CN1508134A - Anti-cance active substance of cyathocline purpurea and separating and purifying method thereof - Google Patents

Anti-cance active substance of cyathocline purpurea and separating and purifying method thereof Download PDF

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CN1508134A
CN1508134A CNA021280703A CN02128070A CN1508134A CN 1508134 A CN1508134 A CN 1508134A CN A021280703 A CNA021280703 A CN A021280703A CN 02128070 A CN02128070 A CN 02128070A CN 1508134 A CN1508134 A CN 1508134A
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ethyl acetate
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CN1232517C (en
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李祖强
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Yunnan University YNU
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Abstract

The present invention relates to anti-cancer active component of Huaiqing chrysanthemum and its separation and purification method. Its anti-cancer compound includes 9 beta-acetyl-costunolide (CP-C) whose molecular formula is C17H22O4 and 9 beta-acetyl-parthenclide (CP-I) whose molecular formula is C17H22O5. Said invention also provides the concrete steps for separation and purification of said anti-cancer components, and discloses three anti-cancer active components of CP-B, CP-C and CP-D.

Description

Anti-cance active substance of cyathocline purpurea and separating and purifying method thereof
Technical field
The present invention relates to the medical technical field, specifically from cup chrysanthemum plant, extract anticancer active constituent and separating and purifying method thereof.
Background technology
The cup chrysanthemum is composite family cup Chrysanthemum plant (Cyathocline Purpurea Buch.-Ham.ex De Don), the cup Chrysanthemum plant whole world has 3 kinds approximately, in state-owned a kind [woods Rong, Chen Yilin. Chinese Plants will (the 74 volume). Beijing: Science Press, 81～82].The cup chrysanthemum mainly is distributed in Southwest China subtropical zone and India.
Cup chrysanthemum complete stool is used as medicine, among the people be used for anti-inflammation hemostasia, control various inflammation and flu, have a high fever etc.
The domestic and international research overview of cup Chrysanthemum plant chemical ingredient:
1981, India scholar Bhimsen A.Nagasampagi etc. produce the over-ground part of cup chrysanthemum from India, separating purifies obtains 5 compositions (compound 1～5), wherein 2 is volatile oil component eudesmane lactone (Eudesmenolide) and derivative, 2 sesquiterpene lactones [Bhimsen A.Nagasampagi, JayantS.Sohoni, and Christa Zdero.A evdesmanolide and a gvaianolide from CyathoclinePurpurea.1981, Phytochemistry, 20 (8): 2034～2036].
1991, India scholar G.J.Chintawar etc. separate from the cup chrysanthemum again and obtain 1 activeconstituents with plant growth regulation: 6 Alpha-hydroxies-4 (14), 10 (15)-guaianolides (compound 6), and the crystal and molecular structure [the G.J.Chintawar andV.M.Pmdmanabhan.The cxystal and molecular structure of a guaianolidefrom Cyathocline Purpurea.1991 of this material have been studied with the X-diffraction method, J of Natural products, 54 (5): 1397～1399].
1993, usefulness high resolution 2D NMR such as India scholar P.Pradhan spectrum has been studied the stereochemistry [P.Pradhan of 2 guaianolides (compound 5 and 6), G.J.Chintalwar and A.Banerji.Stereochemistry of a guaianolide from Cyathocline Purpurea by highresolution NMR spectroscopy.1993, Spectroscopy letters, 26 (6): 997～1004].
In addition, India scholar Jayant S.Sohoni separates from other 2 plant speciess of cup Chrysanthemum (C.Lutea and C.Lyrata) with S.R.Rojatkar etc. and obtains 4 Sesquiterpenes (compound 7～10) [Jayant S.Sohoni, Bhimsen A.Nagasampagi, Jvrgen Ziesche, et al.A germacranolide from Cyathocline Lutea 1984, Phytochemistry, 23 (5): 1181～1183 and S.R.Rojatkar, M.Banerjee, D.D.Sawaikar, et al.Agermacranolide from Cyathocline Lutea.1994, Phytochemistry, 37 (4): 1211～1212].
