CN112876362B - Extraction and separation method and application of macrocyclic diterpenoid compound components in euphorbia lobata fruits - Google Patents
Extraction and separation method and application of macrocyclic diterpenoid compound components in euphorbia lobata fruits Download PDFInfo
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Abstract
The invention relates to an extraction and separation method and application of macrocyclic diterpenoid compound components in euphorbia foliosa fruits, wherein the compound is prepared from euphorbia foliosa (euphorbia foliosa)Euphorbia sororia Schrenk) mature fruits are used as raw materials, and are separated by solvent extraction, solvent extraction and silica gel column chromatography to obtain macrocyclic diterpenoid compounds with the components of 1-8, and the macrocyclic diterpenoid compounds in the components have the content of more than 60 percent through high performance liquid chromatography analysis. Through activity tests of the components 1-8, the composition shows stronger multidrug resistance reversal activity, can reverse the drug resistance of drug-resistant cells to antitumor drugs to a certain degree when being used together with antitumor drugs, and can be used as multidrug resistance reversal drugs.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a method for extracting and separating macrocyclic diterpenoid components and application thereof in preparation of multidrug resistance reversal medicines or medicines combined with antitumor medicines.
Background
Euphorbia sororii A.Schrenk is an annual herbaceous plant of the genus Euphorbia (Euphorbiaceae) of the family Euphorbiaceae, is produced in the middle of the mainland Asia, is mainly distributed in Xinjiang Hotan, changji and other places in China, and mature fruits of the Euphorbia sororiana are common medicinal materials of Uygur nationality medicines. The plant is used as a main component in more than ten mature national medical compound medicines such as Suzi Afu tablet, compound Suzi Afu honey cream, jiafraxi Cuinui, yilititi honey cream, suzi Afu ointment, suzafu powder, sailani powder and the like. At present, sesquiterpenes, diterpenes, triterpenes, sterols, fatty acids, flavones, phenols, saccharides and the like are mainly used as the compounds isolated and identified from the plant.
At present, diterpenoid compounds separated from euphorbia pekinensis kok and fruits are mainly pseudoelemene (Jatrophane) type macrocyclic diterpenoid compounds, and the structural characteristics of the pseudoelemene type macrocyclic diterpenoid compounds are that the framework is formed by fusing a five-membered ring and a twelve-membered ring, namely a 5/12 ring system. The compound has the characteristics of complex structure, most high esterification, various substituent groups such as acetoxyl (-OAc), propionyloxy (-OPr), isobutyryloxy (-OiBu), benzoyloxy (-OBz), pyridinyloxy-3-yl (-ONic) and the like, and large number of substituent groups (usually 3-8) and large molecular weight. Moreover, most of the pseudoolivine-type macrocyclic diterpenoid compounds have been reported to have various degrees of tumor multidrug resistance reversal activity.
In the prior art, the yield of macrocyclic diterpenoid components of euphorbia foliosa fruits of medicinal materials is low, and the total content of macrocyclic diterpenoid compounds in the components is not high, so that the current situation restricts the development of industrial production, the requirement of modern industrial production cannot be met, and the deep development of the medicinal materials is hindered.
The tumor multidrug resistance is the phenomenon that after a drug acts on a tumor to generate drug resistance, the tumor has cross drug resistance to various antineoplastic drugs which are never contacted, have irrelevant structure, different targets and different mechanisms. Multidrug resistance has multiple formation mechanisms, such as topoisomerase abnormality, protein kinase C overexpression, ABC family transporter overexpression and the like, wherein the most important and most extensive research is the overexpression of P-glycoprotein encoded by ABCB1 gene, so that the drug efflux is increased, and the drug resistance is formed. Therefore, the multidrug resistance of the tumor is one of the main reasons for the failure of the chemotherapy to treat the tumor. The multidrug resistance reversal medicine has appeared for three generations, and the reversal activity is weak and the high toxicity is the reason for not being popularized and used on a large scale. In recent years, due to the characteristic of low toxicity of plant medicines, a plurality of pseudo-elemene type macrocyclic diterpenoid compounds separated from plants are found to have remarkable MDR reversal activity, so that the research hotspot is realized. The research researches whether the macrocyclic diterpenoid compound component can be combined with the traditional antitumor drug to serve as a substrate by competitively combining with a transport protein on a drug-resistant cell membrane so as to achieve the aim of reversing the drug resistance of the drug-resistant cell, so that the traditional antitumor drug can play a role in killing tumor cells in the cell and is used for evaluating the activity and the effectiveness of the compound component.
