CN102949385A - Yew branch and leaf extract, extraction method and applications thereof - Google Patents
Yew branch and leaf extract, extraction method and applications thereof Download PDFInfo
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- CN102949385A CN102949385A CN2012104354858A CN201210435485A CN102949385A CN 102949385 A CN102949385 A CN 102949385A CN 2012104354858 A CN2012104354858 A CN 2012104354858A CN 201210435485 A CN201210435485 A CN 201210435485A CN 102949385 A CN102949385 A CN 102949385A
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- ramulus
- folium taxi
- taxi cuspidatae
- pure liquid
- extract
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Abstract
The present invention relates to a yew branch and leaf extract, an extraction method and applications thereof. The yew branch and leaf extract is extracted by adopting branches and leaves of Taxus chinensis var. mairei as raw materials, and comprises cephalomannine and 7-epitaxol, wherein the active ingredients provide an inhabitation effect for squamous cells in skin cancers, and especially provide significant treatment effects for anal squamous cell carcinoma, esophagus squamous carcinoma and tonsil squamous cell carcinoma. According to the present invention, resource advantages of the yew are utilized to deep develop, utilize and research the Taxus chinensis var. mairei; and a yew anti-skin cancer component extraction method with a characteristic of simple operation is provided, and an effect of the component in anti-skin cancer application is determined.
Description
Technical field
The present invention relates to a kind of Ramulus et folium taxi cuspidatae extract, its extracting method and application, extract the treatment that active component is used for skin carcinoma.
Background technology
The paclitaxel that extracts from the bark of Ramulus et folium taxi cuspidatae and branch and leaf is the universally acknowledged native compound with anticancer function, Ramulus et folium taxi cuspidatae be integrate view and admire, material is with, medicinal high seeds of economic worth.Up to the present, chemists have extracted from this platymiscium and have obtained more than 300 taxane diterpene-kind compound and many alkaloid compounds, wherein some composition such as Ramulus et folium taxi cuspidatae time alkali also has the active anticancer of similar paclitaxel, and other compositions such as 1-hydroxyl bar card booth I can come taxol biosynthesis by chemical modification.In addition, some scholars also separate from this platymiscium and obtain 14 kinds in a kind of ecdysterone, 5 kinds of triterpenes, 7 kinds of lignanoids, 3 kinds of glycosides, 8 kinds of bisflavones, flavone compound kaempferol-4-methyl ether, other organic acid and aldehyde etc., glucosides class and polysaccharide compound etc.Scientist finds that Ramulus et folium taxi cuspidatae can also improve body immunity, and the natural functions of human body is recovered.It not only has remarkable result to hypertension, hyperglycemia, hyperlipidemia, nephropathy and dysmenorrhoea etc., can also reduce urine protein and run off.Mainly be to extract paclitaxel as anticarcinogen to the research of Ramulus et folium taxi cuspidatae at present, emphasis is applied to the treatment of colon cancer, tumor of head and neck, gastric cancer, ovarian cancer etc., the almost seldom practical application of other medicinal purposes.
Have not yet to see the relevant report of using at the medicine of preparation treatment anus squamous cell carcinoma, esophagus squamous cell carcinoma, tonsil squamous cell carcinoma etc. about Ramulus et folium taxi cuspidatae extract, in view of this, special proposition the present invention.
Summary of the invention
One of purpose of the present invention is to overcome above-mentioned weak point of the prior art, utilizes the resources advantage of Ramulus et folium taxi cuspidatae, and Taxus mairei is carried out deep development and use research; Anti-skin carcinoma active component in the research Ramulus et folium taxi cuspidatae, find to can be used for cutaneous squamous cell carcinoma anus squamous cell carcinoma, esophagus squamous cell carcinoma, the branch and leaf extract of the multiple squamous cell carcinoma treatment such as tonsil squamous cell carcinoma makes taxus resource can fully play its potential advantages.Simultaneously, accumulate experience for the comprehensive utilization taxus resource, open up new application and lay the foundation.
For achieving the above object, the present invention adopts following technical scheme:
A kind of Ramulus et folium taxi cuspidatae extract is prepared from as raw material take branch and/or the leaf of Taxus mairei, and described Ramulus et folium taxi cuspidatae extract comprises Cephalomannine and 7-Epitaxol.
The extracting method of above-mentioned Ramulus et folium taxi cuspidatae extract comprises the steps:
(1) preparation extractum: get branch of Ramulus et folium taxi cuspidatae and/or leaf, impregnated in the alcoholic solution that mass concentration is 70%-75%, extract with alcoholic solution, the residue medicinal residues continue to use water extraction, merge water and alcoholic solution, reflux with petroleum ether, and Soxhlet is extracted and reclaimed solvent, obtains extractum;
(2) gradient elution: with the extractum water dissolution, upper HP20 macroporous resin, the pure liquid of the pure liquid of the pure liquid of water, 15-25%, 35-45%, 55-65% in order, the pure liquid of 75-85%, the pure liquid of 90-98% and acetone solution gradient elution, collect and merge the eluent of 90-98% alcohol liquid and acetone solution section, namely get Ramulus et folium taxi cuspidatae extract.
Wherein, each eluent preferably is respectively water, 20% pure liquid, 40% pure liquid, 60% pure liquid, 80% pure liquid, 95% pure liquid and acetone solution in the step 2.Collect 95% pure liquid and the eluent of acetone solution section this moment, namely gets the optimal Ramulus et folium taxi cuspidatae extract of purity and productive rate.
Wherein, the consumption of each eluent is 8-10 times of column volume in the step 2, and flow velocity is 1-1.5mL/min.The consumption of preferred each eluent is 9 times of column volumes, and flow velocity is 1.2mL/min.
Wherein, the alcohol in described step 1 and the step 2 is preferably ethanol.
Wherein, described extracting method also comprises Ramulus et folium taxi cuspidatae extract concentrated and desolventizes the step that obtains extract extractum.Described concentrated desolventizing as those skilled in the art grasped.The present invention is not particularly limited this, and those skilled in the art can adopt disclosed any method for concentration of prior art to remove solvent in the eluent, obtain Ramulus et folium taxi cuspidatae Extract extractum or further obtain powder.
Another object of the present invention is to provide the application of Ramulus et folium taxi cuspidatae Extract in the anti-skin carcinoma medicine of preparation treatment.
