CN1821262A - Lonicera macranthoides active total saponin extract and its preparing method and use - Google Patents
Lonicera macranthoides active total saponin extract and its preparing method and use Download PDFInfo
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- CN1821262A CN1821262A CN 200610039294 CN200610039294A CN1821262A CN 1821262 A CN1821262 A CN 1821262A CN 200610039294 CN200610039294 CN 200610039294 CN 200610039294 A CN200610039294 A CN 200610039294A CN 1821262 A CN1821262 A CN 1821262A
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- total saponin
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- lonicera macranthoides
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- 229930182490 saponin Natural products 0.000 title claims abstract description 87
- 150000007949 saponins Chemical class 0.000 title claims abstract description 87
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 84
- 239000000284 extract Substances 0.000 title claims abstract description 58
- 241001170076 Lonicera macranthoides Species 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000000605 extraction Methods 0.000 claims abstract description 20
- 238000002360 preparation method Methods 0.000 claims abstract description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000001556 precipitation Methods 0.000 claims abstract description 9
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- 229910052799 carbon Inorganic materials 0.000 claims abstract description 4
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- 239000003814 drug Substances 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
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- KPDVIKCSTOVVEA-KZFFKTSUSA-N (4aS,6aR,6aS,6bR,8aR,9R,10S,12aR,14bS)-10-[(2S,3R,4S,5S)-3-[(2S,3R,4R,5S,6S)-4-[(2S,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-3,5-dihydroxy-6-methyloxan-2-yl]oxy-4,5-dihydroxyoxan-2-yl]oxy-9-(hydroxymethyl)-2,2,6a,6b,9,12a-hexamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylic acid Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O)O[C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H](OC[C@H](O)[C@@H]2O)O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(CC[C@]6(CCC(C)(C)C[C@H]6C5=CC4)C(O)=O)C)(C)CC3)(C)CC2)(C)CO)O[C@H]([C@@H]1O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O KPDVIKCSTOVVEA-KZFFKTSUSA-N 0.000 claims description 6
- AMXYFWUYMQOLRN-BHGPNLNISA-N (4as,6ar,6as,6br,8ar,9r,10s,12ar,14bs)-10-[(2s,3r,4s,5s)-3-[(2s,3r,4r,5s,6s)-3,5-dihydroxy-6-methyl-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-4,5-dihydroxyoxan-2-yl]oxy-9-(hydroxymethyl)-2,2,6a,6b,9,12a-hexamethyl-1, Chemical compound O([C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H](OC[C@H](O)[C@@H]2O)O[C@@H]2[C@@]([C@H]3[C@]([C@@H]4[C@@]([C@@]5(CC[C@]6(CCC(C)(C)C[C@H]6C5=CC4)C(O)=O)C)(C)CC3)(C)CC2)(C)CO)O[C@H]([C@@H]1O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O AMXYFWUYMQOLRN-BHGPNLNISA-N 0.000 claims description 6
- ORIMHMILXLTJPC-UHFFFAOYSA-N Macranthoside A Natural products OC1C(O)C(C)OC(OC2C(C(OCC3(C)C4C(C5C(C6(CCC7(CCC(C)(C)CC7C6=CC5)C(O)=O)C)(C)CC4)(C)CCC3O)OC(CO)C2O)O)C1OC1OCC(O)C(O)C1O ORIMHMILXLTJPC-UHFFFAOYSA-N 0.000 claims description 6
- ZLIROJWQOROKCU-UHFFFAOYSA-N Macranthoside B Natural products CC1OC(OC2C(O)C(CO)OC(OC3C(O)C(O)C(OCC4(C)C(O)CCC5(C)C4CCC6(C)C5CC=C7C8CC(C)(C)CCC8(CCC67C)C(=O)O)OC3CO)C2O)C(OC9OCC(O)C(O)C9O)C(O)C1O ZLIROJWQOROKCU-UHFFFAOYSA-N 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 5
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- 239000012141 concentrate Substances 0.000 claims description 4
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- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 claims description 2
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- 206010028980 Neoplasm Diseases 0.000 abstract description 25
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- 239000003513 alkali Substances 0.000 abstract description 9
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
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- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 description 2
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 description 2