Disclosure of Invention
The invention aims to provide an extraction and separation method and application of macrocyclic diterpenoid components in euphorbia lobata fruits. The compound is prepared by taking mature fruits of Euphorbia nervosa (Euphorbia sororia A. Schrenk) as a raw material, extracting by using a solvent, extracting by using the solvent, and separating by using silica gel column chromatography to obtain macrocyclic diterpenoid compound components 1-8, wherein the macrocyclic diterpenoid compound content in the components exceeds 60% through high performance liquid chromatography analysis. Through activity tests of the components 1-8, the composition shows stronger multidrug resistance reversal activity, can reverse the drug resistance of drug-resistant cells to antitumor drugs to a certain degree when being used together with antitumor drugs, and can be used as multidrug resistance reversal drugs.
The invention relates to an extraction and separation method of macrocyclic diterpenoid components in euphorbia lobata fruits, which comprises the following steps:
a. taking Euphorbia pekinensis fruit as a raw material, naturally airing, crushing, percolating, cold soaking or heating reflux extraction with 5-10 times volume of 50-99% ethanol aqueous solution, absolute ethyl alcohol, acetone, 50-99% methanol aqueous solution or absolute methanol at room temperature, and concentrating to obtain Euphorbia pekinensis fruit extract;
b. b, dispersing the extract obtained in the step a by using water with the volume of 3-10 times, adding n-hexane, ethyl acetate or a mixed solvent of n-hexane and ethyl acetate with the volume of 3-10 times, extracting for 3-10 times, or dispersing the extract by using n-hexane, ethyl acetate or a mixed solvent of n-hexane and ethyl acetate with the volume of 3-10 times, adding water with the volume of 3-10 times, extracting for 3-10 times, combining the extract, and concentrating under reduced pressure to obtain an extract;
c. dispersing the extract obtained in the step b by using 3-10 times of volume of acetonitrile or 50% methanol aqueous solution, adding 3-10 times of volume of n-hexane, petroleum ether or cyclohexane for extraction for 3-10 times, or dispersing the extract by using 3-10 times of volume of n-hexane, petroleum ether or cyclohexane, adding 3-10 times of volume of acetonitrile or 50% methanol aqueous solution for extraction for 3-10 times, and concentrating to obtain an extract;
d. separating the extract of the acetonitrile or 50% methanol aqueous solution extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using petroleum ether-ethyl acetate or n-hexane-acetone with the volume ratio of 100-0; evaporating the fraction Fr.2 at 38-40 deg.C under reduced pressure to obtain yellow brown viscous liquid, drying the yellow brown viscous liquid at 50 deg.C in water bath to constant weight to obtain macrocyclic diterpene compounds with the following components: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudowhite-olivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudoolivine-6 (17), 11E-diene; compound 8: tylosin A.
The macrocyclic diterpenoid compound component is applied to the preparation of multidrug resistance reversal medicines.
The macrocyclic diterpenoid component can be used for preparing anti-tumor drug combination medicines.
The invention relates to an extraction and separation method of macrocyclic diterpenoid components in euphorbia lobata fruits, which comprises the following steps: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudoolivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudoolivine-6 (17), 11E-diene; compound 8: the tylosin-Zener euphorbia-pekinensis A is determined by high performance liquid chromatography, the mass content of macrocyclic diterpenoid compounds in the component exceeds 60%, and a liquid chromatogram map is shown in figure 1.
The application of the macrocyclic diterpenoid compound components 1-8 in the preparation of multidrug resistance reversal medicines or the combined medicines with antineoplastic medicines is carried out in vitro cytotoxicity activity determination and multidrug resistance reversal activity determination, and the results show that: the macrocyclic diterpenoid compound component has weaker cytotoxic activity on a human breast cancer cell strain MCF-7 and a human breast cancer adriamycin drug-resistant cell strain MCF-7/ADR, and has tumor multidrug resistance reversal activity of different degrees.
Drawings
FIG. 1 is a high performance liquid chromatography contrast map of macrocyclic diterpenoid components of the invention
FIG. 2 is a high performance liquid chromatography overlay chart of 10 batches of macrocyclic diterpenoid component of the invention
Detailed Description
The used reagents n-hexane, cyclohexane, petroleum ether, acetonitrile, ethyl acetate, methanol, ethanol and acetone are analytically pure and are produced from Tianjin Kemiou chemical reagent Limited; acetonitrile in high performance liquid chromatography is HPLC grade (Thermo Fisher company in USA); column chromatography silica gel (100-200 mesh): marine industrial production of Qingdao; the thin-layer chromatography silica gel is HSGF254, produced by yellow silica gel development test factories of cigarette stand city; high performance liquid chromatography (DIONEX corporation, usa): a P680HPLC pump, an ASI-100 autosampler, a TCC-100 column incubator, a UVD170U ultraviolet detector (four wavelength), a quaternary solvent system, an online degasser, and a Chromeleon chromatography workstation.
Euphorbia lobata fruits were collected from the Jimusala county, the Uygur autonomous region of Xinjiang, and identified as European sororia A.Schrenk by Von tassel co-investigator, the institute of ecological geography, xinjiang, academy of sciences of China.
Example 1
a. Taking 10kg of naturally dried euphorbia pekinensis fruits, crushing, percolating and extracting by using 45L of 90% ethanol-water solution at room temperature, and evaporating the solvent under reduced pressure to obtain euphorbia pekinensis fruit extracts;
b. dispersing the extract obtained in the step a by 6 times of volume of water, adding 5 times of volume of ethyl acetate for extraction for 5 times, stirring for 5min by a paddle stirrer each time, standing for 15min for layering, combining ethyl acetate layers, and concentrating under reduced pressure to obtain an ethyl acetate extract;
c. dispersing the ethyl acetate extract obtained in the step b by using 5 times of methanol with the concentration of 50%, adding 5 times of petroleum ether for extraction for 5 times, stirring for 5min by using a paddle stirrer each time, standing for 20min for layering, combining 50% methanol layers, and concentrating under reduced pressure to obtain a 50% methanol extract;
d. separating the 50% methanol extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using petroleum ether-ethyl acetate with the volume ratio of 100-1 to 0, analyzing each fraction by using silica gel Thin Layer Chromatography (TLC), combining the same fractions to obtain 3 components Fr.1-Fr.3, evaporating the fraction Fr.2 at the temperature of 38-40 ℃ under reduced pressure to dryness to obtain a yellowish-brown viscous liquid, drying the yellowish-brown viscous liquid at the temperature of 50 ℃ in a water bath to constant weight, and obtaining the macrocyclic diterpenoid compound with the components: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudowhite-olivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudo-white-olivine-6 (17), 11E-diene; compound 8: tylosin A.
Example 2
a. Taking 10kg of naturally dried euphorbia pekinensis fruit, crushing, extracting with 40L of absolute ethanol solution at room temperature for 4 times by cold immersion, combining the extracting solutions, and evaporating the solvent to dryness under reduced pressure to obtain an euphorbia pekinensis fruit extract;
b. dispersing the extract obtained in the step a by using water with the volume 5 times, adding a mixed solvent of n-hexane and ethyl acetate with the volume 5 times, extracting for 5 times, stirring for 5min by using a paddle stirrer each time, standing for 10min for layering, combining mixed solvent layers of n-hexane and ethyl acetate, and concentrating under reduced pressure to obtain an extract of the mixed solvent of n-hexane and ethyl acetate;
c. dispersing the mixed solvent extract of the normal hexane and the ethyl acetate obtained in the step b by using acetonitrile with the volume 5 times, adding cyclohexane with the volume 5 times, extracting for 8 times, stirring for 5min by using a paddle stirrer each time, standing for 25min for layering, combining acetonitrile layers, and concentrating under reduced pressure to obtain an acetonitrile extract;
d. separating the acetonitrile extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using n-hexane-acetone with the volume ratio of 100-0: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudowhite-olivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudo-white-olivine-6 (17), 11E-diene; compound 8: tylosin A.
Example 3
a. Taking 10kg of naturally dried euphorbia pekinensis fruit, crushing, heating and extracting for 12h by using 70L of acetone, combining extracting solutions, and concentrating a solvent under reduced pressure to obtain an euphorbia pekinensis fruit extract;
b. dispersing the extract obtained in the step a by using water with the volume being 10 times that of the extract, adding n-hexane with the volume being 5 times that of the extract to extract for 10 times, stirring for 10min by using a paddle stirrer each time, standing for 30min for layering, combining n-hexane layers, and concentrating under reduced pressure to obtain an n-hexane extract;
c. dispersing the n-hexane extract obtained in the step b by using acetonitrile with the volume of 10 times, adding petroleum ether with the volume of 5 times for extraction for 10 times, stirring for 5min by using a paddle stirrer each time, standing for 20min for layering, combining acetonitrile layers, and concentrating under reduced pressure to obtain acetonitrile extract;
d. separating the acetonitrile extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using n-hexane-acetone with the volume ratio of 100-0: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudowhite-olivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudo-white-olivine-6 (17), 11E-diene; compound 8: tylosin A.
Example 4
a. Crushing 10kg of naturally dried Euphorbia pekinensis fruit, cold-soaking and extracting with 60L of 80% methanol-water solution at room temperature for 4 times, mixing cold-soaking extractive solutions, and concentrating the solvent under reduced pressure to obtain Euphorbia pekinensis fruit extract;
b. dispersing the extract obtained in the step a by using water with the volume being 7 times that of the extract, adding ethyl acetate with the volume being 5 times that of the extract to extract for 5 times, stirring for 10min by using a paddle stirrer each time, standing for 20min for layering, combining ethyl acetate layers, and concentrating under reduced pressure to obtain an ethyl acetate extract;
c. dispersing the ethyl acetate extract obtained in the step b by using acetonitrile with the volume 6 times, adding n-hexane with the volume 5 times for extraction for 5 times, stirring for 5min by using a paddle stirrer each time, standing for 15min for layering, combining acetonitrile layers, and performing reduced pressure evaporation to obtain an acetonitrile extract;
d. separating the acetonitrile extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using petroleum ether-ethyl acetate with the volume ratio of 100-0: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudocanary-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudoolivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudo-white-olivine-6 (17), 11E-diene; compound 8: tylosin A.
Example 5
a. Taking 10kg of naturally dried euphorbia pekinensis fruits, crushing, percolating and extracting at room temperature for 12h by using 65L of anhydrous methanol, combining the percolate and the solvent, and evaporating the solvent under reduced pressure to obtain an euphorbia pekinensis fruit extract;
b. b, dispersing the extract obtained in the step a by 6 times of volume of water, adding 6 times of volume of ethyl acetate for extraction for 5 times, stirring for 10min by a paddle stirrer each time, standing for 25min for layering, combining ethyl acetate layers, and performing reduced pressure evaporation to obtain an ethyl acetate extract;
c. dispersing the ethyl acetate extract obtained in the step b by using 50% methanol with the volume 6 times that of the ethyl acetate extract, adding n-hexane with the volume 5 times that of the ethyl acetate extract to extract for 8 times, stirring for 5min by using a paddle stirrer each time, standing for 15min for layering, combining acetonitrile layers, and concentrating under reduced pressure to obtain an acetonitrile extract;
d. separating the acetonitrile extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using petroleum ether-ethyl acetate with the volume ratio of 100-0: compound 1: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoCanane-6 (17), 11E-diene; compound 2: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudovitinane-6 (17), 11E-diene; compound 3:14 α -benzoyloxy-15 β -hydroxy-5 α,7 β -diisobutyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-dienetetra; compound 4:14 α -benzoyloxy-15 β -hydroxy-5 α -isobutyryloxy-7 β -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 5:14 α -benzoyloxy-15 β -hydroxy-7 β -isobutyryloxy-5 α -propionyloxy-2 α,3 β,8 α,9 α -tetraacetoxy-pseudoolivine-6 (17), 11E-diene; compound 6: (2S, 3S,4R,5R,7S,8R,13S, 15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudowhite-olivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R, 3R,4S,5R,7S,8S,9S,13S,14S, 15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudo-white-olivine-6 (17), 11E-diene; compound 8: tylosin A.
