CN1821262A - Active total saponin extract of Lonicerae argentina and its preparation method and use - Google Patents
Active total saponin extract of Lonicerae argentina and its preparation 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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- lonicera
- extract
- cinerea
- saponin
- active
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- 229930182490 saponin Natural products 0.000 title claims abstract description 88
- 150000007949 saponins Chemical class 0.000 title claims abstract description 88
- 239000001397 quillaja saponaria molina bark Substances 0.000 title claims abstract description 75
- 239000000284 extract Substances 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 235000017709 saponins Nutrition 0.000 claims abstract description 87
- 241000245240 Lonicera Species 0.000 claims abstract description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 10
- 201000007270 liver cancer Diseases 0.000 claims abstract description 9
- 208000014018 liver neoplasm Diseases 0.000 claims abstract description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229920005989 resin Polymers 0.000 claims abstract description 6
- 239000011347 resin Substances 0.000 claims abstract description 6
- 239000004952 Polyamide Substances 0.000 claims abstract description 4
- 229920002647 polyamide Polymers 0.000 claims abstract description 4
- 238000005119 centrifugation Methods 0.000 claims abstract 3
- 238000002481 ethanol extraction Methods 0.000 claims abstract 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 239000003814 drug Substances 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
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- SMRPGWBDLOQHOS-UHFFFAOYSA-N 5-[4,5-dihydroxy-6-(hydroxymethyl)-3-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxy-3,4-dihydroxy-6-[[9-hydroxy-4-(hydroxymethyl)-4,6a,6b,8a,11,11,14b-heptamethyl-14-oxo-2,3,4a,5,6,7,8,9,10,12,12a,14a-dodecahydro-1H-picen-3-yl]oxy]oxane-2-carboxylic acid Chemical compound OC1C(O)C(O)C(C)OC1OC1C(OC2C(OC(C(O)C2O)C(O)=O)OC2C(C3C(C4C(C5(CCC6(C)C(O)CC(C)(C)CC6C5=CC4=O)C)(C)CC3)(C)CC2)(C)CO)OC(CO)C(O)C1O SMRPGWBDLOQHOS-UHFFFAOYSA-N 0.000 claims 3
- BMWPBKOFJSHJAW-UHFFFAOYSA-N Saponin B Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(CO)C(O)C(OC7OC(CO)C(O)C(O)C7O)C6=O)C(C)(C)C5CCC34C)C2C1)C(=O)O BMWPBKOFJSHJAW-UHFFFAOYSA-N 0.000 claims 3
- PPRSVUXPYPBULA-UHFFFAOYSA-N saponin A Natural products CC1(C)CCC2(CCC3(C)C(=CCC4C5(C)CCC(OC6OC(CO)C(O)C(O)C6=O)C(C)(C)C5CCC34C)C2C1)C(=O)O PPRSVUXPYPBULA-UHFFFAOYSA-N 0.000 claims 3
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- 241001170076 Lonicera macranthoides Species 0.000 description 33
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- 241001570521 Lonicera periclymenum Species 0.000 description 12
- 230000000259 anti-tumor effect Effects 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
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- 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 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 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 description 3
- 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 description 3
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- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 description 2
- 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
- 229960002949 fluorouracil Drugs 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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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-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
本发明涉及一种灰毡毛忍冬活性总皂苷提取物及其制备方法和用途。提取物为灰毡毛忍冬活性总皂苷，制备方法包括50－90％乙醇提取，低极性溶剂萃取，乙酸乙酯萃取，碱水解，丙酮沉淀，离心，大孔树脂或聚酰胺柱层析，活性炭脱色，得到灰毡毛忍冬活性总皂苷。本发明的提取工艺及方法获得的有效成分得率及纯度高，已明确了灰毡毛忍冬活性总皂苷的药理、药效作用及其中的有效成分和它们的含量比例，易于掌握有效剂量。本发明获得的有效成分的有效剂量对体外培养的人癌细胞株有细胞毒活性，对小鼠肝癌H22实体瘤和Lewis肺癌实体瘤具有抑制作用。The invention relates to an active total saponin extract of Lonicera cinerea and its preparation method and application. The extract is the active total saponins of Lonicera cinerea, and the preparation method includes 50-90% ethanol extraction, low polarity solvent extraction, ethyl acetate extraction, alkali hydrolysis, acetone precipitation, centrifugation, macroporous resin or polyamide column chromatography, Decolorize with activated carbon to obtain active total saponins of Lonicerae cinerea. The extraction process and method of the present invention have high yield and high purity of active ingredients, and the pharmacology and drug effects of active total saponins of Lonicera cinerea have been clarified, as well as the active ingredients and their content ratios, and the effective dosage is easy to grasp. The effective dose of the active ingredient obtained by the invention has cytotoxic activity on human cancer cell lines cultured in vitro, and has inhibitory effect 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)
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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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| 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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