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CN116178463B - Anthracene ketone glycoside compound and preparation method and application thereof - Google Patents

Anthracene ketone glycoside compound and preparation method and application thereof Download PDF

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CN116178463B
CN116178463B CN202211695503.6A CN202211695503A CN116178463B CN 116178463 B CN116178463 B CN 116178463B CN 202211695503 A CN202211695503 A CN 202211695503A CN 116178463 B CN116178463 B CN 116178463B
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human
anthrone
cancer cells
oxanthromicina
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CN116178463A (en
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何江波
曹艳茹
王莹
王纪爱
陈秀
陈冰
刘杰阳
王涛
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Kunming University
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Abstract

The invention discloses an anthrone glycoside compound, and a preparation method and application thereof. The compound provided by the invention is separated from fermentation liquor of actinomadurasp.KCC248, and is named oxanthromicinA after identification. The actinomycetes are preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) for 25 days in 1 month 2021, and the preservation address is number 3 of West Song No. 1 of North Star in the Korean area of Beijing city, and the preservation number is CGMCC No.21710. The compound oxanthromicin A provided by the invention can obviously inhibit human acute promyelocytic leukemia cells, human non-small cell lung cancer cells, human liver cancer cells, human breast cancer cells and human colon cancer cells, and the medicine prepared by using the compound can be used for treating related tumor diseases.

Description

Anthracene ketone glycoside compound and preparation method and application thereof
Technical Field
The invention relates to the technical field of medicines, in particular to an anthrone glycoside compound, a preparation method and application thereof.
Background
Tumors are important diseases affecting human health, and the morbidity and mortality of the tumors have continuously increased for a long time, and the tumors become the second leading cause of death worldwide and seriously endanger the physical and mental health of human beings.
Microorganisms are important natural product resources. The microorganism has extremely wide variety and diversified metabolism, and the chemical complexity and diversity of the metabolites are the treasury for developing new medicines. Microbial-derived drugs play an important role in healthcare. Finding lead compounds with antitumor activity from microorganisms has become one of the hot spots of research in the medical field. There are a great deal of literature reports that screening and separating antitumor active ingredients from microbial metabolites has broad prospects. Aiming at the tumor formation mechanism, the anti-cancer active ingredients are searched from the natural products through pharmacological screening or action mechanism and biological activity screening, which is an advanced, rapid and effective screening way for searching novel natural medicines from the natural products at present and is one of strategic transformation strategies of the anti-tumor medicines mainly including innovative medicines.
Disclosure of Invention
The invention aims to provide an anthrone glycoside compound, and a preparation method and application thereof. The compound provided by the invention is oxanthromicinA, has the characteristic of inhibiting various tumor cells, and the medicament prepared by the compound can be widely used for treating various tumor diseases.
In order to achieve the above object, the present invention provides an anthrone glycoside compound, named oxanthromicinA, which is isolated from fermentation broth of actinomycetes actinomadurasp.kcc248, and has a structural formula shown in formula (a):
The anthrone glycoside compound oxanthromicinA provided by the invention has an inhibition effect on human acute promyelocytic leukemia cells, human non-small cell lung cancer cells, human liver cancer cells, human breast cancer cells and human colon cancer cells, and the IC50 of the anthrone glycoside compound oxanthromicinA is 8.7, 17.5, 15.0, 17.8 and 14.6 mu M respectively.
The invention also provides actinomycetes Actinomadirasp.KCC248 which is preserved in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 25 days in 2021, wherein the preservation address is number 3 of West Song No. 1 in the Korean area North Star of Beijing, and the preservation number is CGMCC No.21710.
The invention provides a preparation method of the anthrone glycoside compound, which comprises the following steps:
fermenting actinomycetes actinomadurasp.KCC248;
Concentrating the obtained fermentation liquor, and extracting by adopting ethyl acetate to obtain extract;
separating the extract with macroporous resin at normal pressure and silica gel column chromatography to obtain anthrone glycoside compound.
Preferably, the preparation method comprises the following conditions:
The fermentation medium is as follows: 200g/L potato, 20g/L glucose, 1000mL tap water; the fermentation condition is that the temperature is 28 ℃, the rotating speed is 180 rpm, and the aseptic fermentation is carried out for 8 days;
The separation parameters of the atmospheric macroporous resin separation comprise column signals: the filler is as follows: MCIGELCHP20P; the elution process is as follows: eluting with 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100% methanol water gradient, each gradient eluting with 1000mL, collecting eluate components with gradient of 80%, 90% and 100%, and subjecting to silica gel column chromatography;
the separation parameters of the silica gel column chromatography include column type: The filler is as follows: 200-300 mesh silica gel powder; the mobile phase is: 10 parts of chloroform and 3 parts of methanol, wherein the flow rate is 1mL/min;
collecting eluent after passing through the column with a retention volume of 100-150mL to obtain the compound oxanthromicinA.