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- GNTDGMZSJNCJKK-UHFFFAOYSA-N divanadium pentaoxide Chemical compound O=[V](=O)O[V](=O)=O GNTDGMZSJNCJKK-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 229960002949 fluorouracil Drugs 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
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- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 description 2
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- BDXJANJAHYKTMI-UHFFFAOYSA-N 2,3,4,5-tetramethyl-1h-pyrrole Chemical class CC=1NC(C)=C(C)C=1C BDXJANJAHYKTMI-UHFFFAOYSA-N 0.000 description 1
- ASFAFOSQXBRFMV-LJQANCHMSA-N 3-n-(2-benzyl-1,3-dihydroxypropan-2-yl)-1-n-[(1r)-1-(4-fluorophenyl)ethyl]-5-[methyl(methylsulfonyl)amino]benzene-1,3-dicarboxamide Chemical compound N([C@H](C)C=1C=CC(F)=CC=1)C(=O)C(C=1)=CC(N(C)S(C)(=O)=O)=CC=1C(=O)NC(CO)(CO)CC1=CC=CC=C1 ASFAFOSQXBRFMV-LJQANCHMSA-N 0.000 description 1
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- GCGBHJLBFAPRDB-UHFFFAOYSA-N Hederagenin Natural products CC1(C)CCC2(CCC3(C)C4CCC5C(C)(CO)C(O)CCC5(C)C4CC=C3C2C1)C(=O)O GCGBHJLBFAPRDB-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to active total saponin extract of Lonicera macranthoides and its preparation process and use. The preparation process of active total saponin extract of Lonicera macranthoides includes the steps of extraction with 50-90 % ethyl alcohol solution, extraction with low polar solvent, extraction with ethyl acetate, alkali hydrolysis, acetone precipitation, centrifugation, macro porous resin or polyamide column chromatography, and active carbon decolorizing to obtain active total saponin extract of Lonicera macranthoides. The extraction process of the present invention has high effective component yield and purity, and the product has controllable effective dosage. The effective components the present invention obtains has cytotoxicity on extracorporeally cultured human cancer cell strain, and inhibition on mouse liver cancer H22 solid tumor and Lewis lung cancer solid tumor.
Description
Technical field
The present invention relates to Lonicera macranthoides active total saponin extract and preparation method thereof, and the application of this active total saponin in preparation treatment cancer drug.In particular, the present invention relates to comprise in the largeflower-like honeysuckle flower flower and alabastrum composition that contains antitumour activity sugar chain structure saponin(e and preparation method thereof, by the hydrolysis of total saponins elder generation is divided again, thus obtained extract is directly become to be grouped into by activated saponin, main saponin constituent ratio is clear and definite, be easy to grasp its effective dose, and it is safer, more stable to be used for preparation treatment cancer drug.
Background technology
Largeflower-like honeysuckle flower (Lonicera macranthoides Hand.-Mazz.) is a Caprifoliaceae Caprfoliaceae woodbine, and 2005 editions Pharmacopoeias of People's Republic of China record, and list under the Lonicera confusa DC. item.Woodbine contains a large amount of saponin(es, is mainly ivy type and oleanolic acid type saponin(e.Hederagenin and oleanolic acid type saponin(e have and protect the liver (Shi Jingzhen etc.; several compositions of Lonicera fulvotnetosa Hsu et S. C. Cheng. Ms and largeflower-like honeysuckle flower are to the provide protection of large and small mouse chemical damage; CHINA JOURNAL OF CHINESE MATERIA MEDICA; 1999; 24 (6): 363-364), anticancer (Cytotoxic saponins from the root of Dipsacus asper Wall.Arch Pharm Res.2005,28 (9): 1053-6.) activity.In the separation of Jiangsu Province Chinese Academy of Sciences medicinal plants study development centre main saponin component in, two main saponin components have been obtained, clear and definite composition and the character and the purposes of its saponin component to the largeflower-like honeysuckle flower flower and alabastrum.The present invention takes to separate after the basic hydrolysis, has both simplified processing step, is fit to industrial mass production, has improved the yield of activated saponin again greatly, and the antitumour activity of the Lonicera macranthoides active total saponin extract that obtains thus compares not hydrolysis total saponins height.
Summary of the invention
Technical problem to be solved by this invention is, providing a kind of the extraction with Lonicera macranthoides active total saponin from the largeflower-like honeysuckle flower flower and alabastrum is the composition of main reactive site and the preparation method of said composition, make preparation raw material (active total saponin) yield and purity height, technology is simple and easy to do, is suitable for suitability for industrialized production.
The technical problem that the present invention further will solve is that providing with the Lonicera macranthoides active total saponin is the medicinal use that efficient part prepares medicine.
The technical solution adopted in the present invention is as follows:
The present invention relates to a kind of Lonicera macranthoides active total saponin extract, main reactive site active total saponin content is 50%-90% in the extract, and contained saponin component meets claim 1 formula of (1).Wherein main contained saponin(e and their weight ratio is: Macranthoside B: Macranthoside A=12～6: 1.
Lonicera macranthoides active total saponin extract of the present invention is the extract that obtains by following method: the largeflower-like honeysuckle flower flower and alabastrum, with concentration is the extraction using alcohol of 50%-90%, reclaim solvent, use sherwood oil, gasoline, the extraction of low polar solvents such as benzene, water layer is used ethyl acetate extraction again, water layer reclaims solvent to concentrated extract, add 1%-10%NaOH or KOH solution, 40-100 degraded 2-4 hour down, cold back adds acetone precipitation, and is centrifugal, and centrifugal gained precipitation washes with water to neutrality, with rare alcohol dissolving, the adjusting pH value is 4-7, last macroporous resin column (HP20, D101, AB-8, HPD100, HPD300 etc.) or polyamide column, first water and 20% washing with alcohol post, use the 30%-90% ethanol elution again, collect elutriant, add the medical active carbon decoloring of 3%-5%, filter, concentrate drying promptly gets Lonicera macranthoides active total saponin.
The said Lonicera macranthoides active total saponin extract of the present invention is meant through aforesaid method and extracts the efficient part that obtains.
Lonicera macranthoides active total saponin extract of the present invention proves to have antitumor action through pharmacology, pharmacodynamics and a large amount of animal experiment studies; In experiment in vitro, significantly suppress people's liver cancer HepG
2Cell and people's lung cancer A549 cell propagation; In vivo in the experiment, to H
22Liver cancer cell tumor-bearing mice and Lewis lung cancer cell tumor-bearing mice can suppress growth of tumor, increase tumor-bearing mice immunizing power and lifetime.Ingredient is clear and definite, and dosage is easy to grasp control, therefore can use in the medicine of preparation treatment cancer.
Extract Lonicera macranthoides active total saponin of the present invention can be prepared into tablet, capsule, granule, electuary, powder, pill, oral liquid, syrup, suspensoid, sprays, solution, decoction, gelifying agent, creme, ointment, drops etc.In above-mentioned preparation, the extract of the present invention of effective dose can be separately or with pharmacy acceptable carrier still: composition mixtures such as vehicle, disintegrating agent, lubricant, tinting material.
Embodiment
The saponin component that embodiment one largeflower-like honeysuckle flower bud water section obtains
The largeflower-like honeysuckle flower dry flower, 90% alcohol reflux three times, united extraction liquid, use sherwood oil, ethyl acetate extraction successively, macroporous resin column on the remaining water liquid, through the water-ethanol gradient elution, reversed-phase column purifying decolouring repeatedly obtains two main saponin components, and structure all contains antitumour activity unit.
Two saponin(es are respectively:
Macranthoside B (A), i.e. 3-O-β-D-glucopyranosyl (14)-β-D-glucopyranosyl (1-3)-α-L-rhamnopyranosyl (1-2)-α-L-arabopyranose base-hederagenin;
Macranthoside A (B), i.e. 3-O-β-D-glucopyranosyl (1-3)-α-L-rhamnopyranosyl (1-2)-α-L-arabopyranose base-hederagenin;
The assay of saponin(e
1. instrument and reagent
High performance liquid chromatograph Aglient 1100 series, quaternary pump, automatic sampler, detector: Alltech ELSD2000 (Alltech, Deerfield, IL, USA), chromatographic column: Aglient Zorbax SB-C
18Post (150 * 4.6mm, 5.0 μ m), acetonitrile is a chromatographically pure, phosphoric acid is analytical pure.