In sum, the India scholar has obtained 6 sesquiterpene constituents from the overhead stem portion in cup chrysanthemum ground, separates obtaining 4 Sesquiterpenes from other 2 kinds of cup Chrysanthemum.And by molecular structure and sterie configuration thereof, prophesy 6 Alpha-hydroxies-4 (14) wherein, 10 (15)-more create diene-8 β, and the 12-lactone has plant growth regulating activity.The structural formula of above-mentioned chemical ingredients is:
Figure A0212807000051
Document shows that external above-mentioned work only is confined to chemical ingredients, does not relate to any applied research.
Do not see at present other report about cup Chrysanthemum plant chemical ingredient and applied research thereof.
Summary of the invention
The objective of the invention is from the cup chrysanthemum, to separate the chemical ingredients of purifying and making new advances, it is carried out antitumour activity system pharmacological research, seek, develop the anticancer active constituent of new texture, new active function mechanism.
Another object of the present invention provides the separating and purifying method that extracts anticancer active constituent in a kind of glass of chrysanthemum plant.
Research of the present invention is according to new drug research---screening active ingredients, chemical constitution study, reason research approximately, clinical trial---four step, on the screening active ingredients basis, the anticancer active constituent of from cup chrysanthemum plant, purifying.The new compound that the present invention separates purification is:
(1)
Figure A0212807000061
CP-C (using re-crystallizing in ethyl acetate), white needle, mp152～154 ℃ (the different crystal forms compound that from different solvents, obtains, fusing point has difference); Get molecular formula C by HRFAB-MS (291.1607) 17H 22O 4(calculated value: 291.1596) (table 1,2).By 1HNMR, 1H- 1Hcosy, 13CNMR and DEPT spectrum (table 3) identify that comprehensively CP-C is 9 β-ethanoyl-Costus speciosus lactone, and this is a new compound.
The high resolution of table 1 CP-C and CP-D bombards mass spectrum (HRFABMS) data fast
CP-C CP-D
Mol.formula C 17H 22O 4 C 17H 22O 5
m/z?ofMH +calculated 291.1596 307.1546
m/z?ofMH +observed 291.1607 307.1559
deviation?in?ppm +3.66 +4.51
The chemical ionization mass spectrometry of table 2 CP-C (CIMS) data
m/z Abundance(％) Assignment
C-H 4CI NH 3CI FAB
331 1 - - M+C 3H 5 +
319 1 - - M+C 2H 5 +
308c - 100 - M+NH 4 +
291 25 b 60 MH +
271 5 - - M′+C 3H 5 +
259 10 - - M′+C 2H 5 +
231 100 b 100 (M-HOAc)H +orM′H
Table 3 CP-C's 13CNMR, 1HNMR and 1H- 1Hcosy composes data
Position? 13C?chem.? 1H?chem. 1H 1H- 1H?splittings COSY?to
shifts shifts multiplicity (or?W 1/2)in?Hz positions：
1 131.4 5.19 bd ～11 14
2 25.5 2.0-2.35 b，c m (atδ～1.3)
3 39.2 2.0-2.35 b，c m (atδ～1.3)
4 141.6 - - - -
5 126.9 4.66c bd ～9.8 6.15
6 81.2 4.57c dd 8.7，9.8 5，7
7 47.3 2.73 m (W 1/2?22) 6，8a，13E，13Z
8 33.7 2.16 d(a) ddd ～2，～2，～14.5 8b，9
1.95 d(b) ddd ～10.5，～10.5，14.4 7，8a，9
9 80.8 5.21 dd 2.6，10.9 8a，8b，14
10 134.9 - - - -
11 138.9 - - - -
12 170.0 - - - -
13 120.4 5.55?(E) d 3.2 7
6.29(Z) d 3.5 7
14 11.6 1.45 d 0.8 1.9
15 17.5 1.72 d 1.3 5
Ac?CH 3 21.4 2.05 s - -
Ac?CO 170.5 - - - -
CP-D (ethyl acetate-acetone recrystallization), white crystal, mp164～166 ℃ (the different crystal forms compound that from different solvents, obtains, fusing point has difference); Get molecular formula C by HRFAB-MS (307.1559) 17H 22O 5(calculated value: 307.1546) (table 1,4).By 1HNMR, 1H- 1Hcosy, 13CNMR and DEPT spectrum (table 5) identify that comprehensively CP-D is 9 β-ethanoyl-parthenclide, and this is a new compound.
