CN101352448B - Use of Liriope muscari Bailey C in pharmacy - Google Patents
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- CN101352448B CN101352448B CN2008101960440A CN200810196044A CN101352448B CN 101352448 B CN101352448 B CN 101352448B CN 2008101960440 A CN2008101960440 A CN 2008101960440A CN 200810196044 A CN200810196044 A CN 200810196044A CN 101352448 B CN101352448 B CN 101352448B
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- 241001532026 Liriope muscari Species 0.000 title claims description 18
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- 206010027476 Metastases Diseases 0.000 claims abstract description 16
- 230000009401 metastasis Effects 0.000 claims abstract description 16
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- 201000001441 melanoma Diseases 0.000 claims abstract description 3
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- 208000026310 Breast neoplasm Diseases 0.000 claims description 14
- LVTJOONKWUXEFR-UEZXSUPNSA-N protodioscin Chemical compound O([C@@H]1[C@@H](CO)O[C@H]([C@@H]([C@H]1O)O[C@H]1[C@@H]([C@H](O)[C@@H](O)[C@H](C)O1)O)O[C@@H]1CC2=CC[C@H]3[C@@H]4C[C@@H]5O[C@]([C@H]([C@@H]5[C@@]4(C)CC[C@@H]3[C@@]2(C)CC1)C)(O)CC[C@@H](C)CO[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@@H](C)[C@H](O)[C@@H](O)[C@H]1O LVTJOONKWUXEFR-UEZXSUPNSA-N 0.000 claims description 11
- QDQWGYLCDZBAMD-UHFFFAOYSA-N saponin C Natural products CC1C2C3CCC4C5(C)CCC(O)C(C)(COC6OC(CO)C(O)C(O)C6O)C5CCC4(C)C3(C)CCC27C8OC8C1(C)OC7=O QDQWGYLCDZBAMD-UHFFFAOYSA-N 0.000 claims description 11
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- 201000008275 breast carcinoma Diseases 0.000 claims description 2
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- 210000004027 cell Anatomy 0.000 description 23
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 108010010803 Gelatin Proteins 0.000 description 6
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- 239000008273 gelatin Substances 0.000 description 6
- 229920000159 gelatin Polymers 0.000 description 6
- 235000019322 gelatine Nutrition 0.000 description 6
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- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 4
- 230000033115 angiogenesis Effects 0.000 description 4
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- 230000002401 inhibitory effect Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000012679 serum free medium Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
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- 238000002347 injection Methods 0.000 description 3
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- 239000001397 quillaja saponaria molina bark Substances 0.000 description 3
- 229930182490 saponin Natural products 0.000 description 3
- 150000007949 saponins Chemical class 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241001521474 Liriope <hydrozoan> Species 0.000 description 2
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- 239000008862 fructus schizandrae, radix ginseng, radix ophiopogonis drug combination Substances 0.000 description 2
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- 235000010894 Artemisia argyi Nutrition 0.000 description 1
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- 102000002322 Egg Proteins Human genes 0.000 description 1
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- 241000196324 Embryophyta Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241001529246 Platymiscium Species 0.000 description 1
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- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
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- 244000030166 artemisia Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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Images
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the field of pharmacy, and discloses the application of rusogenin fucopyranoside C in the pharmacy. The invention also provides the application of the rusogenin fucopyranoside C in the preparation of the medicine for resisting tumor metastasis, and more particularly relates to the application in the preparation of the medicine for generating tumour new blood vessels and inhibition of relative tumor metastasis. Being proved by the study of the invention, the rusogenin fucopyranoside C can effectively inhibit the growth, migration and adhesive ability, can inhibit lung metastasis of melanoma B16, and provides new selection in the application of preparing the medicine for resisting tumor metastasis.
Description
Technical field
The invention belongs to pharmaceutical field, relate to the application of liriope muscari Baily saponin C in pharmacy.
Background technology
Liriope muscari Baily Liriope muscari (Decne.) Baily is as " one of former plant of 1995 editions newly-increased kind-Radix Liriopes of Chinese pharmacopoeia demonstrates its distinctive curative effect in secular traditional tcm clinical practice practice.But, liriope muscari Baily still concentrates on decoction pieces or compound preparation in Clinical Application, as SHENGMAI SAN, SHENMAI ZHUSHEYE etc., separately uses as clinical medicine with its extract or monomeric compound and yet there are no so far, therefore, it is furtherd investigate and develops significant.
Aspect chemical constitution study, mainly concentrate on the separation and the discriminating of the saponin component of liriope muscari Baily, since the total saponin content in the Radix Liriopes platymiscium fibrous root in quite long trophophase far above its tuber, therefore with the fibrous root of liriope muscari Baily object as research, separate from liriope muscari Baily so far and differentiated six monomer saponin constituents, one of them is liriope muscari Baily saponin C (DT13).
Up to now, go back the ideal medicine for anti transfer of tumor of neither one in the world.Tumor patient is in case occur to shift, even undergo surgery, put, chemotherapy, the existence time limit still can't be prolonged preferably.Liriope muscari Baily saponin C is not reported in the application aspect the preparation medicine for treating tumor metastasis yet.
Summary of the invention
The purpose of this invention is to provide the application of liriope muscari Baily saponin C aspect the preparation medicine for treating tumor metastasis.
Technical scheme of the present invention is the application of liriope muscari Baily saponin C aspect the preparation medicine for treating tumor metastasis, particularly the application in the preparation medicine for treating tumor metastasis relevant with tumor neogenetic blood vessels generation inhibition.Studies show that liriope muscari Baily saponin C shifts kinds of tumors such as breast carcinoma, melanomas obvious inhibitory action.
Liriope muscari Baily saponin C can make pharmaceutical composition with any one adjuvant that pharmaceutically allows among the present invention, also can with liriope muscari Baily saponin C the medicine for treating tumor metastasis composition compound preparation of antagonism not take place with other.These preparations can be any one dosage forms that pharmaceutically allows, and include but not limited to tablet, granule, pill, oral liquid, injection, membrane, capsule, liposome etc.The consumption of liriope muscari Baily saponin C can be according to variations such as route of administration, patient age, body weight, body surface area, the disease type of being treated and the orders of severity, and its daily dose can be 1mg/kg, can use by one or many.
Beneficial effect of the present invention:
The present invention discovers that DT13 can obviously suppress the growth of human breast cancer cell, move, sticks ability, finds in the body that DT13 can suppress the lung transfer of melanin tumour b16.DT13 is as a drug candidate with obvious antimetastatic activity, research is illustrated the metastasis effect of DT13 and is generated inhibiting mutual relation with new vessels, can offer reference for the development DT13 medicine for anti transfer of tumor of starting with from the tumor neogenetic blood vessels formation inhibitor, for the preparation medicine for treating tumor metastasis provides new selection.
Description of drawings
Fig. 1 is the influence of DT-13 to anaerobic condition human breast cancer cell MDA-MB-435 transfer ability.
Fig. 2 is the influence of DT-13 to anaerobic condition human breast cancer cell MCF-7 transfer ability.
Fig. 3 is the influence of DT-13 to anaerobic condition human breast cancer cell MDA-MB-231 transfer ability.
Fig. 4 is the inhibition experiment photo of DT-13 to angiogenesis.
Fig. 5 is the inhibition experiment photo of DT-13 to angiogenesis.
The specific embodiment
The invention will be further elaborated by the following examples.
The toxicity of embodiment 1DT-13 pair cell detects
(1) material:
Instrument: superclean bench (Chinese mugwort Kelin, Suzhou cleaning equipment company limited), constant temperature CO
2Incubator (German Heraeus company), enzyme-linked immunosorbent assay instrument (U.S. BIO-RAD company), inverted biological microscope (Japanese OLYMPUS company), dull and stereotyped shaking table (the bright experimental apparatus in Jiangsu Province factory).
Reagent: RPMI1640, F12 culture medium (GIBCO), trypsin SIGMA), new-born calf serum (HYCLONE), MTT (SIGMA), DMSO (SIGMA).
Cell strain: human breast cancer cell MDA-MB-435, MDA-MB-231, MCF-7.
(2) method:
1, get and be in each one bottle in cell in good condition exponential phase of growth, add 0.25% tryptic digestive juice, digestion comes off attached cell, counting 2～4 * 10
4Individual/ml, make cell suspension.
2, obtained cell suspension is inoculated on 96 orifice plates, and constant temperature CO is put in 180 μ l/ holes
2Cultivated 24 hours in the incubator.
3, change liquid, adding is subjected to reagent thing DT-13, and cultivated 48 hours in 20 μ l/ holes.
4, MTT is added in 96 orifice plates, 20 μ l/ holes, reaction is 4 hours in the incubator.
5, supernatant is removed in suction, adds DMSO, 150 μ l/ holes, and jolting is 5 minutes on the dull and stereotyped shaking table.
6, be the light absorption value that the 570nm place measures every hole with enzyme-linked immunosorbent assay instrument at wavelength, and calculate cell and suppress.
(3) result
Human breast cancer cell 435
Human breast cancer cell MCF-7
Human breast cancer cell 231
Embodiment 2DT-13 is to the influence of anaerobic condition cell adhesion ability
Purpose: under the vitro detection anaerobic condition, human breast cancer cell is to the variation of basement membrane adhesive capacity
Material: six orifice plates, 96 orifice plates, PBS, 1% gelatin, 2%BSA, serum-free medium, 10% calf serum culture fluid, Digestive system (0.25% trypsin), MTT reagent, DMSO
Method:
One, the preparation of gelatin plate:
1, take by weighing a certain amount of jelly powder, the PBS that adds heat according to the 1g/100ml amount dissolves, and clicks and enters while hot in 96 orifice plates, and 30 μ l/ holes keep flat and rock, and makes at the bottom of the gelatin tiling hole, notes not having bubble.
2, placement is spent the night, and with the gelatin sucking-off, PBS washes one time.
3, inhale after 1 hour with 2%BSA sealing and go.
4, add serum-free medium, 20～30 μ l/ holes, balance is more than 1 hour, and the back is inhaled and is gone.
Two, cell culture
1, eugonic human breast cancer cell is incubated in six orifice plates, counting 7 * 10
4～9 * 10
4, the 2ml/ hole.
2, be cultured to cell and grow to and be paved with at the bottom of the hole at 80% o'clock, remove culture medium, PBS washes one time, changes serum-free medium into, and adds and be subjected to reagent thing DT-13, and normal oxygen is cultivated.
Three, the ability of sticking is measured
1, the cell after the cultivation goes culture medium, and PBS washes one time, and digestion is centrifugal, makes serum-free cell suspension.
2, serum-free cell suspension is added in the gelatin plate of handling well, 200 μ l/ holes, each dosage is established 10 parallel holes.Blank group only adds serum-free medium.
3, cultivate after 1 hour, with syringe culture fluid is inhaled and gone, PBS washes one time, and method is for adding PBS200 μ l/ hole, and the syringe sucking-off is used in the careful piping and druming of rifle head 10 times.
4, add MTT40 μ l/ hole, hatched 4 hours.
5, discard MTT, add DMSO, 150 μ l/ holes, jolting is 5 minutes on the dull and stereotyped shaking table.
6, be the light absorption value that the 570nm place measures every hole with enzyme-linked immunosorbent assay instrument at wavelength, and calculate the cell adhesion suppression ratio:
The result:
MDA-MB-231 cell adhesion suppression ratio (%)
DT-13 10
-8mol/l 20.98
DT-13 10
-7mol/l 30.26
DT-13 10
-6mol/l 50.12
DT-13 10
-5mol/l 55.94
MDA-MB-435
DT13 10μM 58
DT13 1μM 63
DT13 0.01μM 43
MCF-7
DT13 1μM 28.6
DT13 0.1μM 37.1
DT13 10nM 59.1
DT13 1nM 81.6
Embodiment 3DT-13 is to the influence of anaerobic condition cell migration ability
Detect the cell migration ability with Boyden Chamber Transwell.This device divides inside and outside two Room.The inner room bottom is poly carbonate filter membrane (aperture 8 μ m), earlier with 1% gelatin bag quilt, the culture fluid balance 1h of reuse serum-free.Human breast cancer cell (1.5 * 10
4Individual cell/100 μ l) is suspended from the culture fluid kind that contains 2%FBS and variable concentrations test-compound and goes into inner room.The mistress adds then that 0.6ml contains or does not contain the culture fluid of VEGF (20ng/ml).37 ℃ hatch 6h after, discard culture fluid in the hole, with 90% ethanol fixed cell 10min.Wipe the cell that does not move on the inner room upper strata gently with cotton swab, 0.1% crystal violet room temperature dyeing 10min, rinse excess dyestuff with PBS, use 10% acetic acid, 100 μ l/ hole extracting 10min at last, with wavelengthtunable decline orifice plate microplate reader (VERSAmaxTM, Molecular Device Corporation, Sunnyvale, CA USA) measures the OD value under 600nm.
The results are shown in Figure 1,2,3, illustrate that DT13 can obviously suppress the migration of three kinds of breast cancer cells.
Embodiment 4DT-13 is to the inhibition experiment of angiogenesis
Experiment material: 6 age in days eggs purchase in Pukou, Nanjing, through disinfectant operating theater instruments, thick needle, curved tweezer, incubator, through filter paper (pharmacy film), the DT-13 of sterilization.
Operation:
The preparation of medicine film: the DT-13 drug level is 1*10
-3M joins on the diameter 0.5cm sterilization filter paper film, adds 30 μ l respectively, 10 μ l, and 1 μ l is with the medicine film of preparation variable concentrations.
1, with greatly head-up fixing after the 6 age in days fertilized eggs wipings;
2, puncture air chamber place eggshell with thick needle;
3, window with curved tweezer;
4, skin covering of the surface is carefully divested, choose the minimum position of blood vessel and place the medicine film;
5, wide tape seal is put in incubator behind the labelling, the observed result of taking pictures after 2 days.
The results are shown in Figure 4, the arrow place is for placing medicine film place.
Experimental result is seen Fig. 5 for the second time.Drug level is 30nmol, 20nmol, and 10nmol, 1nmol, operating process is with the experimental procedure first time.
Twice experimental result all found, compares with blank, and DT-13 just has tangible angiogenesis suppression action at 10nmol.
Embodiment 5DT-13 is to the inhibitory action of the black mice metastatic tumor of B16
Mouse melanin tumor cell B16 tail vein injection C57 mice, injection cell concentration 1*10
6Individual, the DT-13 of administration simultaneously puts to death animal in two weeks back, and number pulmonary metastasis number is added up.
| Group | Dosage mg/kg | Number of animals (beginning) | Number of animals (end) | Beginning body weight (gram) | Finish body weight (gram) | Lung weight/body weight | Lung metastasis (number) |
| Solvent | ? | 10 | 10 | 18.5±1.0 | 19.2±1.0 | 0.32±0.08 | 32.3±11.9 |
| 5—Fu | 25 | 10 | 10 | 18.4±0.8 | 17.4±0.8 | 0.15±0.02 ** | 11.0±1.1 ** |
| |
10 | 10 | 10 | 18.5±0.9 | 19.3±0.8 | 0.16±0.02 ** | 10.6±1.3 ** |
| DT13 | 5 | 9 | 9 | 18.4±0.8 | 19.8±0.7 | 0.18±0.03 * | 16.3±1.8 * |
| DT13 | 2.5 | 10 | 10 | 18.0±0.8 | 19.6±0.6 | 0.21±0.05 | 27.9±2.8 |
*: P＜0.05
*: P＜0.01, compare with the solvent control group.
DT-13 height, middle dosage group are compared with the solvent control group, and the ratio of lung weight/body weight obviously descends, and pulmonary's metastasis quantity also obviously reduces.The result shows that DT-13 has good inhibitory effect to the black mice metastatic tumor of B16.
Claims (2)
1. the application of liriope muscari Baily saponin C in the preparation medicine for treating tumor metastasis, described tumor is breast carcinoma, melanoma.
2. application according to claim 1 is characterized in that medicine for treating tumor metastasis is the tumor neogenetic blood vessels formation inhibitor.
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| CN2008101960440A CN101352448B (en) | 2008-09-11 | 2008-09-11 | Use of Liriope muscari Bailey C in pharmacy |
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|---|---|---|---|
| CN2008101960440A CN101352448B (en) | 2008-09-11 | 2008-09-11 | Use of Liriope muscari Bailey C in pharmacy |
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| CN101352448B true CN101352448B (en) | 2010-12-22 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104523738A (en) * | 2015-01-20 | 2015-04-22 | 泉州东南中药材种植有限公司 | Application of liriope muscari in preparation of radiation sensitizer |
| CN107519494B (en) * | 2016-06-16 | 2020-12-01 | 天士力医药集团股份有限公司 | Medicinal composition containing liriope muscari baily saponins C |
| CN106943419A (en) * | 2017-05-18 | 2017-07-14 | 天津医科大学 | DT-13 is preparing the application for the treatment of prostate cancer medicine |
| CN110652521A (en) * | 2019-10-28 | 2020-01-07 | 西南大学 | Application of liriope muscari baily saponins C in preparation of acute myeloid leukemia resisting medicine |
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2008
- 2008-09-11 CN CN2008101960440A patent/CN101352448B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| 培奇.抗肿瘤转移药物的研究现状.《上海医药情报研究》.1996,(第1期),1-6. * |
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