The invention also provides a pharmaceutical preparation containing the compound oxanthromicinA.
Preferably, the formulation may be in the form of a tablet, granule, pill, capsule, paste, injection, suppository or patch.
The pharmaceutical preparation provided by the invention can be used for preparing medicines for treating tumor diseases, and especially can be used for treating diseases such as leukemia, lung cancer, liver cancer, breast cancer or colon cancer.
The anthrone glycoside compound oxanthromicinA has the inhibition activity of various tumor cells, has the present applicability in the treatment of various tumor diseases, and has the following advantages:
The invention discloses actinomycetes actinomadurasp.KCC248 for the first time, and an antitumor component oxanthromicinA can be obtained from fermentation broth of actinomycetes.
The invention discloses a method for extracting and separating a compound oxanthromicinA from fermentation liquor of actinomycetes actinomadurasp.KCC248 for the first time.
The novel compound oxanthromicinA provided by the invention provides effective resources for the research and development of lead compounds of novel antitumor drugs, and can also be used for treating tumor-related diseases.
Drawings
FIG. 1 is a high resolution mass spectrum of compound oxanthromicinA in the present invention.
FIG. 2 shows nuclear magnetic resonance 1 HNMR spectra of compound oxanthromicinA of the present invention.
FIG. 3 is a nuclear magnetic resonance 13 CNMR spectrum of compound oxanthromicinA of the present invention.
FIG. 4 is a nuclear magnetic resonance HSQC spectrum of compound oxanthromicinA in the present invention.
FIG. 5 is a nuclear magnetic resonance HMBC spectrum of compound oxanthromicinA in the present invention.
FIG. 6 shows WB results of compound oxanthromicinA of the present invention on the effects of matrix metalloproteinase 9, matrix metalloproteinase 2, cyclin E1 and cyclin D1 on liver cancer cell SMMC-7721.
FIG. 7 shows the WB and quantitative results of the effect of compound oxanthromicinA of the present invention on liver cancer LM-3 cells CyclinD1 and CyclinE.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention aims to provide a compound screened from bacteria, which can effectively inhibit various tumor cells, and the medicine prepared from the compound has potential application effect in tumor treatment.
The actinomycetes Actinomadirasp.KC C248 provided by the invention belongs to a new actinomycetes madurae strain, and the structure of quinone compounds produced by metabolism of the actinomycetes strain has anti-tumor activity. The chemical composition of actinomycetes KC C248 is researched, 1 new anthrone glycoside compound oxanthromicinA is found, and the compound is subjected to antitumor activity screening, so that the result shows that the anthrone glycoside compound oxanthromicinA has obvious activity of inhibiting tumor cell proliferation. Therefore, the anthrone glycoside compound can provide effective resources for the research and development of lead compounds of novel anti-tumor drugs, and can be used for treating tumor-related diseases.
The apparatus and materials of the lanes in the present invention are shown below, and the reagents not specifically described are conventional in the art, and the methods not specifically described are conventional in the art.
A Jasco-P-1020 polarimeter; horiba SEPA-300 polarimeter; brukerTensor27 FT-IR spectrometer; bruker AV-600MHz nuclear magnetic resonance spectrometer; waters Autospec Primier P776 spectrometer; API QSTAR pulsar l spectrometer; silica gel (200-300 mesh, qingdao ocean chemical Co., ltd.); sephadex LH-20 gel (Amerson biosciences, sweden), RP-18 silica gel (40-75 μm, fuji Siemens chemical Co., ltd.). The fractions were monitored by TLC and spots were visualized by heating a silica gel plate in vanillin-H 2SO4.
