CN104208073A - Application of protopanaxadiol to prepare tumor multidrug resistance reversers - Google Patents
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Abstract
The invention belongs to the field of medicines, relates to application of protopanaxadiol to prepare anti-tumor medicines, and especially relates to application of protopanaxadiol to prepare a tumor multidrug resistance reverser. Protopanaxadiol is a natural product, has effect of reversing the multidrug resistance of tumor cells, and can be used as a reverser of tumor multidrug resistance. Protopanaxadiol also has effects of increasing sensitivity of tumor multidrug resistant cells to anti-tumor medicines, and can be used as a chemotherapy sensitizer. The invention also provides a usage method for a pharmaceutical composition formed by combined usage of an antitumor medicine and protopanaxadiol and used for inhibiting proliferation of tumor multidrug resistant cells. The small molecule compound protopanaxadiol, which is developed as a new antitumor medicine or an auxiliary composition of a new antitumor medicine, is obvious in tumor-inhibiting effect, green and environment-friendly, and provides a new approach and means for treating and curing tumors.
Description
Technical field
The invention belongs to the medicine of reverse multiple drug resistance of tumor and enhanced sensitivity antitumor drug, be specifically related to triterpene glucoside unit---the novelty teabag of protopanoxadiol, the i.e. application of protopanoxadiol in preparation tumor multiple medicines inversion agent and antitumor drug sensitizer.
Background technology
The life and health of the malignant tumor serious harm mankind, has become the lethal one of the main reasons of dying of current disease.The essential therapeutic arsenals of current malignant tumor has operative treatment, radiation and chemotherapy etc.For being diagnosed as the patient of late tumor and the patient of transfer having occurred, chemotherapy is one and selects preferably.Clinician obtains maximum curative effect and minimum toxicity by regulating dosage to optimize, and helps patient extend life span and improve prognosis quality of life.But the generation of the drug resistance of tumor cell causes tumor cell to decline to the sensitivity of chemotherapeutics, greatly limit the treatment of chemotherapy effect [Dean, M., T. Fojo, and S. Bates,
tumour stem cells and drug resistance.nat Rev Cancer, 2005.
5(4): p. 275-84.].Tumor cell is to after certain drug resistant, can to the raw crossing drug resistant of never contacted, that structure different, mechanism of action is different Treated with Chemotherapeutic Drugs produce, this phenomenon is called the multidrug resistance (Multiple Drug Resistance, MDR) of tumor.The generation of multidrug resistance be one of great difficult problem faced of current cancer therapies [H, L.,
an overview of cancer multidrug resistance: a still unsolved problem.cell. Mol. Life Sci., 2008.
65: p. 3145 – 3167. Mellor, H.R. and R. Callaghan,
resistance to chemotherapy in cancer:a complex and integrated cellular response.pharmacology, 2008.
81(4): p. 275-300. Mimeault, M., R. Hauke, and S. Batra,
recent Advances on the Molecular Mechanisms Involved in the Drug Resistance of Cancer Cells and Novel Targeting Therapies.clin Pharmacol Ther., 2008.
83: p. 673 – 691.].MDR, once produce, is invalid by strengthening drug dose, can produce stronger toxicity and further inducibly resistant generation [Akan, I., et al.,
n-acetylcysteine enhances multidrug resistance-associated protein 1 mediated doxorubicin resistance.eur J Clin Invest, 2004.
34(10): p. 683-9. Akan, I., et al.,
multidrug resistance-associated protein 1 (MRP1) mediated vincristine resistance:effects of N-acetylcysteine and Buthionine sulfoximine.cancer Cell Int, 2005.
5(1): p. 22. Choi, C.H.,
aBC transporters as multidrug resistance mechanisms and the development of chemosensitizers for their reversal.cancer Cell Int, 2005.
5: p. 30. Liscovitch, M. and Y. Lavie,
cancer multidrug resistance:a review of recent drug discovery research.iDrugs, 2002.
5(4): p. 349-55.].Most medicine relevant with MDR is all hydrophobicity or amphipathic natural product, as taxanes, anthracycline antibiotics, vinca alkaloids etc. [Choi, C.H.,
aBC transporters as multidrug resistance mechanisms and the development of chemosensitizers for their reversal.cancer Cell Int, 2005.
