement steroid compound and preparation method and application thereof |
| CN114573568B (en) * | 2020-12-02 | 2023-06-02 | 广州中医药大学(广州中医药研究院) | 2-quinolinone-citrinin hybrid dimer compound and preparation method and application thereof |
| CN113121557B (en) * | 2021-03-01 | 2022-06-17 | 广州中医药大学(广州中医药研究院) | Marine fungus-derived heterpene compound and application thereof in preparation of anti-inflammatory drugs |
| CN113173969B (en) * | 2021-03-01 | 2022-06-17 | 广州中医药大学(广州中医药研究院) | Marine fungus-derived heteroterpene compound and application thereof in preparation of anti-hepatic fibrosis drugs |
| CN113265337B (en) * | 2021-06-17 | 2022-10-14 | 华南农业大学 | Marine aspergillus versicolor and isolated culture method and application thereof |
| CN115011487B (en) * | 2022-05-12 | 2023-08-18 | 宁波大学 | Sponge coaptation fungus and application thereof in preparation of hetero-terpenoid |
| CN116284035B (en) * | 2023-03-14 | 2024-04-30 | 中国科学院南海海洋研究所 | Marine fungus-derived ring-opening indole diterpenoid compound peniditerpenoid A, and preparation method and application thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100573632B1 (en) * | 2003-10-30 | 2006-04-24 | 한국생명공학연구원 | A novel compound inhibiting acetylcholine esterase produced by Aspergillus terreus Fb501 |
| US8383128B2 (en) * | 2008-03-07 | 2013-02-26 | Indian Institute Of Science | Method of protecting plant(s) and a process thereof |
| CN107721990B (en) * | 2017-08-25 | 2020-10-20 | 中山大学 | Marine fungus-derived isoindolinone compounds, preparation method thereof and application thereof in preparation of anti-inflammatory drugs |
| CN108277164B (en) * | 2017-12-28 | 2019-08-20 | 中山大学 | A kind of indene derivatives derived from endophytic fungi in Excoecaria agallocha and its application in preparing anti-inflammatory drugs |
| CN110117546B (en) * | 2019-04-04 | 2022-03-01 | 广州中医药大学(广州中医药研究院) | Naphthoquinone compound derived from marine fungi and anti-inflammatory application thereof |
-
2020
- 2020-01-08 CN CN202010017838.7A patent/CN111139188B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111139188A (en) | 2020-05-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111139188B (en) | Novel skeleton heteroterpene derivative derived from marine fungi and application of novel skeleton heteroterpene derivative in preparation of anti-inflammatory drugs | |
| CN107298671B (en) | Selenolonic acid H from penicillium oxalicum and application thereof in preparing medicine for resisting human colon cancer | |
| CN107721990B (en) | Marine fungus-derived isoindolinone compounds, preparation method thereof and application thereof in preparation of anti-inflammatory drugs | |
| CN108299462B (en) | Mixed source terpene compound and separation method and application thereof | |
| CN107353274B (en) | Selenolonic acid I from penicillium oxalicum and application thereof in preparation of human esophageal cancer resistant medicine | |
| CN107298672B (en) | Application of seclenic acid I derived from penicillium oxalicum in preparation of anti-human colon cancer drugs | |
| CN113121557B (en) | Marine fungus-derived heterpene compound and application thereof in preparation of anti-inflammatory drugs | |
| CN110117546B (en) | Naphthoquinone compound derived from marine fungi and anti-inflammatory application thereof | |
| CN111321095B (en) | Butenolide dimer with alpha-glucosidase inhibition effect and application thereof | |
| CN111154658B (en) | Marine fungus, novel skeleton heteroterpene derivative prepared from marine fungus, and preparation method and application of novel skeleton heteroterpene derivative | |
| CN107485607B (en) | Application of seclenic acid H derived from penicillium oxalicum in preparation of human esophageal cancer resistant medicine | |
| CN107298670B (en) | Application of medicine derived from penicillium oxalicum seclenum ketonic acid H in preparation of anti-human oral epidermoid carcinoma medicines | |
| CN108315264A (en) | A kind of polyketide in sea paint endogenetic fungus source and its application in preparing anti-inflammatory drug | |
| CN112592350A (en) | Polyketide lithocarpin E-G and preparation method and application thereof | |
| CN113801032B (en) | Long-chain fatty acid glycerol alcohol compound Rubracin B, preparation method and application thereof | |
| CN107298669B (en) | Selenolonic acid I from penicillium oxalicum and application of medicine for resisting human oral epidermoid carcinoma | |
| CN111407764A (en) | Application of novel skeleton heteroterpene derivative in preparation of medicine for resisting non-alcoholic steatohepatitis and hepatic fibrosis | |
| CN110407792B (en) | Secalonic acid J compound derived from penicillium oxalicum and preparation method thereof | |
| CN109456899B (en) | Penicillium and method for producing penicillic acid by fermenting penicillium | |
| CN109134416B (en) | Application of seclenic acid H derived from penicillium oxalicum in preparation of human cervical cancer drugs | |
| CN113173969B (en) | Marine fungus-derived heteroterpene compound and application thereof in preparation of anti-hepatic fibrosis drugs | |
| CN111217878B (en) | Andrastone compounds, preparation method thereof and application thereof in preparing antiallergic drugs | |
| CN109456196B (en) | Quinone compound from marine fungi as well as preparation method and application thereof | |
| CN110407794B (en) | Selenolonic acid K derived from penicillium oxalicum and application thereof in inhibiting cancer cell proliferation | |
| CN110407797B (en) | Secalonic acid K compound derived from penicillium oxalicum and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |