CN1981749B - Use of beta-elemene for inhibiting cerebrovascular endothelial cell - Google Patents
Use of beta-elemene for inhibiting cerebrovascular endothelial cell Download PDFInfo
- Publication number
- CN1981749B CN1981749B CN 200510130367 CN200510130367A CN1981749B CN 1981749 B CN1981749 B CN 1981749B CN 200510130367 CN200510130367 CN 200510130367 CN 200510130367 A CN200510130367 A CN 200510130367A CN 1981749 B CN1981749 B CN 1981749B
- Authority
- CN
- China
- Prior art keywords
- elemene
- beta
- tumor
- cell
- endothelial cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
An application of beta-elemene in suppressing the cerebrovascular endothelial cells with high selectivity and in turn the cerebral tumor and cerebral metastatic tumor, and treating the cerebral tumor and cerebral metastatic tumor is disclosed.
Description
Technical field
The present invention relates to the pharmaceutical chemistry field, particularly, the present invention relates to the new medicinal usage of plant extract beta-elemene.
Background technology
The cerebral tumor at the cerebral glioma the most commonly of being grown up, is the highest malignant tumor of sickness rate in the intracranial tumor wherein; Because it is low to chemosensitivity; In the clinical treatment, mainly adopt excision, the postoperative NACT; Have only patient's muscle power situation better, then can adopt medicines such as chloroethene nitrous gland, hexamethylene nitrous gland, first ring nitrous gland.In recent years, with Rhizoma Dysosmae Versipellis class medicine of new generation, for example the VM-26 treatment also obtains certain curative effect, and still, generally speaking, normally 3-6 month catabasis, prognosis is very abominable.
As for metastatic encephaloma for example the vertigo of tumor such as pulmonary carcinoma, breast carcinoma move, generally can regard the performance of advanced tumor as.Obtain certain curative effect with radiotherapy now, and chemotherapy only is as auxiliary treatment, under the general situation in order of patient, carries out just now that curative effect is not remarkable, effective percentage is low, and the catabasis is short.
Elemene is existing for many years in clinical practice, and have the clinical trial of strict contrast to prove: except that ascites pleural fluid and the shallow table property tumor intratumor injection, elemene is not obvious to the solid tumor curative effect at other each positions.Clinical research in recent years shows, elemene is finished up to 46% the effective percentage (CR+PR) of cerebral glioma glioma, and metastatic encephaloma has also been obtained similar result.And after patient uses repeatedly, there is not to show drug resistance to this medicine yet.And beta-elemene is the main active ingredient of elemene, and its preparation method and Anticancer Activities be existing report in document.
Summary of the invention
The inventor discovers that beta-elemene has the good restraining effect to cerebrovascular endothelial cell, and has high selectivity; Discover that further beta-elemene is through the high selectivity inhibitory action to cerebrovascular endothelial cell, the growth and the vertigo that can effectively suppress the cerebral tumor move growth of tumor.Therefore, we find that beta-elemene can be used as medicine, is used to suppress the growth of cerebrovascular endothelial cell, and then the growth and the vertigo that are used to suppress the cerebral tumor move growth of tumor, so have accomplished the present invention.
A kind of new purposes that the purpose of this invention is to provide beta-elemene promptly is used to suppress the purposes of cerebrovascular endothelial cell;
Further aim of the present invention has provided the purposes that beta-elemene is used to prepare the medicine of treating the cerebral tumor;
Further aim of the present invention has provided beta-elemene and has been used to prepare the purposes that the treatment vertigo moves the medicine of tumor.
Beta-elemene of the present invention can use separately also can with the other medicines compatibility, process pharmaceutical composition, be used for the cerebral tumor, especially glioma brain tumour, and metastatic encephaloma, especially lung cancer with brain metastasis tumor.This pharmaceutical composition can adopt common dosage form, like forms such as injections.
The specific embodiment
Further specify the present invention through embodiment below.It should be understood that embodiments of the invention are to be used to explain the present invention rather than limitation of the present invention.The simple modifications that essence according to the present invention is carried out the present invention all belongs to the present invention and requires the scope protected.
One, the extracorporeal anti-tumor function of beta-elemene
1, experiment purpose
Mtt assay measuring beta-elemene is to the cytotoxicity of multiple animal and human body tumour cell strain
2, instrument and equipment
2.1 CO
2Cell culture incubator, PHARMA SCIENTIFIC
2.2 enzyme-linked immunosorbent assay instrument, Labsystems, wellscan, MK.2
2.3 the vertical double superclean bench of single face, Suzhou Decontamination Equipment Plant
2.4 inverted biological microscope, 37XB/37 * BTV, Shanghai optical instrument six factories
3, experiment material and preparation
3.1 human tumor cranial glia oncocyte (SHG44), stomach cancer cell (MGC), human liver cancer cell (QGY), leukaemia (HL-60), lung carcinoma cell (LAX), murine melanoma (K111), human breast cancer cell strain (Bcap-37), the former leukemia of the chronic marrow of people (K562), National People's Congress's cell lung cancer (NCI-H460).
3.2RPMI (1640) cell culture medium (GIBCO): contain 10% deactivation newborn calf serum (Saida biological Pharmaceutical Co., Ltd., Shanghai city); The L-glutaminase (the import packing, SANGON); Sodium Pyruvate; 1 * 10
5U.L
-1Penicillin, 100mg.L
-1Streptomycin; Aseptic filtration, 4 ℃ of preservations.
3.3 0.25% trypsin solution (Trypsin): available from Invitrogen company ,-20 ℃ of preservations.
3.4 phosphate buffer (PBS): NaCl8g, KCl0.2g, Na
2HPO
41.15g, KH
2PO
40.2g, be dissolved in the 1L distilled water, 121 ℃ of autoclave sterilization 20min, 4 ℃ of preservations.
3.5MTT (AMRESCO) solution: be made into 5mg/ml solution with PBS.
3.6 lysate: every 100ml deionization distilled water contains SDS10g, isobutanol 5ml, concentrated sulphuric acid 0.12ml.
4, experimental technique
Beta-elemene records through mtt assay the cytotoxicity of above-mentioned tumor cell line.Concrete steps are following:
4.1 sample preparation: beta-elemene is dissolved in the dimethyl sulfoxine, obtains the solution that concentration is 10mg/ml.Reuse PBS makes gradient dilution, obtains the dilute sample that concentration is respectively 500 μ g/ml, 50 μ g/ml, 5 μ g/ml, 0.5 μ g/ml, 0.05 μ g/ml.
Add in flat 96 orifice plates 4.2 will dilute good sample, every hole 10 μ l make two parallel testings at every.The corresponding work of DMSO added in the entering plate behind the gradient dilution, as contrast.
4.3 get the cell that is in exponential phase, cell is suspended in the RPMI-1640 culture medium that contains 10% calf serum after trypsinization and washing, through the blue dyeing of Placenta Hominis exclusive method meter viable count, and regulates cell suspending liquid density to 2 * 105 cells/ml.
4.4 in flat 96 orifice plates, every hole adds 90 microlitre cells, overnight incubation in 37 ℃, 5%CO2 cell culture incubator.
4.5 flat 96 orifice plates that will add cell insulation 48 hours in 37 ℃, 5%CO2 cell culture incubator.
4.6 add 10 μ l 5mg/mlMTT solution in every hole, continue in incubator, to be incubated 3～4 hours.
4.7 every hole adds 100 μ l DMSO, continues incubated overnight in incubator, and the first hairpin crystal of generation is fully dissolved.Measure the 492nm absorbance value.
Handle back cell relative survival rate 4.8 calculate beta-elemene according to absorbance value.Computing formula is following:
4.9 through the IC50 of Xlift computed in software beta-elemene to each tumor cell.
5, experimental result
As shown in table 1, the IC of tumor cell that beta-elemene is tested
50All be equal to or greater than 50 μ g/ml, to the IC of periphery vascular endothelial cell
50All at 30--60 μ g/ml, to the IC of cerebrovascular endothelial cell
50At 0.6--0.8 μ g/ml.
Table 1, beta-elemene are to the growth in vitro inhibitory action of various tumor cell strains
| Cell strain | IC50(μg/ml) |
| Bcap37 | ＞100 |
| Bcap37 | ＞100 |
| K111 | 50.58 |
| K562 | ＞100 |
| K562 | ＞100 |
| H460 | ＞100 |
| H460 | ＞100 |
| SHG-44 | 96.56 |
| SHG-44 | 64.59 |
| MGC | ＞100 |
| MGC | ＞100 |
| QGY | 63.69 |
| QGY | 70.16 |
| HL-60 | 51.13 |
| HL-60 | 68.73 |
| LAX | 88.38 |
| LAX | 91.40 |
6, conclusion
Beta-elemene shows there is not obvious antineoplastic basically external to the animal and human's body tumor result who is tested.
Two, beta-elemene antitumor zoopery result
1, receive the reagent thing:
1.1 title: beta-elemene emulsion injection.Corresponding blank Emulsion.
1.2 the unit of providing: Dalian Yuanda Pharmaceutical Co., Ltd
1.3 lot number: 021120
1.4 content, preparation labelled amount: contain beta-elemene 10mg/ml.
1.5 compound method: accurately measure beta-elemene breast and add corresponding blank Emulsion and be diluted to desired concn and get final product.Administration volume 0.5ml/20g Mus.
2, experiment material:
2.1 solvent: corresponding blank Emulsion.
2.2 positive reference substance: Cyclophosphamide for injection, Hua Lian, Shanghai pharmacy group produces, every day intraperitoneal administration once, continuous seven days.
2.3 tumor source: human tumor pulmonary carcinoma A549, human hepatocellular QGY, human tumor ovarian cancer ao10/17, human body carcinoma of prostate PC-3M, human tumor intestinal cancer HCT-8, human body breast carcinoma Bcap-37, human tumor glioma brain tumour SHG44 nude mouse inner model and mouse brain glioma G422 model; Human tumor glioma brain tumour SHG44 cell, MGC stomach cancer cell, QGY HCC, HL-60 leukaemia and LAX lung carcinoma cell go down to posterity by pharmacological room of Shanghai Institute of Pharmaceutical Industry and keep.
3 laboratory animals:
3.1 the source: nude mouse is provided by Shanghai Chinese Academy of Sciences Experimental Animal Center, the quality certification number: Shanghai is moving closes the card word No. 005.Kunming mice the court Animal House is supplied with, the quality certification number: Shanghai is moving closes the card word No. 107.
3.2 body weight: nude mouse was 6 ages in week.Mice 18-20 gram.
3.3 sex: male and female all can, same sex is used in every batch of experiment.
3.4 number of animals: every group of 6 mices of test group and positive controls nude mouse, every group of 10 mices of kunming mice, negative control respectively is two groups.
4 dosage settings: Elemenum Emulsion 50,25 and 12.5mg/kg time.
5 dosage regimens: intravenous administration (i.v), every day 2 times (B.i.d), continuous 10 days, administration was 20 times altogether.
6 experimental control: negative control is given and the isocyatic corresponding blank Emulsion of test group high dose equal-volume, and dosage regimen is intravenous route, every day 2 times, continuous 10 days.Positive control cyclophosphamide CTX 30mg/kg, (q.d) once a day, lumbar injection (i.p), continuous seven days.In vitro tests positive control mitomycin for inj, (Kyowa Hakkokogyo Co., Ltd's product).
7 test key steps:
7.1 axil subcutaneous vaccination model: get eugonic tumor source under the aseptic condition, be equipped with written treaty 1 * 10 with the homogenate legal system
7/ ml cell suspension in the every Mus of nude mouse axil subcutaneous vaccination 0.2ml/, is pressed the administration of experimental design scheme next day, puts to death each treated animal about three weeks, cuts open to get tumor and weigh, and calculates tumor control rate by following formula:
Tumor control rate %=[(the average tumor of the average tumor weight-administration of matched group group is heavy)/average tumor of matched group is heavy] * 100
Human tumor heteroplastic transplantation model operation is identical, but the apparatus of used feedstuff, bedding and padding, cage tool and contact etc. all should use behind the autoclave sterilization, and nude mice places laminar-flow rack to raise.
7.2 intracranial inoculation animal tumor model: get eugonic G-422 or SHG44 tumor source, the homogenate legal system is equipped with written treaty 2 * 10 under the aseptic condition
7/ ml cell suspension, the every Mus intracranial of corresponding host inoculation 0.05ml press the administration of experimental design scheme next day, observes life cycle in each treated animal 30 days, tries to achieve the The average survival time natural law of each group, with the negative control group comparison, obtains increase in life span by following formula.
Increase in life span=[(matched group The average survival time natural law-administration group The average survival time natural law)/matched group The average survival time natural law] * 100
8 result of the tests:
In the body experiment antitumor with 50,25,12.5mg/kg/ time, high, medium and low three dosed administrations of iv * 10 bid schemes are 43.08%, 40.00%, 31.28% to A549 lung cancer model subcutaneous vaccination experimental result.To nude mouse glioma SHG-44G cancer model subcutaneous vaccination experimental result is 44.89%, 39.02%, 32.20%.To QGY liver cancer model subcutaneous vaccination experimental result is 45.89%, 37.20,30.92%.To mice glioma G422 model subcutaneous vaccination experimental result is 44.62%, 40.00%, 32.31%.To human tumor ovarian cancer ao10/17 model subcutaneous vaccination experimental result is 46.42%, 36.25%, 31.08%.To carcinoma of prostate PC-3M model subcutaneous vaccination experimental result is 46.58%, 34.78%, 27.95%%.To intestinal cancer HCT-8 model subcutaneous vaccination experimental result is 54.20%, 46.01%, 35.10%.To breast carcinoma Bcap-37 model subcutaneous vaccination experimental result is 48.08%, 43.23%, 33.08%.
With 60,30,15,7.5mg/kg/d, high, medium and low three dosed administrations of iv * 10qd scheme, to mice glioma G422 in-situ inoculating model, the increase in life span experimental result is 162.84%, 154.10%, 143.17%, 134.97%.To nude mouse glioma SHG-44 in-situ inoculating model, the increase in life span experimental result is 160.36%, 148.69%, 139.31%, 128.72%.With 60,30,15,7.5mg/kg/d, high, medium and low three dosed administrations of it * 10qd scheme, to mice glioma G422 subcutaneous vaccination model, the tumour inhibiting rate experimental result is 60.89%, 47.11%, 32.00%.To nude mouse glioma SHG-44 subcutaneous vaccination model, the tumour inhibiting rate experimental result is 63.24%, 51.35%, 37.83%.To murine sarcoma S180 subcutaneous vaccination model, the tumour inhibiting rate experimental result is 42.35%, 23.47%, 4.59%.To Mice Bearing Lewis Lung Cancer model subcutaneous vaccination model, the tumour inhibiting rate experimental result is 41.74%, 19.92%, 10.59%.
Table 2 beta-elemene breast is to the clinical trial of human body pulmonary carcinoma A549 (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 3 beta-elemene breast is to the clinical trial of human hepatocellular QGY (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 4 beta-elemene breast is to the clinical trial of human body ovarian cancer ao10/17 (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 5 beta-elemene breast is to the clinical trial of human body carcinoma of prostate PC-3M (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 6 beta-elemene breast is to the clinical trial of human body intestinal cancer HCT-8 (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 7 beta-elemene breast is to the clinical trial of human body breast carcinoma Bcap-37 (subcutaneous vaccination)
* *P value＜0.01 is compared with negative control group, and following table together
Table 8 beta-elemene breast is to the clinical trial of nude mouse glioma SHG-44 (subcutaneous vaccination)
Table 9 beta-elemene breast is to the clinical trial of mouse brain glioma G422 (subcutaneous vaccination)
Table 10 β--elemene emulsion injection is to the curative effect of S180 (axil subcutaneous vaccination)
Compare with negative control group
* *P＜0.01
Table 11 β--elemene emulsion injection is to the curative effect of Lewis (axil subcutaneous vaccination)
Compare with negative control group
* *P＜0.01
Table 12 β--elemene emulsion injection is to the clinical trial of G422 (axil subcutaneous vaccination)
Compare with negative control group
* *P＜0.01
Table 13 β--elemene emulsion injection is to the curative effect of SHG-44G (axil subcutaneous vaccination)
Compare with negative control group
* *P＜0.01
The above results shows that tumor inoculation is subcutaneous in axil, intravenous administration; Need very high dosage just to show the antitumor action that it is slight, even, the axil subcutaneous vaccination; When topical promptly produces very high concentration in the tumor in tumor, also only when 60mg/kg.d, just produce light to moderate antitumor action, detect the result of beta-elemene intravenous injection bleeding from anus Chinese medicine concentration and tissue distribution with gas chromatography; Show that all beta-elemene is no matter in blood or in tissue; Drug concentrations all is very low, and promptly in this concentration, beta-elemene is the concentration that is far from reaching its antitumous effect.
Table 14 beta-elemene breast is to the clinical trial of mouse brain glioma G422 (intracranial inoculation)
Annotate:
* *P value＜0.01 is compared with negative control group, and following table together
By the requirement of antitumor drug effect rules, increase in life span is effective more than 125%
Table 15 beta-elemene breast is to the clinical trial of people's glioma brain tumour SHG44 (intracranial inoculation)
Annotate:
* *P value＜0.01 is compared with negative control group, and following table together
By the requirement of antitumor drug effect rules, increase in life span is effective more than 125%
Table 16 beta-elemene is to the curative effect of LEWIS pulmonary carcinoma intracranial inoculation
Table 17 beta-elemene is to the curative effect of S180 tumor intracranial inoculation
Relatively The above results can obtain:
The inoculation of mouse brain glioma G-422 intracranial, intravenously administrable totally 10 days, low dosage 7.5mg/kg increase in life span reaches 134%, the inoculation of human glioma SHG-44 nude mice intracranial, intravenously administrable totally 10 days still is effective.All kinds of tumors comprise the subcutaneous vaccination of glioma nude mice axil, intravenously administrable one day twice totally ten days, in its low dosage dosage actual be 25mg/kg, 50mg/kg.Effect only just appears when high dose (100mgkg/d).It is thus clear that the beta-elemene olive to the solid tumor at other each positions comprise the cerebral tumor in the curative effect of the inoculation of non-brain far away from the effect of beta-elemene to the cerebral tumor and metastatic encephaloma.
For this peculiar phenomenon, at other cancer therapy drugs, for example cyclophosphamide amycin etc. never occurs.And those fat-soluble higher cancer therapy drugs, VM-26 etc. for example, although intracranial tumor is produced effect preferably, to the tumor efficiency of axil subcutaneous vaccination even better, and toxicity is very big.Therefore, the effect of the anti-brain tumor of beta-elemene is very unique.
Three, the anti-cerebral tumor pharmacokinetics of beta-elemene experimental result
(X ± S) (n=5) of the pharmacokinetic parameter of beta-elemene after the administration of table 18 rat intravenous injection
((the n=5) (dosage: 100mg/kg) of X ± S) of the pharmacokinetic parameter of beta-elemene after the administration of table 19 rats by intraperitoneal injection
Tissue/the blood plasma of beta-elemene is than (concentration) (n=5) (dosage: 100mg/kg) after the administration of table 20 rat vein
It is lower that pharmacokinetics inspection is found behind beta-elemene vein or the lumbar injection at blood level; It is also very low in brain, to distribute; Although after in the entering brain, checkout time is longer, no matter at tissue or in blood; Concentration does not far reach the antitumous effect at testing in vitro, and this can explain that the beta-elemene intravenously administrable is to the relatively poor reason of the tumor efficiency of axil subcutaneous vaccination.
Four, beta-elemene rats by intraperitoneal injection 1 month influences brain
Rats by intraperitoneal injection was established three dosage in one month; Promptly 7.5,15 and 30mg/Kg; Three dose groups compare with blank breast, 10% glucose group, and the result finds that 30mg/Kg and 15mg/Kg group respectively have 6 and 1 rats death, reason during administration be that ascites and abdominal adhesions appear in high dose group all animals and 3 rats of middle dose groups; 7.5mg/Kg toxic reaction does not appear in dose groups; Each dose groups hematology, serum biochemistry and urine all do not have obvious variation, explain that beta-elemene does not all have obvious influence to internal organs such as liver, kidneys under certain dosage, but local excitation are more serious.
Rat abdominal cavity administration 1 month, in the dose groups, flaky denatured areas appears in brain in high dose and minority, and doubting is that cerebrovascular disease is caused.Repeat to study for this phenomenon and still obtain identical result.Cerebral tissue to pathological changes carries out electron microscopic examination, and it is caused by cerebrovascular disease further pointing out these flaky denatured areas.
These pathological changes that come across brain all do not occur at each internal organs of whole body, do not see that the blood vessel of other each internal organs has similar pathological change yet.
We have carried out section of successive arrow and crown section to one month brain of beta-elemene high dose rats by intraperitoneal injection again, study and have been found that beta-elemene has region characteristic in IC influence, and XIAONAO, Hippocampus are unaffected.The left side brain is affected than right side brain more easily.