Example 6
The macrocyclic diterpenoid components obtained in the examples 1 to 5 are analyzed by high performance liquid chromatography, the content of each macrocyclic diterpenoid in the components is shown in the table 1, and the reference substance is prepared by separation in the laboratory;
TABLE 1 1g macrocyclic diterpenoid component content
Ten batches of macrocyclic diterpenoid components (G1-G10) were prepared continuously according to the example method, and the utility of the extraction and separation process of macrocyclic diterpenoid components from euphorbia lobata fruits was evaluated by analyzing the total macrocyclic diterpenoid content in the components by high performance liquid chromatography and examining the quality stability of the macrocyclic diterpenoid components (G1-G10), the experimental results are shown in table 2, and the high performance liquid chromatography analysis chart of the macrocyclic diterpenoid components (G1-G10) is shown in fig. 2:
TABLE 2 Total macrocyclic diterpenoid content of ten macrocyclic diterpenoid components
As can be seen from Table 2, the total macrocyclic diterpenoid content in ten consecutive batches of macrocyclic diterpenoid components is higher than 60%, which indicates that the macrocyclic diterpenoid components extracted and separated by the process have good quality stability, completely meet the quality control requirements and can be industrially applied.
Example 7
The application of the macrocyclic diterpenoid components in the euphorbia foliosa fruits in preparing multidrug resistance reversal medicines or preparing antitumor medicines by combining with antitumor medicines takes a human breast cancer cell line (MCF 7) and an adriamycin resistance strain (MCF 7/ADR) thereof as examples;
cytotoxicity testing of macrocyclic diterpenoid components:
materials and reagents: RPMI 1640 medium was purchased from Hyclone; double antibody and fetal bovine serum were purchased from Hyclone; trypsin was purchased from Gibco; thiazole blue (MTT) was purchased from Biosharp; dimethyl sulfoxide (DMSO) was purchased from Amresco; rhodamine 123 is available from Sigma; verapamil hydrochloride was purchased from Sigma; doxorubicin hcl was purchased from shanghai bio ltd; human breast cancer cell lines (MCF 7) and human breast cancer adriamycin resistant cell lines (MCF 7/ADR) were purchased from Shanghai cell banks of Chinese academy of sciences;
cell culture: human breast cancer cell line MCF7 and its adriamycin-resistant cell line MCF7/ADR are cultured in RPMI 1640 complete medium (RPMI 1640 medium +10% FBS +1% double antibody), and all cells are placed in CO 2 Incubator (temperature 37 ℃, concentration 5% CO) 2 ) Maintaining subculture, carrying out drug-resistant culture on the drug-resistant strain cells (MCF 7/ADR) in a complete culture medium with the final concentration of adriamycin of 500ng/mL for one week, carrying out drug-resistant culture in a complete culture medium with the final concentration of 1000ng/mL for one week, and then carrying out culture in a complete culture medium without an anti-tumor drug for two weeks for experiment;
the experimental method comprises the following steps: the MTT method comprises the steps of inoculating MCF7 or MCF7/ADR cells in a logarithmic growth phase into a 96-well microplate at the density of 5000/well, incubating in an incubator at the temperature of 37 ℃ for 24 hours, adding a test monomeric compound, setting 6 concentration gradients and 6 multiple wells for 100 mu L of each well; a cell-free zero-adjusting group, a solvent dimethyl sulfoxide (DMSO) control group and a positive drug control group are additionally arranged; tumor cells at a temperature of 37 ℃ and a concentration of 5% CO 2 After culturing for 48 hours under the conditions, the supernatant was discarded, and thiazole blue (MTT) solution (5 mg/mL, prepared with physiological saline, mixed with complete medium at a ratio of 1 2 Continuously culturing for 4h under the condition; discarding the supernatant, adding 150 mu L of dimethyl sulfoxide (DMSO) into each hole, and detecting the absorbance value (A) of each hole at 570nm by using an enzyme labeling instrument after the formazan is dissolved; the inhibition rate (half inhibition IC) of the macrocyclic diterpenoid component on the growth of tumor cells was calculated according to the following formula 50 Values were calculated using GraphPad Prism software) and macrocyclic diterpenoid component toxicities: survival rate (%) = a drug administration group/a blank group × 100%, antitumor drug adriamycin Resistance Index (RI) = IC 50(MCF7) /IC 50(MCF7/ADR) ；
The experimental results are as follows: the survival rate for the macrocyclic diterpenoid component in the euphorbia folica fruit is shown in table 3:
TABLE 3 survival rates of human breast cancer cell line MCF7 and human breast cancer adriamycin-resistant cell line MCF7/ADR in macrocyclic diterpene compound component environments
As can be seen from Table 3, the macrocyclic diterpenoid component in the Euphorbia lobata fruit does not show cytotoxicity (cell survival rate is more than 70%) to the human breast cancer cell line MCF7 and the human breast cancer adriamycin-resistant cell line MCF7/ADR at the concentration of 12.5mg/mL, which indicates that the cytotoxic activity of the component is low.
Example 8
The compounds 1-8 and macrocyclic diterpenoid compounds are used for testing the activity of reversing the multidrug resistance of the tumor:
in the experiment, macrocyclic diterpenoid compound components in euphorbia foliosa fruits are combined with antitumor drug adriamycin (DOX), growth inhibition on drug-resistant cells is detected before and after the combination, and multi-drug resistance reversal activity test is carried out;
the experimental method comprises the following steps: inoculating MCF7/ADR cells in logarithmic growth phase to a 96-well microplate (100 mu L per well) at a density of 5000 cells/well, incubating for 24h in an incubator at a temperature of 37 ℃, adding adriamycin and a macrocyclic diterpene compound component in the fruits of Euphorbia lobata to be tested or a positive control drug verapamil, setting a concentration gradient of 7 cells per well at 100 mu L per well, setting 6 multiple wells, and setting a blank control group and a solvent dimethyl sulfoxide (DMSO) control group; tumor cells at 37 deg.C and a concentration of 5% 2 After culturing for 48 hours under the conditions, the supernatant was discarded, and thiazole blue (MTT) solution (5 mg/mL, prepared with physiological saline, mixed with complete medium at a ratio of 1 2 Continuously culturing for 4h under the condition, removing the supernatant, adding 150 mu L of dimethyl sulfoxide (DMSO) into each hole, detecting the absorbance value (A) of 570nm of each hole by using an enzyme-labeling instrument after formazan is dissolved, and calculating the inhibition rate of the monomer compound to be tested on the growth of tumor cells according to the following formula: inhibition (%) = (a control group-a administration group)/a control group × 100%; inverse multiple (RF) = IC 50 (Adriamycin)/IC 50 (doxorubicin + compounds);
the experimental results are as follows: the half-growth inhibition rate and the reversal factor of MCF7/ADR cells of macrocyclic diterpenoid compounds 1-8 in Euphorbia lobata fruits when combined with adriamycin are shown in Table 4, and the half-growth inhibition rate and the reversal factor of MCF7/ADR cells of macrocyclic diterpenoid compounds 1-8 in Euphorbia lobata fruits when combined with adriamycin are shown in Table 5:
TABLE 4 half-growth inhibition and fold reversal RF values for doxorubicin-resistant cells MCF7/ADR in human breast cancer in combination with doxorubicin for each compound in the macrocyclic diterpenoid fraction of the Euphorbia folioides fruit
TABLE 5 half growth inhibition and fold reversal RF values for the macrocyclic diterpenoid component of Euphorbia lobata fruits in combination with Adriamycin in human breast cancer Adriamycin resistant cells MCF7/ADR
As can be seen from tables 4 and 5, the macrocyclic diterpene compounds contained in the macrocyclic diterpene compound fractions 1-8 of the Euphorbia lobata fruit have multidrug resistance reversal activities to different degrees, and compared with the individual action of adriamycin after doxorubicin is combined with the macrocyclic diterpene compound fraction contained in the Euphorbia lobata fruit and the verapamil VRP as a positive drug, IC is shown 50 The value is significantly reduced; under the concentration of 50mg \\ \ mL, the reversing multiple of the compound component is as high as 66.2 times, although the reversing multiple is weaker than that of a positive control drug verapamil, the compound component is used as a macrocyclic diterpenoid compound component, has better multidrug resistance reversing activity and is used as a multidrug resistance reversing drug to assist tumor chemotherapy.
Claims (1)
1. A method for extracting and separating macrocyclic diterpenoid components from euphorbia pekinensis fruits is characterized by comprising the following steps:
a. taking Euphorbia pekinensis fruit as a raw material, naturally airing, crushing, percolating, cold soaking or heating reflux extraction with 5-10 times volume of 50-99% ethanol aqueous solution, absolute ethyl alcohol, acetone, 50-99% methanol aqueous solution or absolute methanol at room temperature, and concentrating to obtain Euphorbia pekinensis fruit extract;
b. b, dispersing the extract obtained in the step a by using water with the volume 5-10 times that of the extract, adding n-hexane, ethyl acetate or a mixed solvent of n-hexane and ethyl acetate with the volume 5-10 times that of the extract to extract for 5-10 times, combining the extract, and concentrating under reduced pressure to obtain an extract;
c. dispersing the extract obtained in the step b by using acetonitrile with the volume 5-10 times of that of the extract or 50% methanol aqueous solution, adding n-hexane, petroleum ether or cyclohexane with the volume 5-10 times of that of the extract for extraction for 5-10 times, and concentrating to obtain an extract;
d. separating the extract of the acetonitrile or 50% methanol aqueous solution extract obtained in the step c by using a normal phase silica gel column, performing gradient elution by using petroleum ether-ethyl acetate or n-hexane-acetone with the volume ratio of 100-0; evaporating fraction Fr.2 at 38-40 deg.C under reduced pressure to obtain yellow-brown viscous liquid, drying at 50 deg.C in water bath to constant weight to obtain macrocyclic diterpene compounds: compound 1: (2R,3R,4S,5R,7S,8S,9S,13S,14S,15R) -5-benzoyloxy-15-hydroxy-7-isobutyryloxy-2, 3,8,9, 14-tetraacetoxy-pseudoolivine-6 (17), 11E-a diene; compound 2: (2R,3R,4S,5R,7S,8S,9S,13S,14S,15R) 2,3,8, 9-tetraacetoxy-5, 14-dibenzoyloxy-15-hydroxy-7-isobutyryloxy-pseudoolivine-6 (17), 11E-a diene; compound 3:14α-benzoyloxy-15β-hydroxy-5α,7β-diisobutyryloxy-2α,3β,8α,9α-tetraacetoxy-pseudocanary-6 (17), 11E-diene four; compound 4:14α-benzoyloxy-15β-hydroxy-5α-isobutyryloxy-7β-propionyloxy-2α,3β,8α,9α-tetraacetoxy-pseudocanavanine-6 (17), 11E-a diene; compound 5:14α-benzoyloxy-15β-hydroxy-7β-butyl acetateacyloxy-5-Compoundsα-propionyloxy-2α,3β,8α,9α-tetraacetoxy-pseudocanary-6 (17), 11E-a diene; compound 6: (2S,3S,4R,5R,7S,8R,13S,15R) -3,5,8, 15-tetraacetoxy-7-isobutyryloxy-pseudoolivine-6 (17), 11E-diene-9, 14-dione; compound 7: (2R,3R,4S,5R,7S,8S,9S,13S,14S,15R) -2,3,8, 9-tetraacetoxy-14-benzoyloxy-15-hydroxy-7-isobutyryloxy-5- (2-methyl-butyryloxy) -pseudoolivine-6 (17), 11E-a diene; compound 8: tyla zener euphorbia.
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| CN105820044A (en) * | 2016-01-12 | 2016-08-03 | 云南民族大学 | Novel C22 diterpene compound, preparation method thereof and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| "Jatrophane diterpenoid esters from Euphorbia sororia serving as multidrug resistance reversal agents";Lu Dongli 等;《Fitoterapia》;20131127;第92卷;第244-251页 * |
| "Jatrophane diterpenoids as multidrug resistance modulators from Euphorbia sororia";Hequn Yang 等;《Bioorganic Chemistry》;20210731;第112卷;第104989页 * |
| "Jatrophane diterpenoids from Euphorbia sororia as potent modulators against P-glycoprotein-based multidrug resistance";Hu Rui 等;《European Journal of Medicinal Chemistry》;20180111;第146卷;第157-170页 * |
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