CN105330636A - New flavonoid compound and preparation method and medical application thereof - Google Patents
New flavonoid compound and preparation method and medical application thereof Download PDFInfo
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Abstract
The invention discloses a new flavonoid compound and a preparation method and medical application thereof. The compound is reported for the first time, is the flavonoid compound novel in structure and can be extracted, separated and purified from dry honeysuckle. It is proved through in-vitro tests that the compound can reduce the proliferation activity of breast cancer cells and remarkably restrain the proliferation of breast cancer cells, shows strong in-vitro cell toxicity and has the in-vitro breast cancer resisting effect. The compound (I) is probably the effective targeted treatment medicine for treating breast cancer and can be used for developing medicine for treating breast cancer.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to from the Japanese Honeysuckle of drying, be separated obtain a kind of and there is flavonoid compound for the treatment of mammary cancer effect and preparation method thereof.
Background technology
The dry flower that Japanese Honeysuckle (having another name called honeysuckle, Flos Lonicerae etc.) is caprifoliaceae plant honeysuckle LonicerajaponicaThunb. or the flower just opened.Early summer the flowers are in blossom put before gather, dry.Property sweet, cold, return lung, the heart, stomach warp, have effect that is clearing heat and detoxicating, dispelling wind and heat pathogens, can be used for treatment carbuncle swells furunculosis, larynx numbness, erysipelas, toxic-heat and blood stasis, common cold due to wind-heat, warm morbidity, and medicinal history is long, has the title of " microbiotic in Chinese medicine ".Japanese Honeysuckle is widely distributed in china natural resources, contained complex chemical composition, diverse in function, except medicinal, is also widely used in the industries such as food, makeup and healthcare products.This medicine is traditional Chinese medical science common medicine, has the function of clearing heat and detoxicating, wind-heat dissipating, antiviral, hepatic cholagogic.
Since the sixties in 20th century, the chemical composition of domestic and international researchist to Japanese Honeysuckle has done large quantity research, and result shows: be rich in polytype chemical compositions such as volatile oil, organic acid, iridoid, flavones and triterpenoid saponin in Japanese Honeysuckle.Iridoid glycoside compounds is the compounds that in Japanese Honeysuckle, structure is changeable, finds more than 40 in recent years altogether, mainly comprise closed loop iridoid and open loop iridoid two class from Japanese Honeysuckle.
Japanese Honeysuckle is just extensive and famous with its pharmaceutical use since ancient times.Its effect is mainly clearing heat and detoxicating, cures mainly warm disease heating, toxic-heat and blood stasis, ulcer serious case of furuncle etc.Modern study proves, Japanese Honeysuckle contains the pharmacological component such as chlorogenic acid, luteolin glycosides, stronger restraint is had to the various pathogens such as Hemolytic streptococcus, staphylococcus aureus and upper respiratory tract infection Causative virus etc., in addition also can strengthening immunity, antiearly pregnancy, protect liver, antitumor, anti-inflammatory, antipyretic, hemostasis (blood coagulation), suppress intestinal absorption cholesterol etc., its clinical application widely, can be used for the treatment of respiratory tract infection, bacillary dysentery, acute urinary, hypertension etc. more than 40 with other medicines compatibility and plant illness.
Summary of the invention
The object of this invention is to provide and a kind ofly from the Japanese Honeysuckle of drying, be separated obtain a kind of there is flavonoid compound for the treatment of mammary cancer effect and preparation method thereof.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
There is the compound (I) of following structural formula,
The preparation method of described compound (I), comprise following operation steps: the Japanese Honeysuckle that (a) is dry is pulverized, extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 8 column volumes, then use 70% ethanol elution, 12 column volumes, collect 70% ethanol eluate, concentrating under reduced pressure obtains 70% ethanol elution thing medicinal extract; C in () step (b), 70% ethanol elution thing medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,50:1,30:1,10:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 15:1,10:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 10 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
Further, in step (a), extract with 75% alcohol heat reflux, united extraction liquid.
Further, described macroporous resin is AB-8 type macroporous adsorbent resin.