The chemical ionization mass spectrometry of table 4 CP-D (CIMS) data
n/z Abundance(％) Assignment
CH 4CI NH 3CI FAB
347 3 - - M+C 3H 5 +
335 3 - - M+C 2H 5 +
324 - 30 - M+NH 4 +
307 55 15 100 MH +
287 3 - M’+C 3H 5 +
275 6 - M′+C 2H 5 +
265 10 5 MH-CH 2CO
264 15 M +-CH 2CO
247 100 35 90 (MH-HOAc)H +orM′H
229 65 100 65 M′H-H 2O
Table 5 CP-D's 13CNMR, 1HNMR and 1H- 1H cosy composes data
Position? 13C?chem.? 1H?chem. 1H 1H- 1H?splittings COSY?to
shifts shifts multiplicity (or?W 1/2)in?Hz protons：
1 127.9 5.50 ddq 3.8，12.0；1.3 2a，2b，14
2 23.8 ～2.3 b(a) m 1，2b，
2.45(b) dddd 5.2，12.2，～13，～13 1，2a，3a
3 36.0 * 1.25(a) bddd 5.3，～13，～13 2b，3b
～2.2 b(b) m 3a
4 61.3 - - - -
5 66.1 2.70 d 8.9 6
6 81.6 3.85 t 8.6 5，7
7 44.2 2.91 ddddd 1.7，3.5，3.5，8.5，8.5 6，8a，8b，13E，13Z
8 36.2 2.18 b(a) bd ～14(W 1/2?6) 7，8b，9
2.01(b) ddd 8.5，10.8，14.6 7，8a，9
9 80.8 5.20 dd 2.3，10.9 8a，8b
10 133.0 - - - -
11 138.0 - - - -
12 170.0 - - - -
13 122.1 5.69(E) d 3.2 7
6.36(Z) d 3.6 7
14 11.8 1.73 t 1.3 1
15 17.4 1.32 s - -
Ac?CH 3 21.3 2.09 s - -
Ac?CO 168.7 - - - -
Extracting method of the present invention also comprises the separation purification of following compound:
Figure A0212807000091
CP-B (CHCl 3), leucoplastid crystalline substance, mp134～136 ℃; Get molecular formula C by CI-MS (table 6) 15H 20O 3By 1HNMR, 1H- 1H cosy, 13CNMR and DEPT (table 7) spectrum are confirmed.Data contrast (table 8) basically identical of above-mentioned data and santamarine (Santamarin), pointing out CP-B is santamarine.
The chemical ionization mass spectrometry of table 6 CP-B (CIMS) data
(M’＝M-H 2O)
Abundance (%) Abundance (%) chemi-ionization is arranged
m/z
Assignment
CH 4CI NH 3?CI
289 3 - M+C 3H 5 +
277 2 - M+C 2H 5 +
271 4 - M′+C 3H 5 +
266 - 100 M+NH 4 +
259 8 - M′+C 2H 5 +
249 45 MH +
231 100 M′H
213 45 M′-H 2O
185 10
141 10
Table 7 CP-B's 13CNMR, 1HNMR and 1H- 1H cosy composes data
13CNMR 1HMR
Carbon?or 13C?chem.shifts? 1H?chem.shifts COSy c multiplicity 1H- 1H?splittings
hydrogen (δCDCl 3?77.1) (δCHCl 3?7.25)?to?protons： (or?W 1/2)
1 75.3 3.67 2α，2β dd 6.5，9.9
2 32.9 ～2.1 b(α) 1，2β，3 m
～2.4 b(β) 1，2α，3，14 m
3 121.3 5.35 2α，2β，14 dddq ～1.5，～1.5，～1.5，～1.5
4 133.5 * -
5 51.2 2.34 b 6 bd ～12
6 81.5 3.94 5，7 dd 10.7，11.2
7 51.1 2.49 6，8α，8β，13E，13Z dddd 3.4，3.4，10.8，12.2
8 21.3 ～2.05 b(α) 7，8β，9α m
1.65(β) 7，8α，9α，9β dddd 4.0，12.4，13.5，～13.5
9 34.3 ～2.05 b(α) 8β，9 m
1.30(β) 8α，8β，9α，15 ddd 5.0，～13.5，～13.5
10 40.9 -
11 139.0 * -
12 170.8 -
13 116.9 5.40(E) 7 d 3.0
6.07(Z) 7 d 3.2
14 23.4 1.84 2β，3 m (W h/2?5.5)
15 11.1 0.87 9β s
a?Measured?in?CDCl 3?at?75?MHz?for? 13C?and?300?MHz?for? 1H.All?signals?of?protonatedcarbon?atoms?have?been?analysed?by?DEPT?and?have?been?correlated?with?thecorresponding? 1H?signals?by?Heteronuclear?Correlation(HETCOR).
b?Overlapping?with?other?signals.
c?Homonuclear?Correlated?Spectroscopy.When?signals?overl?ap，COSY?results?may?be?indoubt.
Above-mentioned data and CP-B, CP-C, CP-D nuclear-magnetism spectrum data are provided by Sydney University department of pharmacy.
13CNMR(75MHz)， 1HNMR(300MHz)。
The carbon of all band protons all is confirmed by DEPT spectrum and the relevant spectrum of C-H heteronuclear.
Table 8 CP-B and santamarine (Santamarin) 1HNMR composes contrast
Hydrogens Santamarin(e)in?CDCl 3 CP-B?in?CDCl 3
(δTMS＝0) (δCHCl 3＝7.26)
1 3.68dd(6.5，9.7) 3.67dd(6.5，9.9)
3 5.36bs b 5.35m(W h/2?9.5)
6 3.95dd(10.8，11.1) 3.94dd(10.7，11.2)
13(E) 5.41d(3.0) b 5.40d(3.0)
13(Z) 6.02d(3.3) 6.07d(3.2)
14 1.85bs 1.84m(W h/2?5.5)
15 0.89s 0.87s
Nuclear-magnetism spectrum data and the mass-spectrometric data of CP-B, CP-C, CP-D are provided by Sydney University department of pharmacy and Yunnan University.
The method that the present invention separates the purification above-claimed cpd is:
After cup chrysanthemum complete stool drying and crushing, with 95% alcohol immersion 30～40 days, extract medicinal extract, medicinal extract CP 0Get this medicinal extract, use medicinal extract: diatomite (weight ratio)=added diatomite (chemical pure) and mix in 1: 1～1: 2 with 95% ethanol, in water-bath, fry and do, weigh, use sherwood oil, chloroform respectively: ethyl acetate=1: 0～0: 1, ethyl acetate, interior ketone, methyl alcohol extract successively, get thick component CP 1, CP 2, CP 3, CP 4, CP 5
Use L 1210Cell is respectively to CP 1～CP 5Carry out the cytotoxic activity screening, the result, the antitumour activity size is followed successively by CP 2＞CP 3＞CP 1＞CP 4＞CP 5
To CP 2Carry out column chromatography, make sorbent material with 40～63 μ m silica gel, make eluent with petroleum ether-ethyl acetate-methyl alcohol, wet method dress post is got 11 component: A 1～A 11, through cytotoxic activity test determination, wherein A 4, A 5, A 7 Deng 3 components activity is arranged.
Respectively to A 4, A 5, A 7Make sorbent material Deng 3 components with TLC level silica gel, make eluent with normal hexane-ethyl acetate-methyl alcohol, the vacuum column chromatography is got 10 (C respectively 1～C 10), 8 (B 1～B 8), 8 (D 1～D 8) component.Recrystallization gets 10 pure crystal, and wherein 3 pure crystal are CP-B, CP-C, CP-D.