2. chromatographic condition
Moving phase: 0-10min, acetonitrile: 0.5% phosphoric acid solution is kept ratio (29: 71), 10-25min, the moving phase ratio changes to (46: 54) by (29: 71) are linear, 25-30min, moving phase is kept ratio (46: 54).ELSD manages temperature: 106.Nitrogen flow rate: 2.6L/min.
3. the preparation of reference substance solution
Accurate respectively Macranthoside B and the Macranthoside A reference substance that takes by weighing through the Vanadium Pentoxide in FLAKES dried overnight respectively adds methyl alcohol and makes the solution that every 1ml contains 0.03mg respectively, solution in contrast.
4. sample determination
Get the about 20mg of powder in this product (get in addition simultaneously this product powder measure moisture), accurately claim surely, put in the apparatus,Soxhlet's, add methanol eddy and extracted 4 hours, put cold after, be transferred in the 100ml volumetric flask, methanol constant volume promptly.Each sample detects through high performance liquid phase, and the content ratio that calculates two kinds of prosapogenins is 12～6: 1, sees embodiment three to ten for details.Provide 8 embodiments that extract the assay of Lonicera macranthoides active total saponin and saponin(e by the preparation method of aforementioned techniques scheme below, see following table for details:
Embodiment two
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 50％ | Sherwood oil | 1％ | 40℃ | 2 hours | 6 | HP- 20 | 3％ | 1.4％ | 88％ | 12∶ 1 |
Embodiment three
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Flower | 60％ | Gasoline | 3％ | 60℃ | 3 hours | 7 | HP- 20 | 4％ | 1.4％ | 72％ | 10.4 ∶1 |
Embodiment four
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 70％ | Sherwood oil | 4％ | 60℃ | 2 hours | 6 | D 101 | 3％ | 1.8％ | 80％ | 8.2∶ 1 |
Embodiment five
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Flower | 80％ | Gasoline | 5％ | 80℃ | 4 hours | 4 | D 101 | 4％ | 1.6％ | 76％ | 6.4∶ 1 |
Embodiment six
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 90％ | Sherwood oil | 2％ | 80℃ | 2 hours | 7 | AB-8 | 5％ | 1.7％ | 64％ | 8.7∶ 1 |
Embodiment seven
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 80％ | Sherwood oil | 4％ | 100 ℃ | 2 hours | 7 | HPD1 00 | 4％ | 1.9％ | 69％ | 7.9∶ 1 |
Embodiment eight
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 70％ | Sherwood oil | 6％ | 100 ℃ | 2 hours | 6 | HPD3 00 | 3％ | 2.00％ | 71％ | 7.5∶ 1 |
Embodiment nine
| Material | Extract concentration | Extraction liquid | Alkali concn | Hydrolysis temperature | Hydrolysis time | PH value | Adsorption column | Active C | Yield | Total saponin content | The saponin(e ratio |
| Bud | 60％ | Benzene | 10％ | 60℃ | 2 hours | 5 | Polymeric amide | 4％ | 1.7％ | 60％ | 6∶1 |
The inside and outside anti-tumor activity evaluation experimental of embodiment ten Lonicera macranthoides active total saponin extracts
1. Lonicera macranthoides active total saponin extract is to human liver cancer cell HepG
2And human lung cancer cell A549's restraining effect
1.1 purpose: the research Lonicera macranthoides active total saponin extract is to human liver cancer cell HepG
2And human lung cancer cell A549's growth in vitro restraining effect.
1.2 material: tetramethyl-azoles nitroblue (MTT); Lonicera macranthoides active total saponin extract: face with before being made into respective concentration; People's liver cancer HepG
2Cell strain or the strain of people's lung cancer A549 cell place RPMI-1 640 substratum that contain 10% calf serum, at 5%CO
2, 37 cell culture incubator in go down to posterity stand-by.
1.3 method and result: people's liver cancer HepG that will be in logarithmic phase
2It is 2 * 10 that concentration is adjusted in cell strain or the strain of people's lung cancer A549 cell
5Ml
-1, cultivate after 24 hours, join 5 medicine groups respectively by listed drug level in the table, each drug level is all established 4 multiple holes, the calculating of averaging, other establishes a blank hole.Cell is at 37,5%CO
2Continue under the condition to cultivate after 48 hours, add the MTT liquid 20M1 of 5mg/ml, continue to cultivate 4 hours again, add DMSO dissolving back and measure the OD value at the 570nm place with microplate reader rapidly, calculate inhibitory rate of cell growth, experimental result sees Table 1.
Growth inhibition ratio (%)=(1-dosing hole average A value/blank hole average A value) * 100%
Table 1 couple HepG
2The cytotoxicity of cell and A549 cell
| Adding consistency (μ g/ml) | To HepG 2Cell inhibitory rate | To the A549 cell inhibitory rate |
| 16 32 64 128 200 IC 50 | 8.34％ 32.43％ 52.43％ 67.20％ 77.25％ 58.80μg/ml | 9.90％ 29.34％ 59.41％ 70.20％ 84.85％ 55.84μ g/ml |
Lonicera macranthoides active total saponin extract with different concns is handled HepG
2Cell and A549 cell, the growth of visible cancer cells all is suppressed, and is concentration and time-dependent manner.The result shows that Lonicera macranthoides active total saponin extract has stronger cytotoxicity to tumour cell.
2. Lonicera macranthoides active total saponin extract and the not comparative studies of hydrolysis total saponin extracts antitumour activity
2.1 purpose: relatively Lonicera macranthoides active total saponin extract and the not antitumour activity of hydrolysis total saponin extracts.
2.2 material: Lonicera macranthoides active total saponin extract: face with before being made into respective concentration; Hydrolysis total saponin extracts not: the largeflower-like honeysuckle flower bud is 90% extraction using alcohol with concentration, reclaims solvent, uses sherwood oil, ethyl acetate extraction successively, water layer reclaims solvent to concentrated extract, adds acetone precipitation, and is centrifugal, precipitation is dissolved with rare alcohol, and regulating pH value is 6, last D
101Macroporous resin column, first water and 20% washing with alcohol post are used the 30%-90% ethanol elution again, and elutriant decolouring, filtration, concentrate drying are promptly.
2.3 method and result: the MTT colorimetry is the same, is divided into blank group, active group and not hydrolysis group, and wherein for the hydrolysis component is five concentration groups, experimental result sees Table 2.
Table 2 Lonicera macranthoides active total saponin extract and the not comparison of the antitumour activity of hydrolysis total saponin extracts
| Adding consistency (μ g/ml) | To HepG 2Cell inhibitory rate | To the A549 cell inhibitory rate |
| Active group-60 not hydrolysis groups-20 not hydrolysis group-60 not hydrolysis group-120 not hydrolysis group-250 IC 50 | 50.90％ 3.40％ 5.11％ 7.91％ 15.77％ ＞300μg/ml | 54.71％ 3.17％ 5.39％ 8.34％ 16.86％ ＞300μg/ml |
From experimental result as can be seen, hydrolysis largeflower-like honeysuckle flower total saponine anti-tumor is not active reduces IC
50＞300 μ g/ml.Hydrolysis reaction has improved the anti-tumor activity of extract greatly.
3. Lonicera macranthoides active total saponin extract is to H
22The anti-tumor experiment of tumor-bearing mice
3.1 purpose: the research Lonicera macranthoides active total saponin extract is to rat liver cancer H
22Restraining effect.
3.2 material: 40 of Male Kunming strain mice, body weight (20 ± 2g); 5 FU 5 fluorouracil, the general medicine company product in the rising sun East Sea, Shanghai; Lonicera macranthoides active total saponin extract: face with preceding and be made into respective concentration with physiological saline.Laboratory temperature 20 ± 3, relative humidity: 75%.
3.3 method and result: strip the liver cancer H that went down to posterity 10 days under the aseptic condition
22The cancer piece is made homogenate, and with physiological saline dilution in 1: 3, it is 2 * 10 that counting is adjusted cell concn
6L
-1Every 0.2ml, it is subcutaneous to be inoculated in the right armpit of mouse, be divided into 5 groups after 24 hours at random, promptly dosage group, medicine small dose group and positive controls (5 FU 5 fluorouracil) in negative control group (physiological saline), the heavy dose of group of medicine, the medicine are all irritated stomach respective concentration medicine or physiological saline for every group, once a day, the execution of weighing after ten days, the tumor mass calculating inhibition rate of tumor growth of weighing the results are shown in Table 3.