Described Ramulus et folium taxi cuspidatae Extract can adopt commercially available Cephalomannine and/or 7-Epitaxol as the active component in the medicine, the preferred Ramulus et folium taxi cuspidatae Extract that adopts said extracted method of the present invention to obtain, be equipped with pharmaceutically acceptable carrier, form of medication mainly comprises oral administration and percutaneous dosing, includes but not limited to liquid preparation, granule, tablet, electuary, soft capsule, soft capsule, drop pill, ointment; Contain Cephalomannine and/or 7-Epitaxol 10mg～300mg in each unit formulation, tablet or capsule preparations conventional amount used are taken 1～12 for each person every day; External preparation contains by weight mark meter 0.005%～0.05% of Cephalomannine and/or 7-Epitaxol.
Adopt technique scheme, the present invention has following beneficial effect:
The present invention utilizes the resources advantage of Ramulus et folium taxi cuspidatae, and Taxus mairei is carried out deep development and use research; A kind of extracting method of the anti-skin carcinoma composition of Ramulus et folium taxi cuspidatae simple to operation is provided, and has measured its effect on anti-skin carcinoma is used.
Description of drawings
Fig. 1 is the apoptotic index (blank group) after flow cytometer detects the A431 drug effect;
Fig. 2 is the apoptotic index (Y-2) after flow cytometer detects the A431 drug effect;
Fig. 3 is the apoptotic index (Y-7) after flow cytometer detects the A431 drug effect;
Fig. 4 is the apoptotic index (Y-8) after flow cytometer detects the A431 drug effect;
Fig. 5 is the apoptotic index (Z-1) after flow cytometer detects the A431 drug effect;
Fig. 6 is the apoptotic index (Z-2) after flow cytometer detects the A431 drug effect;
Fig. 7 is the apoptotic index (Z-7) after flow cytometer detects the A431 drug effect;
Fig. 8 is the apoptotic index (Z-8) after flow cytometer detects the A431 drug effect.
The specific embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
(1) preparation of extractum 1: get branch of Ramulus et folium taxi cuspidatae, impregnated in the alcoholic solution that mass concentration is 70%-75%, extract with alcoholic solution, the residue medicinal residues continue to use water extraction, merge water and alcoholic solution, reflux with petroleum ether, Soxhlet is extracted and is reclaimed solvent, obtains extractum Z-1;
Or Taxus leaf, the same branch of Ramulus et folium taxi cuspidatae of operating procedure, corresponding extractum is Y-1;
(2) gradient elution: by mass concentration: with the extractum water dissolution of preparation, upper HP20 macroporous resin, in order water liquid, 20% pure liquid, 40% pure liquid, 60% pure liquid, 80% pure liquid, 95% pure liquid and, the acetone solution gradient elution, the eluent that obtains successively is Z-2, Z-3, Z-4, Z-5, Z-6, collect 95% ethanol and acetone eluent, obtain eluent Z-7, Z-8, be branch of Ramulus et folium taxi cuspidatae and get thing; Eluent is 9 column volumes, and flow velocity is 1.2L/min;
Or Taxus leaf, the same branch of Ramulus et folium taxi cuspidatae of operating procedure, corresponding eluent is followed successively by Y-7 and Y-8, is the Taxus leaf extract.(water liquid, 20% pure liquid, 40% pure liquid, 60% pure liquid, 80% pure liquid, 95% pure liquid and, the acetone solution gradient elution, the eluent that obtains successively is Y-2, Y-3, Y-4, Y-5, Y-6)
Collect Z-7, Z-8 or Y-7, Y-8, concentrate eluant obtains extractum behind the evaporating solvent.
Compare with embodiment 1, distinctive points only is, will extract simultaneously after branch of Ramulus et folium taxi cuspidatae and the leaf mixing in the present embodiment, and eluent is 8 times of column volumes, and flow velocity is 1mL/min.
Compare with embodiment 1, distinctive points only is, only Taxus leaf is extracted in the present embodiment, and eluent is 10 times of column volumes, and flow velocity is 1.5mL/min.
The separation of each composition in test example 1 Ramulus et folium taxi cuspidatae extract
Separating resulting is measured
(1) experiment material, instrument and reagent: hydrogen is composed with Bruker Avance500 type nmr determination (CDC13, TMS are interior mark); Mass spectrum is measured with Vacuum Generators ZAB-HS type mass spectrograph.The post layer chromatography is Haiyang Chemical Plant, Qingdao with silica gel (300～400 order) and thin layer chromatography with silica gel and produces, and other reagent is analytical pure.
(2) extract separation: Ramulus et folium taxi cuspidatae dry root and Ye Ge 2.5kg, adopt the concentrated extractum that obtains of embodiment 1 method, or merge with 95% alcohol heat reflux extraction three times, each three hours, filter, filtrate concentrates to get extractum, extractum with water dissolution after upper HP-20 macroporous resin column, gradient elution, collect 95% ethanol and the acetone position (is Y-7, Y-8, Z-7, Z-8), reclaim solvent extractum with dissolve with methanol after upper silica gel (300～400 order) post, successively use petroleum ether, petroleum ether-ethyl acetate (5:1,3:1,1:1,1:2,1:4), chloroform-methanol (50:1,20:1,10:1,5:1), the methanol gradient elution, every part of 100mL, spray developer TLC checks identical merging, further upper gel LH-20 post, therefrom obtain at last isolation identification and go out 7 kinds of chemical compounds, be respectively taxusin J(taxinine J), baccatin VI, taxicin, China's Ramulus et folium taxi cuspidatae triolefin first element (taxachitriene A), paclitaxel taxol, Cephalomannine (cephalomannine), 7-epi-taxol (7-epitaxol).
(3) structure and evaluation:
Chemical compound (1): white blocks of solid, mp.249-251 ℃.
ESI-MS m/ z : 731 [M + Na ] + 。
1H-NMR(δ，ppm，CDCl3)：1.08(3H，s)，1.13(3H，s)，1.76(3H，s)，2.32(3H，br s)，3.34(1H，d，6.2)，5.94(1H，d，11)，6.22(1H，d，11)，6.64(1H，d，16)，7.78(1H，d，16)，5.45(1H，s)，5.00(1H，s)，7.43(SH，m)，5.40-5.82(4H，m)。
13C-NMR (δ，ppm，CDCl3)：l18.94，35.30，28.32，13.68，15.89，27.19，31.66，20.79，21.36，21.44，21.02，20.79，146.04， 118.40， 128.19，129.02，130.65，169.24，169.43，169.82，170.71，133.68，134.20， 137.12，140.44，37.67，47.10。
Determine that according to document chemical compound is taxusin J(taxinine J), its structural formula is as follows
Taxusin J(taxinine J)
Chemical compound (2): ESI-MS
M/ z: 737 [M+Na]+;
1H-NMR (δ, ppm, CDCl
3): 1.96 (3H, s), 2.06 (3H, s), 2.11 (3H, s), (2.18 3H, s), 2.31 (3H, s), 1.20 (3H, s), (1.60 3H, s), 1.75 (3H, s), 2.37 (2H, m), (5.92 1H, d, 6.0), 3.24 (1H, d, 6.0), (4.94 1H, d, 7.9), 2.46 (1H, m), (1.84 1H, m), 5.53 (1H, dd, 9.7,7.9), 5.97 (1H, d, 11.3), 6.17 (1H, d, 11.3), 6.14 (1H, t, 8.7), (1.98 3H, d, 1.0), 4.18 (1H, d, 7.9), 4.07 (1H, d, 7.9), 8.09 (2H, dd, 8.5,1.4), 7.52 (2H, dd, 8.5,7.4), 7.62 (1H, t, 7.4).
13C-NMR (δ，ppm，CDCl
3)：20.7，20.9，21.2，21.4，22.7，169.1，170.2, 170.4, 169.8, 168.9, 133.7, 129.2, 130.1, 129.2, 166.9, 76.6, 12.7, 15.0, 22.7, 28.3, 42.8, 35.1, 70.4, 141.3, 133.6, 71.8, 75.0, 47.3, 72.5, 35.1, 83.9, 81.5, 47.3,73.3,78.9。
Determine that according to document chemical compound is baccatin VI, its structural formula is as follows
baccatin VI
Chemical compound (3): colourless crystallization, mp 285-287 ℃ (petroleum ether-ethyl acetate).
ESI-MS
m/ z : 839 [M + Na ] + ;
1H-NMR(δ，ppm，CDCl
3)：2.45 (1H , dd ,
J = 2.2, 11.6 ,C1-H) ,6.13 (1H ,dd ,
J = 2.6 ,10.2 ,C2-H) ,3.53 (1H ,d ,
J = 10.2 Hz ,C3-H) ,5.56 (1H ,t ,
J = 4.2 Hz ,C5-H) ,2.34 (1H , ddd ,
J = 2.3 ,6.3 ,14.5 Hz ,C6-H) ,1.79 (1H ,ddd ,
J= 4.0 ,10.6 ,14.6 Hz ,C6-H) ,5.56 (1H ,dd ,
J = 6.3 ,10.8 Hz ,C7-H) ,5.37 (1H , s ,C9-H) , 5137 (1H , s ,C10-H) ,2.95 (1H , dd ,
J = 12.0 ,19.0 Hz ,C14-Ha ) ,2.58 (1H ,d ,
J = 19.0 Hz ,C14-Hb) ,4.08 (1H ,d ,
J =9.1 Hz ,C16-H), 3.61 (1H , d ,
J = 8.1 Hz ,C16-H) ,1.28 (3H , s ,C17-CH3 ) ,1121 (3H , s ,C18-CH3) ,5.19(1H ,d ,
J = 12.3 Hz ,C19-H) ,4.46 (1H ,d ,
J = 12.3Hz ,C19-H) ,5.160 (1H , s ,C20-Ha ) ,4.182 (1H , s ,C20-Hb) , 2.15 , 2.14 , 2.03 , 1.99 ( 4 ×3H , S , 4 ×OAc-CH3) , 1.56 ( 1H , br s , C11-OH) 。
13C-NMR (δ，ppm，CDCl
3)：203.83，172.46，169.69，168.54，167.94，166.66，165.90，146.23，150.00，134.71,133.60,130.23,130.01,129.06, 128.73, 128.78, 117.71, 115.74, 91.44, 82.13, 80.30, 73.84, 70.14, 69.77, 68.59, 63.93,61.24, 49.63, 48.50,40.15,36.92,34.09,21,28,21.26,20.83,20.74,15.51,12.10.
Determine that according to document chemical compound is taxicin, its chemical structural formula is:
taxicin
Chemical compound (4): MS, 1H-NMR conform to taxachitriene A in the document with the 1H-NMR data, therefore determine that chemical compound is taxachitriene A, its chemical structural formula and carbon spectrum data are as follows:
taxachitriene A
Change platform thing (5): white needle-like crystals, mp.207-210 ℃.
ESI-MS m/ z : 876 [M + Na ] + ;
1H-NMR(δ，CDC13，ppm)：1.15(3H．s)，1.24(3H，s)，1.30(3H．s)，1.80(3H，brs)，1.87(IH，m)．2.28(4H，brt)，2.38(4H，brt)，2.50(1H m)，3.80(1H，d，7.2)，4．24(1H，d，8.2)，4．28(1H，d，8.2)，4．40 (1H,m)，4.80(1H，m), 4.90(IH，m)，5.86(1H，d，7.2)，6.23(1H，brt，8)，6.27(1H，s)，7.02(1H，d，8.5)。
13C-NMR (δ，CDC13，ppm)：9.55，14.84，20.84，21.77, 21.81，26.88，35.7，35.6,45.6，55.0，58.6，72.2，72.4，73.2，75.0，75.6，76.6，84.4, 133.7。
Physical constant and spectral data are consistent with the document description of paclitaxel taxol.Its chemical constitution is as follows:
Paclitaxel taxol
Change platform thing (6): clear crystal, mp.179-182 ℃.
ESI-MS m/ z : 832 [M -H ] + ;
1H-NMR (δ, CDC13, ppm): 1.17 (3H, S), 1.28 (3H, S), 1.70 (3H, S), 1.73 (3H, dd, J=1.1,6.9), 1.81 (IH, d, J=1.4), (1.82 3H, d, J=1.2), 1.88 (IH, brt), 2.25 and 2.37 (each 3H, S, 2 * OAC), 2.26 (IH, m), (2.33 IH, J=9.0,15.4.), 2.54 (IH, ddd, J=6.7,9.7,14.8), 3.79 (IH, d, J=7.3.), 4.18 and 4.28 (each IH, d, J=8.5), 4.39 (IH, cld, J=6.7,10.9), 4.71 (IH, d, J=2.8), 5.67 (IH, d, J=7.3), 6.22 (IH, dlf, J=8.3), 6.28 (IH, S), 6.42 (IH, elg, J=1.2,6.9), 6.49 (IH, d, J=8.7NH), 7.36 (IH, m, ph-H), (8.11 2H, d, J=7.9, Ph-H).
Above data are consistent with known compound Cephalomannine, therefore be accredited as Cephalomannine, its chemical constitution is as follows:
Cephalomannine
Change platform thing (7): white block crystallization, mp.187-189 ℃, the LiebermannBurchard reaction is aeruginous, and thin layer is aobvious purple under ultraviolet light.
ESI-MS m/ z: 877 [M+Na]+; In conjunction with 1H-NMR, 13C-NMR data, determine that its molecular formula: C47H51NO14. is with the contrast of its 1H-NMR data and paclitaxel, the hydrogen spectrum of finding both is quite similar, the main distinction is that the H-7 of chemical compound (5) paclitaxel is multiplet 4.40, and the H-7 of this chemical compound then shows as wide unimodal 3.70; The H-20 of chemical compound (5), 20, be the AB quartet 4.19 and 4.30, and the H-20 of chemical compound (7), 20, then show as two hydrogen 4.39 wide unimodal; The H-10 of chemical compound (5) appears at 6.27, and the H-10 of chemical compound (7) then appears at 6.70 relatively low.
According to document, authenticating compound (7) is 7-epi-taxol (7-epitaxol).Its chemical constitution is as follows:
7-epi-taxol (7-epitaxol).
The determination of pharmacological activity of the anti-skin carcinoma activity of test example 2 Ramulus et folium taxi cuspidatae
The present invention is take A431 application on human skin squamous cell JEG-3 as object of study, take present active anticancer to measure more mtt assay, the trypan blue living cells is refused the method for dying, the cellular morphology observational method is come Ramulus et folium taxi cuspidatae component to be measured is carried out the research of anti-skin carcinoma activity before and after Flow Cytometry Assay apoptosis and the medication, but whether preliminary study Ramulus et folium taxi cuspidatae extract part has the effect of In Vitro Anti cutaneous squamous cell carcinoma cell-proliferation activity, the size of its effectiveness, pharmaceutically-active concentration relationship, and whether induce the A431 apoptosis, determine whereby the activity group (one-tenth) of cutaneous squamous cell carcinoma effect sensitivity is divided, lay the foundation for the medicinal function cosmetics of further developing anti-skin carcinoma become potential ancillary drug as cutaneous squamous cell carcinoma.
Test method:
1.1.1 cell culture
Cell: people's epidermis squamous cell carcinoma strain A431.
Cellar culture: be incubated in the DMEM culture medium that contains 10% hyclone (FBS), place 37 ℃, 5%CO
2Cell culture incubator in, changed liquid once in per two days, observe under the inverted microscope, when cell fusion reaches 80% left and right sides, go down to posterity, 0.25% pancreatin solution digestion, the cell counting count board counting is with every hole inoculation 2 * 10
5Individual/ml cell is inoculated in 6 orifice plates, when it enters the logarithmic growth after date, adds respectively different pharmaceutical and processes, and detects corresponding index.
Method is observed medicine to the growth inhibited effect of tumor cell
1.1.2.1 experiment grouping
The Ramulus et folium taxi cuspidatae active component is to the lethal effect of A431:
Each component of Ramulus et folium taxi cuspidatae adopts respectively 1.25 * 10 in this test example
-2, 3 * 10
-3, 1 * 10
-4, 5 * 10
-5, 2.5 * 10
-55 concentration levels of mg/ml, be 48h action time.
1.1.2.2 inoculating cell:
The trophophase cell of taking the logarithm after 0.25% trypsinization, is diluted to single cell suspension with the DMEM culture fluid that contains 10% FBS, with every hole 5 * 10
3Individual being inoculated in 96 well culture plates, every hole 200 μ l are in 37 ℃, 5% CO
2Incubator in cultivate.
After 1.1.2.3 cell is fully adherent, serum-free culture 24h synchronization, the every hole of experimental group add respectively contain different pharmaceutical culture fluid 100 μ l, make final concentration be respectively preset concentration, adjusting every hole final volume is 200 μ l, establishes 6 multiple holes for every group, every culture plate stays a hole not add cell, only add culture fluid, be made as the zeroing hole, continue in incubator, to cultivate.
1.1.2.4 every hole adds 5mg/ml MTT solution 20 μ l, 37 ℃ hatch 4 hours after, every hole adds three liquid of 80 μ L, incubated overnight.
1.1.2.5 with the zeroing of zeroing hole, enzyme-linked immunosorbent assay instrument detects every hole absorbance (Absorbance, A value) at 570nm wavelength place, calculates cells survival rate (Survivil Rate, SR) and suppression ratio (Inhibitory Rate, IR).
Survival rate (SR)=experimental port A value/control wells A value * 100%
Suppression ratio (IR)=(control wells A value-experimental port A value)/control wells A value * 100%.
1.1.2.6 the suppression ratio that the time is abscissa, respectively organize cell is vertical coordinate, draws cell growth curve.
It is active that the trypan blue living cells refuses to dye the method counting cells
1.1.3.1 experiment grouping:
Each component of Ramulus et folium taxi cuspidatae adopts respectively 1.25 * 10 in this test
-2, 3 * 10
-3, 1 * 10
-4, 5 * 10
-5, 2.5 * 10
-55 concentration levels of mg/ml.
Relatively each component of Ramulus et folium taxi cuspidatae is to the A431 effect.
1.1.3.2 collect the cell through drug effect 48h, after removing supernatant, trypsinization with 0.25%, after being diluted to single cell suspension with culture fluid, the centrifugal supernatant of abandoning of 1000rpm, re-suspended cell after the PBS washing, triplicate, according to ratio and the 0.4% trypan blue mixing of 1:1, dyeing 1-2min finishes the dead cell counting in the 2min after dyeing is finished.
1.1.3.3 interpretation of result: the blue transfect cell of microscopically is dead cell, and counting uses the blood cell counting plate counting.
Cellular morphology is observed
Work as 25cm
2During cell enlargement to 60% in the culture bottle-70%, add respectively different separation components, establish simultaneously the solvent control group, observation of cell metamorphosis under the phase contrast microscope behind the cultivation 48h.
Flow cytometry inspection apoptosis rate
1.1.5.1 experiment grouping:
Each component of Ramulus et folium taxi cuspidatae adopts respectively 1.25 * 10 in this test
-2, 3 * 10
-3, 1 * 10
-4, 5 * 10
-5, 2.5 * 10
-55 concentration levels of mg/ml.
1.1.5.2 collect the cell through the exponential phase of drug effect 48h, keep supernatant, 0.25% trypsin digestion cell is diluted to single cell suspension with culture fluid, the centrifugal supernatant of abandoning of 1200rpm, re-suspended cell after 4 ℃ of PBS washings, triplicate.
1.1.5.3 preparation binding buffer liquid (deionized water: binding buffer liquid is 1:4), adjusting cell concentration is 1 * 10
6Individual/ml.
1.1.5.4 get the cell suspension of 100 μ l in the streaming pipe of 5 ml, add iodate the third ingot solution of 5 μ l nnexin-V/FITC and 10 μ l, 100 μ g/ml.
1.1.5.5 hatch 15min. in the room temperature lucifuge after mixing
Dilute good binding buffer liquid 1.1.5.6 add 400 μ l in reaction tube, in time up flow type cell instrument (FACS) is analyzed, and finishes to control the high PI false positive that the cytotoxicity of PI produces in 30min.
Statistical analysis
Experimental data adopts SPSS 8.0 software t methods of inspection to carry out statistical analysis, represents with means standard deviation.
Result of the test:
1, the Ramulus et folium taxi cuspidatae component is to the growth inhibited effect of people's epidermis squamous cell carcinoma A431
Adopt three generations A431 cell, per generation is selected 6 porocytes, amounts to 18 groups of cell experiment numerical value and adds up, and MTT result's (table 1, table 2) shows: Y-7, Y-8, Z-7, Z-8 suppress active strong, and these 4 positions suppress active to be increased with inhibition concentration.
Table 1 Ramulus et folium taxi cuspidatae component to the growth inhibited effect of people's epidermis squamous cell carcinoma A431 (suppression ratio (%), n=18)
| Sample number | 1.25×10 -2mg/ml | 3×10 -3 mg/ml | 1×10 -4 mg/ml | 5×10 -5 mg/ml | 2.5×10 -5 mg/ml |
| Y-1 | -2.34±0.12 | -1.18±0.10 | -0.11±0.04 | 2.44±0.01 | 3.02±0.15 |
| Y-2 | -5.17±0.16 | -5.67±0.13 | -5.81±0.08 | -5.86±0.11 | -6.60±0.07 |
| Y-3 | -38.66±0.34 | -3.33±0.14 | -1.37±0.05 | -0.01±0.20 | 1.40±0.10 |
| Y-4 | -30.22±0.18 | -7.85±0.13 | -7.46±0.15 | -4.74±0.03 | -0.93±0.10 |
| Y-5 | -48.71±0.35 | -5.13±0.08 | -4.71±0.18 | -2.35±0.07 | -0.60±0.16 |
| Y-6 | -19.96±0.21 | -9.16±0.13 | -7.76±0.09 | -5.54±0.08 | -2.16±0.07 |
| Y-7 | 8.28±0.16 | 6.91±0.26 | 5.94±0.12 | 1.93±0.21 | 0.63±0.09 |
| Y-8 | 16.49±0.09 | 15.78±0.07 | 13.79±0.21 | 11.71±0.08 | 4.00±0.18 |
| Z-1 | -4.58±0.12 | -2.47±0.16 | 0.05±0.03 | 2.79±0.12 | 4.44±0.16 |
| Z-2 | -8.05±0.07 | -6.53±0.13 | -3.80±0.09 | -2.65±0.09 | -2.60±0.09 |
| Z-3 | -72.98±0.49 | -15.53±0.19 | -4.60±0.07 | -2.90±0.07 | -2.73±0.12 |
| Z-4 | -28.29±0.22 | -8.03±0.08 | -0.50±0.07 | -0.48±0.16 | 0.33±0.09 |
| Z-5 | -8.59±0.07 | -6.41±0.06 | -3.98±0.23 | 1.71±0.09 | 1.51±0.05 |
| Z-6 | -9.19±0.09 | -9.13±0.05 | -7.43±0.18 | -6.21±0.12 | -5.81±0.07 |
| Z-7 | 19.28±0.11 | 6.92±0.12 | 6.69±0.09 | 6.07±0.06 | 5.43±0.09 |
| Z-8 | 20.75±0.09 | 7.17±0.04 | 6.24±0.09 | 5.34±0.11 | 4.19±0.12 |
Table 2 Ramulus et folium taxi cuspidatae component to the growth inhibited effect of people's epidermis squamous cell carcinoma A431 (survival rate (%), n=18)
| Sample number | 1.25×10 -2mg/ml | 3×10 -3 mg/ml | 1×10 -4 mg/ml | 5×10 -5 mg/ml | 2.5×10 -5 mg/ml |
| Y-1 | 102.34±0.12 | 101.18±0.10 | 100.11±0.04 | 97.56±0.01 | 96.98±0.15 |
| Y-2 | 105.17±0.16 | 105.67±0.13 | 105.81±0.08 | 105.86±0.11 | 106.40±0.07 |
| Y-3 | 138.66±0.34 | 103.67±0.14 | 101.37±0.05 | 100.01±0.20 | 98.60±0.10 |
| Y-4 | 130.22±0.18 | 107.15±0.13 | 106.54±0.15 | 104.74±0.03 | 100.93±0.10 |
| Y-5 | 148.71±0.35 | 105.87±0.08 | 104.71±0.18 | 102.35±0.07 | 100.60±0.16 |
| Y-6 | 119.96±0.21 | 110.84±0.13 | 106.24±0.09 | 105.46±0.08 | 102.16±0.07 |
| Y-7 | 91.72±0.16 | 93.91±0.26 | 94.06±0.12 | 98.07±0.21 | 99.63±0.09 |
| Y-8 | 83.51±0.09 | 84.22±0.07 | 86.21±0.21 | 88.29±0.08 | 96.00±0.18 |
| Z-1 | 104.58±0.12 | 102.47±0.16 | 99.95±0.03 | 97.21±0.12 | 95.56±0.16 |
| Z-2 | 110.95±0.07 | 106.53±0.13 | 103.80±0.09 | 102.45±0.09 | 102.40±0.09 |
| Z-3 | 172.98±0.49 | 115.53±0.19 | 104.60±0.07 | 103.10±0.07 | 102.73±0.12 |
| Z-4 | 128.29±0.22 | 110.97±0.08 | 100.50±0.07 | 100.48±0.16 | 99.67±0.09 |
| Z-5 | 110.41±0.07 | 106.59±0.06 | 103.98±0.23 | 98.29±0.09 | 98.49±0.05 |
| Z-6 | 110.81±0.09 | 110.87±0.05 | 107.43±0.18 | 106.79±0.12 | 105.81±0.07 |
| Z-7 | 80.72±0.11 | 93.08±0.12 | 93.31±0.09 | 93.93±0.06 | 94.57±0.09 |
| Z-8 | 79.25±0.09 | 92.83±0.04 | 93.76±0.09 | 94.66±0.11 | 95.81±0.12 |
2, the trypan blue living cells refuses to dye method counting cells activity
According to the MTT experimental result, A431 has inhibiting active component (Y-7, Y-8, Z-7, Z-8 for people's epidermis squamous cell carcinoma, getting 1.25 * 10-2mg/ml concentration studies) carry out cell counting, the result shows: cell quantity decreases drastically behind the drug effect, show that these medicines have the growth that suppresses skin cancer cell strain A431 cell, through visible all the group with blank of t check significant difference is arranged.