Use L 1210Clone is carried out the antitumour activity screening to crude extract, component and 10 pure crystal (compound), and the result shows: component A 4, A 5, A 7Have antitumour activity, CP-B, CP-C, CP-D in 10 pure crystal are anticancer active constituents.
According to international standard: (half casualty-producing concentrations) IC 50≤ 4 μ g/ml, promptly dosage is less than or equal to 4 μ g/ml, can kill or suppress the chemical ingredients of 50% cancer cells, is regarded as activeconstituents.
The evaluation of above-mentioned 3 anticancer active constituents:
Under the cytotoxicity test result instructs, from A 4, A 5, A 7Separate in the component and obtain CP-B, CP-C, these 3 anticancer active constituents of CP-D.By CIMS, HRFABMS, 1HNMR, 13CNMR, DEPT reach 1H- 1The Hcosy spectrum, analysis-by-synthesis has been identified structure, CP-B, CP-C, CP-D are respectively: santamarine (Santamarin), 9 β-ethanoyl-Costus speciosus lactone (new compound) and 9 β-ethanoyl-parthenclide (new compound).
The pharmacological research of three activeconstituentss and antitumour activity thereof:
Select 5 kinds of cancerous cell line: KB (human nasopharyngeal carcinoma), MCF-7 (human breast carcinoma), CCRF-CEM (human leukemia), L 1210(mouse leukemia), LS174T (human colon carcinoma); Respectively CP-B, CP-C, CP-D are carried out the cytotoxicity test of system.
The result shows: 1. CP-B, CP-C, CP-D have antitumour activity to 5 kinds of cancer cells; Activity is all above international standard (IC 50≤ 4 μ g/ml).2. the KB cell is the most responsive to 3 compounds, IC 50Between 0.16～0.29 μ g/ml, secondly be L 1210, MCF-7, CCRF-CEM, LS174T; To CP-B antitumour activity size is KB＞L successively 1210＞MCF-7＞CCRF-CEM＞LS174T is KB 〉=MCF-7＞CCRF-CEM＞L to CP-C successively 1210＞LS174T is KB＞CCRF-CEM＞MCF-7＞L to CP-D successively 1210＞LS174T.Detailed results sees the following form and Fig. 6, Fig. 7, Fig. 8.
CP-B, CP-C, CP-I) to L 1210, CCRF-CEM, KB, LS174T
With five kinds of cell growth inhibitory effects of MCF-7 (after 48 hours)
Cellline Origin source Estimated IC 50(μ g/mL) calculates
CP-B CP-C CP-D
L1210 murine?leukaemia 0.41 0.89 0.59
CCRF-CEM human?leukaemia 0.59 0.65 0.49
KB human?nasopharyngeal?cancer 0.16 0.25 0.29
LS174T human?colon?adenocarcinoma 0.92 1.28 1.08
MCF-7 human?breast?adenocarcinoma 0.53 0.63 0.50
CP-B, CP-C, CP-D are to L 1210Inhibitory rate of cell growth (%)
Compound Dose Growth?inhibition(％±SD) IC 50
(μg/mL) 48?h?after?drug?treatment a (ug/mL)
Control 0 0
CP-B 0.1 14.4±5.3
0.3 39.3±6.7
1.0 80.8±2.3 0.41
3.0 98.9±1.1
10.0 100.0±0.0
CP-C 0.1 10.7±2.3
0.3 28.2±5.7
1.0 51.1±2.5 0.89
3.0 98.5±0.4
10.0 100.0±0.0
CP-D 0.1 13.8±4.6
0.3 30.3±10.4
1.0 79.3±10.1 0.49
3.0 98.2±0.3
10.0 100.0±0.0
Above-mentioned pharmacological testing is finished in Sydney University's cancer department of medial science pharmacological evaluation chamber.
The separating and purifying method simple and feasible that the present invention adopts, the yield height has one's own knack.Three anticancer active constituents (CP-B, CP-C, CP-D) activity of gained is stronger, has surpassed international standard, and a cup chrysanthemum raw material is easy to get (can artificial culture), has the prospect of the PTS of being developed as.