Inhibition rate of tumor growth inhibiting rate (%)=(it is heavy that the average knurl of average knurl weight/control group is organized in the 1-treatment) * 100%
Table 3 Lonicera macranthoides active total saponin is to H
22The influence of tumor-bearing mice and Lewis tumor-bearing mice tumour inhibiting rate
| Group | n | To H 22The tumour inhibiting rate of tumor-bearing mice | To Lewis tumor-bearing mice moral tumour inhibiting rate |
| Dosage group 40mg/kg medicine small dose group 20mg/kg positive controls in the heavy dose of group of the medicine 80m/kg medicine | 8 8 8 8 | 50.6％ 39.7％ 27.4％ 56.7％ | 46.61％ 37.56％ 26.24％ - |
Lonicera macranthoides active total saponin is to H
22The influence of tumor-bearing mice lifetime the results are shown in Table 4.
Table 4 Lonicera macranthoides active total saponin is to H
22The influence of tumor-bearing mice lifetime
| Group | n | Living phases |
| Dosage group positive controls in the negative control group medicine | 8 8 8 | 14.1±2.13 29.3±1.36 29.1±2.15 |
The result shows: sample sets and negative control group be the survival time of mice phenomenal growth relatively, highly significant difference is arranged, p＜0.01.
4. Lonicera macranthoides active total saponin extract is to the restraining effect of Mice Bearing Lewis Lung Cancer
4.1 purpose: the research Lonicera macranthoides active total saponin extract is to the restraining effect of Mice Bearing Lewis Lung Cancer.
4.2 material: 32 of Male Kunming strain mice, body weight (20 ± 2g); Lonicera macranthoides active total saponin extract: face with preceding and be made into respective concentration with physiological saline.Laboratory temperature 20 ± 3, relative humidity: 75%.
4.3 method and result: strip the Lewis lung cancer cancer piece that went down to posterity 10 days under the aseptic condition, make homogenate, with physiological saline dilution in 1: 3, it is 1 * 10 that counting is adjusted cell concn
7Ml
-1, every 0.2ml, it is subcutaneous to be inoculated in the right armpit of mouse, be divided into 5 groups after 24 hours at random, promptly dosage group and medicine small dose group in negative control group (physiological saline), the heavy dose of group of medicine, the medicine are all irritated stomach respective concentration medicine or physiological saline for every group, once a day, the execution of weighing after ten days.Weighing tumor mass weight is calculated inhibition rate of tumor growth, sees Table 3.
Inhibition rate of tumor growth inhibiting rate (%)=(it is heavy that the average knurl of average knurl weight/control group is organized in the 1-treatment) * 100%
The result shows: Lonicera macranthoides active total saponin extract can significantly suppress H
22It is heavy that the tumor-bearing mice knurl heavily reaches Lewis tumor-bearing mice knurl, has antitumour activity.
Claims (5)
1. Lonicera macranthoides active total saponin extract, it is characterized in that main reactive site active total saponin content is 50%-90% in the extract, contained saponin component meets following general formula (1), wherein main contained saponin(e and their weight ratio is: Macranthoside B: Macranthoside A=12～6: 1
R
3＝H，OH
R
2＝H
R
1=H, sugar chain (1).
2. extract according to claim 1, it is characterized in that said extract is the extract that obtains by following method: the largeflower-like honeysuckle flower flower and alabastrum, with concentration is the extraction using alcohol of 50%-90%, reclaim solvent, with the low polar solvent extraction, water layer is used ethyl acetate extraction again, water layer reclaims solvent to concentrated extract, add 1%-10%NaOH or KOH solution, 40-100 degraded 2-4 hour down, and cold back adds acetone precipitation, centrifugal, centrifugal gained precipitation washes with water to neutrality, and with rare alcohol dissolving, the adjusting pH value is 4-7, last macroporous resin column or polyamide column, elder generation's water and 20% washing with alcohol post are used the 30%-90% ethanol elution again, collect elutriant, the medical active carbon decoloring that adds 3%-5%, filter, concentrate drying promptly gets Lonicera macranthoides active total saponin.
