CN111592462B - Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof - Google Patents
Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN111592462B CN111592462B CN202010516611.7A CN202010516611A CN111592462B CN 111592462 B CN111592462 B CN 111592462B CN 202010516611 A CN202010516611 A CN 202010516611A CN 111592462 B CN111592462 B CN 111592462B
- Authority
- CN
- China
- Prior art keywords
- column chromatography
- formula
- acetonitrile
- methanol
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
- C07C69/78—Benzoic acid esters
-
- 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
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/58—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/14—All rings being cycloaliphatic
- C07C2602/32—All rings being cycloaliphatic the ring system containing at least eleven carbon atoms
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to macrocyclic diterpenoid compounds isolated from euphorbia multocida, a preparation method and application thereof, namely euphorbia multocida (A)E.glomerulans) The whole grass of (1) is taken as a raw material, is extracted by an organic solvent, is separated by three to four methods of a solvent extraction method, a normal phase silica gel column chromatography, a reverse phase silica gel column chromatography and a Sephadex LH-20 gel column chromatography to obtain three novel pseudo-bruane macrocyclic diterpenoid compounds of a formula (I), a formula (II) and a formula (III), and is subjected to anti-tumor multi-drug resistance reversal activity determination, and the result shows that: the three new pseudo-bruane macrocyclic diterpenoid compounds shown in the formula (I), the formula (II) and the formula (III) have antitumor multi-drug resistance reversing activities in different degrees, can reverse the drug resistance of tumor drug-resistant cells to antitumor drugs in different degrees by combining with the antitumor drugs, and can be used for preparing the antitumor multi-drug resistance reversing drugs.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to three novel macrocyclic diterpenoid compounds in euphorbia pekinensis and application thereof in preparing tumor multidrug resistance reversal medicines or preparing anti-tumor combination medicines by combining with anti-tumor medicines.
Background
Euphorbia (Euphorbia L.) plants have significant medicinal value and are widely used in the treatment of tumors. The compounds isolated from Euphorbia plants mainly include sesquiterpene, diterpene, triterpene, etc., and the diterpene components are most reported in research. The diterpenoid compounds have the activities of resisting tumors, viruses, bacteria and the like, and have important value in the research of anticancer drugs. Euphorbia mulberosa (Euphorbia Glomulums) is a perennial herb of the genus Euphorbia of the family Euphorbiaceae. It is mainly distributed in Zhongya and Xinjiang Qitai, wuluqiqi, hodgkin, and Jiang Cheng. The material basis for the medicinal effect of the euphorbia pekinensis is not elucidated so far. Therefore, the systematic research on the macrocyclic diterpenoid ingredients in the euphorbia pekinensis is carried out, the substance basis of the pharmacological activity of the euphorbia pekinensis is determined, and the discovery of a diterpenoid new compound with specific activity is of great significance.
Euphorbia (Euphorbia L.) plants are rich in macrocyclic diterpenoid components. The macrocyclic diterpenoid compounds are diterpenoid compounds with more than six-membered ring structures in molecules, the skeleton structures are rich and changeable, and the biological activities are various and comprise the activities of cell toxicity, multidrug resistance reversion, antibiosis, HIV virus resistance and the like. The highest number of pseudoolivine (Jatrophane) diterpenes has been found, which are structurally characterized by a skeleton fused from a five-membered ring and a twelve-membered ring, i.e. a 5/12 ring system. Previous research shows that the pseudo-elemene diterpene is usually in a high oxidation state and generally has a larger molecular structure with a molecular weight of above 700, and the structure is rich because various substituents such as acetoxyl, propionyloxy, benzoyloxy, isobutyryloxy, pyridine acyloxy-3-yl and the like are usually arranged on the skeleton. These substituents differ in ring position, number and configuration, resulting in a rich diversity of such compounds. The research on the structure-activity relationship shows that the influence of the substituent on the skeleton on the pharmacological action is large, so that the substitution mode of the pseudo-bruane type compound determines the novelty of the compound.
Tumor multidrug resistance (MDR), which is a phenomenon that after a drug acts on a tumor to cause drug resistance, the tumor has cross-resistance to multiple antineoplastic drugs which are never contacted, unrelated in structure, different in target and different in mechanism. MDR has multiple formation mechanisms, one of the most important mechanisms is the overexpression of ABC family transporter (the P-glycoprotein, P-gp, coded by ABCB1 gene, which is the most widely and deeply studied at present) so as to increase drug efflux and form drug resistance. In recent years, a plurality of macrocyclic diterpenoid compounds isolated from euphorbia plants are found to have remarkable anti-tumor multi-drug resistance reversing activity, so that the macrocyclic diterpenoid compounds become one of research hotspots.