A kind of pharmaceutical composition, this pharmaceutical composition contains the compound according to claim 1 (I) for the treatment of significant quantity and pharmaceutically acceptable carrier.
The application of described compound (I) in the medicine of preparation treatment mammary cancer.
The application of described pharmaceutical composition in the medicine of preparation treatment mammary cancer.
When the compounds of this invention is used as medicine, directly can uses, or use with the form of pharmaceutical composition.
This pharmaceutical composition contains the compounds of this invention (I) for the treatment of significant quantity, and all the other are acceptable on pharmacology, nontoxic to humans and animals and pharmaceutically acceptable carrier of inertia and/or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, dripping pill, micropill, suspensoid, emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into water-based or oily solution, aseptic powder injection, liposome or the emulsion etc. of sterilizing.
Accompanying drawing explanation
Fig. 1 is compound (I) structural formula;
Fig. 2 is that the theoretical ECD value of compound (I) compares with experiment ECD value;
Fig. 3 is that various dose compound (I) is on the impact of Cells Proliferation of Human Breast Cancer vigor;
Fig. 4 is that various dose compound (I) is on the impact of Apoptosis of Breast Cancer.
Embodiment
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Embodiment 1: compound (I) is separated preparation and structural identification
Reagent source: ethanol, sherwood oil, ethyl acetate, propyl carbinol, methylene dichloride are analytical pure, purchased from Shanghai Ling Feng chemical reagent company limited, methyl alcohol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.
Preparation method: the Japanese Honeysuckle (8kg) of drying is pulverized by (a), (25L × 3 time) are extracted with 75% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste (3L), use sherwood oil (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated propyl carbinol (3L × 3 time) to extract successively, obtain petroleum ether extract, acetic acid ethyl ester extract (347g) and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract AB-8 type macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 8 column volumes, use 70% ethanol elution, 12 column volumes again, collect 70% ethanol eluate, concentrating under reduced pressure obtains 70% ethanol elution thing medicinal extract (129g); C in () step (b), 70% ethanol elution medicinal extract purification on normal-phase silica gel is separated, successively with volume ratio be 80:1 (8 column volumes), the methylene chloride-methanol gradient elution of 50:1 (8 column volumes), 30:1 (6 column volumes), 10:1 (8 column volumes) and 1:1 (5 column volumes) obtains 5 components; D component 4 (31g) is separated further by purification on normal-phase silica gel in () step (c), successively with volume ratio be 15:1 (8 column volumes), the methylene chloride-methanol gradient elution of 10:1 (10 column volumes) and 5:1 (6 column volumes) obtains 3 components; E in () step (d), component 2 (14g) reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8-10 column volume elutriant, elutriant concentrating under reduced pressure obtains pure compound (I) (28mg).
Structural identification: HR-ESIMS shows [M+Na]
+for m/z375.1214, can obtain molecular formula in conjunction with nuclear-magnetism feature is C
21h
20o
5, degree of unsaturation is 12.Hydrogen nuclear magnetic resonance modal data δ
h(ppm, DMSO-d
6, 400MHz): H-2 (5.87, s), H-5 (7.86, d, J=8.8), and H-6 (6.61, m), H-2 ', 6 ' (7.32, m), H-3 ', 5 ' (7.34, m), H-4 ' (7.38, m), H-7 " (6.17, d, J=12.5), and H-8 " (5.96, d, J=12.5), 9 "-Me (1.08, s), 9 "-Me (1.11, s), 9 "-OMe (3.24, s), carbon-13 nmr spectra data δ
c(ppm, DMSO-d
6, 100MHz): 92.8 (CH, 2-C), 197.1 (C, 3-C), 191.3 (C, 4-C), 115.1 (C, 4a-C), 127.6 (CH, 5-C), 105.8 (CH, 6-C), 162.6 (C, 7-C), 115.1 (C, 8-C), 158.9 (C, 8a-C), 138.1 (C, 1 '-C), 127.3 (CH, 2 ', 6 '-C), 129.7 (CH, 3 ', 5 '-C), 126.7 (CH, 4 '-C), 116.9 (CH, 7 "-C), 141.5 (CH, 8 "-C), 72.2 (C, 9 "-C), 29.1 (CH
3, 9 " and-Me), 29.1 (CH
3, 9 " and-Me), 55.8 (CH
3, 9 " and-OMe), carbon atom mark is see Fig. 1.Infrared spectra shows that this compound contains aromatic carbon (2965cm
-1, 1679cm
-1and 1591cm
-1).
1the bimodal signal δ H5.96 of HNMR spectrum display two (d, J=12.5Hz, H-8 ") and 6.17 (d, J=12.5Hz, H-7 "), they are two cis-form olefin protons.H-7 " and H-8 " coupling constant (J=12.5Hz) demonstrate above-mentioned inference.
13cNMR spectrum shows 21 carbon signals, comprise three methyl (δ C29.1,9 "-Me, 29.1,9 "-Me and 55.8,9 "-OMe), ten methynes (δ C92.8,2-C; 127.6,5-C; 105.8,6-C; 127.3,2 ', 6 '-C; 129.7,3 ', 5 '-C; 126.7,4 '-C; 116.9,7 "-C; 141.5,8 "-C), eight quaternary carbons (δ C197.1,3-C; 191.3,4-C; 115.1,4a-C; 162.6,7-C; 115.1,8-C; 158.9,8a-C; 138.1,1 '-C; 72.2,9 "-C).Nuclear magnetic data shows that this compound is flavonoid compound.H-8 in NOESY spectrum " with methoxyl group proton (δ H3.24, s), and H-8 " and H-7 " dependency, show C-9 " (δ C72.2) position is connected with a methoxyl group.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is about correlation type nuclear magnetic data, can substantially determine this compound as shown in Figure 1, steric configuration is determined further by ECD test, theoretical value and experimental value basically identical (Fig. 2).
Embodiment 2: compound (I) pharmacological action is tested
One, material and instrument
DMEM/F12 is high, and sugared nutrient solution is purchased from Beijing Hyclone company.Modified form RPMI-l640 substratum is purchased from Beijing Hyclone company.Standard foetal calf serum is purchased from Tianjin TBD company.CCK-8 cell viability detection reagent is purchased from Amada Co., Ltd. colleague chemistry institute.AnnexinV-FITC/PI apoptosis detection kit is purchased from Shanghai Yan Bin Chemical Industry Science Co., Ltd.JC-1 mitochondrial membrane potential probe is purchased from the green skies, Jiangsu biotechnology research institute.Dimethyl alum (DMSO) and phosphate buffer soln (PBS) are purchased from Sigma Products.Compound (I) is made by oneself, and preparation method is shown in that embodiment 1, HPLC normalization method purity is greater than 98%.
-70 DEG C of cryogenic refrigerator (FormaScientificInc. companies, the U.S.), electronic balance (Shimadzu AEL-200 type, Japan), Bechtop (SW-CJ-IC type, Suzhou), constant temperature oscillator (THZ-C type), disinfection with high pressure steam pot (YX-400B type), CO
2cell culture incubator (FormaSeries II, Thermalelectroncorp., the U.S.), thermostat water bath (BHW2-I type, domestic), inverted phase contrast microscope (CK40 type, Olympus, Japan), microscope (BX51 type, Olympus, Japan), micro-digit collecting system (DP71 type, Olympus, Japan), multi-functional microplate reader (InfiniteM200 type, TECAN, Switzerland), desk-top room temperature supercentrifuge (TGL-16G type, Beijing), desk-top low-temperature and high-speed whizzer (2K15 type, sigma company, the U.S.), BL60 electronic balance (Beijing Sai Duolisi instrument system company limited), LD4-40 Large Copacity whizzer (system in Beijing Jing founds whizzer company limited), 2K15 high temperature low speed refrigerated centrifuge (sigma company), UV765 ultraviolet spectrophotometer (Shanghai Precision Scientific Apparatus Co., Ltd), laser confocal scanning microscope (MRC-1024 type, Bio-Rad, Britain).
Two, test method
1, cell strain and cultivation
Select MCF-7 Breast Cancer Cell (estrogen receptor positive ER
+, the negative HERZ/neu of human epidermal growth factor acceptor-2
-), mammary cancer MDA-MB-231 cell (ER
-, HERZ/neu
-) and mammary cancer MDA-MB-453 (ER
-, HERZ/neu
+) cell strain, three kinds of cell strains all come from Chinese Sciences Academy Biochemistry And Cell Biology Institute.MCF-7 cell and MDA-MB-231 cell cultures are in the high sugared nutrient solution of the DMEM/F12 containing 10% standard foetal calf serum, and MDA-MB-453 cell cultures is in the RPMI-1640 nutrient solution containing 10% standard foetal calf serum, and above cell is all in 37 DEG C, 5%CO
2under condition, cellar culture goes down to posterity.Three kinds of breast cancer cells are attached cell, going down to posterity of attached cell: when cell attachment growth 2 ~ 3d to 80% merges, discard old nutrient solution, with the 0.1mol/LPBS preparation adding the pancreatin (pH7.4) of appropriate 0.25% in appropriate 0.1mol/LPBS (pH7.4) rinsing cell 2 backward bottles, filtration sterilization.Digestion, be advisable just to cover bottom Tissue Culture Flask, light Microscopic observation, treats that tenuigenin bounces back, and after intercellular substance increases, adds fresh nutrient solution immediately and stops digestion, and repeatedly blow and beat gently with suction pipe, makes to form cell suspension at the bottom of cell detachment bottle; Adjustment cell concn, draws appropriate cell suspension and adds in new culturing bottle, add 6mL fresh culture, basis of microscopic observation, be positioned over cell culture incubator.
2, cell proliferation viability examination
Utilize CCK-8 method detection compound (I) on the impact of three kinds of Cells Proliferation of Human Breast Cancer vigor.CCK-8 test kit (CellCountingKit-8) is the test kit of the detection cell proliferation of being developed by Japanese colleague's chemistry institute, containing WST-8 [2-(2-methoxyl group-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2 in CCK-8 reagent, 4-disulfonic acid benzene)-2H-tetrazolium monosodium salt], it is reduced to the yellow formazan product with high water soluble under the effect of electron carrier 1-methoxyl group-5-toluphenazine methyl-sulfate (1-MethoxyPMS) by the desaturase in cell mitochondrial, the quantity generating formazan thing is directly proportional to the quantity of viable cell.Measure its absorbance value by microplate reader at 450nm wavelength place, can indirectly reflect viable cell quantity.
Concrete operations are: by breast cancer cell MCF-7, MDA-MB-231, MDA-MB-453 routine passage of logarithmic phase, cell counting, with 2.5 × 10
5/ mL cell concn is inoculated in 96 porocyte culture plates, and every hole 200 μ L, overnight incubation, treats cell attachment and well-grown.Experiment divides 5 groups, often organize work 6 parallel holes, add the compound (I) of different amount respectively, make its final concentration be respectively 200,150,100,50mg/mL, control group adds equivalent distilled water, cultivate 24h, discard nutrient solution, wash 3 times with PBS, add fresh medium every hole 200 μ L, then every hole adds 10 μ LCCK-8, cultivate 4h harvested cell, detect 450nm place optical density value [D (450)] with microplate reader, with the zeroing of distilled water blank, calculate the proliferation inhibition rate that different concns compound (I) acts on.
Cell proliferation inhibition rate=[contrast D (450) experimental group D (450)]/contrast D (450) × 100%
3, apoptotic detection
Utilize AnnexinV/PI two dye Fluorescein activated cell sorter detection compound (I) on the impact of breast cancer cell MCF-7, MDA-MB-231, MDA-MB-453 apoptosis.Experiment is carried out in strict accordance with AnnexinV apoptosis detection kit operation instructions.Concrete operations are: by breast cancer cell MCF-7, MDA-MB-231, MDA-MB-453 routine passage of logarithmic phase, cell counting, with 10
5/ mL cell concn is inoculated in 6 porocyte culture plates, every hole 5mL, and overnight incubation, treats cell attachment and well-grown.Experiment divides 5 groups, often organizes work 3 parallel holes, adds the compound (I) of various dose respectively, make its final concentration be respectively 150,100,50mg/mL, control group adds equivalent distilled water, cultivates 24h, discards nutrient solution, cell is washed 3 times with the PBS (pH7.2) of 4 DEG C of precoolings, conventional trysinization, makes single cell suspension, centrifugal, with the binding damping fluid Eddy diffusion cell that 250 μ L dilute, and its concentration is made to be 1 × 10
6/ mL, the cell suspension getting 100 μ L, in 5mL streaming pipe, adds 5 μ LAnnexinV/FITC solution, leave standstill 5min, add the PI solution 10 μ L of 20 μ g/mL, after mixing, place 10min in room temperature lucifuge, in reaction tubes, add 400 μ LPBS, utilize machine testing on flow cytometer respectively to organize fluorescence situation.Cell divides to 4 phases according to AnnexinV and PI staining conditions respectively, AnnexinV and the PI equal negative cells that dyes is survive cell, PI stained positive, AnnexinV are negative staining is physical abuse or ruptured cell, AnnexinV and the PI dyeing all positive is downright bad or non-viable apoptotic cell, and AnnexinV stained positive, PI are negative staining is judged to be viable apoptotic cell.
4, the detection of mitochondrial membrane potential in anoxic
Utilize mitochondrial membrane potential in anoxic molecular probe JC-1 and laser confocal scanning microscope detection compound (I) on the impact of breast cancer cell MBA-MD-453 mitochondrial membrane potential.Concrete operations are: by the breast cancer cell MDA-MB-453 routine passage of logarithmic phase, cell counting, with 10
5/ mL cell concn is inoculated in 6 porocyte culture plates, every hole 5mL, overnight incubation, treat cell attachment and well-grown experiment points 5 groups, often organize work 3 parallel holes, add the compound (I) of various dose respectively, make its final concentration be respectively 150,100,50mg/mL, control group adds equivalent distilled water, cultivates 24h, discard nutrient solution, wash cell 3 times with the PBS (pH7.2) of 4 DEG C of precoolings, add 1.0mLPBS, add the JC-l of 10 μ L2.5mmol/L, its final concentration is made to be 25 μm of ol/L, at 37 DEG C, 5%CO
2in incubator, lucifuge hatches 20min, washs 3 times with PBS, and laser confocal scanning microscope detects to be analyzed.
5, the statistical study of data
All data are all with mean number scholar standard deviation
represent, adopt SPSS12.0 statistical software, carry out one-way analysis of variance to experimental result, P<0.05 thinks to there is significant difference between the two.
Three, result and conclusion
1, cell proliferation viability examination result
CCK-8 method is utilized to detect the impact of different concns compound (I) effect on three kinds of Cells Proliferation of Human Breast Cancer vigor, result shows, compound (I) effect can make three kinds of Cells Proliferation of Human Breast Cancer activity all reduce, and reduce degree increase with the increase of activity, demonstrate dose-effect relationship, 100, 150 all there is significant difference (P<0.05 with 200mg/mL group compared with control group, P<0.01), compound (I) effect can significantly suppress breast cancer cell MCF-7, MDA-MB-231, the propagation of MDA-MB-453, demonstrate stronger vitro cytotoxicity, show that compound (I) has stronger In Vitro Anti mammary cancer effect.The results are shown in Figure 3 and table 1.
2, apoptotic detected result
Utilize AnnexinV/PI two dye Fluorescein activated cell sorter detection compound (I) on the impact of breast cancer cell MCF-7, MDA-MB-231, MDA-MB-453 apoptosis, apoptosis analytical results shows, compound (I) effect 24h can make the apoptosis rate of three kinds of breast cancer cells all raise, demonstrate short apoptosis effect, and there is dose-effect relationship, 100 have significant difference (P<0.05) with 150mg/mL group compared with control group shows that compound (I) can remarkable inducing mammary cancer cell-apoptosis, plays antitumor action.The results are shown in Figure 4.
Sum up, this research, by compound (I) anti-breast cancer Effect study, finds that compound (I) can make three kinds of Cells Proliferation of Human Breast Cancer activity reduce, significantly suppresses its propagation, demonstrate stronger vitro cytotoxicity, there is In Vitro Anti mammary cancer effect.
Table 1 various dose compound (I) is to the inhibiting rate (%) of Cells Proliferation of Human Breast Cancer
| 50mg/mL | 100mg/mL | 150mg/mL | 200mg/mL | |
| MDA-MB-453 cell | 14.80 | 29.80 | 33.10 | 44.30 |
| MDA-MB-231 cell | 11.40 | 24.00 | 30.60 | 37.50 |
| MCF-7 cell | 10.30 | 20.20 | 26.20 | 35.10 |
Embodiment 3
The preparation of tablet: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, vehicle is added, pelletizing press sheet than the ratio for 1:9 in itself and excipient weight.
Embodiment 4
Prepared by oral liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, oral liquid method for making makes oral liquid routinely.
Embodiment 5
The preparation of capsule or granule: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, add vehicle in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 6
The preparation of injection liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Embodiment 7
The preparation of aseptic powder injection: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, be dissolved in sterile water for injection, stirring makes molten, filter with aseptic suction funnel, aseptic essence filter again, be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains powder injection.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.Those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the protection domain of technical solution of the present invention.
Claims (7)
1. there is the compound (I) of following structural formula,
2. the preparation method of compound according to claim 1 (I), it is characterized in that comprising following operation steps: the Japanese Honeysuckle of drying is pulverized by (a), extract with 70 ~ 80% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 8 column volumes, then use 70% ethanol elution, 12 column volumes, collect 70% ethanol eluate, concentrating under reduced pressure obtains 70% ethanol elution thing medicinal extract; C in () step (b), 70% ethanol elution thing medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,50:1,30:1,10:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 15:1,10:1 and 5:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 8 ~ 10 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
3. the preparation method of compound according to claim 2 (I), is characterized in that: in step (a), extracts, united extraction liquid with 75% alcohol heat reflux.
4. the preparation method of compound according to claim 2 (I), is characterized in that: described macroporous resin is AB-8 type macroporous adsorbent resin.
5. a pharmaceutical composition, is characterized in that: this pharmaceutical composition contains the compound according to claim 1 (I) for the treatment of significant quantity and pharmaceutically acceptable carrier.
6. the application of compound according to claim 1 (I) in the medicine of preparation treatment mammary cancer.
7. the application of pharmaceutical composition according to claim 5 in the medicine of preparation treatment mammary cancer.
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| CN107158063A (en) * | 2017-05-11 | 2017-09-15 | 临沂大学 | Enteric-coated micro-pill that a kind of utilization honeysuckle effective part composition is made and preparation method thereof |
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2015
- 2015-12-07 CN CN201510887475.1A patent/CN105330636A/en not_active Withdrawn
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| CN105294819A (en) * | 2015-10-20 | 2016-02-03 | 淄博夸克医药技术有限公司 | Novel limonin compound for treating breast cancer |
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| CN105481874A (en) * | 2015-12-22 | 2016-04-13 | 陈杰 | Novel diterpene compound for treating ovarian cancer |
| CN105566342A (en) * | 2015-12-22 | 2016-05-11 | 潘豪杰 | Novel diterpenoid for treating melanoma and preparation method thereof |
| CN105461732A (en) * | 2016-01-04 | 2016-04-06 | 范素琴 | Novel diterpenoid compound and preparation method and medical application thereof |
| CN105663118A (en) * | 2016-02-26 | 2016-06-15 | 温州统益生物医药科技有限公司 | Application of flavonoids compound to preparation of neuroprotection medicament |
| CN106243065A (en) * | 2016-09-09 | 2016-12-21 | 中国科学院西北高原生物研究所 | A kind of new sesquiterpenoid and biomedical uses thereof |
| CN107158063A (en) * | 2017-05-11 | 2017-09-15 | 临沂大学 | Enteric-coated micro-pill that a kind of utilization honeysuckle effective part composition is made and preparation method thereof |
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Application publication date: 20160217 |