CN110818728A - Preparation method and application of polythiodiketopiperazine compound Secoemestrin C - Google Patents
Preparation method and application of polythiodiketopiperazine compound Secoemestrin C Download PDFInfo
- Publication number
- CN110818728A CN110818728A CN201911160563.6A CN201911160563A CN110818728A CN 110818728 A CN110818728 A CN 110818728A CN 201911160563 A CN201911160563 A CN 201911160563A CN 110818728 A CN110818728 A CN 110818728A
- Authority
- CN
- China
- Prior art keywords
- acid
- compound
- methanol
- dichloromethane
- polythiodiketopiperazine
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
- C07D513/22—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains four or more hetero rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
-
- 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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biomedical Technology (AREA)
- Virology (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 belongs to the technical field of natural medicines, and particularly relates to a polysulphide diketopiperazine compound Secoemestrin C obtained from microorganisms, particularly fungal fermentation products, and medicinal salts thereof in preparation and treatment of diseases related to cancers, such as lung cancer, pancreatic cancer, colon cancer, breast cancer, gastric cancer, liver cancer and the like. The structure of the polythiodiketopiperazine compound or the medicinal salt thereof is shown as follows:
Description
Technical Field
The invention belongs to the technical field of natural medicines, and particularly relates to a polysulphide diketopiperazine compound Secoemestrin C obtained from microorganisms, particularly fungal fermentation products, and medicinal salts thereof in preparation and treatment of diseases related to cancers, such as lung cancer, gastric cancer, pancreatic cancer, colon cancer, breast cancer, liver cancer and the like.
Background
Malignant tumors (cancer) are the second leading cause of death worldwide, and nearly one sixth of deaths worldwide are caused by cancer. According to statistics, about 880 million people die from various cancers in 2015, wherein 169 ten thousands of lung cancers, 78.8 thousands of liver cancers, 77.4 thousands of colorectal cancers and 75.4 thousands of gastric cancers die, and the cancers become an important factor seriously threatening human health and influencing family happiness, thereby bringing huge burden to families and society.
At present, clinically, anti-tumor drugs can be mainly classified into the following categories according to their action mechanisms: cytotoxic drugs, monoclonal antibodies, hormones, biological response modifiers and the like. The cytotoxic antitumor drugs mainly comprise: alkaloids such as paclitaxel, vinblastine, hydroxycamptothecin, and docetaxel; biological alkylating agents such as cyclophosphamide and ifosfamide; antibacterial agents such as adriamycin and daunorubicin; cisplatin, carboplatin and other platinum compounds, gemcitabine, methotrexate, tegafur and the like. The monoclonal antibodies mainly comprise trastuzumab, alemtuzumab cetuximab, rituximab and the like. The hormones mainly comprise tamoxifen, moxifene, exemestane, flutamide, anastrozole, formestane, norrex, goserelin, megestrol, dydromedrogesterone and the like; the biological response regulator mainly comprises interferon, interleukin-2 and thymosin.
Although the application of the medicines brings great gospel to cancer patients to a certain extent, with the long-term application of the medicines, some medicines have secondary drug resistance in clinic, wherein the medicines comprise the anti-tumor medicines of gefitinib and pemetrexed for treating lung cancer; imatinib for treating gastrointestinal stromal tumors; fulvestrant, an anti-tumor drug for the treatment of breast cancer; and the antineoplastic agent teniposide for treating glioma. In addition, the problems of poor selectivity, large adverse reaction and the like of the traditional antitumor drugs are always the motivation of the world scientists to search for new antitumor drugs.
The natural medicine has rich resources, and microorganisms, medicinal plants and marine organisms are always important sources of medicine leads and innovative medicines. The secondary metabolite of the microorganism is more popular with researchers due to the advantages of diverse chemical structures, wide biological activity, strong repeatability, easy mass acquisition of samples by fermentation and modern synthetic biology technology, and the like.
The invention separates and purifies the anti-tumor active component of a naked cell fungus Emericella sp.1454 rice fermentation product, identifies a multi-sulfur diketopiperazine compound Secoemestrin C with the activities of resisting lung cancer, pancreatic cancer, colon cancer, breast cancer, liver cancer and the like, and the activity test result shows that the compound has stronger toxicity to various tumor cell strains for the first time and also shows stronger anti-tumor activity in animal bodies. Therefore, the compound has a good application prospect of antitumor activity. The invention relates to a Secoemestrin C producing strain, a preparation method, an anti-tumor cell strain and in-vivo anti-tumor activity, which have not been reported so far.
Disclosure of Invention
One of the objects of the present invention is to provide a strain of a fungus, chaetomium nudum (fungus 1454).
The strain of the invention has a preservation number: CGMCC No. 17765.
The strain is delivered to China general microbiological culture Collection center for preservation in 2019, 05 and 30 months, and the preservation unit address is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, on Beijing, with a deposit number: CGMCC No.17765, and the classification and naming are as follows: emericella sp.
The strain is used for preparing a polythiodiketopiperazine compound (Secoremestrin C).
Another object of the present invention is to provide a polythiodiketopiperazine compound (Secoremestrin C) or a pharmaceutically acceptable salt thereof.
The structure of the compound is shown as formula I:
the molecular formula of the compound is as follows: c27H22N2O10S4Molecular weight of：662。
The medicinal salt of the polythiodiketopiperazine compound comprises a salt formed by the compound and an inorganic acid, such as hydrochloric acid and sulfuric acid, a salt formed by an organic acid, such as acetic acid, trifluoroacetic acid, citric acid, maleic acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid or malic acid, and a salt formed by an amino acid, such as alanine, aspartic acid and lysine, or a salt formed by a sulfonic acid, such as methanesulfonic acid and p-toluenesulfonic acid.
The invention also provides a preparation method of the polythiodiketopiperazine compound or the medicinal salt thereof.
The invention is numbered from the deposit for the first time: separating and extracting the compound shown in the formula I from a strain (fungus 1454) of CGMCC No. 17765.
The preparation method of the polythiodiketopiperazine compound or the medicinal salt thereof comprises the following steps:
fermenting fungi 1454 with rice to obtain rice fermented product, ultrasonically extracting the rice fermented product with 10L 95% ethanol/water for 3 times, extracting with 10L 50% ethanol/water for 3 times, mixing extractive solutions, concentrating under reduced pressure to obtain water suspension, and extracting with ethyl acetate to obtain ethyl acetate extract; separating the obtained extract by normal phase silica gel chromatographic column chromatography, sequentially eluting with dichloromethane, dichloromethane-methanol and methanol in a gradient manner, wherein dichloromethane-methanol fraction is an active component, and purifying the active component by gel column chromatography and HPLC semi-preparative to obtain the compound shown in formula I.
Preferably, the preparation method of the polythiodiketopiperazine compound or the pharmaceutically acceptable salt thereof comprises the following steps:
fermenting fungi 1454 with rice, ultrasonically extracting fermented rice product with 10L 95% ethanol for 3 times, extracting with 10L 50% ethanol for 3 times, mixing extractive solutions, concentrating under reduced pressure to obtain water suspension, and extracting with ethyl acetate to obtain ethyl acetate extract; separating the obtained extract with normal phase silica gel column chromatography, sequentially eluting with dichloromethane, dichloromethane-methanol (40:1), dichloromethane-methanol (30:1), and dichloromethane-methanol (20:1)Dichloromethane-methanol (15:1), dichloromethane-methanol (10:1) and dichloromethane-methanol (5:1), methanol gradient elution, wherein dichloromethane-methanol (15:1) fraction J is the active component, then eluting fraction J with Sephadex LH-20 gel column methanol, wherein fraction J-F is semi-preparative purified (Capcell-Pak C)185μm,10×250mm,46％CH3CN/H2O,1.5ml/min) to obtain the compound shown in the formula I.
Wherein, the fungus 1454, the strain is delivered to China general microbiological culture Collection center for preservation in 2019, 05 and 30 months, and the preservation unit address is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, on Beijing, with a deposit number: CGMCC No. 17765.
The fourth purpose of the invention is to provide the pharmaceutical application of the compound shown in the formula I or the pharmaceutical salt thereof.
The invention relates to the application of the polysulfur diketopiperazine compound or the medicinal salt thereof in preparing medicaments for treating various tumor cell line toxicities and antitumor medicaments. The compound or the medicinal salt thereof is applied to preparing the medicines for resisting lung cancer, pancreatic cancer, colon cancer, breast cancer and liver cancer.
The invention adopts a cell proliferation and cytotoxicity detection method (CellCounting Kit) based on water-soluble tetrazolium salt to screen the activity of the fermentation product, the extracted fraction and the separated polysulfidedione piperazine compound shown in the formula I for resisting various human tumor cell strains of fungi 1454. Experimental results show that the compound shown in the formula I has strong cytotoxicity on A549 (lung cancer cells), HCT8 (colon cancer cells), MDA-MB-231 (breast cancer cells), MCF-7 (breast cancer cells), AsPC-1 (pancreatic cancer cells), BxPC-3 (pancreatic cancer cells), MIA-paca-2 (pancreatic cancer cells) and SU86.86 (pancreatic cancer cells). The polysulfur diketopiperazine compound can be used for preparing medicaments for resisting tumor-related diseases; the polysulfidedione piperazine compound is used as an active ingredient and is compatible with one or more pharmaceutically acceptable carriers, excipients or auxiliary materials to prepare the antitumor pharmaceutical composition. The medicine and the medicine composition can be used for clinical treatment of tumor resistance. The compound can also be combined with known medicaments to form a compound preparation for treating cancer diseases.
The fifth purpose of the invention is to provide the application of the pharmaceutical composition containing the compound shown in the formula I or the pharmaceutical salt thereof.
A pharmaceutical composition containing a compound shown as a formula I or a medicinal salt thereof is used for preparing toxicity on various tumor cell strains and an application of the pharmaceutical composition in antitumor drugs.
The application of the invention comprises resisting lung cancer, pancreatic cancer, colon cancer, gastric cancer, breast cancer and liver cancer.
The invention also provides a pharmaceutical composition containing the compound shown in the formula I or the medicinal salt thereof.
The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
The pharmaceutical composition provided by the invention takes the compound shown as the formula (I) and the medicinal salt thereof as the active pharmaceutical ingredients.
The pharmaceutical composition contains 0.1-99.9% of the compound shown in the formula (I) or the pharmaceutical salt thereof in the composition, and 0.1-99.9% of the pharmaceutically acceptable carrier in the composition.
The pharmaceutical composition is in the form of a formulation suitable for pharmaceutical use.
The medicinal preparation is tablet, capsule, granule, pill, powder, unguent, suspension, injection, powder for injection, suppository, cream, drop or patch. Wherein the tablet is a sugar-coated tablet, a film-coated tablet, an enteric-coated tablet or a sustained-release tablet; the capsule is hard capsule, soft capsule or slow release capsule; the powder injection is freeze-dried powder injection.
The pharmaceutical composition of the present invention is in the form of a preparation, wherein each preparation contains the compound of the present invention in an effective amount of 0.1-1000 mg, and each preparation unit, such as each tablet of a tablet, each capsule, or each dose, such as 100mg per dose.
The pharmaceutical compositions of the present invention may be formulated as solid or semi-solid pharmaceutical preparations in the form of powders, tablets, dispersible powders, capsules, cachets, suppositories, and ointments, using a solid carrier. The solid carrier which may be used is preferably one or more substances selected from diluents, flavouring agents, solubilising agents, lubricants, suspending agents, binders, bulking agents and the like, or may be an encapsulating substance. In the powdery preparation, 5-70% of micronized active ingredients are contained in a carrier. Suitable solid carriers include magnesium carbonate, magnesium stearate, talc, sucrose, lactose, pectin, dextrin, starch, gelatin, methylcellulose, sodium carboxymethylcellulose, low boiling waxes, cocoa butter, and the like. Because of their ease of administration, tablets, powders, cachets, capsules and the like represent the most advantageous oral solid dosage forms.
Liquid formulations of the present invention include solutions, suspensions and emulsions. For example, parenteral injection preparations may be in the form of water or water-propylene glycol solutions, which are adjusted in isotonicity, pH, etc. to suit the physiological conditions of the living body. The liquid preparation can also be prepared into solution in polyethylene glycol or water solution. Aqueous solutions for oral administration can be prepared by dissolving the active ingredient in water, followed by the addition of suitable amounts of coloring, flavoring, stabilizing and thickening agents. Aqueous suspensions suitable for oral administration can be prepared by dispersing the micronized active ingredient in viscous materials such as natural and synthetic gums, methylcellulose, sodium carboxymethylcellulose, and other known suspending agents.
It is particularly advantageous to formulate the above pharmaceutical preparations in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form of a formulation refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect. Such dosage unit forms may be in the form of a pack, such as a tablet, capsule or powder in a small tube or vial, or an ointment, gel or cream in a tube or bottle.
Although the amount of active ingredient contained in the dosage unit form may vary, it is generally adjusted within the range of 1 to 800mg, depending on the potency of the active ingredient selected.
The preferred dosage for a given situation can be determined by one skilled in the art in a routine manner. Generally, the amount of the active ingredient to be initially treated is lower than the optimum dose of the active ingredient, and then the dose to be administered is gradually increased until the optimum therapeutic effect is achieved. The total daily dose may be administered once or in divided doses for therapeutic purposes.
The invention separates and prepares the multi-sulfo diketopiperazine compound shown in the formula I from the fermentation product of a fungus 1454, has strong inhibitory activity to a plurality of human tumor cell strains and tumors in vivo, and is expected to be developed into a novel clinical anti-tumor medicament.
Compared with the existing similar antitumor drugs, the compound shown in the formula (I) has more excellent treatment effect and lower toxicity. Meanwhile, the source of the invention is a microbial natural product, the invention is easy to obtain, the repeatability is good, the preparation method has simple process and short time, the cost of the medicine is greatly reduced, and the invention is suitable for large-scale production.
Drawings
FIG. 1: high resolution Mass Spectrometry of Compound 1
FIG. 2: process for preparation of Compound 11H NMR spectrum
FIG. 3: process for preparation of Compound 113C NMR spectra
FIG. 4: concentration-survival rate curve for killing 8 tumor cell strains by using compound 1
FIG. 5: growth inhibition effect of compound 1 on human breast cancer MCF-7 nude mouse transplantation tumor
Detailed description of the preferred embodiments：
Embodiments of the present invention are applicable to the preparation of polythiodiketopiperazine based compounds from any microorganism, and are not limited to fungal fermentations. The following examples are set forth to aid those skilled in the art in a better understanding of the present invention and are not intended to limit the invention in any way.
< example 1> fermentation for fungi 1454:
inoculating activated fungus 1454 strain to PDA slant, and culturing at 25 deg.C in incubator for one week. The seeds were harvested by slant cutting, crushed, inoculated into 3 500mL Erlenmeyer flasks containing 100mL of PDB medium, and cultured at 25 ℃ for 5 days with shaking to obtain a seed solution. Placing 100g rice and 100ml distilled water into 500ml triangular flask, sealing, sterilizing at 121 deg.C for 15min, cooling, adding 10ml seed solution (30 bottles in total) into each flask, and culturing at 25 deg.C for 30 days to obtain fungus 1454 rice fermented product.
< example 2> extraction of fungal 1454 fermentate and obtaining of extract:
ultrasonically extracting the rice fermented product in example 1 with 10L 95% ethanol/water for 3 times, and extracting with 10L 50% ethanol/water for 3 times, mixing extractive solutions, concentrating under reduced pressure to obtain water suspension, and extracting with ethyl acetate to obtain ethyl acetate part extract.
< example 3> isolation, preparation and structural characterization of compound 1:
separating the extract obtained in example 2 by normal phase silica gel chromatography, sequentially eluting with dichloromethane, dichloromethane-methanol (40:1), dichloromethane-methanol (30:1), dichloromethane-methanol (20:1), dichloromethane-methanol (15:1), dichloromethane-methanol (10:1), dichloromethane-methanol (5:1) and methanol in gradient manner, wherein dichloromethane-methanol (15:1) fraction is active component, eluting the component with Sephadex LH-20 gel column methanol, and purifying with semi-preparative method (Capcell-Pak C)185μm,10×250mm,46％CH3CN/H2O,1.5ml/min) to obtain the compound shown in the formula I.
Structural characterization of Compound 1 of formula I
(1) Compound 1 is a white powder, readily soluble in methanol solution. High resolution electrospray mass spectrometry (HRESIMS) gives the excimer ion peak M/z 685.0053[ M + H ]]+It is suggested that the molecular composition thereof is C27H22N2O10S4(calcd for C27H22N2O10S4Na, 685.0054). Process for preparation of Compound 113C NMR(Acetone-d6150MHz) is: 166.8(C-1),32.3 (C-NCH)3-2),68.7(C-3),164.4(C-4),60.2(C-5a),72.7(C-6),108.5(C-7),140.1(C-8),142.4(C-10),113.2(C-10a),78.5(C-11),81.2(C-11a),123.8(C-1'),122.9(C-2'),145.5(C-3'),156.3(C-4'),56.6(C-4-OMe '),113.2(C-5'),129.0(C-6'),165.9(C-7'),146.5(C-1 "), 154.7 (C-2"), 117.7(C-3 "), 128.6 (C-4"), 130.8(C-5 "), 118.5 (C-6"), 191.0(C-7 "). The hydrogen and carbon spectra data of the compound were compared with Secoemestrin C [ Oike M, Nozawa K, Kawai K I. intumescent to emist from Emericella foveolate. phytochemistry,1997,46(1):123-.]The data reported in the literature are substantially consistent, and compound 1 was therefore identified as secoemestin C.
The structural formula of the compound of the invention is:
< example 4> test for ability of Compound of formula I to kill tumor cells
Cell Counting Kit, which is a Cell proliferation and cytotoxicity detection method based on water-soluble tetrazolium salt. It can be reduced to soluble orange yellow formazan (formazan) by the dehydrogenase in mitochondria in the presence of the electron coupling reagent 1-MethoxPMS. The number of formazans is proportional to the number of viable cells. The faster the cell proliferation, the less cytotoxic and the greater the number of cells, the darker the color, and the more linear the shade of the color is with the number of cells.
Procedure for the preparation of the
HCT8 (colon cancer cells), MDA-MB-231 (breast cancer cells), MCF-7 (breast cancer cells), AsPC-1 (pancreatic cancer cells), BxPC-3 (pancreatic cancer cells), MIA-paca-2 (pancreatic cancer cells), PANC-1 (pancreatic cancer cells), SU86.86 (pancreatic cancer cells), and SW1990 (pancreatic cancer cells) in logarithmic growth phase were digested with 0.25% pancreatic enzyme to adjust the cell concentration to 2.5X 104one/mL. And (4) plating, adding the cell culture solution with the adjusted concentration into the wells of a 96-well plate, wherein each well is 200 mu l, the periphery of each well is sealed by serum-free culture medium, and the cells are placed in an incubator to be cultured to a logarithmic phase. 1mg of a test compound 1, Gemcitabine (Gemcitabine), Adriamycin (Dox) and Oxaliplatin (Oxaliplatin) are respectively prepared into a 10mM DMSO stock solution for later use. The test compounds were diluted in complete medium to give final concentrations of 5. mu.M, 4. mu.M, 3. mu.M, 2. mu.M, 1. mu.M, 0.5. mu.M, respectively. The original culture medium was discarded and 200. mu.l of the drug solution was added to each well. 3 duplicate wells were set for each concentration, and a blank control and a solvent control were set. The culture was continued in the cell incubator for 48 h. Taking out 96-well plate, discarding supernatant, adding 100 μ l complete culture medium containing 10% CCK Solution into each well, culturing in cell culture box for 1-4 hr, and labeling with enzyme readerAbsorbance at 450nm was measured.
The survival rate of tumor cells is (fluorescence absorption value of administration well-fluorescence absorption value of blank well)/(fluorescence absorption value of solvent control well-fluorescence absorption value of blank well). times.100%
Determining the survival rate under each concentration, and calculating the IC of the sample to be detected by using SigmaPlut software50The value is obtained.
The experimental results are as follows: IC of compound of formula I for killing 8 tumor cells50The values are shown in table 1 and the concentration-survival curves are shown in figure 1.
TABLE 1 toxicity of Compounds of formula I and Positive controls on 9 tumor cell lines
< example 5> growth inhibitory Effect of the Compound of formula I on human Breast cancer MCF-7 nude mouse transplanted tumor
25 female BALB/c nude mice with the weight of 18-22g are taken, human breast cancer MCF-7 cells are inoculated on the shoulders of the nude mice, and each mouse is inoculated with 1 multiplied by 107One cell, the tumor length is 300mm3When the size is large, the tumor mass is taken for transfer. The length of the tumor to be 100mm3The nude mice are divided into groups according to the tumor size and the body weight during the size, so that the average value of the tumor size of each group is about 100mm3And the average body weight of each group is close to that of each group, and each group contains 5 nude mice. The compound of formula i and doxorubicin were administered intraperitoneally once every other day, respectively. Each nude mouse was injected with 200 μ l, and the control group was given a blank solvent control. Tumor diameter and body weight were measured every 2 days during the experiment according to the formula V ═ ab2And/2 calculating the tumor volume (a: the tumor major diameter, b: the tumor minor diameter), drawing a tumor growth curve, and observing the weight change. Animals were sacrificed on day 24 of the experiment, tumors were isolated and weighed, and tumor inhibition rate was calculated (fig. 2, table 2).
TABLE 2 growth inhibition of IMB-122 compound on human breast cancer MCF-7 nude mouse transplantable tumors
| Group of | Number of nude mice | Tumor weight (g) Mean ± SD | Inhibition rate% (tumor weight) ean + -SD |
| Control group | 5 | 0.99±0.38 | - |
| 2.5mg/kg of a Compound of formula I | 5 | 0.30±0.09 | 69.6±8.8** |
| 5.0mg/kg of a Compound of formula I | 5 | 0.37±0.01 | 63.2±10.7 |
| 10.0mg/kg of a Compound of formula I | 5 | 0.26±0.11 | 73.8±11.2** |
| Adriamycin 3.0mg/kg | 5 | 0.42±0.16 | 57.9±15.9 |
P <0.01 compared to control group
Experimental results show that the compound shown in the formula I can obviously inhibit the growth of human breast cancer MCF-7 nude mouse transplantation tumor, wherein the tumor inhibition rates of 2.5mg/kg and 10.0mg/kg dose groups are 69.6% +/-8.8% and 73.8% +/-11.2%, respectively, and both show stronger tumor growth inhibition than that of adriamycin (the tumor inhibition rate is 57.9% +/-15.9%) at 3.0mg/kg (tolerance dose).