Experimental example 1 isolation and identification of Strain
The microorganism in the Bayan jaw rare earth ore from the Chinese inner Mongolia Baotou is subjected to high-flux screening, a pure cultured strain is obtained by separation, and the strain is named as Actinomadura sp.KCC 248 by morphology and 16S rRNA identification. The actinomycetes are preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) for 25 days in 1 month 2021, and the preservation address is number 3 of West Song No.1 of North Star in the Korean area of Beijing city, and the preservation number is CGMCC No.21710. Wherein the nucleotide sequence of the 16S rRNA of the bacterium is shown as SEQ ID No.1, and the specific sequence is as follows (5 '. Fwdarw.3'):
TAGAGTTTGATCCTGGCTCAGGACGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAGCGGAAAGGCCCCTTCGGGGGTACTCGAGCGGCGAACGGGTGAGTAACACGTGAGCAACCTGCCCCTGACTTCGGGATAAGCCTGGGAAACCGGGTCTAATACCGGATACGACTGCACCGGGCATGTGGTGTGGTGGAAAGATTTTTCGGTTGGGGATGGGCTCGCGGCCTATCAGCTTGTTGGTGGGGTGATGGCCTACCAAGGCGACGACGGGTAACCGGCCTGAGAGGGCGACCGGTCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCGCAATGGGCGGAAGCCTGACGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGACGAAGCGGAAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGTTTGTCGCGTCTGTCGTGAAAGCCCACGGCTTAACCGTGGGTCTGCGGTGGATACGGGCAGACTGGAGGCAGGTAGGGGAGAACGGAATTCCCGGTGTAGCGGTGAAATGCGCAGATATCGGGAGGAACACCGGTGGCGAAGGCGGTTCTCTGGGCCTGTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGCGCTAGGTGTGGGGTTCTTCCACGGATTCCGCGCCGTAGCTAACGCATTAAGCGCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTTGCTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATCGCCGGAAAACTGTTGGAGACAGCAGGTCCTTTTTGGGCCGGTGACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGTTCCATGTTGCCAGCACTTCGGGTGGGGACTCATGGGAGACCGCCGGGGTCAACTCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGCCCCTTATGTCTTGGGCTGCAAACATGCTACAATGGCCGGTACAGAGGGCTGCGATATCGTGAGGTGGAGCGAATCCCTTAAAGCCGGTCTCAGTTCGGATCGAAGTCTGCAACTCGACTTCGTGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCAACGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGCAACACCCGAAGCCCGTGGCCCAACCTTTTTGGGGGGAGCGGTCGAAGGTGGGGCCGGCGATTGGGACGAAGTCGTAACAAGGTAGCCGTACCGGAAGGTGCGGCTGGATCACCTCCTA.
Experimental example 2 fermentation of KCC248 and extraction and separation of anthrone glycoside metabolite from bacterial liquid-fermentation of actinomycetes Actinomadirasp. KCC248
The strain was inoculated into a 500mL flask containing 100mL of a seed medium (the solvent was water, the solute consisted of 4g/L of yeast extract, 4g/L of glucose, 5g/L of malt extract and 1g/L of soybean peptone), and cultured at 28℃for 2 days at a rotational speed of 180 rpm to obtain a seed culture solution. 5mL of the seed culture was inoculated into a 500mL flask containing 100mL of a fermentation medium (containing 200g/L of potato, 20g/L of glucose, pH 7.2), and cultured at 28℃for 8 days at a rotation speed of 180 rpm, to obtain a fermentation broth. Filtering the fermentation broth with gauze, collecting filtrate, and concentrating for use.
2. Extraction and separation of anthrone glycoside metabolite oxanthromicinA in bacterial liquid
Concentrating the fermentation broth of actinomycetes actinomadurasp.KCC248 from 30L to 2L, extracting the concentrated solution, adding 3 times of ethyl acetate, extracting for 3 times respectively, mixing the extracts, and concentrating to obtain ethyl acetate extract.
The specific culture method of the fermentation broth comprises the following steps: potato culture medium (200 g/L potato, 20g/L glucose, 1000mL tap water) was used, and the fermentation was performed aseptically at 28℃and 180 rpm for 8 days.
The method also comprises normal pressure macroporous resin separation, silica gel column chromatography separation and the like.
Wherein, the specific parameters of atmospheric macroporous resin separation are as follows:
Column model:
The filler is as follows: MCIGELCHP20P;
the elution process is as follows: the eluate was eluted by (10, 20, 30, 40, 50, 60, 70, 80, 90, 100%) methanol-water gradients, each gradient eluting 1000mL, and fractions of 80%, 90% and 100% were collected and then separated by silica gel column chromatography.
The parameters for silica gel column chromatography are specifically as follows:
Column model:
the filler is as follows: 200-300 mesh silica gel powder.