Under the long-time beta-elemene effect of heavy dose, think that according to histology and electron microscopic examination variation has taken place the rat intracranial vessel, this variation can cause blood supply disorder, so cause characteristic pathological changes in the brain.
Five, the extracorporeal blood vessel endotheliocyte is cultivated and to beta-elemene sensitivity
The result show cerebrovascular endothelial cell to beta-elemene than responsive tens of times of the vascular endothelial cell at other positions, and more responsive more than 100 times than general tumor cell.
1, beta-elemene experiment in vitro
1.1 experiment purpose: mtt assay measuring B-elemene is to the cytotoxicity of multiple animal and human body tumour cell strain
1.2 instrument and equipment
(1) CO
2Cell culture incubator, PHARMA SCIENTIFIC
(2) enzyme-linked immunosorbent assay instrument, Labsystems, wellscan, MK.2
(3) the vertical double superclean bench of single face, Suzhou Decontamination Equipment Plant
(4) inverted biological microscope, 37XB/37 * BTV, Shanghai optical instrument six factories
1.3 experiment material and preparation
(1) human umbilical vein endothelial cell (ECV-304)
Rat brain vascular endothelial cell and rat aorta endotheliocyte, by this laboratory former be commissioned to train foster.
(2) RPMI (1640) cell culture medium (GIBCO): contain 10% inactivated fetal bovine serum; The L-glutaminase (the import packing, SANGON); Sodium Pyruvate; 1 * 10
5U.L
-1Penicillin, 100mg.L
-1Streptomycin; Aseptic filtration, 4 ℃ of preservations.
(3) 0.25% trypsin solutions (Trypsin): available from Invitrogen company ,-20 ℃ of preservations.
(4) phosphate buffer (PBS): NaCl8g, KCl0.2g, Na
2HPO
41.15g, KH
2PO
40.2g, be dissolved in the 1L distilled water, 121 ℃ of autoclave sterilization 20min, 4 ℃ of preservations.
(5) MTT (AMRESCO) solution: be made into 5mg/ml solution with PBS.
(6) lysate: every 100ml deionization distilled water contains SDS10g, isobutanol 5ml, concentrated sulphuric acid 0.12ml.
1.4 experimental technique
Beta-elemene records through mtt assay the cytotoxicity of above-mentioned vascular endothelial cell.Concrete steps are following:
(1) sample preparation: beta-elemene is dissolved in the dimethyl sulfoxine, obtains the solution that concentration is 10mg/ml.Reuse PBS makes gradient dilution, obtains the dilute sample that concentration is respectively 500 μ g/ml, 50 μ g/ml, 5 μ g/ml, 0.5 μ g/ml, 0.05 μ g/ml.
(2) will dilute good sample and add in flat 96 orifice plates, every hole 10 μ l make two parallel testings at every.The corresponding work of DMSO added in the entering plate behind the gradient dilution, as contrast.
(3) get cultured rat aorta endotheliocyte of former generation, cerebrovascular endothelial cell and people's huve cell strain EV304; After trypsinization and washing, be suspended in the RPMI-1640 culture medium that contains 10% peptide Ox blood serum; Through trypan blue dyeing exclusive method meter viable count, and regulate cell suspending liquid density to 2 * 10
5Cell/ml.
(4) in flat 96 orifice plates, every hole adds 90 microlitre cells, overnight incubation in 37 ℃, 5%CO2 cell culture incubator.
Flat 96 orifice plates that (5) will add cell insulation 48 hours in 37 ℃, 5%CO2 cell culture incubator.
(6) add 10 μ l 5mg/mlMTT solution in every hole, continue in incubator, to be incubated 3～4 hours.
(7) every hole adds 100 μ l DMSO, continues incubated overnight in incubator, and the first hairpin crystal of generation is fully dissolved.Measure the 492nm absorbance value.
(8) calculate beta-elemene according to absorbance value and handle back cell relative survival rate.Computing formula is following:
(9) through the IC50 of Xlift computed in software beta-elemene to each tumor cell.