Claims (9)
1. A polysulfide diketopiperazine compound Secoemestrin C or a medicinal salt thereof is characterized in that the structure of the compound is shown as a formula I:
2. the polythiodiketopiperazine compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt of the compound comprises a salt with an inorganic acid such as hydrochloric acid or sulfuric acid, a salt with an organic acid such as acetic acid, trifluoroacetic acid, citric acid, maleic acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid or malic acid, and a salt with an amino acid such as alanine, aspartic acid, lysine or a salt with a sulfonic acid such as methanesulfonic acid or p-toluenesulfonic acid.
3. The polythiodiketopiperazine compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound represented by formula i is isolated from a rice fermentation product of fungus 1454.
4. The process for preparing a polythiodiketopiperazine compound or a pharmaceutically acceptable salt thereof according to claim 1, comprising the steps of:
fermenting fungi 1454 with rice, ultrasonically extracting fermented rice product with 10L 95% ethanol for 3 times, extracting with 10L 50% ethanol for 3 times, mixing extractive solutions, concentrating under reduced pressure to obtain water suspension, extracting with ethyl acetate,obtaining ethyl acetate part extract; separating the obtained extract with normal phase silica gel chromatographic column, sequentially eluting with dichloromethane, dichloromethane-methanol (40:1), dichloromethane-methanol (30:1), dichloromethane-methanol (20:1), dichloromethane-methanol (15:1), dichloromethane-methanol (10:1), dichloromethane-methanol (5:1), and methanol, wherein dichloromethane-methanol (15:1) fraction J is active component, eluting component J with Sephadex LH-20 gel column methanol, wherein component J-F is semi-preparative purified (Capcell-Pak C)185μm,10×250mm,46％CH3CN/H2O,1.5ml/min) to obtain the compound shown in the formula I.
5. The use of the polythiodiketopiperazine compound of claim 1 or the pharmaceutically acceptable salt thereof in the preparation of drugs for treating various tumor cell line toxicities and antitumor drugs.
6. The use of a pharmaceutical composition comprising the polythiodiketopiperazine compound of claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of tumor cell toxicity and for the treatment of tumors.
7. Use of a compound of claim 6 or a pharmaceutical composition of claim 7 for the manufacture of a medicament against lung, stomach, pancreas, colon, breast and liver cancers.
8. A pharmaceutical composition comprising the polythiodiketopiperazine compound of claim 1 or a pharmaceutically acceptable salt thereof.
9. A naked cell fungus strain, the preservation number is: CGMCC No. 17765.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911160563.6A CN110818728B (en) | 2019-11-23 | 2019-11-23 | Preparation method and application of polythiodiketopiperazine compound Secoemestrin C |
| PCT/CN2020/128569 WO2021098593A1 (en) | 2019-11-23 | 2020-11-13 | Secoemestrin c preparation method and use therof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911160563.6A CN110818728B (en) | 2019-11-23 | 2019-11-23 | Preparation method and application of polythiodiketopiperazine compound Secoemestrin C |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110818728A true CN110818728A (en) | 2020-02-21 |
| CN110818728B CN110818728B (en) | 2021-02-23 |
Family
ID=69558644
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911160563.6A Active CN110818728B (en) | 2019-11-23 | 2019-11-23 | Preparation method and application of polythiodiketopiperazine compound Secoemestrin C |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN110818728B (en) |
| WO (1) | WO2021098593A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112094330A (en) * | 2020-09-22 | 2020-12-18 | 中国科学院微生物研究所 | Polythiodiketopiperazine synthesis related protein and related biological material and application thereof |
| WO2021098593A1 (en) * | 2019-11-23 | 2021-05-27 | 中国医学科学院医药生物技术研究所 | Secoemestrin c preparation method and use therof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002026744A1 (en) * | 2000-09-27 | 2002-04-04 | Kyowa Hakko Kogyo Co., Ltd. | Mpc1001 derivatives |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110818728B (en) * | 2019-11-23 | 2021-02-23 | 中国医学科学院医药生物技术研究所 | Preparation method and application of polythiodiketopiperazine compound Secoemestrin C |
-
2019
- 2019-11-23 CN CN201911160563.6A patent/CN110818728B/en active Active
-
2020
- 2020-11-13 WO PCT/CN2020/128569 patent/WO2021098593A1/en active Application Filing
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002026744A1 (en) * | 2000-09-27 | 2002-04-04 | Kyowa Hakko Kogyo Co., Ltd. | Mpc1001 derivatives |
Non-Patent Citations (2)
| Title |
|---|
| HIDEYUKI ONODERA ET AL.: "MPC1001 and Its Analogues: New Antitumor Agents from the Fungus Cladorrhinum Species", 《ORGANIC LETTERS》 * |
| MIKIKO OOIKE ET AL.: "AN EPITETRATHIODIOXOPIPERAZINE RELATED TO EMESTRIN FROM EMERICELLA FO VEOLA TA", 《PHYTOCHEMISTRY》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021098593A1 (en) * | 2019-11-23 | 2021-05-27 | 中国医学科学院医药生物技术研究所 | Secoemestrin c preparation method and use therof |
| CN112094330A (en) * | 2020-09-22 | 2020-12-18 | 中国科学院微生物研究所 | Polythiodiketopiperazine synthesis related protein and related biological material and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2021098593A1 (en) | 2021-05-27 |
| CN110818728B (en) | 2021-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6358957B1 (en) | Phenylahistin and the phenylahistin analogs, a new class of anti-tumor compounds | |
| CN101045046B (en) | Use of Brazil hemoatoxy type compound for preparing antineoplastic | |
| CN110818728B (en) | Preparation method and application of polythiodiketopiperazine compound Secoemestrin C | |
| CN113402509B (en) | Meishadazole compounds and preparation method and application thereof | |
| KR0169536B1 (en) | Novel ginseng saponins, process for preparation thereof and anti-tumor agents containing the same as an active ingredient | |
| CN112062738B (en) | Artemisinol A-B, pharmaceutical composition thereof, and preparation method and application thereof | |
| CN110218200B (en) | Cyclic depsipeptide compound in mangrove endophytic fungi and preparation method and application thereof | |
| CN113061124B (en) | Sesquiterpene dimer compound, and preparation method, application and pharmaceutical composition thereof | |
| CN1190497C (en) | Preparation technology of geldanamycin and its derivative | |
| CN115677471B (en) | Rose alkyl diterpenoid compound, preparation method, pharmaceutical composition and anti-tumor application | |
| CN105085221B (en) | Compound with antifungal and anti-tumor activity and preparation method and application | |
| CN113582863B (en) | Aminoethyl biphenyl compound and preparation method and application thereof | |
| CN115124543B (en) | Tricyclic pyridone compounds with antiviral activity and application thereof | |
| CN109705183A (en) | Smelly seven secondary metabolites and its pharmaceutical composition and preparation method and its application | |
| CN113582878B (en) | Derivative SGP-17-52 of natural product enantiomer-kaurane diterpenoid SGP-17 | |
| CN117486897A (en) | Marine source gamma-pyrone compound Aspergillus oryzae A and preparation method and application thereof | |
| CN111808155B (en) | Ginseng coumarin compound and application thereof in medicine | |
| CN116115602B (en) | New application of compound chaetomougilin O in preparation of medicaments for preventing or treating thyroid cancer | |
| CN113321631B (en) | Biandrographolide G, preparation method and application thereof in medicines | |
| CN103275139A (en) | C16:1DLSL and application thereof | |
| CN109575089B (en) | Acylated glucose compounds, pharmaceutical composition, preparation method and application thereof | |
| CN112979741B (en) | Withanolide II compound and extraction method and application thereof | |
| CN110218174B (en) | Compound and preparation method and application thereof | |
| CN118027063A (en) | Novel lactam compound DIDYLACTAM A derived from marine mangrove fungi, and preparation method and application thereof | |
| CN117820273A (en) | Linderane type sesquiterpene oligomer, preparation method thereof and application thereof in preparation of anti-leukemia drugs |
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 |