The mobile phase is: 10 parts of chloroform and 3 parts of methanol, wherein the flow rate is 1mL/min;
The eluted solution after passing through the column with a retention volume of 100-150mL was collected to obtain compound oxanthromicinA.
Experimental example 3 structural identification of Compound oxanthromicinA
Mass spectrometry and nuclear magnetic resonance analysis are carried out on the compound, the compound oxanthromicinA (1), and conventional physicochemical data comprise: the high resolution mass spectrum of amorphous powder ,[α]20D+207.5(c0.0012,DMSO);UV(MeOH)λmax(logε):256(4.40),314(4.34);HR-ESI-MSm/z:977.2721[M–H]–(calcd.for C23H49O11,977.2716). compound oxanthromicinA is shown in figure 1; 1 H and 13 CNMR data for compound oxanthromicinA are shown in Table 1 below, with their nuclear magnetic resonance 1 HNMR spectra shown in FIG. 2 and their nuclear magnetic resonance 13 CNMR spectra shown in FIG. 3. The compound was found to be an anthrone glycoside compound containing 3 methyl groups (δ C 20.9,15.7,34.5), 1 ketocarbonyl group (δ C 189.9) and 1 carboxycarbonyl group (δ C 169.0). Wherein 3 of the anthraceneaglycones are substituted H signals, delta H 7.01,6.75,7.35. Which also contains a sugar signal: delta H 5.06,3.95,3.61,3.49,3.14,3.37. Wherein, according to the C-10 signal (. Delta. C 80.6) and binding to HR-MS:977.2721[ M-H ] – speculates that the compound is a dimer and that symmetry is better. The compound was determined to contain D-mannose by hydrolyzing the compound with trifluoroacetic acid and detecting a sugar signal by HPLC.
1 H and 13 CNMR data (δin ppm, J in Hz) for compound oxanthromicin A of Table 1
Wherein, the sugar identification comprises the following specific steps: 10mg of the sample was precisely weighed into an ampoule, 3M TFA10mL was added and hydrolyzed at 120℃for 3 hours. Accurately sucking the acid hydrolysis solution, transferring to a tube, drying by adopting nitrogen, adding 10mL of water, mixing uniformly by vortex, sucking 100 mu L of deionized water, adding 900 mu L of deionized water, and centrifuging at 12000rpm for 5min. The supernatant was analyzed by IC. Chromatographic column: dionexCarbopac TM PA20 (3 x 150); mobile phase: h 2 O; b15 mM NaOH; 15mMNaOH&100mM NaOAC; flow rate: 0.3mL/min; sample injection amount: 5. Mu.L; column temperature: 30 ℃; a detector: an electrochemical detector. D-mannose was determined by comparison with standard sugar.
Meanwhile, by measuring the two-dimensional spectrum of the compound oxanthromicinA, wherein the nuclear magnetic resonance HSQC spectrum of the compound oxanthromicinA is shown in fig. 4, and the nuclear magnetic resonance HMBC spectrum is shown in fig. 5, it can be known that signals of all carbon atoms and hydrogen atoms of the compound belong to the chemical structure thereof, and the chemical structure of the compound oxanthromicinA is shown in formula (a):
Description: wherein, delta chemical shift units are ppm, 1 H-NMR and 13 C-NMR test solvents are dimethyl sulfoxide (DMSO), and hydrocarbon signals of the compounds are attributed through HSQC and HMBC spectrograms.
Experimental example 4 anti-tumor Activity test of Compounds isolated from KCC248
Drugs used in this experiment: positive control: cisplatin (cis-platinum) and Taxol (Taxol).
Sample to be measured: the oxanthromicinA and other 9 compounds separated from fermentation broth of actinomycetes Actinomadirasp. KC C248 have the following structural formulas:
the experiment adopts MTT to detect cytotoxic activity, and the experimental principle is as follows: the presence of dehydrogenase in the mitochondria of living cells reduces yellow thiazole blue MTT to insoluble blue purple formazan, and the enzyme disappears in dead cells, and MTT is not reduced. Formazan was dissolved in DMSO (dimethyl sulfoxide), and absorbance (OD) was measured at 570nm using a microplate reader, with OD values proportional to the number of living cells.
Tumor cell lines used in the present invention: human acute promyelocytic leukemia cells (HL-60), human non-small cell lung cancer cells (A-549), human liver cancer cells (SMMC-7721), human breast cancer cells (MCF-7) and human colon cancer cells (SW 480), human liver cancer cells LM-3.
