CN113121561B - Sulfur-containing diene compound and preparation method and application thereof - Google Patents
Sulfur-containing diene compound and preparation method and application thereof Download PDFInfo
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- CN113121561B CN113121561B CN202110366990.0A CN202110366990A CN113121561B CN 113121561 B CN113121561 B CN 113121561B CN 202110366990 A CN202110366990 A CN 202110366990A CN 113121561 B CN113121561 B CN 113121561B
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- 229910052717 sulfur Inorganic materials 0.000 title claims abstract description 42
- 239000011593 sulfur Substances 0.000 title claims abstract description 42
- -1 diene compound Chemical class 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
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- 238000000605 extraction Methods 0.000 claims abstract description 13
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
- C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
- C07D495/04—Ortho-condensed systems
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
Abstract
The invention discloses a sulfur-containing diene compound, a preparation method and application thereof; the molecular formula of the sulfur-containing diene compound is C 17 H 16 O5S; the extraction method of the compound comprises the following steps: leaching fructus Rubi in organic solvent to obtain leaching solution, concentrating the leaching solution, and adding water to prepare water suspension; extracting the water suspension, concentrating the extract to obtain an extract; separating and purifying the extract to obtain the sulfur-containing diene compound; the invention separates and identifies a sulfur-containing diene compound with a novel structure from rubus suavissimus, and the compound has anti-tumor pharmacological activity and can be used for preparing anti-tumor drugs.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical active ingredients, and particularly relates to a sulfur-containing diene compound, and a preparation method and application thereof.
Background
The pomace (Rubus hirsutus thunder.) is a plant of the genus Rubus (Rubus l.) of the family Rosaceae (rosacea), also called wild Du Li, march, is perennial deciduous shrubs, has stronger root-bark seedling capability, wide distribution and large resource amount; the fluffy mallow has wide application in folk, the whole herb can be used as a medicine, the fruit part contains chemical components such as phenols, flavonoid, anthocyanin and the like, has the functions of dispelling wind and removing dampness, clearing heat and stopping bleeding, and is generally used for treating diseases such as rheumatic arthralgia, acute Huang Danxing jaundice and the like; at present, researches on the awning at home and abroad are relatively few in particular to separation and identification of monomer components and pharmacological activity of the monomer components, and the monomer components are not related to the pharmacological activity of the anti-tumor drugs, so that the application range of the anti-tumor drugs is still to be further expanded.
Disclosure of Invention
The invention aims to provide a sulfur-containing diene compound, a preparation method and application thereof, so as to solve the problems in the background art.
In order to achieve the above purpose, the present invention provides the following technical solutions:
a sulfur-containing diene compound, which is a sulfur-containing diene hirsutupyran a, the sulfur-containing diene compound (sulfur-containing diene hirsutupyran a) having the structural formula:
the preparation method of the sulfur-containing diene compound comprises the following steps:
(1) Placing the rubus suavissimus in an organic solvent, and leaching by a cold leaching method or a hot reflux extraction method to obtain leaching liquor;
(2) Concentrating the leaching solution, adding water to prepare an aqueous suspension, extracting the aqueous suspension, and concentrating the extract to obtain an extract;
the cold soaking method is to put the awning into a sealed container, add a certain volume of organic solvent according to the formula, and soak for about 2 weeks; shaking frequently during the process, and filtering after the active ingredients in the fructus Foeniculi are fully dissolved in the organic solvent to obtain leaching solution;
the thermal reflux extraction method is to use an organic solvent for heating extraction, and a reflux heating device is needed to avoid solvent volatilization loss; compared with the cold leaching method, the hot reflux extraction method has the advantages of complex operation and high extraction efficiency;
(3) Separating and purifying the extract by normal phase silica gel column chromatography to obtain the sulfur-containing diene compound.
Before the step (1), the method further comprises a crushing step of crushing the fluffy cloth.
In the step (1), the ratio of the weight (kg) of the cotton-padded jacket to the volume (L) of the organic solvent is 1:1-2:1.