5: p. 30., Ambudkar, S.V., et al.,
biochemical, cellular, and pharmacological aspects of the multidrug transporter.annu Rev Pharmacol Toxicol, 1999.
39: p. 361-98. Krishna, R. and L.D. Mayer,
multidrug resistance (MDR) in cancer. Mechanisms, reversal using modulators of MDR and the role of MDR modulators in influencing the pharmacokinetics of anticancer drugs.eur J Pharm Sci, 2000.
11(4): p. 265-83.].The generation of MDR has number of mechanisms, comprise the minimizing that cell absorbs some water soluble drugs, the change of cell cycle, the enhancing of DNA repair function, the apoptosis of T suppression cell, medicine in the change of intracellular metabolic process, to hydrophobic drug and the [Stavrovskaya such as increase being easy to some drug effluxes entering cell, A.A.
cellular mechanisms of multidrug resistance of tumor cells.biochemistry (Mosc), 2000. 65 (1): p. 95-106.].The forming process of tumor MDR is complicated, and these resistance mechanisms often exist simultaneously, but based on a kind of mechanism, interacts between different mechanisms, may work alone or in combination [Szakacs, G., et al.,
targeting multidrug resistance in cancer.nat Rev Drug Discov, 2006. 5 (3): p. 219-34].
In order to overcome the MDR characteristic of tumor cell to chemotherapeutics, researcheres are finding the inhibitor of effective abc transport albumen always, these compounds are otherwise known as the inhibitor of MDR, regulator, inversion agent or chemotherapeutic sensitizer, they can regulate the function [Choi of multiple abc transport albumen usually, C.H.
aBC transporters as multidrug resistance mechanisms and the development of chemosensitizers for their reversal.cancer Cell Int, 2005.
5: p. 30. Liscovitch, M. and Y. Lavie,
cancer multidrug resistance:a review of recent drug discovery research.iDrugs, 2002.
5(4): p. 349-55.].1981, Tsuruo reported first calcium channel blocker verapamil (Verapamil) and pherylarsin oxide trifluoperazine (TFP) all can strengthen tolerant mice leukemia P388/VCR cell to vincristine (vincristine in testing in vitro and in vivo, VCR) sensitivity [Ozben, T.
mechanisms and strategies to overcome multiple drug resistance in cancer.fEBS Lett, 2006. 580 (12): p. 2903-9.].In the research of 30 years subsequently, being constantly found and studying of new MDR reversal agents, but higher cytotoxicity is a large problem of puzzlement MDR inhibitor research and development always.Therefore, the MDR inhibitor that exploitation toxicity is less, reverse effect is stronger or inversion agent remain one of focus of tumour medicine research.
In the research process of MDR inhibitor of new generation, the MDR reversal agents of screening high-efficiency low-toxicity from natural product day by day pay attention to by medical circle and become the focus of research gradually.Multiple natural product is found to have the effect of reversing tumor cell MDR.Fructus Momordicae (
siraitia grosvenorii Swingle) be Cucurbitaceae herbaceos perennial, be the distinctive plant of China, mainly originate in the ground such as Yongfu, Guangxi, Lingui and Long Sheng.The fruit of Fructus Momordicae is edible, also can be used as medicine, existing more than 300 year of the medicinal history among the people in Guangxi.Fructus Momordicae is cool in nature, sweet in the mouth, has clearing heat and moistening lung, effect of laxation relieving constipation [Tsuruo, T., et al.,
overcoming of vincristine resistance in P388 leukemia in vivo and in vitro through enhanced cytotoxicity of vincristine and vinblastine by verapamil.cancer Res, 1981. 41 (5): p. 1967-72.].Fructus Momordicae because its sugariness is high, heat is low and widely used as sweeting agent, the aspartame of Chang Zuowei overweight people and diabetics.One of main chemical compositions of Fructus Momordicae is triterpene glucoside, such chemical composition has antitussive, eliminates the phlegm, relievings asthma, the pharmacological action such as immunity moderation, reduction blood glucose, antiviral and inhibition tumor cell propagation [QI YP, T.M.,
chemical constituent of Luohanguo and applied studies.food Research and Development, 2001. 23 (5): p. 158-159.].
