CN112479887B - Ester compound in purslane and extraction and separation method and application thereof - Google Patents
Ester compound in purslane and extraction and separation method and application thereof Download PDFInfo
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- CN112479887B CN112479887B CN202011351224.9A CN202011351224A CN112479887B CN 112479887 B CN112479887 B CN 112479887B CN 202011351224 A CN202011351224 A CN 202011351224A CN 112479887 B CN112479887 B CN 112479887B
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- purslane
- elution
- methanol
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/66—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety
- C07C69/73—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of unsaturated acids
- C07C69/738—Esters of keto-carboxylic acids or aldehydo-carboxylic acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
- A23L33/12—Fatty acids or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1892—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns the sorbent material moving as a whole, e.g. continuous annular chromatography, true moving beds
-
- 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
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- 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
- C07C67/52—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/09—Geometrical isomers
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- 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- General Chemical & Material Sciences (AREA)
- Botany (AREA)
- Crystallography & Structural Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
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Abstract
The invention relates to the field of extraction and separation of traditional Chinese medicines, in particular to a novel compound extracted, separated and identified from a purslane medicinal material, and an extraction and separation method and application thereof. The invention provides (7E, 9E) -6-oxooctadecyl-7, 9-ethyl dienoate extracted from purslane, an extraction and separation method and application thereof, the ester compound is prepared by sequentially adopting alcohol decoction extraction, silica gel column chromatography, polyamide column chromatography, ODS (oxide dispersion strengthened) medium-pressure column and Sephadex LH-20 purification and liquid phase separation, the extraction and separation method is simple, convenient, rapid and environment-friendly, the purity of the compound separated by the method is higher, and pharmacological experiments prove that the obtained compound has the effects of resisting tumors and cholinesterase, so the novel ester compound extracted from purslane, and salts and derivatives thereof can be used as natural products to develop new traditional Chinese medicines, and have wide medicine application prospects.
Description
Technical Field
The invention relates to the field of extraction and separation of traditional Chinese medicines, in particular to a new compound extracted, separated and identified from a purslane medicinal material and an extraction and separation method thereof, and specifically relates to an ester compound in purslane and an extraction and separation method and application thereof.
Background
Herba Portulacae (Portulaca oleracea L.), also called herba Portulacae and herba Portulacae, is a plant of Portulacaceae. Purslane has drought and waterlogging resistance, light and yin resistance, wide distribution and rich resources, is taken as a medicinal and edible wild plant, takes dry overground parts of purslane as a medicament in 2015 edition pharmacopoeia of the people's republic of China, has the effects of clearing heat and detoxicating, cooling blood and stopping bleeding, stopping dysentery and the like, and is used for treating heat toxin and bloody dysentery, carbuncle swelling and furuncle, eczema, erysipelas, snake and insect bite, hematochezia, hemorrhoidal bleeding, metrorrhagia and metrostaxis and the like.
Modern pharmacological research of purslane shows that the purslane has the effects of resisting inflammation, relieving pain, resisting bacteria and viruses, reducing blood pressure, reducing blood fat, resisting oxidation and cancers, relaxing skeletal muscles and smooth muscles, regulating immune function and the like. Research shows that numerous chemical components of purslane provide material basis for various pharmacological actions of purslane, and the main chemical components of purslane comprise flavonoids, coumarins, terpenes, steroids, alkaloids, amino acids, various pigments, minerals and the like. In recent years, many researchers have focused on the content measurement of chemical components of purslane, pharmacodynamics, pharmacokinetics and the like, however, no research on the separation and in-vitro analysis of a novel ester compound has been reported in purslane.
Most of the chemical components separated from purslane are known and have low structural novelty, so the development and separation of new compounds in purslane are urgently needed.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a novel ester compound (7E, 9E) -6-oxooctadecyl-7, 9-ethyl dienoate extracted from purslane, an extraction and separation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme.
A health food prepared from herba PortulacaeAn isolated ester compound having the formula C20H34O3The chemical name is (7E, 9E) -6-oxooctadecyl-7, 9-ethyl dienoate, and the chemical structural formula is as follows:
further, the extraction and separation method for extracting and separating the ester compounds from the purslane comprises the following steps.
And 2, evaporating the liquid medicine obtained in the step 1 to dryness, putting the liquid medicine on a silica gel column, eluting the liquid medicine by using ethyl acetate, and recovering the ethyl acetate under reduced pressure to obtain an extract so as to obtain an ethyl acetate extract.
And 3, separating the ethyl acetate extract obtained in the step 2 by using a polyamide column, eluting by using ethanol, evaporating ethanol part to dryness, then loading the ethanol part to a silica gel column, sequentially performing gradient elution by using ethyl acetate-methanol to obtain a plurality of elution parts, detecting by using a thin-layer chromatography, developing, combining the developed elution parts, and concentrating the combined elution parts under reduced pressure until the combined elution parts are dry for later use.
And 4, carrying out chromatographic separation on the product obtained in the step 3 by using a pretreated ODS (octadecylsilane chemically bonded silica) column, carrying out gradient elution by using methanol-water to obtain a plurality of elution parts, detecting by using a thin-layer chromatography, developing, and concentrating each developed elution part under reduced pressure until the elution parts are dried to obtain a concentrate for later use.
And 5, carrying out chromatographic separation on the product obtained in the step 4 by using a pretreated Sephadex LH-20 column, eluting by using methanol to obtain a plurality of elution parts, detecting by using a thin-layer chromatography, developing, combining the developed elution parts, and concentrating the combined elution parts under reduced pressure until the combined elution parts are dried for later use.
And 6, performing HPLC (high performance liquid phase) separation preparation on the concentrate obtained in the step 5, wherein the volume ratio of methanol to 0.1% formic acid is 80: 20 as a mobile phase to prepare an ester compound.
Further, the alcohol solvent in the step 1 is 50% ethanol, the extraction times are 2 times, each time of extraction and reflux is 2 hours, and the dosage of the ethanol is 8-16 times of that of the medicinal materials.
Further, the mobile phase elution procedure used in step 2 is isocratic elution.
Further, the volume ratio of water to ethanol in the step 3 is 5: and (4) performing 95 isocratic elution.
Further, the volume ratio of ethyl acetate to methanol in the step 3 is 5:1 and 2: gradient elution 1.
Further, the volume ratio of methanol to water in the step 4 is 60: 40. 70: 30. 80: 20. 90: 10 and 100: elution was performed with a gradient of 0.
Further, the pretreatment process of the ODS column and the sephadex comprises soaking in methanol for 24 hours, loading on the column, washing with methanol until no turbidity is formed in the dripped water, and then balancing with an initial mobile phase.
Further, the methanol elution procedure in step 5 is isocratic elution.
Furthermore, the separated ester compounds extracted from purslane and the application of the salts and derivatives thereof in preparing anti-tumor and anti-cholinesterase medicines are provided.
The anti-tumor and anti-cholinesterase products include, but are not limited to, pharmaceuticals.
Compared with the prior art, the invention has the following beneficial effects.
The separation and pharmacological activity research of the new purslane compound is not reported in the journal of the prior paper, the invention provides a new compound derived from purslane and an extraction and separation method aiming at the new compound, the separation, purification and preparation are carried out by adopting alcohol extraction, a polyamide column, silica gel column chromatography, ODS medium-pressure column and HPLC, the new compound is successfully extracted and separated, the operation steps of the method are only six steps, the operation method is simple and rapid, the extraction and separation process mainly adopts alcohol extraction and ethyl acetate elution, the process method is environment-friendly, the purity of the compound separated by the method is higher than 90 percent, in addition, the research shows that the compounds have obvious antitumor and cholinesterase resisting activity, therefore, the new compound and the derivatives thereof can be used as other compounds for synthesizing a guide substance and a raw material for new medicine development and pharmacological activity research, can also be used for preparing anti-tumor and anti-cholinesterase drugs.
Drawings
FIG. 1 shows that the ester compounds with anti-tumor and anti-cholinesterase activities in purslane of the present invention1H-NMR spectrum chart.
FIG. 2 shows that the purslane of the present invention has anti-tumor and anti-cholinesterase activity13C-NMR spectrum chart.
FIG. 3 is the nuclear magnetic resonance carbon spectrum (DEPT) spectrum of the new esters with anti-tumor and anti-cholinesterase activities in purslane of the present invention.
FIG. 4 shows NMR of novel esters with antitumor and anticholinesterase activity in Portulaca oleracea according to the invention1H-1HCOSY spectrum.
FIG. 5 is a nuclear magnetic resonance HMBC spectrogram of the novel ester compounds with anti-tumor and anti-cholinesterase activities in the purslane of the present invention.
FIG. 6 is a nuclear magnetic resonance HSQC spectrum of the new ester compounds with anti-tumor and anti-cholinesterase activities in purslane of the present invention.
FIG. 7 is the NOESY spectrum of the ester compounds with antitumor and anticholinesterase activity in purslane of the present invention.
FIG. 8 is a high resolution mass spectrum of the novel esters with anti-tumor and anti-cholinesterase activities in purslane of the present invention.
Detailed Description
Example 1 an ester compound extracted and separated from purslane and a method for extracting and separating the same.
An ester compound extracted from herba Portulacae with molecular formula of C20H34O3Named as (7E, 9E) -6-oxooctadecyl-7, 9-dienoic acid ethyl ester, and has the chemical structural formula:
table 1 shows the nuclear magnetic data of the new compounds:1H-NMR of13C-NMR in DMSO.
TABLE 1 Nuclear magnetic data for novel compounds of the invention
The compound is light yellow oily substance, is easily soluble in methanol, and is insoluble in water. HRESI (+) TOFMS gave M/z 323.2584[ M + H]+Has an excimer ion peak of 323.2586 molecular weight. Bonding of1H-NMR,13C-NMR and DEPT data, presuming that the possible molecular formula of the compound is C20H34O3The unsaturation degree is 4.13The C-NMR spectrum and the DEPT spectrum showed 20 carbon signals of 2 CH3 (delta: 13.82; 14.10), 12 CH2 (delta: 21.92; 23.53; 24.40; 24.56; 28.08; 28.08; 28.42; 30.84; 32.38; 33.45; 39.24; 59.58), 4 CH (delta 128.05; 128.96; 142.74; 145.31), 2 quaternary carbons (delta: 172.53; 200.10), respectively.
In the presence of deuterated DMSO as solvent1The H-NMR spectrum shows two methyl signals, δ 0.85(3H, t, J ═ 6.90), δ 1.17(3H, t, J ═ 7.20); 12 methylene signals, δ 01.25(8H, m), δ 1.48(8H, m), δ 2.17(2H, m), δ 2.25(2H, t, J ═ 7.02), δ 2.54(2H, t, J ═ 7.50), δ 4.04(2H, t, J ═ 7.20); the 4 methine signals are each δ 6.10(1H, d, J ═ 15.54); δ 6.26(2H, m); δ 7.18(1H, m); according to13C and HSQC spectra indicated the presence of 2 methyl groups, 12 aliphatic methylene groups and 4 methine groups;13c and HMBC spectra show that H-2 and C-1, C-3, C-4; h-3 and C-1, C-2, C-5; h-4 and C-2, C-5, C-6; h-5 and C-3, C-4, C-6; h-7 and C-6, C-9; h-8 and C-6, C-10; h-9 and C-7, C-8, C-11; h-10 and C-8, C-11; h-11 and C-9, C-10, C-12, C-14; h-12 and C-11, C-14, C-15; h-13 and C-14, C-15, C-16; h-14 and C-15, C-16; h-15 and C-14, C-16, C-17; h-16 and C-14, C-15, C-17, C-18; h-17 and C-15, C-16, C-18; h-18 is related to C-16, C-17. 4 methine signals δ 6.10(1H, d, J ═ 15.54); δ 6.26(2H, m), which illustrates the presence of a double bond and further demonstrates that attachment to the carbonyl carbon results in a high chemical shift, and from the above information, the novel compound can be identified as above and designated (7E, 9E) -6-oxooctadecyl-7, 9-dienoic acid ethyl ester.
The invention also provides an extraction and separation method for extracting the separated ester compounds from the purslane, which comprises the following specific steps.
And 2, evaporating the liquid medicine obtained in the step 1 to dryness, performing chromatographic separation by using a silica gel column, isocratically eluting by using ethyl acetate (115L), wherein the silica gel is 100-200 meshes, and recovering the ethyl acetate to obtain an extract under reduced pressure at the temperature of below 40 ℃ to obtain an ethyl acetate extract.
And 3, separating the ethyl acetate extract obtained in the step 2 by using a polyamide column, performing gradient elution by using ethanol, evaporating the ethanol to dryness, performing chromatographic separation by using a silica gel column, wherein the silica gel is 200-300 meshes, performing gradient elution by using ethyl acetate-methanol (5:1, 2:1, v: v) in sequence to obtain 20 parts (namely 15 bottles with 500mL in each bottle), detecting by using a thin-layer chromatography, developing, combining the developed 1-9 elution parts, and concentrating the combined 1-9 parts at the temperature of below 40 ℃ under reduced pressure until the parts are dry for later use.
And 4, performing pretreated ODS (octadecylsilane chemically bonded silica) medium-pressure column chromatography separation on the product obtained in the step 3, wherein the granularity of the filler is 20-40 mu m, performing gradient elution (pressurizing to enable the flow rate to be 1mL/min and the temperature to be room temperature) by using methanol-water (60/40, 70/30, 80/20, 90/10, 100/0, v/v) to obtain 25 parts (namely, performing gradient elution to obtain 25 bottles, wherein each bottle is 100mL), detecting by using thin-layer chromatography, developing, reserving the developed 20-25 parts, and concentrating under reduced pressure below 50 ℃ until the parts are dry for later use.
And 5, subjecting the product obtained in the step 4 to chromatography separation by a pretreated Sephadex LH-20 column, eluting by methanol to obtain 20 elution parts (namely 20 bottles are obtained in total and each bottle is 50mL), detecting by thin-layer chromatography, developing, retaining the developed 8-11 parts, and concentrating under reduced pressure below 50 ℃ until the parts are dried for later use to obtain the new compound. The pretreatment process of the ODS and the sephadex comprises the steps of soaking in methanol for 24 hours, loading on a column, washing with methanol until no turbidity exists in dropping water, and then balancing with an initial mobile phase.
And 6, separating and preparing the new compound obtained in the step 5 by HPLC, wherein the ratio of methanol: 0.1% formic acid (80: 20, v/v) is used as a mobile phase, the detection wavelength is 210nm and 280nm, the new compound is obtained by separation and preparation, and the purity measured by a normalization method is 90-99%.
Example 2 antitumor effect of the novel ester compounds of the present invention.
1 main material.
1.1 drugs and reagents.
The new ester compound used in the experiment is prepared by the method, the purity of the new ester compound is 90-99%, the new ester compound is precisely weighed and diluted by DMSO to be a solution required by each dosage group. DMEM high-glucose medium, fetal bovine serum (Hyclone, usa); penicillin and streptomycin (Hangzhou Sijiqing Co., Ltd.).
1.2 cell lines.
Human colon cancer cell Caco-2, human breast cancer cell MCF-7, human gastric cancer cell BGC-823, human lung adenocarcinoma cell SPC-A1, human liver cancer cell BEL-7402, human cervical cancer cell Hela-229, ovarian cancer cell Ho-8910, and human oral epidermoid carcinoma cell KB (Shanghai cell Bank of China academy of sciences).
1.3 grouping.
Divided into a control group, an experimental group and a zero-adjustment group (culture solution containing DMSO solvent).
2 experimental methods.
2.1 cell culture.
DMEM high-sugar medium, added with 0% fetal calf serum, l% antibiotics (100U/mL penicillin and 100. mu.g/mL streptomycin), and placed at 37 ℃ with 5% CO2Culturing in an incubator.
2.2MTT method for detecting cell proliferation.
Taking cells in logarithmic growth phase for inoculationCultured in 96-well culture plate with cell density of 1 × 104one/mL, 100. mu.L per well, temperature 37 ℃, 5% CO2After overnight culture under the conditions, the experimental groups were added with the novel compounds of the present invention at different concentrations, each group was provided with 3 multiple wells, and after adding the drug, the mixture was placed at 37 ℃ and 5% CO2Culturing in an incubator for 48 h. Absorbing the culture solution containing the medicine, and adding the mixture into the culture solution in a volume ratio of 4: 1 and MTT (5 mg/mL) for 4 hours, carefully absorbing the supernatant, adding 150 mu L of DMSO into each hole, placing the hole on a shaker to shake so as to completely dissolve crystals (5min), and detecting the absorbance (A) value of each hole by a microplate reader at the wavelength of 570 nm. Then, the inhibition rate of each concentration of compound on cell growth is calculated, and the inhibition rate formula is as follows: the inhibition rate of cell growth was (1-a dosing well/a control well) × 100%, and the data was processed using SPSS software, and the inhibition rate was plotted against the drug concentration to calculate the IC50 value.
3, experimental results.
Experimental results show that the two novel alkaloid compounds have an inhibitory effect on proliferation of human colon cancer cells Caco-2, human breast cancer cells MCF-7, human gastric cancer cells BGC-823, human lung adenocarcinoma cells SPC-A1, human liver cancer cells BEL-7402, human cervical cancer cells Hela-229, ovarian cancer cells Ho-8910 and human oral epidermoid cancer cells KB, and the inhibitory rate is also obviously increased along with the increase of the drug concentration, namely the inhibitory rate is concentration dependent. The IC50 values for the above eight tumor cells for the two novel alkaloid compounds of the present invention are shown in Table 2.
TABLE 2 inhibition of tumor cells by the novel compounds of the present invention
Example 3 anticholinesterase action of the novel compounds of the invention.
1. The main material.
1.1 drugs and reagents.
The new compound used in the experiment is prepared by the method, the purity of the new compound is 90-99%, and the new compound is sodium dihydrogen phosphate, disodium hydrogen phosphate (national medicine group chemical reagent limited), physostigmine (Vast. Biotechnology), phosphorus 5, 5' -dithiobis (2-nitrobenzoic acid) (dithiobistriitrobenzoic acid, DTNB, Shanghai jin Cui Biotechnology limited), acetylcholinesterase (AChE) and thioacetylcholine iodide (Acetylthiochromoline iodide, ATCI, great even America biotechnology limited).
1.2 grouping.
The test group is divided into a negative control group, a positive control group and an experimental group.
2 experimental methods.
2.1 sample preparation.
Accurately weighing 0.11mg of the sample and physostigmine respectively, and preparing into five gradient concentrations of 2.5, 5.0, 10.0, 20.0 and 40uM by taking methanol as a solvent respectively. 7.8005g of sodium dihydrogen phosphate and 17.907g of disodium hydrogen phosphate were weighed precisely, and the volume was adjusted to 500mL with distilled water, and 26.5mL of sodium dihydrogen phosphate and 473.5mL of disodium hydrogen phosphate were taken to prepare 500mL of PBS (0.1M pH 8.0); 0.0594g of DTNB is precisely weighed, 10mL of PBS is added to prepare a DTNB solution (15 mmol/L); accurately weighing 0.01g of AChE, adding 10mL of PBS, and preparing AChE solution (0.2 u/mL); 0.044mg of ATCI was precisely weighed, and the volume was adjusted to 10mL with distilled water to prepare an ATCI solution (15 mmol/L).
2.2 modified Ellman method for determination of anticholinesterase activity.
To a 96-well plate were added 140. mu.L of PBS (0.1M pH 8.0), 10. mu.L of DTNB (15mmol/L), 15. mu.L of AChE (0.2u/mL), and 20. mu.L of the sample solution in this order. The negative control group experiment uses methanol to replace the sample, and the positive control group experiment uses physostigmine to replace the sample. After incubation at 37 ℃ for 10min, 10uL of ATCI (15mmol/L) was added. After incubation at 20 ℃ for 10min, the absorbance was measured at 410nm using a microplate reader. The inhibition was calculated according to the following formula: inhibition (%) - (blank-sample)/blank × 100%.
3, experimental results.
The results of the experiment are shown in table 3. The experimental result shows that the compound has the function of resisting cholinesterase.
TABLE 3 anticholinesterase inhibitory Activity of the invention
In conclusion, the invention provides a new ester compound extracted from purslane and an extraction and separation method thereof, the ester compound is prepared by sequentially adopting alcohol decoction extraction, silica gel column chromatography, polyamide column chromatography, ODS medium-pressure column and Sephadex LH-20 purification and liquid phase separation, the method is simple, convenient, rapid and environment-friendly, the purity of the compound obtained by the separation method is high, and the obtained compound is extracted from the common traditional Chinese medicine purslane and has the functions of anti-tumor and anti-cholinesterase, so the new ester compound, the salt and the derivative thereof can be used as a natural product to develop new traditional Chinese medicines, and the new ester compound has wide prospects.
Claims (9)
2. the method for extracting and separating the ester compounds from the purslane according to claim 1, which comprises the following steps:
step 1, extracting dry purslane medicinal materials by adopting an alcohol solvent, filtering alcohol extract, combining filtrates, directly heating and concentrating, and cooling to room temperature to obtain liquid medicine for later use;
step 2, evaporating the liquid medicine obtained in the step 1 to dryness, putting the liquid medicine on a silica gel column, eluting the liquid medicine by using ethyl acetate, and recovering the ethyl acetate under reduced pressure to obtain an extract so as to obtain an ethyl acetate extract;
step 3, separating the ethyl acetate extract obtained in the step 2 by using a polyamide column, isocratically eluting by using a mixture of water and ethanol with a volume ratio of 5:95, evaporating the ethanol part to dryness, then feeding the evaporated ethanol part to a silica gel column, sequentially carrying out gradient elution by using ethyl acetate-methanol to obtain a plurality of elution parts, detecting by using a thin-layer chromatography, developing, combining the developed elution parts, and concentrating the combined elution parts under reduced pressure to dryness for later use;
step 4, separating the product obtained in the step 3 by pretreated ODS column chromatography, performing gradient elution by using methanol-water to obtain a plurality of elution parts, detecting by using thin-layer chromatography, developing, and concentrating the developed elution parts under reduced pressure to dryness to obtain a concentrate for later use;
step 5, subjecting the product obtained in the step 4 to chromatography separation by a pretreated sephadex column, eluting by methanol to obtain a plurality of elution parts, detecting by a thin-layer chromatography, developing, combining the developed elution parts, and concentrating the combined elution parts under reduced pressure to dryness for later use;
and 6, performing HPLC separation preparation on the concentrate obtained in the step 5, wherein the volume ratio of methanol to 0.1% formic acid is 80: 20 as a mobile phase to prepare the ester compound.
3. The method for extracting and separating the ester compounds from the purslane according to claim 2, wherein the alcohol solvent in the step 1 is 50% ethanol, the extraction times are 2 times, the reflux is performed for 2 hours each time, and the dosage of the ethanol is 8-16 times of that of the medicinal materials.
4. The method for extracting and separating the ester compounds from the purslane according to claim 2, wherein the ethyl acetate used in the step 2 is eluted at an isocratic elution.
5. The method for extracting and separating the ester compounds from the purslane according to claim 2, wherein the volume ratio of ethyl acetate to methanol in gradient elution in the step 3 is 5:1 and 2: 1.
6. the method for extracting and separating the ester compounds from the purslane according to claim 2, wherein the volume ratio of methanol to water eluted in the gradient in the step 4 is 60: 40. 70: 30. 80: 20. 90: 10 and 100: 0.
7. the method of claim 2, wherein the ODS column and sephadex column are pretreated by soaking in methanol for 24 hr, loading on the column, washing with methanol until the drop water is free from turbidity, and balancing with initial mobile phase.
8. The method for extracting and separating the ester compounds from the purslane according to claim 2, wherein the methanol elution in the step 5 is isocratic elution.
9. The use of the isolated ester compounds extracted from purslane as claimed in claim 1 in the preparation of anti-tumor and anti-cholinesterase drugs.
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101643403A (en) * | 2009-04-10 | 2010-02-10 | 陈海燕 | Method for extracting linolenic acid from purslane |
CN102173987A (en) * | 2011-03-07 | 2011-09-07 | 东北师范大学 | New manchurian walnut bark acid and derivatives thereof, and preparation methods and medical application thereof |
CN102516066A (en) * | 2011-12-22 | 2012-06-27 | 东北师范大学 | Ostopanic acid, ostopanic acid analogues and their preparation method and use |
CN110194755A (en) * | 2019-04-03 | 2019-09-03 | 辽宁中医药大学 | Compound Oleracone H in purslane, extraction and separation method and application thereof |
CN110294733A (en) * | 2019-04-03 | 2019-10-01 | 辽宁中医药大学 | One kind Oleracone I of key compound containing peroxide and its extraction separation method and application in purslane |
Family Cites Families (1)
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WO2011139067A2 (en) * | 2010-05-04 | 2011-11-10 | 한국생명공학연구원 | Method for preparing extract containing omega fatty acids from plant by using supercritical carbon dioxide extraction |
-
2020
- 2020-11-26 CN CN202011351224.9A patent/CN112479887B/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101643403A (en) * | 2009-04-10 | 2010-02-10 | 陈海燕 | Method for extracting linolenic acid from purslane |
CN102173987A (en) * | 2011-03-07 | 2011-09-07 | 东北师范大学 | New manchurian walnut bark acid and derivatives thereof, and preparation methods and medical application thereof |
CN102516066A (en) * | 2011-12-22 | 2012-06-27 | 东北师范大学 | Ostopanic acid, ostopanic acid analogues and their preparation method and use |
CN110194755A (en) * | 2019-04-03 | 2019-09-03 | 辽宁中医药大学 | Compound Oleracone H in purslane, extraction and separation method and application thereof |
CN110294733A (en) * | 2019-04-03 | 2019-10-01 | 辽宁中医药大学 | One kind Oleracone I of key compound containing peroxide and its extraction separation method and application in purslane |
Non-Patent Citations (1)
Title |
---|
JA, a new type of polyunsaturated fatty acid isolated from Juglans;Xiu-Li Gao, et al.;《Apoptosis》;20151130;第21卷;第340-350页 * |
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