Description of drawings
Fig. 1 slightly mentions solvent division process schema for cup chrysanthemum chemical ingredients of the present invention (CP);
Fig. 2 is component CP of the present invention 2Separate the process flow sheet of purifying;
Fig. 3 is of the present invention from component A 4Separating purifies obtains anticancer active constituent CP-C process flow sheet;
Fig. 4 is of the present invention from component A 5Separating purifies obtains anticancer active constituent CP-B process flow sheet;
Fig. 5 is of the present invention from component A 7Separating purifies obtains anticancer active constituent CP-D process flow sheet;
Fig. 6 is that CP-B of the present invention is to L 1210, CCRF-CEM, KB, LS174T and five kinds of growth of cancer cells of MCF-7 influence (curve) figure;
Fig. 7 is that CP-C of the present invention is to L 1210, CCRF-CEM, KB, LS174T and five kinds of growth of cancer cells of MCF-7 influence (curve) figure;
Fig. 8 is that CP-D of the present invention is to L 1210, CCRF-CEM, KB, LS174T and five kinds of growth of cancer cells of MCF-7 influence (curve) figure.
Among the figure, CP 1～CP 5Be sherwood oil, chloroform: ethyl acetate, ethyl acetate, acetone, methyl alcohol are successively to medicinal extract extraction (being that solvent is cut apart) obtained component; A 1～A 11Be CP 211 components that column chromatography obtains.
Embodiment
Embodiment 1:
1, with after the cup chrysanthemum complete stool drying and crushing, get and pulverize 10 kilograms in sample, with 95% alcohol immersion 30 days, extract medicinal extract, must medicinal extract CP 0≈ 800 grams;
2, get this medicinal extract 800 grams, press 1: 1 mixed, mix, in water-bath, fry and do, weigh W with 95% ethanol with diatomite Sample=710 grams are used the 1L cable type extractor according, and use sherwood oil, chloroform respectively: ethyl acetate=3: 1, ethyl acetate, acetone, methyl alcohol extract successively, get thick component CP 1(120 gram), CP 2(100 gram), CP 3(100 gram), CP 4(130 gram), CP 5(150 gram), as shown in Figure 1.
3, use L 1210Cell is respectively to CP 1～CP 5Carry out the cytotoxic activity screening, the result, the antitumour activity size is followed successively by CP 2＞CP 3＞CP 1＞CP 4＞CP 5
4, to CP 2Carry out column chromatography, make sorbent material with 40～63 μ m silica gel, make eluent with petroleum ether-ethyl acetate-methyl alcohol, wet method dress post is got 11 component A 1～A 11, as shown in Figure 2, through cytotoxic activity test determination, wherein A 4, A 5, A 7 Deng 3 components activity is arranged.
5, respectively to A 4, A 5, A 7Make sorbent material Deng 3 components with TLC level silica gel, make eluent with normal hexane-ethyl acetate-methyl alcohol, the vacuum column chromatography is got 10 (C respectively 1～C 10), 8 (B 1～B 8), 8 (D 1～D 8) component, recrystallization gets 10 pure crystal, and wherein 3 crystal are CP-B, CP-C, CP-D.Technical process such as Fig. 3, Fig. 4, shown in Figure 5.
6, use L 1210Cancerous cell line is to crude extract CP 0, component A 1～A 11, C 1～C 10, B 1～B 8, D 1～D 8Reach 10 pure compounds and carry out the antitumour activity screening, the result shows: component A 4, A 5, A 7Have antitumour activity, 3 Compound C P-B, CP-C, CP-D are anticancer active constituents in 10 pure compounds.
Embodiment 2:
1, with embodiment 1;
2, get this medicinal extract 800 grams, press 1: 2 mixed, mix, in water-bath, fry and do, weigh W with 95% ethanol with diatomite Sample=710 grams are used the 1L cable type extractor according, and use sherwood oil, chloroform respectively: ethyl acetate=1: 0, ethyl acetate, acetone, methyl alcohol extract successively, get thick component CP 1(120 gram), CP 2(80 gram), CP 3(120 gram), CP 4(130 gram), CP 5(150 gram), as shown in Figure 1.