3. extract according to claim 2, it is characterized in that using sherwood oil, gasoline, the extraction of low polar solvents such as benzene, water layer is used ethyl acetate extraction again, water layer reclaims solvent to concentrated extract, add 1%-10%NaOH or KOH solution, 40-100 degraded 2-4 hour down, cold back adds acetone precipitation, and is centrifugal, and centrifugal gained precipitation washes with water to neutrality, with rare alcohol dissolving, the adjusting pH value is 4-7, mixes sample or directly goes up macroporous resin column (HP20, D101, AB-8, HPD100, HPD300) or polyamide column, first water and 20% washing with alcohol post, use the 30%-90% ethanol elution again, collect elutriant, add the medicinal granular active carbon decolouring of 3%-5%, filter, concentrate drying promptly gets Lonicera macranthoides active total saponin.
4. the process for extracting of the largeflower-like honeysuckle flower flower and alabastrum efficient part Lonicera macranthoides active total saponin that is extracted according to claim 1-3 is characterized in that the Lonicera macranthoides active total saponin of being carried is: Macranthoside B: Macranthoside A.
5. the Macranthoside B that is extracted according to claim 1-3: Macranthoside A,
The application of its feature in the medicine of preparation treatment or prevention human hepatocellular.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101406509B (en) * | 2008-11-28 | 2011-08-31 | 江苏省中国科学院植物研究所 | Lonicera confusa extract and preparation method and application thereof |
| CN102408465A (en) * | 2011-09-23 | 2012-04-11 | 江苏省中国科学院植物研究所 | Lonicera macranthoides chlorogenic acid ester saponin as well as preparation thereof and application |
| CN103908482A (en) * | 2012-12-28 | 2014-07-09 | 中国医学科学院药用植物研究所 | Ethyl acetate extract of Lonicera macranthoides, preparation method and application thereof |
| CN105311085A (en) * | 2014-06-06 | 2016-02-10 | 天津药物研究院 | Extract of honeysuckle stem or its original plant honeysuckle leaf or flower and preparation method and use thereof |
| CN109678922A (en) * | 2018-11-12 | 2019-04-26 | 广州市香雪制药股份有限公司 | Triterpene compound and preparation method thereof, application drop |
| CN110403971A (en) * | 2018-04-26 | 2019-11-05 | 胤安国际（辽宁）基因科技股份有限公司 | A kind of extracting method and its application of honeysuckle extract |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101406509B (en) * | 2008-11-28 | 2011-08-31 | 江苏省中国科学院植物研究所 | Lonicera confusa extract and preparation method and application thereof |
| CN102408465A (en) * | 2011-09-23 | 2012-04-11 | 江苏省中国科学院植物研究所 | Lonicera macranthoides chlorogenic acid ester saponin as well as preparation thereof and application |
| CN102408465B (en) * | 2011-09-23 | 2015-09-30 | 江苏省中国科学院植物研究所 | A kind of largeflower-like honeysuckle flower chlorogenic acid Ester Saponin and its production and use |
| CN103908482A (en) * | 2012-12-28 | 2014-07-09 | 中国医学科学院药用植物研究所 | Ethyl acetate extract of Lonicera macranthoides, preparation method and application thereof |
| CN105311085A (en) * | 2014-06-06 | 2016-02-10 | 天津药物研究院 | Extract of honeysuckle stem or its original plant honeysuckle leaf or flower and preparation method and use thereof |
| CN105311085B (en) * | 2014-06-06 | 2020-01-14 | 天津药物研究院 | Leaf or flower extract of honeysuckle stem or its base plant honeysuckle, its preparation method and application |
| CN110403971A (en) * | 2018-04-26 | 2019-11-05 | 胤安国际（辽宁）基因科技股份有限公司 | A kind of extracting method and its application of honeysuckle extract |
| CN109678922A (en) * | 2018-11-12 | 2019-04-26 | 广州市香雪制药股份有限公司 | Triterpene compound and preparation method thereof, application drop |
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