Table 3 couple people's epidermis squamous cell carcinoma A431 has inhibiting active component cell counting (n=3)
| Sample number | Z-7 | Z-8 | Blank | Y-7 | Y-8 |
| Cell number | (1.48±0.04)×10 6/ ml** | (1.35±0.02)×10 6/ ml** | (2.25±0.06)×10 6/ml | (1.85±0.05)×10 6/ml** | (1.73±0.04)×10 6/ml** |
With blank group * * p＜0.01, * p＜0.05
3, Flow cytometry Semen Phaseoli component is on the apoptotic impact of A431
According to the MTT experimental result, (Y-7, Y-8, Z-7, Z-8 get 1.25 * 10 for people's epidermis squamous cell carcinoma A431 inhibiting active component
-2Mg/ml concentration is studied) carry out Apoptosis by Flow Cytometry (Fig. 1-8), the result shows: among the A431 apoptosis figure, UL represents dead cell, and UR represents the apoptosis in late period, and LL represents living cells, and LR represents early apoptosis, and UR+LR is the apoptosis total value.Apoptosis rate according to the visible blank group of numerical value is 15.95%, the apoptosis rate that the active group of Ramulus et folium taxi cuspidatae (one-tenth) is divided is respectively 17.63%, 34.17%, 17.43%, 29.18%, as seen all have significance to improve than blank group, wherein Y-8, Z-8 are especially obvious to the apoptotic effect of A431 cell.
The result shows: the stronger position of anti-skin carcinoma activity, and leaf is Y-7, Y-8; Branch is Z-7, Z-8.
And these several positions all are the less positions of polarity, and the Ramulus et folium taxi cuspidatae antioxidant content just in time separates, and investigate for the technique of later on industrialized great production and lay a good foundation.
Test example 3 analytical test examples 1 isolated chemical compound is to the growth inhibited effect of people's epidermis squamous cell carcinoma A431
The determination of pharmacological activity result of the anti-skin carcinoma activity of test example 2 Ramulus et folium taxi cuspidatae shows that the stronger position leaf of anti-skin carcinoma activity is Y-7, Y-8, branch is Z-7, Z-8, in order to investigate further the effective ingredient at these positions, this test example is with Y-7, Y-8, Z-7, Z-8 combines, it is conducted in-depth research, therefrom isolate 7 kinds of chemical compounds, be respectively taxusin J(taxinine J), baccatin VI, taxicin, China's Ramulus et folium taxi cuspidatae triolefin first element (taxachitriene A), paclitaxel taxol, Cephalomannine (cephalomannine), 7-epi-taxol (7-epitaxol).Wherein paclitaxel, Cephalomannine, 7-epi-taxol have stronger inhibitory action to cutaneous squamous cell carcinoma A431.
Adopt three generations A431 cell, in per generation, selected 6 porocytes, amount to 18 groups of cell experiment numerical value and add up, MTT result's (table 4, table 5) shows: paclitaxel taxol, Cephalomannine (cephalomannine), 7-epi-taxol (7-epitaxol) have stronger inhibition activity to A431.
Table 4 Ramulus et folium taxi cuspidatae composition to the growth inhibited effect of people's epidermis squamous cell carcinoma A431 (suppression ratio (%), n=18)
| Sample | 1.25×10
-2mg/ |
3×10 -3 mg/ml | 1×10
-4 mg/ |
5×10 -5 mg/ml | 2.5×10 -5 mg/ml |
| taxinine J | -5.79±0.16 | -3.03±0.14 | -2.43±0.11 | -1.75±0.09 | -0.32±0.04 |
| baccatin VI | -10.51±0.21 | -5.72±0.17 | -1.46±0.08 | -0.24±0.10 | 1.66±0.07 |
| taxicin | -7.86±0.14 | -4.94±0.10 | -2.40±0.06 | -1.53±0.11 | -0.03±0.10 |
| taxachitriene A | -25. 17±0.23 | -8.42±0.16 | -7.33±0.18 | -3.65±0.26 | -1.01±0.04 |
| taxol | 21.52±0.22 | 13.04±0.17 | 7.53±0.11 | 3.68±0.08 | 1.35±0.15 |
| cephalomannine | 17.28±0.12 | 15.08±0.32 | 8.34±0.15 | 2.56±0.22 | 0.60±0.05 |
| 7-epitaxol | 16.32±0.15 | 10.46±0.13 | 5.71±0.18 | 3.79±0.21 | 2.30±0.15 |
Table 5 Ramulus et folium taxi cuspidatae composition to the growth inhibited effect of people's epidermis squamous cell carcinoma A431 (survival rate (%), n=18)
| Sample | 1.25×10
-2mg/ |
3×10 -3 mg/ml | 1×10
-4 mg/ |
5×10 -5 mg/ml | 2.5×10 -5 mg/ml |
| taxinine J | 105.79±0.16 | 103.03±0.14 | 102.43±0.11 | 101.75±0.09 | 100.32±0.04 |
| baccatin VI | 110.51±0.21 | 105.72±0.17 | 101.46±0.08 | 100.24±0.10 | 98.34±0.07 |
| taxicin | 107.86±0.14 | 104.94±0.10 | 102.40±0.06 | 101.53±0.11 | 100.03±0.10 |
| taxachitriene A | 125. 17±0.23 | 108.42±0.16 | 107.33±0.18 | 103.65±0.26 | 101.01±0.04 |
| taxol | 78.48±0.22 | 86.96±0.17 | 92.47±0.11 | 96.32±0.08 | 98.65±0.15 |
| cephalomannine | 82.72±0.12 | 84.92±0.32 | 91.66±0.15 | 97.44±0.22 | 99.40±0.05 |
| 7-epitaxol | 83.68±0.15 | 89.54±0.13 | 94.29±0.18 | 96.21±0.21 | 97.70±0.15 |
Above-mentioned digital proof: the biological activity of paclitaxel and its chemical constitution have substantial connection, and the β of complete taxane-ring, C13 position-carboxylic acid amide esters side chain and expoxy propane structure are necessary by active anticancer; The C13 ester side chain is also essential; The Oxotane ring is the important feature of active anticancer, and open loop is without this activity; The acetyl group of C10 and C7-hydroxyl are unimportant.