The organic solvent is one or more of methanol, ethanol, chloroform, ethyl acetate and acetone.
Preferably, the organic solvent is methanol, ethanol or a mixture thereof; methanol and ethanol have relatively high polarity, and more active ingredients with medium and low polarity can be obtained by leaching with methanol or ethanol.
During extraction, a solvent which is mutually insoluble with the organic solvent adopted during leaching is selected; when methanol is used for leaching, ethyl acetate is used for extraction; when ethanol is used for leaching, chloroform is used for extraction; in order to increase the extraction yield, it is preferable to concentrate the leaching solution, add water to prepare an aqueous suspension, and extract the aqueous suspension.
The eluent in the step (3) is petroleum ether and ethyl acetate mixed solution; gradient elution is carried out on petroleum ether and ethyl acetate according to the volume ratio of 9:1, 7:1, 4:1, 3:1, 1:1, 1:3 and 1:9 in sequence, fractions eluted when the volume ratio of petroleum ether to ethyl acetate is 7:1 are collected, and rotary evaporation is carried out on the fractions, so that the sulfur-containing diene compound is obtained; the polarity of petroleum ether and ethyl acetate is low, and the target compound can be subjected to full chromatography to remove all or most of impurities; compared with other organic solvents, the petroleum ether and the ethyl acetate have lower toxicity and are beneficial to the physical health of researchers.
The method also comprises the steps of high performance liquid chromatography separation and recrystallization of the eluted fraction in sequence before rotary evaporation of the fraction.
Before the high performance liquid chromatography separation step, the method further comprises a reverse silica gel column chromatography step, wherein the eluent used in the reverse silica gel column chromatography step is a mixed solution of methanol and water; gradient elution is sequentially carried out on methanol and water according to the volume ratio of 9:1, 7:1, 4:1, 1:1, 1:3 and 1:9, each gradient elution is carried out for 1L, the fractions are detected by TLC, the fractions with the elution ratio of 7:1 are combined and concentrated, and the target compound is obtained by recrystallization by acetone.
The application of the sulfur-containing diene compound in preparing antitumor drugs.
Compared with the prior art, the invention has the beneficial technical effects that,
(1) The invention extracts and separates sulfur-containing diene compound with novel structure from the awning by utilizing polarity difference of sulfur-containing diene substances, and the method has simple operation, high extraction yield and high product purity, and is suitable for large-scale production.
(2) The sulfur-containing diene compound provided by the invention has wide pharmacological activity and can be used for preparing antitumor drugs.
Drawings
FIG. 1 is a schematic diagram of a sulfur-containing diene compound hirsutupyran A 1 H-NMR spectrum (500 MHz).
FIG. 2 is a schematic representation of a sulfur-containing diene compound hirsutupyran A 13 C-NMR spectrum (125 MHz).
FIG. 3 is a structural formula of sulfur-containing diene hirsutupyran A.
FIG. 4 is a graph of hydrogen correlation and remote hydrocarbon correlation for sulfur-containing diene compound hirsutupyran A.
Detailed Description
The invention will be further described with reference to the accompanying drawings and specific examples.
The preparation of the sulfur-containing diene compound:
example 1
Taking 10kg of the fluffy-fructus kochiae, crushing, leaching with 10L of methanol for 2 weeks, concentrating the methanol leaching solution, suspending with 1L of distilled water, extracting the water suspension with 1L of ethyl acetate for 3 times, and concentrating the ethyl acetate extract to obtain 260g of extract; mixing with silica gel (100 mesh, 100 g), and performing normal phase silica gel column chromatography (200-300 mesh, 1kg; silica gel column)The dimension L is 500mm, and the dimension L is equal to,
120 mm), sequentially carrying out gradient elution by petroleum ether and ethyl acetate mixed solutions with volume ratios of 9:1, 7:1, 4:1, 3:1, 1:1, 3:1:9, wherein each gradient elution is 5L; TLC detection of the fractions, collecting fractions with elution ratio of 7:1 (petroleum ether: ethyl acetate), merging and concentrating to obtain crude product 300mg, purity 60%, yield 3%, and re-crystallizing with acetone (room temperature) to obtain target compound 100mg, purity 99% and yield 1%.
Example 2
Taking 10kg of rubus suaeda, crushing, carrying out reflux extraction with 5L of ethanol, concentrating ethanol extract, suspending with 1L of distilled water, extracting the water suspension with 1L of chloroform for 3 times, and concentrating chloroform extract to obtain 234g of extract; mixing with silica gel (100 mesh, 100 g), subjecting to normal phase silica gel column chromatography (200-300 mesh, 1kg; silica gel column size L500 mm,
120 mm), sequentially carrying out gradient elution by petroleum ether and ethyl acetate mixed solutions with volume ratios of 9:1, 7:1, 4:1, 3:1, 1:1, 3:1:9, wherein each gradient elution is 5L; TLC detection of the fractions, collecting fractions with an elution ratio of 7:1 (petroleum ether: ethyl acetate), merging and concentrating to obtain 260mg of crude product, with a purity of 65%, a yield of 2.6%, and recrystallizing with acetone (at room temperature) to obtain 120mg of target compound, with a purity of 99%, and a yield of 1.2%.
Example 3
Taking 10kg of crushed awning, crushing, extracting with 5L of mixed solution of methanol and ethanol, concentrating the methanol/ethanol leaching solution to obtain 366g of extract, mixing with 100g of diatomite, performing hot dipping with 5L of chloroform for 3 times, performing normal phase silica gel column chromatography (200-300 meshes, 1kg; silica gel column size L500 mm,
120 mm), sequentially carrying out gradient elution by petroleum ether and ethyl acetate mixed solutions with volume ratios of 9:1, 7:1, 4:1, 3:1, 1:1, 3:1:9, wherein each gradient elution is 5L; the fractions were checked by TLC and eluted at a ratio of 10:1 (chloroform: methanol)) And (3) mixing and concentrating the fractions to obtain 350mg of crude product, with purity of 60%, yield of 3.5%, and recrystallizing (room temperature) with acetone/n-hexane (volume ratio of 2:3) to obtain 210mg of target compound with purity of 99% and yield of 2.1%.
Example 4
Extracting 10kg of crushed herba Suaeda salsa with 5L of mixed solution of methanol and ethanol, concentrating the extractive solution to obtain extract 350g, mixing with 100g of diatomite, hot-dipping with 5L of chloroform for 3 times, performing normal phase silica gel column chromatography (200-300 mesh, 1kg; silica gel column size L500 mm,
120 mm), sequentially carrying out gradient elution by using mixed solution of chloroform and methanol in volume ratios of 9:1, 7:1, 4:1, 3:1, 1:1, 1:3 and 1:9, wherein each gradient elution is 5L; TLC detection of the fractions, collecting the fractions with elution ratio of 4:1, combining and concentrating; subjecting the concentrated fractions to reverse phase silica gel column chromatography, eluting with methanol and water mixed solution with volume ratio of 9:1, 7:1, 4:1, 1:1, 1:3, and 1:9 sequentially, performing gradient elution for 1L each time, detecting each fraction by TLC, mixing, concentrating the fractions with elution ratio of 7:5:1 (petroleum ether: chloroform: methanol) to obtain crude product 400mg, purity 65%, yield 4%, and recrystallizing with acetone to obtain target compound 340mg, purity 99%, and purity 3.4%.
Target fraction 7:5:1 (petroleum ether: chloroform: methanol) obtained by normal phase silica gel column chromatography can be purified by high performance liquid chromatography after concentrating; the detection wavelength of the high performance liquid chromatography is 210nm, the eluting agent is 20% -100% methanol in sequence, the eluting peak with the retention time of 45-48min is collected, the eluting solution is combined and concentrated, and the eluting solution is recrystallized by acetone, so that the target compound is obtained, and the yield is 1%.
Structural identification of sulfur-containing diene compounds:
the target compounds obtained in examples 1-4 were subjected to purity identification by HPLC, structural identification by mass spectrometry and nuclear magnetic resonance techniques, nuclear magnetic resonance was measured by Bruker AVANCE DRX-500NMR spectrometer, TMS was used as an internal standard; high resolution mass spectrometry fticlms was measured with Bruker Apex Spectrometer; electrospray mass spectrum ESI-MS was performed using Bruker Esquire 3000 plus Spectrometer assay。
The compound is 1 H-NMR spectra 13 The C-NMR spectra are shown in FIG. 1 and FIG. 2, respectively, and the NMR data are shown in Table 1; the compound structure was resolved by hydrogen-hydrogen correlation and remote hydrocarbon correlation maps (fig. 4).
NMR data of the compounds of Table 1
ESI-TOF-MS shows molecular ion peak m/z333, and by combining mass spectrum data and analyzing NMR spectrum of the compound, the compound is new sulfur-containing diene compound, named hirsutupyran A and molecular formula C 17 H 16 O5S has a structure shown in figure 3.
The application of the sulfur-containing diene compound in preparing antitumor drugs.
Analysis of antitumor Activity of sulfur-containing Diene Compounds:
HeLa cells were cultured in RP-MI 1640 medium containing 10% calf serum, penicillin 100IU/mL and streptomycin 100g/mL, 1 time for 3d and 1 time for 5d passage. Cells were all placed at 37 ℃. Taking logarithmic growth phase cells, diluting to 5×10 with RPMI 1640 medium 4 Per mL single cell suspension, inoculated in 96-well cell culture plate, each concentration was multiplexed in 3 wells, 180. Mu.L per well. After 12h incubation in incubator, 20 μl of test solution with different concentrations is added into each well of the drug group, and a blank control group (equal volume of RPMI 1640 medium is used for replacing test drugs) is arranged in parallel for 48h co-culture; 50 mu L of MTT solution with the concentration of 1mg/mL is added into each hole, after the culture is continued for 4 hours, the supernatant is sucked out, 150 mu L of dimethyl sulfoxide (DMSO) is added into each hole, and MTT reduction products are fully dissolved; the optical density (D) of each drug group and blank group was measured at 492nm wavelength on a microplate reader, and the inhibition ratio (IR,%) and half Inhibition Concentration (IC) of the drug to tumor cell growth were calculated according to the formula 50 ) And preliminary evaluation of the drug effect was performed.
IR (%) = (1-dosing mean D value/control mean D value) ×100%.
From the experimental results, it is known that the IC of the compound 50 =11 μm (Hela cells), indicating that the compound has better antitumor effect.
The antitumor drug is prepared by taking the sulfur-containing diene compound as a main active ingredient and adding pharmaceutically acceptable auxiliary materials, and can be prepared into a preparation according to a preparation method recorded in pharmaceutics; the preparation can be injection, instillation liquid, powder injection, granule, tablet, medicinal granules, powder, oral liquid, sugar-coated tablet, film-coated tablet, enteric-coated tablet, buccal agent, granule, pill, paste, pellet, spray, dripping pill, disintegrating agent, orally disintegrating tablet, micropill and the like.
The preparation method of the sulfur-containing diene compound dripping pill preparation comprises the following steps:
mixing 0.5g of sulfur-containing diene compound R-3-methyl-6, 8-dihydroxyl-7-methyl-3, 4-dihydro sulfur-containing diene-1-ketone and 10.5g of polyethylene glycol-6000 uniformly, heating and melting, transferring into dripping pill drip irrigation, dripping the liquid medicine into liquid paraffin at 6-8 ℃ to remove oil, and preparing 300 dripping pills.
The preparation method of the freeze-dried powder injection containing the sulfur-containing diene compound comprises the following steps:
taking 0.5g of sulfur-containing diene compound R-3-methyl-6, 8-dihydroxyl-7-methyl-3, 4-dihydro sulfur-containing diene-1-ketone, 4.5g of glucose, 0.9g of sodium thiosulfate and 1000mL of distilled water, uniformly mixing the components, freeze-drying, and sub-packaging for 400 to obtain the sulfur-containing diene compound.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention, but rather to enable any modification, equivalent replacement, improvement or the like to be included within the spirit and principles of the invention.
Claims (3)
1. The preparation method of the sulfur-containing diene compound is characterized in that the structural formula of the sulfur-containing diene compound is as follows:
the method comprises the following steps:
(1) Placing the rubus suavissimus in an organic solvent, and leaching by a cold leaching method or a hot reflux extraction method to obtain leaching liquor;
(2) Concentrating the leaching solution, adding water to prepare an aqueous suspension, extracting the aqueous suspension, and concentrating the extract to obtain an extract;
(3) Separating and purifying the extract by normal phase silica gel column chromatography to obtain the sulfur-containing diene compound;
before the step (1), the method further comprises a crushing step of crushing the fluffy cloth;
in the step (1), the ratio of the weight of the said roof and the volume of the organic solvent is 1:1-2:1;
the organic solvent is one or more of methanol, ethanol, chloroform, ethyl acetate and acetone;
the eluent in the step (3) is petroleum ether and ethyl acetate mixed solution; and (3) carrying out gradient elution on petroleum ether and ethyl acetate sequentially according to the volume ratio of 9:1, 7:1, 4:1, 3:1, 1:1, 1:3 and 1:9, collecting fractions eluted when the volume ratio of petroleum ether to ethyl acetate is 7:1, and carrying out rotary evaporation on the fractions to obtain the sulfur-containing diene compound.
2. The method for producing a sulfur-containing diene compound according to claim 1, further comprising a step of subjecting the eluted fraction to high performance liquid chromatography separation and recrystallization in this order, before subjecting the fraction to rotary evaporation.
3. The method for producing a sulfur-containing diene compound according to claim 2, further comprising a reverse-phase silica gel column chromatography step, wherein the eluent used in the reverse-phase silica gel column chromatography step is a mixture of methanol and water, before the high performance liquid chromatography separation step; gradient elution is sequentially carried out on methanol and water according to the volume ratio of 9:1, 7:1, 4:1, 1:1, 1:3 and 1:9, each gradient elution is carried out for 1L, the fractions are detected by TLC, the fractions with the elution ratio of 7:1 are combined and concentrated, and the target compound is obtained by recrystallization by acetone.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015152437A1 (en) * | 2014-04-01 | 2015-10-08 | 경북대학교 산학협력단 | Novel histone methyl transferase inhibitor and use thereof |
| CN106831911A (en) * | 2017-02-08 | 2017-06-13 | 浙江大学 | A kind of method that pelargonin anthocyanin monomer is isolated and purified in the Lei Cong Peng |
| CN111777657A (en) * | 2020-07-21 | 2020-10-16 | 丽水市中医院 | Saponin compound and preparation method and application thereof |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015152437A1 (en) * | 2014-04-01 | 2015-10-08 | 경북대학교 산학협력단 | Novel histone methyl transferase inhibitor and use thereof |
| CN106831911A (en) * | 2017-02-08 | 2017-06-13 | 浙江大学 | A kind of method that pelargonin anthocyanin monomer is isolated and purified in the Lei Cong Peng |
| CN111777657A (en) * | 2020-07-21 | 2020-10-16 | 丽水市中医院 | Saponin compound and preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| Mahmoud, Ahmed A. et al..Xanthienopyran, a novel thienocyclopentapyran in fruits of Xanthium pungens.《Tetrahedron Letters》.1995,第36卷(第49卷),第8985-8986页. * |
| Ying-Hui Dai et al..A new thiaziedione from the fruits of Xanthium sibiricum.《Journal of Asian Natural Products Research》.2008,第10卷(第4期),第303-305页. * |
| Yu-Sheng Shi et al..A new thiophene and two new monoterpenoids from Xanthium sibiricum.《Journal of Asian Natural Products Research》.2015,Figure 1，说明书第6页右栏第二段. * |
| Zhao Xia et al..Bioactive sulfur-containing compounds from Xanthium sibiricum, including a revision of the structure of xanthiazinone.《Phytochemistry》.2020,第173卷第112293页. * |
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