3,4,5,6 is identical with embodiment 1;

Claims (4)

1, a kind of chemical ingredients of extracting from the cup chrysanthemum is characterized in that this compound is 9 β-ethanoyl-Costus speciosus lactone (CP-C), and molecular formula is C 17H 22O 4, its structural formula is:
Figure A0212807000021
2, a kind of chemical ingredients of extracting from the cup chrysanthemum is characterized in that this compound is 9 β-ethanoyl-parthenclide (CP-D), and molecular formula is C 17H 22O 5, its structural formula is:
Figure A0212807000022
3, according to the separating and purifying method of claim 1 and 2 described anti-cance active substance of cyathocline purpurea, it is characterized in that:
(1) with after the cup chrysanthemum complete stool drying and crushing, with 95% alcohol immersion 30～40 days, extract medicinal extract, medicinal extract CP 0
(2) get this medicinal extract medicinal extract: diatomite (weight ratio)=1: 1～1: 2 adding diatomite, mix with 95% ethanol, in water-bath, fry and do, weigh, use sherwood oil, chloroform respectively: ethyl acetate=1: 0～0: 1, ethyl acetate, acetone, methyl alcohol extract successively, get thick component CP 1, CP 2, CP 3, CP 4, CP 5
(3) use L 1210Cell is respectively to CP 1～CP 5Carry out the cytotoxic activity screening, the result, the antitumour activity size is followed successively by CP 2＞CP 3＞CP 1＞CP 4＞CP 5
(4) to CP 2Carry out column chromatography, make sorbent material with 40～63 μ m silica gel, make eluent with petroleum ether-ethyl acetate-methyl alcohol, wet method dress post is got 11 component: A 1～A 11, through cytotoxic activity test determination, wherein A 4, A 5, A 7Deng 3 components activity is arranged;
(5) respectively to A 4, A 5, A 7Make sorbent material Deng 3 components with TLC level silica gel, make eluent with normal hexane-ethyl acetate-methyl alcohol, the vacuum column chromatography is got 10 (C respectively 1～C 10), 8 (B 1～B 8), 8 (D 1～D 8) component; Recrystallization gets 10 pure crystal, and wherein 3 crystal are CP-B, CP-C, CP-D;
(6) use L 1210Clone is carried out the antitumour activity screening to crude extract, component and 10 pure crystal, and the result shows: component A 4, A 5, A 7Have antitumour activity, pure crystal CP-B, CP-C, CP-D are anticancer active constituents.
4,, it is characterized in that Compound C P-C and CP-D have antitumour activity according to the purposes of claim 1 and the 2 described chemical ingredientss of from the cup chrysanthemum, extracting.
CN 02128070 2002-12-19 2002-12-19 Anti-cance active substance of cyathocline purpurea and separating and purifying method thereof Expired - Fee Related CN1232517C (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103524520A (en) * 2013-09-25 2014-01-22 中国人民解放军第四军医大学 Method for extracting parthenolide from plant raw material
WO2014067379A1 (en) * 2012-10-30 2014-05-08 天津尚德药缘科技有限公司 Deuterated dimethyl amine parthenolide, preparation method and use thereof in drug preparation

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014067379A1 (en) * 2012-10-30 2014-05-08 天津尚德药缘科技有限公司 Deuterated dimethyl amine parthenolide, preparation method and use thereof in drug preparation
CN103788103A (en) * 2012-10-30 2014-05-14 天津尚德药缘科技有限公司 Deuterated dimethylamino parthenolide, preparation method thereof and applications thereof in medicine preparation
CN103788103B (en) * 2012-10-30 2016-08-03 天津尚德药缘科技股份有限公司 Deuterated dimethyl amine parthenolide, its preparation method and the purposes in preparing medicine
CN103524520A (en) * 2013-09-25 2014-01-22 中国人民解放军第四军医大学 Method for extracting parthenolide from plant raw material
CN103524520B (en) * 2013-09-25 2015-12-09 中国人民解放军第四军医大学 A kind of method extracting parithenolide from plant material

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