Protopanoxadiol protopanoxadiol, molecular formula is: C30 H52 O3, and to be 460, CA index name be molecular weight: Dammar-24-ene-3,12,20-triol.No. CAS is 6892-79-1.Be generally white powder, in Radix Ginseng, content is more.
Its chemical constitution is as follows:
2004, the Chinese invention patent application (application number: the preparation method disclosing a kind of protopanoxadiol 200410018038.8) that name is called " preparation method of protopanoxadiol and Protopanaxatriol ".Be specially: with panax species or the total saponin extracts of leaf, the Herb Gynostemmae Pentaphylli extract of gynostemma plant for raw material, carry out oxidation Basic fluxing raction in organic solvent, through column chromatography purification, obtained protopanoxadiol.Wherein, the amount ratio of the total saponin extracts of panax species or leaf, the Herb Gynostemmae Pentaphylli extract of gynostemma plant and organic solvent is 40-120g/L, carries out alkaline hydrolysis with sodium alkoxide, and concentration range is 0.2-1.5mol/L, alkaline hydrolysis temperature is 80-140 DEG C, and the response time is 24-90 hour.
2006, Li Xuwen etc. also reported the method being prepared protopanoxadiol by base catalysis degraded.And protopanoxadiol consistent with plant inner structure has been prepared by base catalysis degraded, and it is separated and structural characterization. Quinquefolium saponin and highly basic are dissolved in high boiling organic solvent, degrade under normal pressure and hot conditions. determined the best degradation condition preparing protopanoxadiol by orthogonal test, and degradation product is obtained protopanoxadiol through method separation such as extraction, column chromatography and recrystallization.(" base catalysis edman degradation Edman prepares anti-cancer active compound 20 (S)-protopanoxadiol
"li Xuwen; Kim Yong Il; Gui Mingyu; Zhang Hanqi; Zhang Longqing; SCI, 03 phase in 2006).
About protopanoxadiol, to the adjustment of tumor cell drug resistance, there is not been reported at present.
Summary of the invention
The object of this invention is to provide a kind of micromolecular compound protopanoxadiol, namely protopanoxadiol is preparing the novelty teabag in multidrug resistance reversing agent.
Another object of the present invention is to the application of the antineoplastic pharmaceutical compositions of providing package containing the antitumor drug such as protopanoxadiol and VCR and ADR in the medicine of preparation treatment multidrug resistance of tumor.
The invention provides protopanoxadiol and preparing the application in antitumor drug, described application is the application of protopanoxadiol in the reversal agent of drug resistance preparing antitumor drug.
Wherein, protopanoxadiol can be multidrug resistance reversing agent.
Described tumor comprises oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer, adenocarcinoma of lung, colon cancer or cancer of pancreas.
Described antitumor drug is vincristine, daunorubicin, paclitaxel or 5-fluorouracil.
Protopanoxadiol also can be the sensitizer of antitumor drug.
Present invention also offers a kind of antineoplastic pharmaceutical compositions, the effective ingredient of described antineoplastic pharmaceutical compositions is antitumor drug and protopanoxadiol.
Described antitumor drug is cell cycle specific agents or cell cycle nonspecific agent (CCNSA).
Described antitumor drug is vincristine, daunorubicin, paclitaxel or 5-fluorouracil.
Present invention also offers a kind of method impelling external cells of resistant tumors enhanced sensitivity, namely in the culture medium of cells of resistant tumors, add protopanoxadiol.
Described tumor comprises oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer, adenocarcinoma of lung or cancer of pancreas.
The invention provides protopanoxadiol and prepare the application in antitumor drug.Protopanoxadiol is natural product, can the propagation of obvious inhibition tumor cell when high dose.Experiment shows, protopanoxadiol obviously can overcome the drug resistance of multidrug resistance cell KB/VCR and MCF-7/ADR, the resistant characterization of reverse multiple drug resistance of tumor cell, strengthens chemotherapeutics to the therapeutic effect of cells of resistant tumors.Micromolecular compound of the present invention is developed as new antitumor drug or its auxiliary element, and tumor killing effect is obvious, environmental protection, provides a kind of new approach and means by for treatment and healing tumor.