Disclosure of Invention
The invention aims to provide a macrocyclic diterpenoid compound separated from euphorbia pekinensis and a preparation method and application thereof. Taking whole plant of Euphorbia mulberosa (Euphorbia Glomerula) as a raw material, extracting with a solvent, and separating by two to three methods of a solvent extraction method, a normal phase column chromatography, a reverse phase column chromatography and a Sephadex LH-20 gel column chromatography to obtain 3 novel pseudo-white-elenane macrocyclic diterpenoid compounds of a formula (I), a formula (II) and a formula (III). The invention carries out in vitro cytotoxicity activity determination and multidrug resistance reversal activity determination on the pseudo-brumiane macrocyclic diterpenoid compounds shown in the formula (I), the formula (II) and the formula (III). The results show that: the novel terpenoid compounds of the formula (I), the formula (II) and the formula (III) have antitumor multi-drug resistance reversal activities in different degrees, and can be used for preparing antitumor multi-drug resistance reversal drugs or combined with antitumor drugs to prepare antitumor combined drugs.
The invention relates to a macrocyclic diterpenoid compound separated from euphorbia pekinensis, which has the structural formula:
wherein: the compound of formula (I) is (2S) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy-pseudoelenane-6 (17), 11E-diene-9, 14-dione;
the compound of formula (II) is (2R) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one;
the compound of formula (III) is (2S, 3S,4R,5R,7S,8R,13S, 15R) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione.
The preparation method of the macrocyclic diterpenoid compound separated from the euphorbia pekinensis comprises the following steps:
a. taking the whole plant of euphorbia pekinensis as a raw material, crushing, performing percolation, cold soaking or heating reflux extraction at room temperature by using 5-10 times of 50-99% ethanol solution, absolute ethanol, 50-99% methanol solution, absolute methanol, pure acetone or 50-99% acetone solution at room temperature, and performing vacuum concentration to obtain a crude extract of euphorbia pekinensis;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding cyclohexane, normal hexane or petroleum ether for extraction, adding acetonitrile for extraction for 2-5 times, and evaporating the acetonitrile extract to dryness in vacuum to obtain a solvent for concentration to obtain an extract;
or dispersing the crude extract obtained in the step a by using petroleum ether, n-hexane or cyclohexane, adding acetonitrile for extraction for 2-5 times, evaporating the acetonitrile extract to dryness in vacuum, and concentrating the solvent to obtain an extract;
c. separating three or four of the acetonitrile extract obtained in the step b by normal phase silica gel column chromatography, reverse phase silica gel column chromatography and Sephadex LH-20 gel column chromatography to obtain the compound (2S) of the formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) 3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy-pseudoolivine-6 (17), 11E-diene-9, 14-dione, the compound of formula (II) being (2R) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one and the compounds of formula (III) are (2S, 3S,4R,5R,7S,8R,13S, 15R) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-dien-9, 14-dione.
The normal phase silica gel column chromatography in the step c is normal pressure or pressurized column chromatography, the filler is silica gel, the eluent is a mixture of at least two solvents of petroleum ether, cyclohexane, normal hexane, acetone, chloroform, ethyl acetate and methanol with the volume ratio of 100-0, and isocratic elution or gradient elution is adopted.
The reverse phase column chromatography in step c is normal pressure or pressurized column chromatography, the eluant is 30-99% methanol solution or 10-99% acetonitrile-methanol-water mixed solution, and isocratic elution or gradient elution is adopted.
And c, performing Sephadex LH-20 gel column chromatography in the step c by using normal-pressure column chromatography, wherein an eluant is a mixture of at least two solvents of methanol, dichloromethane and trichloromethane with the volume ratio of 1.
The application of the macrocyclic diterpenoid compound separated from the euphorbia pekinensis in preparing a tumor multidrug resistance reversing medicine.
The macrocyclic diterpenoid compounds in the euphorbia pekinensis, the compounds in the formula (I), the formula (II) and the formula (III) are used for preparing the medicine combined with the anti-tumor medicine.
The macrocyclic diterpenoid compounds separated from euphorbia multocida as well as the preparation method and the application thereof are characterized in that the experimental results show that the 3 new macrocyclic diterpenoid compounds obtained by the method are subjected to tumor multidrug resistance reversal activity determination: the compounds of the formula (I), the formula (II) and the formula (III) can inhibit the drug exclusion effect mediated by P-glycoprotein in drug-resistant cells to different degrees, thereby showing the multidrug resistance reversal activity of breast cancer, and can be used for preparing antitumor multidrug resistance reversal drugs or preparing antitumor combination drugs by combining with antitumor drugs.
The macrocyclic diterpenoid compounds in the euphorbia pekinensis can be obtained by separation and purification from plants or synthesis through chemical modification methods well known to those skilled in the art.
The macrocyclic diterpenoid compounds in the euphorbia pekinensis provided by the invention are structurally determined by modern spectrum means such as high-resolution mass spectrum, one-dimensional and two-dimensional nuclear magnetic resonance spectrum and the like, and the structure identification process is as follows:
the compound of formula (I) is a white amorphous powder, [ alpha ]]25D+87(c 0.2,MeOH)；UV(MeOH)λ max (log ε) 230 (4.06) nm; ECD (MeOH) 214 (. DELTA.. Epsilon. -4.31), 235 (. DELTA.. Epsilon. + 7.61), 255 (. DELTA.. Epsilon. + 0.22), 301 (. DELTA.. Epsilon. + 3.13) nm; HRESI (+) MS gave the peak of the excimer ionm/z 697.2610[M+Na] + (calculated value is C) 38 H 42 O 11 Na 697.2625), its molecular formula is determined to be C 38 H 42 O 11 (ii) a According to 1 H， 13 Determining the structure by C NMR and two-dimensional nuclear magnetic resonance data, wherein the skeleton type is pseudo-elemene (Jatrophane) type and is named as (2S) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy-pseudoelenane-6 (17), 11E-diene-9, 14-dione, which 1 H and 13 c NMR data are shown in Table 1[600MHz ] ( 1 H)，150MHz( 13 C) Solvent (c): CDCl 3 ]。
The compound of formula (II) is a white amorphous powder, [ alpha ]]25D-5(c 0.1,MeOH)；UV(MeOH)λ max (log ε) 230 (4.04) nm; ECD (MeOH) 218 (Δ ε -1.69), 220 (Δ ε -1.77), 237 (Δ ε + 2.08), 254 (Δ ε -0.47), 301 (Δ ε + 0.69) nm; according to HRESI (+) MS (m/z 757.2831 2M + Na] + Calculated value of C 40 H 46 O 13 Na 757.2836) to determine its molecular formula as C 40 H 46 O 13 (ii) a According to 1 H， 13 Determining the structure by C NMR and two-dimensional nuclear magnetic resonance data, wherein the skeleton type is pseudo-elemene (Jatrophane) type and is named as (2R) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-diene. It is provided with 1 H and 13 the C NMR data are shown in Table 1[600MHz ] ( 1 H)，150MHz( 13 C) The solvent: CDCl 3 ]。
The compound of formula (III) is a white amorphous powder, [ alpha ]]25D+91(c 0.1,MeOH)；UV(MeOH)λ max (log ε) 229 (4.26) nm; ECD (MeOH) 203 (Δ ε + 0.81), 217 (Δ ε -12.94), 236 (Δ ε -0.43), 245 (Δ ε -0.83), 299 (Δ ε + 4.71) nm; according to which 13 C NMR and HRESI (+) MS (m/z 663.2771[ M ] +Na)] + Theoretical value C 35 H 44 O 11 Na 663.2781) data to determine its molecular formula as C 35 H 44 O 11 (ii) a According to 1 H， 13 The structure is determined by C NMR and two-dimensional nuclear magnetic resonance data, the framework type is pseudo-brubane (Jatrophane) type, and the name is (2S, 3S,4R,5R,7S,8R,13S, 15R) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-brubane-6 (17), 11E-diene. It is composed of 1 H and 13 the C NMR data are shown in Table 1[400MHz ] (C NMR) 1 H)，100MHz( 13 C) The solvent: CDCl 3 ]。
TABLE 1 of the compounds of formula (I), formula (II) and formula (III) 1 H and 13 c NMR data [ delta (ppm), J (Hz)]
Drawings
FIG. 1 shows the preparation of the compounds of formula (I) according to the invention 1 H NMR(600MHz,CDCl 3 ) A spectrogram;
FIG. 2 shows the preparation of the compounds of formula (I) according to the invention 13 C NMR(150MHz,CDCl 3 ) A spectrogram;
FIG. 3 shows the preparation of the compounds of formula (II) according to the invention 1 H NMR(600MHz,CDCl 3 ) A spectrogram;
FIG. 4 shows the preparation of the compound of formula (II) according to the invention 13 C NMR(150MHz,CDCl 3 ) A spectrogram;
FIG. 5 shows the preparation of the compound of formula (III) according to the invention 1 H NMR(400MHz,CDCl 3 ) A spectrogram;
FIG. 6 shows the preparation of the compound of formula (III) according to the invention 13 C NMR(100MHz,CDCl 3 ) And (4) a spectrogram.
Detailed Description
Example 1
a. Taking 4.7kg of euphorbia multocida whole plant, crushing, extracting by a 30L acetone percolation method at room temperature, and evaporating the solvent to dryness under reduced pressure to obtain a crude extract of euphorbia multocida;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding cyclohexane for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using a petroleum ether-ethyl acetate = 100; performing Sephadex LH-20 column chromatography on the F9E, and separating chloroform-methanol with the volume ratio of 1; wherein the F9E3 section is prepared into a reverse phase column C 18 Separating by 5 mu m of 150 multiplied by 10mm, and carrying out isocratic elution by using a mixed solution of acetonitrile and water with a volume ratio of 48 * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; subjecting the fraction of the F12 section to Sephadex LH-20 column chromatography with the volume ratio of 1; wherein F12BC is subjected to preparation of a reverse phase column C 18 Separating by 5 mu m of 150 multiplied by 10mm, and carrying out isocratic elution by using a mixed solution of acetonitrile and water as an eluent with the volume ratio of 50 * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
Example 2
a. Taking 4.7kg of euphorbia pekinensis whole plant, crushing, extracting by a 50L absolute methanol percolation method at room temperature, and evaporating the solvent to dryness under reduced pressure to obtain a crude extract of euphorbia pekinensis;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding n-hexane for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using a solvent system comprising cyclohexane and ethyl acetate in a volume ratio of 100-0; separating the F9E by Sephadex LH-20 column chromatography, and eluting with methanol to obtain components F9E1-F9E5; wherein the F9E3 section is prepared into a reverse phase column C 18 5 μm 150X 10mm separation, isocratic elution with 48% acetonitrile-water solution to give (2S) as the compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; separating the component of the F12 section by Sephadex LH-20 column chromatography, and eluting with methanol to obtain components F12A-F12E; separating the F12BC component by RP-18 reverse phase column, gradient eluting with 20% -100% methanol-water solution, collecting 50% methanol-water solution eluate, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating by 5 μm10 × 250mm, and isocratically eluting with 57% acetonitrile-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one.
Example 3
a. Taking 4.7kg of euphorbia multocida whole plant, crushing, performing cold-leaching extraction at room temperature by using 60L of 80% methanol-water solution, and evaporating the solvent under reduced pressure to obtain a crude extract of euphorbia multocida;
b. dispersing the crude extract obtained in the step a by using n-hexane, adding acetonitrile for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using normal hexane and ethyl acetate as solvent systems in a volume ratio of 100-0; separating the F9E component by RP-18 reverse phase column, gradient eluting with 30% -100% methanol-water solution, collecting 60% methanol-water solution eluate, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 48% acetonitrile-water solution to obtain (2S) as compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; separating the component in the F12 section by Sephadex LH-20 column chromatography, and eluting with methanol to obtain components F12A-F12E; separating F12BC component by RP-18 reverse phase column, gradient eluting with 30% -100% acetonitrile-water solution, collecting 55% methanol-water solution eluent, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating by 5 μm10 × 250mm, and isocratically eluting with 57% acetonitrile-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
Example 4
a. Taking 4.7kg of euphorbia multocida whole plant, crushing, percolating and extracting with 50L of absolute ethyl alcohol at room temperature, and evaporating the solvent to dryness under reduced pressure to obtain a crude extract of euphorbia multocida;
b. dispersing the crude extract obtained in the step a by using petroleum ether, adding acetonitrile for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using a solvent system comprising petroleum ether and ethyl acetate in a volume ratio of 100-0; separating the F9E component by RP-18 reverse phase column, gradient eluting with 30% -100% acetonitrile-water solution, collecting 50% acetonitrile-water solution eluate, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 48% acetonitrile-water solution to obtain (2S) compound (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; separating the F12 section component by Sephadex LH-20 column chromatography, and eluting with dichloromethane to methanol with the volume ratio of 1; the F12BC component is used for preparing a reverse phase column C 18 Separating by 5 μm10 × 250mm, and isocratically eluting with 50% acetonitrile-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
Example 5
a. Taking 4.7kg of euphorbia multocida whole plant, crushing, performing cold-leaching extraction at room temperature by using 60L of 70% ethanol-water solution, and evaporating the solvent under reduced pressure to obtain a crude extract of euphorbia multocida;
b. b, dispersing the crude extract obtained in the step a by using cyclohexane, adding acetonitrile for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using a solvent system comprising cyclohexane to ethyl acetate in a volume ratio of 100-0; separating the F9E component by RP-18 reverse phase column, gradient eluting with 40% -100% methanol-water solution, collecting 60% methanol-water solution eluate, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 68% methanol-water solution to obtain (2S) compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; separating the component in the F12 section by Sephadex LH-20 column chromatography, and eluting with chloroform-methanol with the volume ratio of 1; the F12BC component is used for preparing a reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 50% acetonitrile-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
Example 6
a. Taking 4.7kg of euphorbia mulberosa, crushing, performing reflux extraction at 80 ℃ by using 70L of 90% ethanol-water solution, and evaporating the solvent under reduced pressure to obtain a euphorbia mulberosa crude extract;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding petroleum ether for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using petroleum ether and ethyl acetate as solvent systems in a volume ratio of 100-0; carrying out Sephadex LH-20 column chromatography on the F9E, and eluting and separating by using anhydrous methanol to obtain components F9E1-F9E5; the F9E3 section is used for preparing a reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 68% methanol-water solution to obtain (2S) compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione; separating the component in the F12 section by Sephadex LH-20 column chromatography, and eluting with chloroform-methanol with the volume ratio of 1; subjecting the F12BC fraction to preparative reverse phase column C 18 Separating by 5 μm10 × 250mm, and isocratically eluting with 60% methanol-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one.
Example 7
a. Taking 4.7kg of euphorbia mulberosa whole plant, crushing, extracting by 40L of acetone-water solution percolation method with the concentration of 80% at room temperature, and evaporating the solvent to dryness under reduced pressure to obtain a euphorbia mulberosa crude extract;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding cyclohexane for extraction, combining acetonitrile layers, and evaporating the acetonitrile under reduced pressure to obtain an acetonitrile extract;
c. separating the acetonitrile extract obtained in the step b by using a normal phase silica gel column, performing gradient elution by using normal hexane and ethyl acetate as solvent systems in a volume ratio of 100-0; carrying out Sephadex LH-20 column chromatography on the F9E, and eluting and separating dichloromethane and methanol in a volume ratio of 1; subjecting the F9E3 section to a preparative reverse phase column C 18 Separating by 5 mu m 150 multiplied by 10mm, and carrying out isocratic elution by using a mixed solution of acetonitrile and water with the volume ratio of 48 * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; and (3) passing the fraction of the F12 section through a Sephadex LH-20 column, wherein the volume ratio of the chloroform to the fraction of the F12 section is 1: separating with methanol chromatography to obtain components F12A-F12E; subjecting F12BC to reverse phase column C preparation 18 Separating by 5 μm 150 × 10mm, and eluting with mixed solution of acetonitrile and water at volume ratio of 50 * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one.
Example 8
The application of the macrocyclic diterpenoid compound separated from the euphorbia pekinensis kom herb in preparing anti-tumor multidrug resistance reversal medicines or preparing anti-tumor medicines by combining with the anti-tumor medicines is shown by taking a human breast cancer cell strain and an adriamycin resistance strain thereof as an example;
the cytotoxicity and resistance index of any one of the compounds of formula (I) -formula (III) of examples 1-7 were tested:
materials and reagents: high-glucose DMEM medium was purchased from HyClone; diabody, fetal bovine serum FBS and trypsin were purchased from Gibco; thiazole blue (MTT) was purchased from Biosharp; dimethyl sulfoxide (DMSO) was purchased from Amresco; rhodamine 123 is available from Sigma; doxorubicin hcl and verapamil hcl were purchased from Sigma-Aldrich; the rest other reagents or solvents are purchased from Tianjin Chengyuan chemical reagent, inc.;
cell lines: human breast cancer cell line MCF-7 and human breast cancer adriamycin resistant cell line MCF-7/ADR (purchased from Shanghai cell bank of Chinese academy of sciences);
and (3) cell culture: culturing human breast cancer cell strain MCF-7 and its adriamycin-resistant cell strain MCF-7/ADR in high-sugar DMEM complete medium (high-sugar DMEM medium +10% FBS +1% double antibody); all cells were incubated at 37 ℃ CO 2 Maintaining subculture in an incubator with the concentration of 5 percent; drug-resistant strains (MCF-7/ADR cells) are subjected to drug-resistant culture for 2 generations in a complete culture medium with the adriamycin concentration of 1 mu g/mL so as to maintain the drug resistance of the adriamycin; the culture was then carried out in complete doxorubicin-free medium for two weeks before use in the experiment;
the experimental method comprises the following steps: namely, MTT method, in which MCF-7 or MCF-7/ADR cells in logarithmic growth phase are grown at 5X 10 4 Inoculating the single compound/mL of the single compound in a 96-well plate (100 mu L/well), incubating the plate in an incubator at the temperature of 37 ℃ for 24 hours, adding the single compound to be tested (20 mu L/well), setting different concentration gradients, and setting 3-6 multiple wells for each gradient; a blank cell-free control group, a solvent dimethyl sulfoxide (DMSO) control group and a positive drug control group are additionally arranged; tumor cells at 37 deg.C, 5% 2 Culturing for 48h under the conditions, discarding the supernatant, adding thiazole blue (MTT) solution (5 mg/mL, prepared with PBS, mixed with complete medium at a ratio of 1 2 Continuously culturing for 2h 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 survival rate and the inhibition rate of the test monomeric compound on the tumor cells are calculated according to the following formulas: cell viability% = (compound OD-blank OD/solvent control OD-blank OD) × 100%; cell inhibition rate% = 1-cell survival rate% = [1- (combination)Object OD-blank OD/solvent control OD-blank OD)]X 100%, according to the inhibition rate under each concentration, fitting by a graphpad software formula to obtain IC 50 (ii) a Inverse multiple (RF) = (DOX-IC used alone) 50 ) /(combination of DOX and Compound IC) 50 )；
The experimental results are as follows: the survival rates of the compounds of formula (I) -formula (III) isolated from the whole herb of Euphorbia meyeriana for human breast cancer cell line MCF-7 and human breast cancer doxorubicin-resistant cell line MCF-7/ADR are shown in Table 2:
TABLE 2 survival rates of compounds of formula (I) -formula (III) from whole plant of Euphorbia mexicana at 10. Mu.M concentration against human breast cancer cell line MCF-7 and human breast cancer doxorubicin-resistant cell line MCF-7/ADR
As shown in the table, the compounds of formula (I) to formula (III) showed no cytotoxicity (survival rate > 80%) at 10. Mu.M concentration against both human breast cancer cell line MCF-7 and human breast cancer doxorubicin-resistant cell line MCF-7/ADR.
Example 9
The compound of formula (I) -formula (III) reverses the activity of tumor multidrug resistance test:
in the experiment, macrocyclic diterpenoid compounds (I) - (III) separated from the euphorbia pekinensis whole plant are selected to be combined with an anti-tumor drug adriamycin (DOX), growth inhibition on drug-resistant cells before and after combination is detected, and a multi-drug resistance reversal activity test is carried out;
the experimental method comprises the following steps: the human breast cancer adriamycin resistant cell line MCF-7/ADR cells in the logarithmic growth phase are divided into 5 multiplied by 10 4 Inoculating to 96-well plate (100 μ L per well), incubating at 37 deg.C for 24 hr, adding adriamycin and monomer compound or verapamil (20 μ L per well), setting different concentration gradients, setting 3-6 multiple wells, setting blank control group and dimethyl sulfoxide (DMS) as solventO) control group; tumor cells at 37 ℃ and 5% CO 2 Culturing for 48h under conditions, discarding the supernatant, adding thiazole blue (MTT) (5 mg/mL, prepared in physiological saline, mixed with complete medium at a ratio of 1 2 Continuing to culture the formazan for 2h under the condition, then discarding 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 the 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: percent cell viability% = (compound OD-blank OD/solvent control OD-blank OD) × 100%; cytostatic% = 1-cell viability% = [1- (compound OD-blank OD/solvent control OD-blank OD)]X 100%, fitting with graphpad to obtain IC 50 (ii) a Inverse multiple (RF) = (DOX-IC used alone) 50 ) /(Combined use of DOX and Compound IC) 50 )；
The experimental results are as follows: half maximal Inhibitory Concentration (IC) of the compound of formula (I) -formula (III) in combination with Adriamycin on MCF-7/ADR cells 50 ) And the fold reversal thereof are shown in table 3:
TABLE 3 half inhibitory concentration and fold reversal of MCF-7/ADR cells for the combination of a Compound of formula (I) with Adriamycin
As shown in Table 3, the compounds of formula (I) -III, when used in combination with doxorubicin, showed an IC value that is comparable to that of doxorubicin alone 50 The values are obviously reduced, the reduction degree is expressed by reversal times, and the reversal times (22.16, 20.80 and 43.63) of the compounds of the formula (I) and the formula (III) have stronger activity of reversing the multidrug resistance of the tumors compared with a positive control drug verapamil (29.29).
Claims (6)
1. A pseudo-olivine macrocyclic diterpenoid compound isolated from Euphorbia pekinensis is characterized in that the compound has the structural formula:
wherein: the compound of formula (I) is (2S) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy-pseudoelenane-6 (17), 11E-diene-9, 14-dione;
the compound of formula (II) is (2R) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticjaban-6 (17), 11E-dien-9-one;
the compound of formula (III) is (2S, 3S,4R,5R,7S,8R,13S, 15R) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione.
2. The method for the preparation of macrocyclic diterpenes isolated from euphorbia pekinensis as claimed in claim 1, characterized by the following steps:
a. taking the whole plant of euphorbia pekinensis as a raw material, crushing, performing percolation, cold soaking or heating reflux extraction at room temperature by using 5-10 times of 50-99% ethanol solution, absolute ethanol, 50-99% methanol solution, absolute methanol, pure acetone or 50-99% acetone solution at room temperature, and performing vacuum concentration to obtain a crude extract of euphorbia pekinensis;
b. dispersing the crude extract obtained in the step a by using acetonitrile, adding cyclohexane, normal hexane or petroleum ether for extraction, adding acetonitrile for extraction for 2-5 times, and evaporating the acetonitrile extract to dryness in vacuum to obtain a solvent for concentration to obtain an extract;
or dispersing the crude extract obtained in the step a by using petroleum ether, n-hexane or cyclohexane, adding acetonitrile for extraction for 2-5 times, evaporating the acetonitrile extract to dryness in vacuum, and concentrating the solvent to obtain an extract;
c. c, subjecting the acetonitrile extract obtained in the step b to normal phase silica gel column chromatography, reverse phase silica gel column chromatography and Sephadex LH-20 gel column chromatography to prepare reverse phaseSeparating three or four of the column chromatography to obtain the compound (2S) of the formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy-pseudoolivine-6 (17), 11E-diene-9, 14-dione, the compound of formula (II) being (2R) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybispseudoolivine-6 (17), 11E-dien-9-one and the compound of formula (iii) are (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy-pseudoolivine-6 (17), 11E-dien-9, 14-dione, wherein the normal phase silica gel column chromatography used in step c is normal pressure or pressurized column chromatography, the filler is silica gel, and the eluent is a mixture of at least two solvents of petroleum ether, cyclohexane, n-hexane, acetone, chloroform, ethyl acetate, methanol in a volume ratio of 100 to 1-0, and isocratic elution or gradient elution is used; the reverse phase column chromatography is normal pressure or pressurized column chromatography, the eluent is 30-99% methanol solution or 10-99% acetonitrile-methanol-water mixed solution, and isocratic elution or gradient elution is adopted; the Sephadex LH-20 gel column chromatography is normal pressure column chromatography, and the eluent is a mixture of at least two solvents of methanol, dichloromethane and trichloromethane with the volume ratio of 1.
3. The method for preparing macrocyclic diterpene compounds isolated from euphorbia multocida as claimed in claim 2, wherein the macrocyclic diterpene compounds isolated from euphorbia multocida are isolated by three of normal phase silica gel column chromatography, sephadex LH-20 gel column chromatography and preparative reverse phase column chromatography in step c, the acetonitrile extract obtained in step b is isolated by normal phase silica gel column chromatography, gradient elution is carried out with petroleum ether-ethyl acetate or n-hexane: ethyl acetate as solvent system with volume ratio of 100Performing gradient elution by using chloroform-acetone as a solvent system to obtain components F9A-F9J; carrying out Sephadex LH-20 column chromatography on the F9E, and separating chloroform-methanol, dichloromethane-methanol or anhydrous methanol with the volume ratio of 1; wherein the F9E3 section is prepared into a reverse phase column C 18 Separating by 5 μm 150 × 10mm, eluting with mixed solution of acetonitrile 52 and water or methanol-water solution with concentration of 68% at volume ratio of 48, and isocratically eluting to obtain (2S) compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudo-elemene-6 (17), 11E-diene-9, 14-dione; separating the component in the F12 section by Sephadex LH-20 column chromatography, and eluting with chloroform to methanol or chloroform to methanol at a volume ratio of 1; wherein F12BC is subjected to preparation of a reverse phase column C 18 Separating at 5 μm 150 × 10mm, and eluting with acetonitrile-water mixed solution at volume ratio of 50 * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxy pseudoolivine-6 (17), 11E-dien-9-one.
4. The method for preparing macrocyclic diterpene compounds isolated from euphorbia multocida as claimed in claim 2, wherein the separation is performed by four of normal phase silica gel column chromatography, reverse phase silica gel column chromatography, sephadex LH-20 gel column chromatography and preparative reverse phase column chromatography in step c, the acetonitrile extract obtained in step b is separated by normal phase silica gel column, gradient elution is performed by using cyclohexane: ethyl acetate as solvent system with volume ratio of 100To compositions F9A-F9J; separating the F9E by Sephadex LH-20 column chromatography, and eluting with methanol to obtain components F9E1-F9E5; wherein the F9E3 section is prepared into a reverse phase column C 18 Separating at 5 μm 150 × 10mm, and isocratic eluting with 48% acetonitrile-water solution to obtain (2S) as compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione; separating the component of the F12 section by Sephadex LH-20 column chromatography, and eluting with methanol to obtain components F12A-F12E; separating F12BC component by RP-18 reverse phase column, gradient eluting with 20% -100% methanol-water solution, collecting 50% methanol-water solution eluate, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating by 5 μm10 × 250mm, and isocratically eluting with 57% acetonitrile-water solution to obtain (2R) compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
5. The method for preparing macrocyclic diterpenoid compounds isolated from euphorbia multocida as claimed in claim 2, characterized in that the separation is performed by four of normal phase silica gel column chromatography, reverse phase silica gel column chromatography, sephadex LH-20 gel column chromatography and preparative reverse phase column chromatography in step c, the acetonitrile extract obtained in step b is separated by normal phase silica gel column, gradient elution is performed by using a solvent system of n-hexane: ethyl acetate, petroleum ether: ethyl acetate or cyclohexane: ethyl acetate in a volume ratio of 100 to 1-0; separating the F9E fraction by RP-18 reversed phase columnGradient eluting with 30% -100% methanol-water solution or 30% -100% acetonitrile-water solution, collecting 60% methanol-water solution eluate or 50% acetonitrile-water solution eluate, evaporating under reduced pressure, and preparing reverse phase column C 18 Separating at 5 μm10 × 250mm, isocratically eluting with 48% acetonitrile-water solution or 68% methanol-water solution to obtain (2S) compound of formula (I) * ,3S * ,4R * ,5R * ,7S * ,8R * ,13S * ,15R * ) -3, 7-dibenzoyloxy-15-hydroxy-5, 8-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione, and the compound of formula (iii) is (2s, 3s,4r,5r,7s,8r,13s, 15r) -3-benzoyloxy-15-hydroxy-8-isobutyryloxy-5, 7-diacetoxy pseudoolivine-6 (17), 11E-diene-9, 14-dione; separating the F12 section component by Sephadex LH-20 column chromatography, eluting with methanol, and eluting with dichloromethane to methanol at a volume ratio of 1; separating F12BC component by RP-18 reverse phase column, gradient eluting with 30% -100% acetonitrile-water solution, collecting 55% methanol-water solution eluent, evaporating to dryness under reduced pressure, and preparing reverse phase column C 18 Separating at 5 μm10 × 250mm, and isocratically eluting with 57% acetonitrile-water solution or 50% acetonitrile-water solution to obtain (2R) as compound of formula (II) * ,3R * ,4R * ,5R * ,7S * ,8R * ,13S * ,14R * ,15R * ) -5, 7-dibenzoyloxy-3, 15-dihydroxy-2, 8, 14-triacetoxybioticpseudoolivine-6 (17), 11E-dien-9-one.
6. Use of macrocyclic diterpenes isolated from euphorbia multocida according to claim 1 for the preparation of anti-tumor multidrug resistance reversing drugs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010516611.7A CN111592462B (en) | 2020-06-09 | 2020-06-09 | Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010516611.7A CN111592462B (en) | 2020-06-09 | 2020-06-09 | Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111592462A CN111592462A (en) | 2020-08-28 |
| CN111592462B true CN111592462B (en) | 2022-10-11 |
Family
ID=72186328
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010516611.7A Active CN111592462B (en) | 2020-06-09 | 2020-06-09 | Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111592462B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113461704B (en) * | 2021-06-02 | 2022-07-26 | 中国医学科学院药用植物研究所 | Monoterpene coumarin compound, preparation method and application thereof |
| CN115160336B (en) * | 2022-07-21 | 2023-10-03 | 复旦大学 | Oxygen-containing five-membered ring derivative of euphorbia lathyris alkane diterpene, preparation method and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106431925B (en) * | 2016-06-20 | 2019-01-18 | 中山大学 | False white olive alkane type macrocyclic diterpene compounds and its application |
| CN107805238B (en) * | 2017-11-02 | 2021-02-05 | 中国科学院新疆理化技术研究所 | Pseudo-elemene diterpene lactone and preparation method and application thereof |
| CN107827752B (en) * | 2017-11-02 | 2020-07-28 | 中国科学院新疆理化技术研究所 | Macrocyclic diterpenoid compound in euphorbia foliosa fruits, preparation method and multi-drug resistance reversing application |
-
2020
- 2020-06-09 CN CN202010516611.7A patent/CN111592462B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111592462A (en) | 2020-08-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Shaker et al. | Secondary metabolites from Calotropis procera (Aiton) | |
| CN107827752B (en) | Macrocyclic diterpenoid compound in euphorbia foliosa fruits, preparation method and multi-drug resistance reversing application | |
| Long et al. | Indole alkaloids from the aerial parts of Kopsia fruticosa and their cytotoxic, antimicrobial and antifungal activities | |
| Dong et al. | Chemical constituents from Brucea javanica | |
| CN111592462B (en) | Macrocyclic diterpenoid compounds separated from euphorbia multocida as well as preparation method and application thereof | |
| Li et al. | New cytotoxic compounds from flowers of Lawsonia inermis L. | |
| Luo et al. | Cytochalasins from mangrove endophytic fungi Phomopsis spp. xy21 and xy22 | |
| Liu et al. | Cytochalasins and polyketides from the fungus Diaporthe sp. GZU-1021 and their anti-inflammatory activity | |
| CN106749107B (en) | Terpenoid in Roots of Euphorbia soongarica and its preparation method and application | |
| Liu et al. | New sesquiterpenoids from Aglaia odorata var. microphyllina and their cytotoxic activity | |
| Zhang et al. | The absolute configurations of hyperilongenols A–C: rare 12, 13-seco-spirocyclic polycyclic polyprenylated acylphloroglucinols with enolizable β, β′-tricarbonyl systems from Hypericum longistylum Oliv. | |
| Yang et al. | Chemical constituents of Abies delavayi | |
| Ghanadian et al. | New jatrophane diterpenes from Euphorbia osyridea with proapoptotic effects on ovarian cancer cells | |
| CN112661644B (en) | Terane diterpenoid compound in euphorbia pekinensis fruit as well as preparation method and application thereof | |
| CN107805238B (en) | Pseudo-elemene diterpene lactone and preparation method and application thereof | |
| Fan et al. | Dihydro-β-agarofuran sesquiterpene polyesters isolated from the stems of Tripterygium regelii | |
| Liu et al. | Monomeric and dimeric ent-kauranoid-type diterpenoids from Rabdosia japonica and their cytotoxicity and anti-HBV activities | |
| Yang et al. | Lutescins A and B, two new ellagitannins from the twigs of Trigonostemon lutescens and their antiproliferative activity | |
| Bang et al. | Colletotrichalactones A-Ca, unusual 5/6/10-fused tricyclic polyketides produced by an endophytic fungus, Colletotrichum sp. JS-0361 | |
| An et al. | Trijugin-and mexicanolide-type limonoids from the fruits of Heynea trijuga that reverse multidrug resistance in MCF-7/DOX cells | |
| Isaka et al. | Comparison of the bioactive secondary metabolites from the scale insect pathogens, anamorph Paecilomyces cinnamomeus, and teleomorph Torrubiella luteorostrata | |
| Kuang et al. | Sesquiterpene Glucosides from Chloranthus japonicus Sieb. | |
| CN115611963B (en) | Preparation and application of steroid compounds in ailanthus altissima leaves | |
| Kang et al. | Iizukines C− E from a saline soil fungus Aspergillus iizukae | |
| CN111548327B (en) | Carbon-reduced kaurane diterpene, preparation method thereof and application thereof in preparation of antitumor 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 |