The experimental method comprises the following steps: tumor cells in the logarithmic growth phase were taken, digested with pancreatin, and then blown into single cell suspensions, diluted to 1X 10 5 cells/mL with medium, inoculated into 96-well plates (100. Mu.L/well), and cultured in a 5% CO 2 saturated humidity incubator at 37℃for 24 hours. Samples oxanthromicinA to be tested with different concentrations are added into an experimental group, cisplatin and taxol solutions are added into a positive control group, the solution to be tested is replaced by DMSO diluted to 1000 times by 1640 culture medium is added into a negative control group, the solution to be tested is diluted to 1000 times by 1640 culture medium is added into wells without cells into a blank control group, and 3 repeated wells are respectively arranged in each group. The cells were incubated at 37℃for 48 hours with 5% CO 2, 10. Mu.L of the ready-prepared MTT solution containing 5mg/mL was added to each well, and incubated at 37℃for 4 hours. The supernatant was discarded, 150 μ LDMSO of formazan was added to each well, and the absorbance value (OD 570) was measured at 570nm with an enzyme-labeled instrument, and the inhibition (%) was calculated. The calculation formula is as follows: inhibition (%)) = (1-OD Sample/ODControl)×100％.IC50 represents drug concentration at 50% inhibition, IC 50 value was calculated by inhibition regression analysis of SPSS26.0 software .
The results show that the compound oxanthromicinA has obvious inhibition effect on human acute promyelocytic leukemia cells, human non-small cell lung cancer cells, human liver cancer cells, human breast cancer cells and human colon cancer cells, the IC 50 is 8.7, 17.5, 15.0, 17.8 and 14.6 mu M respectively, the specific data are shown in the following table 2, and other compounds are inactive.
Cytotoxic Activity of the compounds of Table 2
According to the cytotoxicity detection frying pan of the compound, the IC 50 of the compound on the liver cancer cell SMMC-7721 is found to be 15 mu M, so the concentration of the compound below the IC 50 is selected for subsequent experiments, wherein the concentrations comprise three concentrations of 1 mu M,3 mu M and 10 mu M respectively. Firstly, performing a western blotting experiment (WB), and finding that the compound has obvious inhibition effect on matrix metalloproteinase 9 (MMP 9) and matrix metalloproteinase 2 (MPP 2) of liver cancer cell SMMC-7721; in addition, the Cyclin is detected, and the compound has remarkable inhibition effect on Cyclin E1 (CyclinE 1) and Cyclin D1 (Cyclin D1) of hepatoma cell SMMC-7721, and the WB result is shown in figure 6, wherein A in the figure shows the influence of oxanthromicinA with different concentrations on MMP9 and MPP2, and B in the figure shows the influence of oxanthromicin A with different concentrations on Cyclin D1 and CyclinE 1. Thus, it is presumed that the compound may affect metastasis and invasion of cancer cells, thereby inhibiting proliferation of tumor cells.
In order to further verify the regulation and control effect of the compound on the cycle-related factors, the change of the protein expression quantity of the cycle-related factors is carried out after the liver cancer LM-3 cells are treated by adopting a Westernblot detection compound. The WB results are shown in FIG. 7, wherein A in the figure shows the effect of oxanthromicinA on liver cancer LM-3 cells CyclinD1 and CyclinE1, and B and C in the figure show the quantitative results of CyclinD1 and CyclinE, respectively. It was found that the protein levels of cycled D1 and cycled E1 were significantly reduced in LM-3 cells treated with compound oxanthromicinA. The above results indicate that the compounds can be used to down-regulate the expression levels of G0/G1 phase-related cyclains and CDKs.
The intensive activity research shows that oxanthromicinA has strong anti-tumor activity and can be used for developing tumor treatment medicines.
Experimental example 5 pharmaceutical preparation containing oxanthromicinA
1. Tablet formulation
Formulation recipe: 5 parts of oxanthromicinA, 24 parts of powdered sugar, 35 parts of starch, 6 parts of dextrin and 9 parts of 30% ethanol. Mixing sugar powder with starch dextrin, dissolving oxanthromicinA to 30% ethanol solution, adding into sugar powder, granulating, sieving with 16 # sieve, drying at 60deg.C for 3 hr. The whole granules were mixed with 21 parts of magnesium stearate and then tableted to prepare tablets each weighing 100mg. Wherein, all parts are parts by weight.
2. Freeze-dried preparation
The salt derivative (sodium salt or potassium salt) of oxanthromicinA is dissolved in sterile water with the concentration of 5mg/mL, then is sterilized by an intermittent hot pressing method, and is filtered by a Buchner funnel and a microporous filter membrane of 3.0 mu m to remove heat source, and finally is filtered by a microporous filter membrane of 0.22 mu m. After filtration, the solution was poured into penicillin bottles, 2mL each. Freezing penicillin bottles at-50deg.C, and freeze drying to powder in a freeze dryer after all freezing.
3. Other dosage forms
The oxanthromicinA provided by the invention can also be prepared into other dosage forms including granules, pills, capsules, ointment, injection, suppositories, patches and the like by adopting a conventional preparation method, has the same antitumor activity, has a wider application range, can be used as a good antitumor drug, has a treatment effect on various tumors, and has remarkable application effects and prospects.
While the present invention has been described in detail through the foregoing description of the preferred embodiment, it should be understood that the foregoing description is not to be considered as limiting the invention. Many modifications and substitutions of the present invention will become apparent to those of ordinary skill in the art upon reading the foregoing. Accordingly, the scope of the invention should be limited only by the attached claims.

Claims (8)

1. An anthrone glycoside compound is oxanthromicin A, which is separated from fermentation liquor of actinomycetes Actinomadura sp.KC C248, and the structural formula of the compound is shown as formula (A):
。
2. the anthrone glycoside compound according to claim 1, wherein the compound has inhibitory effect on human acute promyelocytic leukemia cells, human non-small cell lung cancer cells, human liver cancer cells, human breast cancer cells and human colon cancer cells, and has IC 50 of 8.7, 17.5, 15.0, 17.8 and 14.6 μm, respectively.
3. The anthrone glycoside compound according to claim 1, wherein actinomycetes Actinomadura sp.kc C248 is deposited in the China general microbiological culture Collection center (ccm) for 25 th month of 2021, with a deposit address of the institute 3, the west way No. 1, the korean region North Star, and a deposit number of the CGMCC No.21710.
4. A process for the preparation of the anthrone glycoside compounds according to claim 1, comprising:
Fermenting actinomycetes Actinomadura sp.kc C248 as claimed in claim 3;
Concentrating the obtained fermentation liquor, and extracting by adopting ethyl acetate to obtain extract;
separating the extract by normal pressure macroporous resin and silica gel column chromatography to obtain anthrone glycoside compound according to claim 1.
5. The method of claim 4, comprising the steps of:
the fermentation medium is as follows: 200 g/L potato, 20 g/L glucose, 1000: 1000 mL tap water; the fermentation condition is that the temperature is 28 ℃, the rotating speed is 180 rpm, and the aseptic fermentation is carried out for 8 days;
the normal pressure macroporous resin separation comprises the following separation parameters: 10X 50cm;
The filler is as follows: MCI GEL CHP 20P;
the elution process is as follows: eluting with 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100% methanol water gradient, each gradient eluting with 1000mL, collecting eluate components with gradient of 80%, 90% and 100%, and subjecting to silica gel column chromatography;
the separation parameters of the silica gel column chromatographic separation comprise the column type: 5X 50cm;
the filler is as follows: 200-300 mesh silica gel powder;
The mobile phase is: 10 parts of chloroform and 3 parts of methanol, wherein the flow rate is 1mL/min;
Collecting eluent after passing through the column with a retention volume of 100-150mL to obtain the compound oxanthromicin A.
6. A pharmaceutical formulation comprising the anthrone glycoside of claim 1.
7. The pharmaceutical formulation of claim 6, wherein the formulation is in the form of a tablet, granule, pill, capsule, paste, injection, suppository, or patch.
8. The use of the pharmaceutical preparation according to claim 6 for the preparation of a medicament for treating a neoplastic disease, wherein the neoplastic disease is one or more of leukemia, lung cancer, liver cancer, breast cancer and colon cancer.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000103798A (en) * 1998-09-30 2000-04-11 Dai Ichi Seiyaku Co Ltd Cell adhesion inhibitor
CN103288889A (en) * 2013-02-22 2013-09-11 中国科学院昆明植物研究所 Anthraquinone derivatives, their pharmaceutical compositions and application in medicine preparation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000103798A (en) * 1998-09-30 2000-04-11 Dai Ichi Seiyaku Co Ltd Cell adhesion inhibitor
CN103288889A (en) * 2013-02-22 2013-09-11 中国科学院昆明植物研究所 Anthraquinone derivatives, their pharmaceutical compositions and application in medicine preparation

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