Detailed description of the invention
The cytotoxicity of protopanoxadiol is less, to the IC of kinds of tumor cells
50all be greater than 40 μMs, there is good safety.The present invention, by random screening, finds that this natural product has the effect of reversion MDR.In view of the efficient feature of its low toxicity, have a good application prospect.Experiment shows, protopanoxadiol can strengthen the inhibition that chemotherapeutics is bred tumor multi-medicine drug-resistant cell strain, the effect of reverse multidrug drug resistance.Such as, show in embodiment, protopanoxadiol can reverse the drug-resistant effect of KB/VCR cell to VCR, strengthens KB/VCR cell to the sensitivity of chemotherapeutics VCR, promotes that VCR is to the inducing action of the apoptosis of KB/VCR cell.Protopanoxadiol significantly can overcome the drug resistance of multidrug resistance cell KB/VCR and MCF-7/ADR, the multidrug resistance characteristic of reversing tumor cell, strengthens chemotherapeutics to the therapeutic effect of tumor multi-medicine drug-resistant tumor cell.
Micromolecular compound of the present invention (S) protopanoxadiol is purchased from institute of materia medica of the Chinese Academy of Sciences, and high performance liquid chromatography surveys purity, HPLC >=98%.Its structure is as follows:
The invention provides protopanoxadiol and prepare the application in multidrug resistance reversing agent (Reversal agent).
Described tumor can be the various tumors such as oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer or cancer of pancreas to chemotherapeutics generation drug resistance.Described tumor cell can be the tumor cell of oral cancer, breast carcinoma, hepatocarcinoma or cervical cancer to chemotherapeutics generation drug resistance.Such as, multidrug resistance cell KB/VCR and MCF-7/ADR adopted in embodiment.Protopanoxadiol has remarkable result for the drug resistance caused due to drug efflux increase.
Described antitumor drug can be medicines resistant to liver cancer, anti-oral cancer medicine or Antilung gland cancer medicine, can be cell cycle specific agents (CCSA), as vincristine, paclitaxel etc., it also can be the cell cycle nonspecific agent (CCNSA) such as daunorubicin, 5-fluorouracil (CCNSA).
Described antitumor drug reversal agent of drug resistance can be made into the pharmaceutically acceptable dosage form of any one, comprises tablet, capsule, granule, oral liquid, slow releasing preparation, controlled release preparation, nanometer formulation, injection etc.
Present invention also offers the application of protopanoxadiol in the sensitizer (sensitizer) preparing antitumor drug.
Protopanoxadiol can overcome the resistant characterization of tumor multi-medicine drug-resistant cell, and increase tumor drug resistance cell to the sensitivity of medicine, the enhancing G2/M phase of VCR to the multidrug resistance cell caused by the apoptosis-promoting effect of KB/VCR cell and ADR blocks.
The reversal agent of drug resistance of the antitumor drug described in the present invention or the active component of sensitizer are protopanoxadiols.
Present invention also offers a kind of antineoplastic pharmaceutical compositions, said composition comprises protopanoxadiol and antitumor drug.Described antitumor drug can be the cell cycle specific agents such as vincristine, paclitaxel (CCSA), also can be the cell cycle nonspecific agent (CCNSA) such as daunorubicin, 5-fluorouracil (CCNSA).Protopanoxadiol also can with surgical operation conbined usage, with one or more Western medicine conbined usage, with Chinese herbal medicine conbined usage, with radiation treatment conbined usage.
In one embodiment of the present of invention, protopanoxadiol can strengthen the apoptosis-promoting effect of VCR to KB/VCR cell, can reverse the drug-resistant effect of KB/VCR cell to VCR, make VCR again play apoptosis-promoting effect.Because VCR itself is a microtubule inhibitors, the depolymerization of microtubule can be caused, cause apoptosis, make the cytosis being in the sub-G1 phase.Therefore, protopanoxadiol significantly can promote that VCR is to the apoptosis-induced effect of tumor multi-medicine drug-resistant cell KB/VCR, and this effect strengthens along with the increase of protopanoxadiol concentration.In the test to KB cell, do not find similar phenomenon.
On the other hand, the invention provides a kind of method of multidrug resistance tumor cells to chemotherapy drug susceptibility increasing In vitro culture, namely in the culture medium of multidrug resistance tumor cells, add protopanoxadiol.
Described tumor comprises oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer, adenocarcinoma of lung or cancer of pancreas.Described tumor cell can be the various tumor cells such as oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer or cancer of pancreas to chemotherapeutics generation drug resistance.Such as, multidrug resistance cell KB/VCR and MCF-7/ADR adopted in embodiment.
After tumor cell dosing, in culture medium, the concentration of protopanoxadiol can be greater than 20 μMs, is even greater than 50 μMs.But, even if the concentration of protopanoxadiol is lower in culture medium, such as 5-20 μM, even close to but when not comprising 0 μM, protopanoxadiol still can play the effect of multidrug resistance reversing agent or chemotherapeutic sensitizer.
Micromolecular compound protopanoxadiol of the present invention can adopt the preparation method of various routine to prepare.Such as, the method can synthesized from separation and purification in plant (such as, Fructus Momordicae) or artificial chemistry.
Utilize micromolecular compound of the present invention, by various conventional screening assays, can filter out, with protopanoxadiol, interactional material occur, as receptor, inhibitor or antagonist etc.
The present invention and inhibitor, antagonist etc., when carrying out using (administration) on treating, can provide different effects.Usually, but these materials are formulated in nontoxic, inertia with in pharmaceutically acceptable aqueous carrier medium, wherein pH is about 5-8 usually, and preferably pH is about 6-8, although pH value can with being formulated the character of material and disease to be treated and changing to some extent.The pharmaceutical composition prepared can carry out administration by conventional route, comprising (but being not limited to): intramuscular, intraperitoneal, subcutaneous, Intradermal or topical.
For protopanoxadiol of the present invention, can by itself and suitable pharmaceutically acceptable carrier coupling.This kind of pharmaceutical composition contains the compound and pharmaceutically acceptable carrier or excipient for the treatment of effective dose.This kind of carrier comprises (but being not limited to): normal saline, buffer, glucose, water, glycerol, ethanol and combination thereof.Pharmaceutical preparation should match with administering mode.Protopanoxadiol of the present invention can be made into injection form, such as, be prepared by conventional method with normal saline or the aqueous solution containing glucose and other adjuvant.The pharmaceutical composition of such as Tablet and Capsula and so on, is prepared by conventional method.Pharmaceutical composition such as injection, solution, Tablet and Capsula should aseptically manufacture.The dosage of active component is treatment effective dose, such as every day about 1 microgram/kg body weight-Yue 5 mg/kg body weight.In addition, protopanoxadiol of the present invention also can use together with other treatment agent.
When protopanoxadiol of the present invention is used as medicine, the protopanoxadiol for the treatment of effective dose can be applied to mammal, wherein this treatment effective dose is usually at least about 10 micrograms/kg body weight, and be in most of the cases no more than about 8 mg/kg body weight, preferably dosage is about 10 micrograms/kg body weight ~ 1 mg/kg body weight.Certainly, concrete dosage also should consider the factor such as route of administration, patient health situation, and these are all within skilled practitioners skill.
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this.Should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and can combining mutually between specifically described each technical characteristic in below (eg embodiment), thus form new or preferred technical scheme.As space is limited, tiredly no longer one by one to state at this.
implement 1:CCK-8 test kit and detect drug-resistant cell strain to the drug resistance multiple of protopanoxadiol
Experiment material:
Protopanoxadiol extracts from plant Fructus Momordicae (having production firm and article No. thereof), and purity is not less than 95%.Human oral cancer cell strain KB and drug-resistant cell strain KB/VCR thereof provided by Chinese Academy of Sciences's medicine, MCF-7 cell strainHJ2mm/ADR and drug-resistant cell strain MCF-7/ADR thereof, human colon carcinoma HCT-8 and drug-resistant cell strain HCT-8/VCR thereof are all purchased from Nanjing Kai Ji biotech firm.Verapamil (VPL), vincristine (VCR) and amycin (ADR) are purchased from Roche chemical company, and purity is all greater than 99%.CCK-8 test kit is purchased from colleague company.
Experimental technique:
Cell recovery
1) from liquid nitrogen container, take out cryopreservation tube, directly drop in 37 DEG C of warm water, and shake makes it melt as early as possible frequently.
2) from 37 DEG C of water-baths, take out cryopreservation tube, with suction pipe sucking-off cell suspension, inject centrifuge tube and add more than 10 times culture fluid, low-speed centrifugal after mixing, abandons supernatant, then repeats to wash once with culture fluid.
3), after suitably diluting with culture fluid, inoculated and cultured bottle, be placed on 37 DEG C of incubator quiescent culture, next day changes culture fluid, continues to cultivate.Go down to posterity when being cultured to finite concentration.
Passage is cultivated
Human oral cancer cell strain KB and drug-resistant cell strain KB/VCR thereof is incubated in the α-MEM culture medium containing 10% hyclone and 1mM Sodium Pyruvate, and MCF-7 cell strainHJ2mm/ADR and drug-resistant cell strain MCF-7/ADR thereof is incubated in 1640 culture medium containing 10% hyclone.
The situation of observation of cell growth every day, when cell grows to about 90% degree of converging (attached cell) in culture bottle, goes down to posterity according to the ratio of 1:3 to 1:5, about went down to posterity once every 2 ~ 4 days.Method is as follows:
1) with 1 × phosphate buffer wash cell once.
2) add 1 ~ 2 ml 0.25% tryptic digestive juice digestion, be placed in 37 DEG C of incubator numbers minute.Pat Tissue Culture Flask with hands, make cell separation.
3) trypsinization is stopped with the suitable culture medium containing 10%Gibco hyclone.Cell is sub-packed in new culture bottle, continues to cultivate.
Cell cryopreservation
1) get the cell being cultured to exponential phase, trypsinization, to be collected in centrifuge tube and to count, centrifugal.
2) reject trypsin and old culture fluid, add the frozen culture fluid (containing 10% DMSO, 40% DMEM and 50% Gibico hyclone) configured, in cryopreserving liquid, the ultimate density of cell is 0.5-1 × 10
7/ ml.Blowing and beating gently with suction pipe makes cell even, and be then distributed in aseptic cryopreservation tube, often pipe adds 1-1.5ml.
3) cryopreservation tube is put into freezing storing box and be placed in-80 DEG C, move in liquid nitrogen container after 5 hours and preserve.
CCK-8 tests
KB and KB/VCR cell and MCF-7 and MCF-7/ADR cell are all inoculated in 96 orifice plates according to the density in 3500/ hole, the ADR of variable concentrations after 24 h, VCR, is added in each hole after protopanoxadiol and VCR and the protopanoxadiol one α-MEM reinstated containing 10% hyclone prepares.After cultivating 48 h, discard culture fluid, every hole adds 90 μ L not containing culture medium and the 10 μ L CCK-8 reagent of serum.After 37 DEG C of reaction 2 h, microplate reader reads the light absorption value (OD450) of 450 nm wavelength.By calculating the cell proliferation ratio of medication group relative to matched group.Blank group only adds culture medium for not adding cell, and matched group is add the DMSO with medicine same volume, cell survival rate=(experimental group OD450-blank group OD450)/(matched group OD450-blank group OD450).Pass through IC
50value calculates drug resistance multiple (Resistance Fold, RF) again.
The IC of RF=drug-resistant cell strain
50the IC of value/parental cell strain
50value.Each concentration establishes 3 repeating holes, experiment repetition 3 times.
All mdr cells grow 3 days before tying up to Cell suppression test in without pharmaceutical culture medium.Each numerical value is 3 independent experiment results, IC
50represent with " means standard deviation " form.VCR, vincristine; ADR, daunorubicin; RF, drug resistance multiple.
Experimental result:
KB/VCR and MCF-7/ADR is two kinds of conventional multidrug resistance cell strains, and in the present embodiment, these two kinds of cells also show the characteristic of multidrug resistance.As shown in table 1, KB/VCR cell is respectively 81.9 times and 94.4 times relative to the drug resistance multiple of KB cell to VCR and ADR, and the drug resistance multiple that MCF-7/ADR cell compares VCR and ADR with MCF-7 cell is 38.1 times and 20.9 times respectively, display experiment mdr cell used has multidrug resistance, and MDR activity is similar to the result of bibliographical information.
Protopanoxadiol is to the half-inhibition concentration IC of parental cell strain KB and MCF-7
50be respectively 107.6 μ Μ and 112.3 μ Μ, to the IC of drug-resistant cell strain KB/VCR and MCF-7/ADR
50be respectively 121 μ Μ and 131.8 μ Μ, between the two without difference, multidrug resistance cell strain KB/VCR and MCF-7/ADR does not show the crossing drug resistant to compound protopanoxadiol, illustrates that protopanoxadiol can escape the multidrug resistance of mdr cell.Protopanoxadiol under the concentration of 40 below μ Μ, cell growth without obvious suppression, higher than can the propagation of T suppression cell under the concentration of 100 μ Μ.
Conclusion: protopanoxadiol can overcome the drug resistance of drug-resistant cell strain, drug-resistant cell strain is substantially identical to the sensitivity of protopanoxadiol.
embodiment 2:CCK-8 method detects protopanoxadiol to the reverse effect of tumor cell multidrug resistance activity
Experiment material: with embodiment 1.
Experimental technique:
Protopanoxadiol enhanced sensitivity is tested: KB/VCR, MCF-7/ADR and HCT-8/VCR cell is inoculated in 96 orifice plates according to the density in 3500/ hole, the VCR of variable concentrations after 24 h, and the VCR of variable concentrations and the protopanoxadiol one α-MEM reinstated containing 10% hyclone prepare after be added in each hole.After cultivating 48 h, discard culture fluid, with method in embodiment 1, survey drug-resistant cell strain and parental cell thereof to the activity of VCR and VCR+ protopanoxadiol with CCK-8 test kit, plot curve.Each concentration establishes 3 repeating holes, experiment repetition 3 times.
Experimental result:
The drug resistance multiple of KB/VCR cell to VCR is 81.9 times, after adding the protopanoxadiol of 10 μMs, make the sensitivity of KB/VCR cell to VCR add 1.5 times, and ADR is to the IC of mdr cell MCF-7/ADR
50have dropped 4.3 times, VCR is to the IC of HCT-8/VCR
501.2 times have comparatively been fallen under HCT-8.Protopanoxadiol can reverse multiple drug resistance of tumor cell to the drug resistance of chemotherapeutics.And this effect shows in parental cell and not obvious.
Conclusion:
Protopanoxadiol can the drug resistance of reverse multiple drug resistance of tumor cell KB/VCR and HCT-8/VCR.
Protopanoxadiol is natural product, has certain Clinical practice and is worth.That studies the chemistry and biology of this compounds along with people gos deep into, its molecular mechanism of action will be progressively clear and definite, this will promote modifying for chemical structure and the structure activity study of this compounds further, and contribute to the medical value improving this compounds.
The all documents mentioned in the present invention are quoted as a reference all in this application, are just quoted separately as a reference as each section of document.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (10)
1. protopanoxadiol is preparing the application in antitumor drug, it is characterized in that, described application is the application of protopanoxadiol in the reversal agent of drug resistance preparing antitumor drug.
2. apply as claimed in claim 1, it is characterized in that, protopanoxadiol is multidrug resistance reversing agent.
3. apply as claimed in claim 1, it is characterized in that, described tumor comprises oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer, adenocarcinoma of lung, colon cancer or cancer of pancreas.
4. apply as claimed in claim 1, it is characterized in that, described antitumor drug is vincristine, daunorubicin, paclitaxel or 5-fluorouracil.
5. apply as claimed in claim 1, it is characterized in that, protopanoxadiol is the sensitizer of antitumor drug.
6. an antineoplastic pharmaceutical compositions, is characterized in that, the effective ingredient of described antineoplastic pharmaceutical compositions is antitumor drug and protopanoxadiol.
7. antineoplastic pharmaceutical compositions as claimed in claim 6, it is characterized in that, described antitumor drug is cell cycle specific agents or cell cycle nonspecific agent (CCNSA).
8. antineoplastic pharmaceutical compositions as claimed in claim 6, it is characterized in that, described antitumor drug is vincristine, daunorubicin, paclitaxel or 5-fluorouracil.
9. impel a method for external cells of resistant tumors enhanced sensitivity, it is characterized in that, in the culture medium of cells of resistant tumors, add protopanoxadiol.
10. method as claimed in claim 9, it is characterized in that, described tumor comprises oral cancer, breast carcinoma, hepatocarcinoma, pulmonary carcinoma, cervical cancer, adenocarcinoma of lung or cancer of pancreas.
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| CN105399794A (en) * | 2015-08-13 | 2016-03-16 | 北京农学院 | Fructus momordicae triterpene saponin and salt thereof, preparation method and applications of fructus momordicae triterpene saponin and salt thereof, and pharmaceutical composition containing fructus momordicae triterpene saponin and salt thereof |
| CN108314698A (en) * | 2017-01-17 | 2018-07-24 | 刘珂 | Protopanoxadiol sodium phosphate and its preparation and its independent or multiple medicine combine the application in treating nephrotic syndrome |
| US10525064B2 (en) * | 2015-05-22 | 2020-01-07 | Intelligent Synthetic Biology Center | Anticancer adjuvant containing panaxadiol ginsenocide compound |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1418633A (en) * | 2002-10-22 | 2003-05-21 | 海南亚洲制药有限公司 | Anti-cancer assistant medicine contg. 20(S)-protopanaxadiol as effective component, and its application |
| CN1571673A (en) * | 2001-09-21 | 2005-01-26 | 博新药业股份有限公司 | Protopanaxadiol and protopanaxatriol and their use as synergistic anti-cancer agents |
| CN1771042A (en) * | 2002-06-11 | 2006-05-10 | 博新药业有限公司 | Protopanaxadiol and protopanaxatriol and their use as anti-cancer agents |
| CN104116746A (en) * | 2013-04-28 | 2014-10-29 | 祁展楷 | Panaxoside composition as well as preparation method and application thereof |
-
2013
- 2013-06-01 CN CN201310212784.XA patent/CN104208073A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1571673A (en) * | 2001-09-21 | 2005-01-26 | 博新药业股份有限公司 | Protopanaxadiol and protopanaxatriol and their use as synergistic anti-cancer agents |
| CN1771042A (en) * | 2002-06-11 | 2006-05-10 | 博新药业有限公司 | Protopanaxadiol and protopanaxatriol and their use as anti-cancer agents |
| CN1418633A (en) * | 2002-10-22 | 2003-05-21 | 海南亚洲制药有限公司 | Anti-cancer assistant medicine contg. 20(S)-protopanaxadiol as effective component, and its application |
| CN104116746A (en) * | 2013-04-28 | 2014-10-29 | 祁展楷 | Panaxoside composition as well as preparation method and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10525064B2 (en) * | 2015-05-22 | 2020-01-07 | Intelligent Synthetic Biology Center | Anticancer adjuvant containing panaxadiol ginsenocide compound |
| CN105399794A (en) * | 2015-08-13 | 2016-03-16 | 北京农学院 | Fructus momordicae triterpene saponin and salt thereof, preparation method and applications of fructus momordicae triterpene saponin and salt thereof, and pharmaceutical composition containing fructus momordicae triterpene saponin and salt thereof |
| CN105399794B (en) * | 2015-08-13 | 2017-04-12 | 北京农学院 | Fructus momordicae triterpene saponin and salt thereof, preparation method and applications of fructus momordicae triterpene saponin and salt thereof, and pharmaceutical composition containing fructus momordicae triterpene saponin and salt thereof |
| CN108314698A (en) * | 2017-01-17 | 2018-07-24 | 刘珂 | Protopanoxadiol sodium phosphate and its preparation and its independent or multiple medicine combine the application in treating nephrotic syndrome |
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