Taxol, cephalomannine, 7-epitaxol have embodied obvious active anticancer owing to having complete taxanes ring, the β of C13 position-carboxylic acid amide esters side chain and expoxy propane structure; And taxinine J and taxicin all do not have β-carboxylic acid amide esters side chain and expoxy propane structure, baccatin VI not to have β-carboxylic acid amide esters side chain, taxachitriene A not to embody its active anticancer without complete taxane-ring.
1 isolated Cephalomannine of test example 4 test examples is tested the human tumor cells proliferation inhibition activity
Experimental technique: cutaneous T cell lymphoma (CTCL), cutaneous squamous cell carcinoma A431, the strain of S-180 mice anus sarcoma cell, people's esophageal cancer cell strain Eca-109, people's larynx squamous cell carcinoma strain Hep-2 are all available from U.S. American Type Culture Collection.Cell culture adds 5% calf serum (Invitrogene) at DMEM culture medium (Invitrogene), 2mM glutamine, 37 ℃, 10%CO2 incubator.
Tetramethyl azo azoles salt (abbreviation mtt assay) detection of drugs is on the impact of tumor cell proliferation: select the exponential phase cell, density with 7000/ hole is inoculated in 96 orifice plates, put 37 ℃, cultivate the medicine that adds variable concentrations after 24 hours in 10% incubator, each concentration is established 6 in multiple hole, cultivate respectively 24,48 and 72 hours, culture fluid in the sucking-off orifice plate, adding concentration in every hole is the MTT 50 μ l of 1mg/ml, put into incubator and continue to cultivate 3-4 hour, take out 96 orifice plates that added MTT, every hole adds the DMSO of 150 μ l, light shaking orifice plate 30min, after the crystal of bottom, hole dissolves fully, orifice plate is put into microplate reader survey wavelength at the absorbance value in each hole of 570nm, the record result, test at least triplicate, with IC50 computed in software half propagation inhibition concentration IC50.
Cephalomannine was processed 48 hours, half inhibition appreciation rate IC50 to cutaneous T cell lymphoma (CTCL) is about 10.11 ± 1.39 μ M, cutaneous squamous cell carcinoma A431 is 30.05 ± 3.46 μ M, the strain of S-180 mice anus sarcoma cell is 40.75 ± 6.50 μ M, people's esophageal cancer cell strain Eca-109 is 82.68 ± 7.06 μ M, people's larynx squamous cell carcinoma strain Hep-2 is 96.33 ± 8.53 μ M, shows that Cephalomannine can use in the above-mentioned malignant tumor of preparation treatment.
1 isolated Cephalomannine treatment rheumatoid Cutis Bufonis tinea of test example 5 test examples, pernicious psoriasic clinical observation
51 years old-71 years old male patient peasant's 5 example, citizen's 3 examples; 40 years old-69 years old female patient peasant 4 examples, each example is antipruritic through being coated with the obvious decrustation of medicine of the present invention 5-7 day outward, and the surface is moved back and got final product dab 2-3 day after leprosy is seen its tender skin, and each coating applies thoughtfully to connect after medicine is done by its focus size and repastes therapy, namely heal through 7-10 day, cure rate is 100%.
Medicine described herein is that the Y-7 mass percent concentration is 95% aqueous solution.
1 isolated 7-Epitaxol of test example 6 test examples is tested the human tumor cells proliferation inhibition activity
Experimental technique: cutaneous T cell lymphoma (CTCL), cutaneous squamous cell carcinoma A431, the strain of S-180 mice anus sarcoma cell, people's esophageal cancer cell strain Eca-109, people's larynx squamous cell carcinoma strain Hep-2 are all available from U.S. American Type Culture Collection.Cell culture adds 5% calf serum (Invitrogene) at DMEM culture medium (Invitrogene), 2mM glutamine, 37 ℃, 10%CO2 incubator.
Tetramethyl azo azoles salt (abbreviation mtt assay) detection of drugs is on the impact of tumor cell proliferation: select the exponential phase cell, density with 7000/ hole is inoculated in 96 orifice plates, put 37 ℃, cultivate the medicine that adds variable concentrations after 24 hours in 10% incubator, each concentration is established 6 in multiple hole, cultivate respectively 24,48 and 72 hours, culture fluid in the sucking-off orifice plate, adding concentration in every hole is the MTT 50 μ l of 1mg/ml, put into incubator and continue to cultivate 3-4 hour, take out 96 orifice plates that added MTT, every hole adds the DMSO of 150 μ l, light shaking orifice plate 30min, after the crystal of bottom, hole dissolves fully, orifice plate is put into microplate reader survey wavelength at the absorbance value in each hole of 570nm, the record result, test at least triplicate, with IC50 computed in software half propagation inhibition concentration IC50.
The 7-Epitaxol was processed 48 hours, half inhibition appreciation rate IC50 to cutaneous T cell lymphoma (CTCL) is about 9.23 ± 0.86 μ M, cutaneous squamous cell carcinoma A431 is 24.35 ± 2.75 μ M, the strain of S-180 mice anus sarcoma cell is 40.13 ± 4.22 μ M, people's esophageal cancer cell strain Eca-109 is 79.16 ± 7.94 μ M, people's larynx squamous cell carcinoma strain Hep-2 is 81.06 ± 7.51 μ M, shows that the 7-Epitaxol can use in the above-mentioned malignant tumor of preparation treatment.