1.5 experimental result
Table 21, beta-elemene are to the growth in vitro inhibitory action of vascular endothelial cell
| Cell strain | IC50(μg/ml) |
| ECV-304 | 44.69 |
| ECV-304 | 37.80 |
| The rat heart vascular endothelial cell | 56.97 |
| The rat heart vascular endothelial cell | 67.11 |
| The rat brain vascular endothelial cell | 0.82 |
| The rat brain vascular endothelial cell | 0.68 |
Shown in table 21, the beta-elemene test is to the IC of periphery vascular endothelial cell
50All at 30--70 μ g/ml, to the IC of cerebrovascular endothelial cell
50At 0.6--0.8 μ g/ml.
Carry out external medicament sensitivity test with mtt assay and show, cerebrovascular endothelial cell to the sensitivity of beta-elemene much larger than other cells.
Comparative result obtains:
Rat is with 60mg/kg dosage lumbar injection beta-elemene after three days, and 1) tail vein injection AM Bison (5mg/kg), after administration, got brain in 4 hours, the HPLC method is measured its medicament contg; 2) tail vein injection vincristine sulfate liposome (1mg/kg) was got brain in 0.5 hour after administration, the HPLC method is measured its medicament contg.
Mensuration result shows, cerebral tissue Chinese medicine content and relatively not changing without the beta-elemene injection.
The vascular endothelial cell and the vascular endothelial cell in the normal condition undertissue of In vitro culture growth are different, and this growth situation rapidly is similar to the vascular endothelial cell in the tumor tissues.Therefore, the situation of the cerebrovascular endothelial cell during to the similar to a certain extent cerebral tumor of sensitivity (intracranial primary tumor or metastatic tumor) of beta-elemene during the cerebrovascular endothelial cell In vitro culture.We think why the cerebral tumor produces goodish curative effect to beta-elemene; Be because beta-elemene has suppressed the cerebral tumor vascular endothelial cell in the active growth; Thereby reach inhibition, finally cause cerebral tumor blood supply obstacle, reach the effect of gripping the cerebral tumor cerebral tumor blood vessel.
This experiment shows that still beta-elemene after short application use, does not influence blood brain barrier, and vincristine and amphotericin B are not all increased their concentration in brain.Therefore, infer that beta-elemene seems still different to the effect of intracranial malignant tumor vascular endothelial cell and the effect of general cytotoxicity.
We have the responsive especially rake point of a pair of beta-elemene according to above results presumption intracranial malignant tumor blood vessel, and the vascular endothelial cell of this rake point at tumor or other positions is not exist or distribute few.
Claims (1)
1. beta-elemene is used for treating the purposes of the medicine of lung cancer with brain metastasis tumor in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510130367 CN1981749B (en) | 2005-12-13 | 2005-12-13 | Use of beta-elemene for inhibiting cerebrovascular endothelial cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510130367 CN1981749B (en) | 2005-12-13 | 2005-12-13 | Use of beta-elemene for inhibiting cerebrovascular endothelial cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1981749A CN1981749A (en) | 2007-06-20 |
| CN1981749B true CN1981749B (en) | 2012-12-05 |
Family
ID=38164783
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510130367 Active CN1981749B (en) | 2005-12-13 | 2005-12-13 | Use of beta-elemene for inhibiting cerebrovascular endothelial cell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1981749B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105998019A (en) * | 2016-06-29 | 2016-10-12 | 青岛云天生物技术有限公司 | Medicine composition for treating brain glioma and application thereof |
| CN107308140B (en) * | 2017-08-04 | 2019-12-24 | 石药集团远大(大连)制药有限公司 | Application of beta-elemene in preparation of medicine for treating ewing sarcoma and inhibition of phosphorylation of insulin receptor of tumor cells |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1200266A (en) * | 1997-04-14 | 1998-12-02 | 中国科技开发院医药科技开发所 | Method for preparing anti-cancer medicine using beta-elemene as main effective ingredient, and components of the medicine |