1 isolated 7-Epitaxol treatment rheumatoid Cutis Bufonis tinea of test example 7 test examples, pernicious psoriasic clinical observation
51 years old-71 years old male patient peasant's 7 example, citizen's 3 examples; 40 years old-69 years old female patient peasant 3 examples, each example is antipruritic through being coated with the obvious decrustation of medicine of the present invention 5-7 day outward, and the surface is moved back and got final product dab 2-3 day after leprosy is seen its tender skin, and each coating applies thoughtfully to connect after medicine is done by its focus size and repastes therapy, namely heal through 7-10 day, cure rate is 100%.
Medicine described herein is that the Z-7 mass percent concentration is 95% aqueous solution.
Although, above used general explanation, the specific embodiment and test, the present invention is described in detail, on basis of the present invention, can make some modifications or improvements it, and this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. Ramulus et folium taxi cuspidatae extract is characterized in that: be prepared from take the branch of Taxus mairei and/or leaf as raw material, described Ramulus et folium taxi cuspidatae extract comprises Cephalomannine and 7-Epitaxol.
2. the extracting method of a Ramulus et folium taxi cuspidatae extract, it is characterized in that: described extracting method comprises the steps:
(1) preparation extractum: get branch of Ramulus et folium taxi cuspidatae and/or leaf, impregnated in the alcoholic solution that mass concentration is 70%-75%, extract with alcoholic solution, the residue medicinal residues continue to use water extraction, merge water and alcoholic solution, reflux with petroleum ether, and Soxhlet is extracted and reclaimed solvent, obtains extractum;
(2) gradient elution: with the extractum water dissolution, upper HP20 macroporous resin, the pure liquid of the pure liquid of the pure liquid of water, 15-25%, 35-45%, 55-65% in order, the pure liquid of 75-85%, the pure liquid of 90-98% and acetone solution gradient elution, collect and merge the eluent of 90-98% ethanol and acetone solution section, namely get Ramulus et folium taxi cuspidatae extract.
3. extracting method as claimed in claim 2, it is characterized in that: each eluent is respectively water, 20% pure liquid, 40% pure liquid, 60% pure liquid, 80% pure liquid, 95% pure liquid and acetone solution in the step 2.
4. extracting method as claimed in claim 2, it is characterized in that: the consumption of each eluent is 8-10 times of column volume in the step 2, and flow velocity is 1-1.5mL/min.
5. the extracting method of Ramulus et folium taxi cuspidatae extract as claimed in claim 1, it is characterized in that: the alcohol in described step 1 and the step 2 is ethanol.
6. the extracting method of Ramulus et folium taxi cuspidatae extract as claimed in claim 2 is characterized in that: described extracting method also comprises Ramulus et folium taxi cuspidatae extract concentrated and desolventizes the step that obtains extract extractum.
7. contain the application of Ramulus et folium taxi cuspidatae extract in the anti-skin carcinoma medicine of preparation treatment of Cephalomannine and/or 7-Epitaxol.
8. the Ramulus et folium taxi cuspidatae extract that contains Cephalomannine and/or 7-Epitaxol suppresses the application in the squamous cell medicine in the skin carcinoma in preparation.
9. contain the application of Ramulus et folium taxi cuspidatae extract in preparation treatment anus squamous cell carcinoma, esophagus squamous cell carcinoma, tonsil squamous cell cancer drug of Cephalomannine and/or 7-Epitaxol.
10. according to claim 6-8 each described application is characterized in that: described medicine is liquid preparation, granule, tablet, electuary, soft capsule, soft capsule, drop pill, ointment.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191041A (en) * | 2013-04-18 | 2013-07-10 | 江苏红豆杉药业有限公司 | Chinese yew branch and leaf extractive with anti-oxidative effect as well as extraction method and application of yew branch and leaf extractive |
| CN103828619A (en) * | 2014-03-31 | 2014-06-04 | 东北林业大学 | Method for promoting growth of camptotheca acuminate and increasing content of camptothecin |
| CN104825940A (en) * | 2015-05-28 | 2015-08-12 | 张卫民 | TCM (traditional Chinese medicine) preparation for vulvar squamous cell carcinomas and preparing method thereof |
| CN105878302A (en) * | 2015-01-09 | 2016-08-24 | 北京罗瑞生物科技有限公司 | Preparation method of taxus water extract and application of taxus water extract in treating onychomycosis and beriberi |
| CN105963351A (en) * | 2016-05-11 | 2016-09-28 | 张士舜 | Traditional Chinese medicine composition for treating lung squamous carcinoma and preparation method thereof |
| CN110376235A (en) * | 2019-07-18 | 2019-10-25 | 浙江大学 | Test method is composed in the space correlation of Cephalomannine based on nuclear magnetic resonance technique |
| CN111610280A (en) * | 2020-07-02 | 2020-09-01 | 哈尔滨工业大学（威海） | Optimization method of extraction process of taxanes from Taxus chinensis |
| CN112250651A (en) * | 2020-11-12 | 2021-01-22 | 南京林业大学 | Taxus chinensis branch and leaf phenylpropanoids compound and extraction and separation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560197A (en) * | 2009-06-01 | 2009-10-21 | 西北农林科技大学 | Method for extracting paclitaxel from artificially cultivated yew branches and leaves |
-
2012
- 2012-11-05 CN CN2012104354858A patent/CN102949385A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560197A (en) * | 2009-06-01 | 2009-10-21 | 西北农林科技大学 | Method for extracting paclitaxel from artificially cultivated yew branches and leaves |
Non-Patent Citations (3)
| Title |
|---|
| 何法霖: "红豆杉枝叶中紫杉醇及其类似物的提取分离工艺及含量测定研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 张志强等: "树脂层析法脱色和浓缩紫杉醇", 《高校化学工程》 * |
| 王莹: "南方红豆杉中三尖杉宁碱和7-表-10-去乙酰基紫杉醇高效诱导提取纯化工艺研究", 《CNKI网络出版》 * |
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| CN104825940A (en) * | 2015-05-28 | 2015-08-12 | 张卫民 | TCM (traditional Chinese medicine) preparation for vulvar squamous cell carcinomas and preparing method thereof |
| CN105963351A (en) * | 2016-05-11 | 2016-09-28 | 张士舜 | Traditional Chinese medicine composition for treating lung squamous carcinoma and preparation method thereof |
| CN110376235A (en) * | 2019-07-18 | 2019-10-25 | 浙江大学 | Test method is composed in the space correlation of Cephalomannine based on nuclear magnetic resonance technique |
| CN110376235B (en) * | 2019-07-18 | 2020-09-15 | 浙江大学 | Measurement method of spatial correlation spectrum of mannine based on nuclear magnetic resonance technology |
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