| CN1244389A (en) * | 1998-08-12 | 2000-02-16 | 大连科宇药业科技开发有限公司 | Elemi olefine injecta and its preparation |
| CN1413580A (en) * | 2002-09-18 | 2003-04-30 | 中国远大国际集团有限公司 | Elemene antitumor medicine and its preparation method |
-
2005
- 2005-12-13 CN CN 200510130367 patent/CN1981749B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1200266A (en) * | 1997-04-14 | 1998-12-02 | 中国科技开发院医药科技开发所 | Method for preparing anti-cancer medicine using beta-elemene as main effective ingredient, and components of the medicine |
| CN1244389A (en) * | 1998-08-12 | 2000-02-16 | 大连科宇药业科技开发有限公司 | Elemi olefine injecta and its preparation |
| CN1413580A (en) * | 2002-09-18 | 2003-04-30 | 中国远大国际集团有限公司 | Elemene antitumor medicine and its preparation method |
Non-Patent Citations (6)
| Title |
|---|
| 史淑荣等.颈动脉灌注榄香烯治疗脑肿瘤.吉林中医药22 4.2002,22(4),第28页讨论部分. |
| 史淑荣等.颈动脉灌注榄香烯治疗脑肿瘤.吉林中医药22 4.2002,22(4),第28页讨论部分. * |
| 王怀瑾等.榄香烯治疗脑转移癌的临床疗效观察.大连医科大学学报21 2.1999,21(2),第113页表1. |
| 王怀瑾等.榄香烯治疗脑转移癌的临床疗效观察.大连医科大学学报21 2.1999,21(2),第113页表1. * |
| 窦桂荣等.榄香烯对脑神经胶质细胞瘤抑制的实验研究.天津医科大学学报4 1.1998,4(1),第10-11页讨论部分. |
| 窦桂荣等.榄香烯对脑神经胶质细胞瘤抑制的实验研究.天津医科大学学报4 1.1998,4(1),第10-11页讨论部分. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1981749A (en) | 2007-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Sau et al. | Combination of cationic dexamethasone derivative and STAT3 inhibitor (WP1066) for aggressive melanoma: a strategy for repurposing a phase I clinical trial drug | |
| Momose et al. | Mitochondrial inhibitors show preferential cytotoxicity to human pancreatic cancer PANC-1 cells under glucose-deprived conditions | |
| CN108939080A (en) | The purposes of composition containing tyrosine kinase inhibitor and resveratrol in the preparation of antitumor drugs | |
| CN109091480A (en) | Cancer composition 10-hydroxycamptothecine and gram azoles are for Buddhist nun's treatment lung cancer and purposes | |
| CN1981749B (en) | Use of beta-elemene for inhibiting cerebrovascular endothelial cell | |
| CN102614170A (en) | Application of artemisinin B in preparation of antitumor drugs | |
| CN102688489B (en) | Pharmaceutical composition containing triptolide, triptolide derivative and Bc1-2 inhibitor and application thereof | |
| WO2022188491A1 (en) | Tumor chemotherapy pharmaceutical composition | |
| CN103992394B (en) | The cationic peptide of a kind of Prof. Du Yucang and the purposes in preparing antineoplastic thereof | |
| CN104892707B (en) | A kind of controlled syntheses compound CLCN and its application in medicines resistant to liver cancer | |
| CN105213366A (en) | The medical usage of guttiferone compound and pharmaceutical composition thereof | |
| CN107286123A (en) | A kind of preparation method of dibenzofuran class compound and application | |
| CN107951888A (en) | The drug regimen and its application of Afatinib and 10-hydroxycamptothecine | |
| CN107412736B (en) | Anti-tumor combined medicine and application thereof in preparing anti-cancer medicine | |
| CN104055786B (en) | The application in preparation preventing and treating tumour medicine of medicagenic acid-3-O-β-D-pyranglucoside, medicagenic acid and salt thereof | |
| CN110893192B (en) | Pharmaceutical composition for treating nasopharyngeal carcinoma | |
| CN104623215A (en) | Anti-tumor medicine composition | |
| CN102198139B (en) | Pharmaceutical composition with antineoplastic effect, and application method thereof | |
| JI et al. | Nimotuzumab with cisplatin or fluorouracil on human esophageal squamous cell carcinoma EC1 cells. | |
| CN111419836B (en) | Use of 12 alpha-methoxy-germacrane-triene-12, 6 alpha-acetal for combating human gliomas | |
| CN105517558A (en) | Filipendula vulgaris extract and uses thereof | |
| CN103833766B (en) | Shore, Agra dimethylamine fumarate and the purposes in prepared by medicine thereof | |
| CN109200050A (en) | Application of the alkaloid in terms of reversing lung cancer cisplatin-resistant | |
| CN103948590A (en) | NEDD8 (neural precursor cell-expressed developmentally downregulated 8)activating enzyme inhibitor and medical application thereof | |
| CN104003998A (en) | Oridonin 14-0-sustituted nitrogen mustard derivatives, and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |