CN1833636A - Application of biphenyl cyclooctadiene lignin in treatment of tumour medicine - Google Patents

Application of biphenyl cyclooctadiene lignin in treatment of tumour medicine Download PDF

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CN1833636A
CN1833636A CN 200510049382 CN200510049382A CN1833636A CN 1833636 A CN1833636 A CN 1833636A CN 200510049382 CN200510049382 CN 200510049382 CN 200510049382 A CN200510049382 A CN 200510049382A CN 1833636 A CN1833636 A CN 1833636A
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benzene ring
lignanoid
couplet benzene
cell
ring octadiene
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胡汛
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Ningbo Yingnuo Pharmaceutical Technology Co Ltd
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Ningbo Yingnuo Pharmaceutical Technology Co Ltd
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Abstract

An application of the biphenyl cyclooctadiene lignan in preparing the medicine for treating tumor by poisoning the tumor cells and reversing the resistance of tumor cells to multiple medicines is disclosed.

Description

The application of couplet benzene ring octadiene lignanoid in preparation medicine for treating tumor thing
(1) technical field
The present invention relates to the application of application, especially the couplet benzene ring octadiene lignanoid of couplet benzene ring octadiene lignanoid in preparation medicine for treating tumor thing in preparation antitumor cell multidrug resistance inversion agent medicine.
(2) background technology
Tumor is to cause one of human main causes of death, and chemotherapy of tumors is a kind of main means of treatment tumor.Yet chemotherapy of tumors can be failed because of tumor cell obtains Drug resistance.
The reason of tumor cell generation multidrug resistance has multiple, the wherein most importantly bonded transport protein of tumor cells expression ATP [ATP binding cassette (ABC) transporters].P-glycoprotein (P-glycoprotein, P-gp), multidrug-associated protein 1 (multidrug resistant associatedprotein 1, MRP1) and breast carcinoma sorcin (breast cancer resistance protein BCRP, be also referred to as MXR or ABCG2), be several main protein that cause tumor multi-medicine drug-resistant of generally acknowledging at present.These protein are transmembrane protein, and their effect is equivalent to Teat pipette, the medicine that enters in the tumor cell can be discharged the extracellular, make intracellular drug level very low, and therefore tumor cell can escape the attack of antineoplastic agent.(GottesmanMM，Fojo T & Bates SE.Nature Review/cancer.2：48-58，2002)。
At present anticarcinogen that uses clinically such as amycin, mitomycin, epirubicin, daunorubicin, vincristine, high Folium et Ramulus Cephalotaxi ester, rice take off medicine such as anthraquinone and all do not escape the drug-fast destiny of tumor, because these medicines are the substrate of P-glycoprotein or MRP1 or BCRP.Therefore, behind P-glycoprotein or MRP1 in the tumor cell or BCRP high expressed, the Drug resistance that tumor cell obtains is not merely to a certain anticarcinogen, but to multiple anticarcinogen.Make the tumor cell of high expressed P-glycoprotein or MRP1 or BCRP regain sensitivity to anticarcinogen, effective method suppresses these proteinic functions exactly, makes it can not bring into play the function of ' Teat pipette '.Therefore, the medicine of these protein functions of development inhibition has important meaning to the chemotherapy of tumor.At present, a variety of anticarcinogens being arranged clinically, is medicine at P-glycoprotein or MRP1 or BCRP but do not have a kind of.Therefore, it is very urgent to develop a kind of like this medicine.
Phenyl alkylamide derivant calcium antagonist verapamil is to find the earliest and have to reverse the active phenyl alkylamide derivant of MDR that the P-glycoprotein mediates more by force.Its calcium antagonistic activity and its MDR reverse effect to blood vessel, cardiac muscle is irrelevant, and relevant with the cardiovascular system side effect, its side effect seriously hinders clinical practice.
Couplet benzene ring octadiene lignanoid is a Fructus Schisandrae Chinensis: the main component in the dry mature fruit of magnoliaceae schisandra Schisandra chinensis (Turcz.) Baill. or Magnoliacea plant schisandra chinensis Schisandra sphenanthera Rehd.et Wils., Fructus Schisandrae Chinensis is listed in top grade in Shennong's Herbal, effect with convergence, astringent or styptic treatment for spontaneous sweating, supplementing QI for promoting the production of body fluid, kidney calming, it is traditional Chinese medical science strengthening by means of tonics medicine commonly used, has multiple pharmacological effect, but do not see that it has clear and definite antineoplastic report, does not especially see the report that it has artitumor multi-medicine-resistant.
(3) summary of the invention
The present invention is for the application of couplet benzene ring octadiene lignanoid in preparation medicine for treating tumor thing is provided.
For reaching goal of the invention the technical solution used in the present invention be:
Couplet benzene ring octadiene lignanoid is applied to prepare the medicine for treating tumor thing.
Described couplet benzene ring octadiene lignanoid structure is suc as formula shown in the I:
Figure A20051004938200081
In the formula, R 1, R 2, R 5, R 6Be respectively hydroxyl or methoxyl group, perhaps, R 1With R 2, R 5With R 6Ring formation not separately
Perhaps R 1With R 2, R 5With R 6Form the alcoxyl ring separately;
R 3For one of following: 1.-O-CH 32.-OH;
Figure A20051004938200091
R 4For one of following: 1.-O-CH 32.-OH;
Figure A20051004938200092
R 7For one of following: 1.-H; 2.-OH;
Figure A20051004938200093
R 8, R 9One of it is following respectively to do for oneself: 1.-and H; 2.-OH;
R 10For one of following: 1.-H; 2.-OH;
Figure A20051004938200094
Figure A20051004938200095
Perhaps, R 7With R 10Be linked to be oxo bridge, R 1～R 6, R 8, R 9Definition as mentioned above;
Perhaps, R 3With R 7Form the acyl-oxygen ring, R 1, R 2, R 4～R 6, R 8～R 10Definition as mentioned above.
The said structure formula mainly comprises following material according to each substituent group definition:
Schisandrin A Schisandrin B
Figure A20051004938200102
Schisandrol A Schisantherin A
Schisandrin C Schisantherin B
Schisantherin C Schisantherin D
Figure A20051004938200111
Schisantherin E Schisantherin F
Figure A20051004938200112
Schisantherin G Schisantherin H
Schisantherin I Schisantherin J
Figure A20051004938200114
Schisantherin K Schisantherin L：R1＝OH，R2＝OAng
Schisantherin M：R1＝OTig，R2＝Oang
Schisantherin N：R1＝OAc，R2＝Oang
Figure A20051004938200121
Schisantherin O Schisandrol B，Schisantherinol B
Figure A20051004938200122
Schisandrol D，Schisantherinol D Schisandrol E，Schisantherinol E
Methylschisandrol E Angelogomisin P：R＝Ang
Methylschisantherinol E TiglogomisinP：R＝Tig
Gomisin A
Gomisin D Gomisin E
Figure A20051004938200133
Gomisin F Gomisin G
Figure A20051004938200141
Gomisin H Angeloylgomisin H
Figure A20051004938200142
Tigloylgomisin H Benzoylgomisin H
Figure A20051004938200143
Gomisin J Gomisin K1
Gomisin K2 Gomisin K3
Figure A20051004938200151
Gomisin M1 Gomisin M2
Angeloylgomisin M1
angeloylgomisin R
Gomisin N Gomisin O
Figure A20051004938200161
Epigomisin O Gomisin Q
Figure A20051004938200162
Gomisin R Angelogomisin O
Figure A20051004938200163
Angeloisogomisin O Benzoylgomisin O
Figure A20051004938200171
Benzoylgomisin P Benzoylgomisin Q
Figure A20051004938200172
Benzoylisogomisin O kadsurin R＝-COCH 3
Figure A20051004938200175
kadsurarin
R＝-COCH 3
Figure A20051004938200181
kadasutherin isokadsuranin
neokadsuranin 5，8-epoxy-6，7-dimethyl 2′，3′，2″，3″-dimethylenedioxy-4′，1″-dimethoxy-1，2：3，4-dib enzo-1，3-cyclootadiene
Further, described couplet benzene ring octadiene lignanoid structural formula is as follows:
Figure A20051004938200191
In the formula, R 1, R 2, R 5, R 6Be respectively hydroxyl or methoxyl group, perhaps, R 1With R 2Or R 5With R 6Form the alcoxyl ring separately;
R 7For one of following: 1.-H; 2.-OH;
Figure A20051004938200192
R 8, R 9One of it is following respectively to do for oneself: 1.-and H; 2.-OH;
R 10For one of following: 1.-H; 2.-OH;
Figure A20051004938200194
Preferably, described couplet benzene ring octadiene lignanoid is one of following formula:
1. wuweizisu A; 2. wuweizisu B; 3. schisandrin C; 4. schizantherin A; 5. schizantherin B; 6. schizantherin C; 7. schizantherin D; 8. schizantherin E; 9. 10. wuweizi alcohol B of wuweizi alcohol A.
Especially, described couplet benzene ring octadiene lignanoid can be applicable to prepare tumor multidrug-resistance inversion agent medicine.
Described inversion agent medicine also can contain the antitumor drug as tumor multidrug-resistance inversion agent sensitivity to couplet benzene ring octadiene lignanoid, and drug excipient or carrier.
Described antitumor drug to tumor multidrug-resistance inversion agent sensitivity is one of following or the mixing of its arbitrary proportion:
1. the substrate of P-glycoprotein 2. multidrug-associated protein (multidrug resistant associatedprotein, substrate MRP1) be breast carcinoma sorcin (breast cancer resistantprotein, substrate BCRP) 3..
Concrete, described antitumor drug to tumor multidrug-resistance inversion agent sensitivity is one of following or the mixing of its arbitrary proportion:
The Doxorubicin amycin; The Actinomycin D actinomycin D; Altreatamine; The Bleomycin bleomycin; The Busulphan busulfan; The Capecitabine capecitabine; The Carboplatin carboplatin; The Carmustine carmustine; The Chlorambucil chlorambucil; The Cisplatin cisplatin; The cyclophosphamide cyclophosphamide; The cytarbine cytosine arabinoside; Dacarabazine, the daunorubicin daunorubicin; The epirubicin epirubicin; The etoposide etoposide; Etoposide; Etoposide; The acid of fludarbine fluorine vidarabine; The fluorouracil fluorouracil; The gemcitabine gemcitabine; The herceptin Trastuzumab; The hydroxyurea hydroxyurea; The idarubicin idarubicin; The ifosfamide ifosfamide; The irinotecan Irinotecan; The lomustine lomustine; Lomustine; The melphalan melphalan; Alkeran; The mercaptopurine purinethol; The methotrexate methotrexate; The mitomycin mitomycin; The mitozantrone mitoxantrone; Dithranol; The oxaliplatin oxaliplatin; The procarbazine procarbazine; First (base) benzyl hydrazine; The rituxan Mabthera; The steroids steroid; The streptozocin streptozocin; Streptozotocin; The taxol paclitaxel, the taxotere taxotere; Tamozolomide, the thioguanine thioguanine; The thiotepa thio-tepa; Thiotef; Tespamin; Tomudex Raltitrexed (raltitrexed); The topotecan topotecan; The treosulfan treosulfan; The uracil-tegufur uracil; The vinblastine vinblastine; Vincaleucoblastine; The vindesine vindesine; The vinorelbine vinorelbine.
Described medicine for treating tumor thing can be made into one of following dosage form:
1. 2. 3. 4. 5. slow releasing agent of granule of capsule of tablet of injection.
Described couplet benzene ring octadiene lignanoid also can be applicable to prepare improves the sensitizer medicine of tumor to chemotherapeutics.
Described couplet benzene ring octadiene lignanoid also can be applicable to prepare the medicine that promotes apoptosis of tumor cells.
Described couplet benzene ring octadiene lignanoid also can adopt two or more mixture when being applied to the medicine for treating tumor thing, described couplet benzene ring octadiene lignanoid is mixed in varing proportions.
The beneficial effect of the application of couplet benzene ring octadiene lignanoid of the present invention in preparation medicine for treating tumor thing is as follows: (1) has potential applicability in clinical practice, couplet benzene ring octadiene lignanoid is the multidrug resistance of reversing tumor effectively, this compound effects can suppress the function of P-glycoprotein in the P-glycoprotein; (2) couplet benzene ring octadiene lignanoid has lower toxicity, and therefore, couplet benzene ring octadiene lignanoid has the application prospect of good clinical chemotherapy of tumors.
(4) description of drawings
Fig. 1 is the expression percentage ratio of P-glycoprotein in multidrug resistance cell.
Fig. 2 is the expression of P-glycoprotein in multidrug resistance cell.
Fig. 3 is the expression percentage ratio of MRP1 in multidrug resistance cell.
Fig. 4 is the expression of MRP1 in multidrug resistance cell.
Fig. 5 is that 10 kinds of couplet benzene ring octadiene lignanoids increase daunorubicin gathering in the MDR cell K562/Adr of P-glycoprotein high expressed cell.
Fig. 6 is that 10 kinds of couplet benzene ring octadiene lignanoids can increase daunorubicin gathering in the MDR of MRP1 high expressed cell HL60/adr cell.
(5) specific embodiment
The present invention is described further below in conjunction with specific embodiment:
Embodiment 1: the P-glycoprotein (P-gp) of multidrug resistance cell and the detection of MRP1.
(1) experiment material:
Cell strain: K562/adr, MCF7/adr and KBV200 multidrug resistance cell strain are the cell of Pgp albumen high expressed.It is that MCF-7/adr and KBV200 multidrug resistance cell strain are available from Chinese Academy of Medical Sciences's Blood Research Institute that K562/adr is induced to build by the Zhejiang University institute of oncology.HL60/adr is the multidrug resistance tumor cells strain of MRP1 high expressed, available from Chinese Academy of Medical Sciences's Blood Research Institute.
The monoclonal fluorescent-labeled antibody of reagent: P-gp is U.S. company BD R-PE-17F9.MRP1 monoclonal fluorescent antibody is available from U.S. Santa Cruz company.
Instrument: culture bottle, culture plate, CO2 gas incubator, flow cytometer (FACS Calibur, U.S. Becton-Dickinson company).
(2) experimental technique:
Cell P-gp and MRP1 express and measure:
Each the strain cell of trophophase of taking the logarithm, collecting cell makes 1 * 10 6The suspension of/ml adds P-gp or MRP1 monoclonal antibody (1: 400), 4 ℃ of lucifuge reaction 45min, and PBS gives a baby a bath on the third day after its birth inferior, and after overhanging with PBS, flow cytometer detects cell fluorescence intensity.
(3) experimental result:
Fig. 1 is K562/ADR, MCF7/adr, the expression of the P-gp of KBV200 (P-glycoprotein) and its parent's susceptibility cell K562, the comparison of MCF-7 and KBV200.98% cellular expression P-gp is arranged in the K562/adr cell, and the K562 cell has only 1% cell that the expression of P-gp is arranged; 96% cellular expression P-gp is arranged in the MCF7/adr cell, and the MCF-7 cell has only 3% cell that the expression of P-gp is arranged; 92% cellular expression P-gp is arranged in the KBV200 cell, and the KB cell has only 1% cell that the expression of P-gp is arranged.
Fig. 2 is K562/ADR, MCF7/adr, the expression of the P-gp of KBV200 (P-glycoprotein) and its parent's susceptibility cell K562, the comparison of MCF-7 and KBV200.The expression of K562/adr, MCF7/adr KBV200 cell P-gp is respectively its susceptibility parental cell K562, MCF-7 and KB cell 24.4,25.3 and 24.2 times.
Fig. 3 is the expression of MRP1 of HL60/adr and the comparison of its parent's susceptibility cell HL60.97% cellular expression MRP1 is arranged in the HL60 cell, and the HL60 cell has only 1% cell that the expression of MRP1 is arranged;
Fig. 4 is the expression of MRP1 of HL60/adr and the comparison of its parent's susceptibility cell HL60.The expression of HL60/adr cell MRP1 is 23.4 times of its susceptibility parental cell HL60 cell.
(4) conclusion:
K562/ADR, MCF7/adr, KBV200 are the multidrug resistance cell strain of P-gp high expressed.That is: P-gp is the main cause of these cell multidrug resistances.HL60/adr is the multidrug resistance cell strain of MRP1 high expressed, that is: MRP1 is the main cause of these cell multidrug resistances.
Embodiment 2: the multi-drug resistance of the tumor of the reverse P-glycoprotein mediation of couplet benzene ring octadiene lignanoid
All there is the effect of the multi-drug resistance of the tumor that reverses the mediation of P-glycoprotein in the lignanoid that has a couplet benzene ring octadiene for proof, and we study with 10 kinds of different couplet benzene ring octadiene lignanoids of structure.These 10 kinds of couplet benzene ring octadiene lignanoids are respectively: wuweizisu A, wuweizisu B, schisandrin C, schizantherin A, schizantherin B, schizantherin C, schizantherin D, schizantherin E, wuweizi alcohol A and wuweizi alcohol B.These lignanoids are available from Chinese biological goods and drug identification institute and Hua Sheng Bioisystech Co., Ltd of Chengdu Cisco.
Cell strain: K562/adr, MCF7/adr, KBV200 multidrug resistance cell strain are main resistance mechanism with Pgp albumen high expressed.
Reagent: various couplet benzene ring octadiene lignanoid is dissolved in DMSO respectively, is made into 10mg/ml, be made into working solution with RPMI-1640 again as mother solution; The RPMI-1640 culture medium; Daunorubicin.
Instrument: culture bottle, culture plate, FCM.
(2) experimental technique:
Daunorubicin DNR gathers in the FACS method analysis of cells: K562/adr, MCF7/adr, KBV200 cell culture are in the fresh medium of no medicine.Experiment is divided into 2 groups, matched group and reverse group.Matched group adds fresh medium, and it is 10 μ g/ml that the reverse group adds couplet benzene ring octadiene lignanoid to final concentration, adds daunorubicin DNR after hatching 1h, and concentration is respectively 2 μ g/ml, and 37 ℃ of incubations are analyzed medicine with FCM after 90 minutes and gathered intracellular.
(3) experimental result:
Fig. 5 illustrates that 10 kinds of couplet benzene ring octadiene lignanoids all can increase daunorubicin gathering in MDR cell K562/Adr cell.Among the figure: A:K562/adr cell+daunorubicin; B:K562/adr cell+daunorubicin+wuweizisu A; C:K562/adr cell+daunorubicin+wuweizisu B; D:K562/adr cell+daunorubicin+schisandrin C; E:K562/adr cell+daunorubicin+schizantherin A; F:K562/adr cell+daunorubicin+schizantherin B; G:K562/adr cell+daunorubicin+schizantherin C; H:K562/adr cell+daunorubicin+schizantherin D; I:K562/adr cell+daunorubicin+schizantherin E; J:K562/adr cell+daunorubicin+wuweizi alcohol A; K:K562/adr cell+daunorubicin+wuweizi alcohol B.
Conclusion: couplet benzene ring octadiene lignanoid can suppress the function of P-glycoprotein, recovers the drug accumulation in the MDR tumor cell.The MDR's of this 10 kinds of reverse P-of couplet benzene ring octadiene lignanoids glycoproteins mediation is active and inequality.
Embodiment 3: couplet benzene ring octadiene lignanoid suppresses the multi-drug resistance of the tumor of MRP1 mediation
(1) experiment material:
Cell strain: the HL60/adr multidrug resistance cell strain is main resistance mechanism with MRP1 albumen high expressed.
Reagent: various couplet benzene ring octadiene lignanoid is dissolved in DMSO respectively, is made into 10mg/ml, be made into working solution with RPMI-1640 again as mother solution; Daunorubicin.
Instrument: culture bottle, culture plate, FCM.
(2) experimental technique:
Daunorubicin gathers in the FACS method analysis of cells: the HL60/adr cell culture is in the fresh medium of no medicine.Experiment is divided into 2 groups, matched group and reverse group.Matched group adds fresh medium, and it is 10 μ g/ml that the reverse group adds couplet benzene ring octadiene lignanoid to final concentration, adds daunorubicin DNR after hatching 1h, and concentration is respectively 2 μ g/ml, and 37 ℃ of incubations are analyzed medicine with FCM after 90 minutes and gathered intracellular.
(3) experimental result:
Fig. 6 illustrates that 10 kinds of couplet benzene ring octadiene lignanoids all can increase daunorubicin gathering in MDR cell HL60/adr cell.Among the figure: A:HL60/adr cell+daunorubicin; B:HL60/adr cell+daunorubicin+wuweizisu A; C:HL60/adr cell+daunorubicin+wuweizisu B; D:HL60/adr cell+daunorubicin+schisandrin C; E:HL60/adr cell+daunorubicin+schizantherin A; F:HL60/adr cell+daunorubicin+schizantherin B; G:HL60/adr cell+daunorubicin+schizantherin C; H:HL60/adr cell+daunorubicin+schizantherin D; I:HL60/adr cell+daunorubicin+schizantherin E; J:HL60/adr cell+daunorubicin+wuweizi alcohol A; K:HL60/adr cell+daunorubicin+wuweizi alcohol B.
Conclusion: couplet benzene ring octadiene lignanoid can suppress the function of MRP1, increases the drug accumulation in the MDR tumor cell.
Embodiment 4: couplet benzene ring octadiene lignanoid reverses the dose-effect relationship cell of the multi-drug resistance of the tumor of P-gp mediation: the K562/adr multidrug resistance cell strain is main resistance mechanism with Pgp albumen high expressed.
Reagent: wuweizisu A or schizantherin A are dissolved in DMSO, are made into 10mg/ml as mother solution, are made into working solution with RPMI-1640 again; The RPMI-1640 culture medium; Daunorubicin.
Experimental technique:
Cell inoculation is to 96 porocyte culture plates, 8000 cells in every hole after the overnight incubation, add the wuweizisu A of variable concentrations or the daunorubicin of schizantherin A and variable concentrations (1,10,100,1000,10000ng/ml), continue to cultivate after 72 hours, do the MTT experiment.Cell inhibitory rate (IR)=[1-(experimental port A average/control wells A average)] * 100%.Calculating IR adopts Origin 7.0 data processing softwares to obtain half-inhibition concentration (IC with the match of Sigmodel function 50), every experiment triplicate.Couplet benzene ring octadiene lignanoid is IC to the drug resistance reverse multiple of MDR cell 50(add a kind of couplet benzene ring octadiene lignanoid) is divided by IC 50(not adding couplet benzene ring octadiene lignanoid).
Experimental result:
Wuweizisu A and schizantherin A are dose dependent to the reverse of multidrug resistance in concentration is 1 μ g/ml to 20 μ g/ml scope, result of the test sees Table 2:
Table 2: couplet benzene ring octadiene lignanoid wuweizisu A and schizantherin A reverse K562/adr to the drug-fast dose-effect relationship of daunorubicin
Couplet benzene ring octadiene lignanoid IC 50(ng/ml) Drug resistance reverses multiple
Contrast 4780±1327
Wuweizisu A
1μg/ml 568±89 8
10μg/ml 67±23 71
20μg/ml 48±18 100
Schizantherin A
1μg/ml 266±65 18
10μg/ml 71±16 67
20μg/ml 30±8 159
Embodiment 5: couplet benzene ring octadiene lignanoid is to the reverse of the multi-drug resistance of the tumor of P-gp mediation
Cell: K562/adr, MCF7/adr, KBV200 multidrug resistance cell strain are main resistance mechanism with Pgp albumen high expressed.
Reagent: wuweizisu A, wuweizisu B, schisandrin C, schizantherin A, schizantherin B, schizantherin C, schizantherin D, schizantherin E, wuweizi alcohol A and wuweizi alcohol B are dissolved in DMSO respectively, be made into 10mg/ml as mother solution, be made into working solution with RPMI-1640 again; The RPMI--1640 culture medium; Amycin, vincristine, paclitaxel.
Instrument: culture bottle, culture plate, FCM.
Experimental technique:
Cell inoculation is to 96 porocyte culture plates, and 8000 cells in every hole after the overnight incubation, add a kind of couplet benzene ring octadiene lignanoid and anticarcinogen (amycin, vincristine or paclitaxel etc.), continues to cultivate after 72 hours, does the MTT experiment.Cell inhibitory rate (IR)=[1-(experimental port A average/control wells A average)] * 100%.Calculating IR adopts Origin 7.0 data processing softwares to obtain half-inhibition concentration (IC with the match of Sigmodel function 50), every experiment triplicate.It is IC that the drug resistance of the MDR of couplet benzene ring octadiene lignanoid cell reverses multiple 50(add a kind of couplet benzene ring octadiene lignanoid) is divided by IC 50(not adding couplet benzene ring octadiene lignanoid).
Experimental result:
Couplet benzene ring octadiene lignanoid can reverse the Drug resistance of multidrug resistance tumor cells.The results are shown in Table 3, table 4, table 5.
Table 3.10 kind of couplet benzene ring octadiene lignanoid reverses the drug resistance of multidrug resistance tumor cells to amycin
Couplet benzene ring octadiene lignanoid (10 μ g/ml) IC50(ng/ml)
K562/adr KBv200 MCF-7/adr
Contrast 4780±1237 26±6 2556±638
Wuweizisu A 67±23(71) 2.5±0.6(10.4) 245±35(10.4)
Wuweizisu B 167±28(29) 5.3±1.6(4.9) 189±29(14)
Schisandrin C 89±21(53) 3.6±1.1(7.2) 132±23(19)
Schizantherin A 71±16(67) 3.4±0.7(7.6) 59±12(43)
Schizantherin B 39±8(122) 1.8±0.5(14) 157±28(16)
Schizantherin C 83±15(58) 3.6±0.7(7) 98±26(26)
Schizantherin D 93±21(51) 2.9±0.5(9) 163±35(16)
Schizantherin E 67±23(90) 2.8±0.7(9.2) 67±21(38)
Wuweizi alcohol A 860±195(6) 11±2(2.4) 564±162(5)
Wuweizi alcohol B 923±201(5) 12±3(2.2) 642±189(4)
Numerical value in the bracket is represented drug-fast reverse multiple.
Table 4.10 kind of couplet benzene ring octadiene lignanoid reverses the drug resistance of multidrug resistance tumor cells to vincristine
Couplet benzene ring octadiene lignanoid (10 μ g/ml) IC50(ng/ml)
K562/adr KBv200 MCF-7/adr
Contrast 612±139 22±3 501±67
Wuweizisu A 7.3±2.5(84) 3.0±0.6(7) 8.4±2.1(60)
Wuweizisu B 11±3(56) 3.1±0.5(7) 7.2±1.9(70)
Schisandrin C 14±5(44) 4.2±0.8(5) 9.8±2.2(51)
Schizantherin A 7.7±2.1(79) 3.7±0.9(6) 24±5(21)
Schizantherin B 12±3(51) 2.5±0.6(9) 26±7(19)
Schizantherin C 6±2(102) 3.1±0.5(7) 27±8(19)
Schizantherin D 7±3(87) 2.5±0.6(9) 31±5(16)
Schizantherin E 15±3(41) 3±1(7) 21±3(24)
Wuweizi alcohol A 89±13(7) 12±4(2) 126±32(4)
Wuweizi alcohol B 102±27(6) 9±2(2) 112±23(4)
Numerical value in the bracket is represented drug-fast reverse multiple.
Table 5.10 kind of couplet benzene ring octadiene lignanoid reverses the drug resistance of multidrug resistance tumor cells to paclitaxel
Couplet benzene ring octadiene lignanoid (10 μ g/ml) IC50(ng/ml)
K562/adr KBv200 MCF-7/adr
Contrast 38±5 20±2 41±5
Wuweizisu A 6.4±1.3(6) 1.9±0.2(11) 6.1±2.0(7)
Wuweizisu B 7.9±2.1(5) 2.5±0.4(8) 7.9±2.5(5)
Schisandrin C 8.6±2.3(4) 3.6±0.8(6) 5.5±1.5(7)
Schizantherin A 1.1±0.4(35) 2.4±0.3(8) 6.3±1.5(7)
Schizantherin B 0.9±0.2(42) 1.9±0.4(11) 5.6±1.9(7)
Schizantherin C 1.5±0.4(25) 1.6±0.3(13) 5.5±2.1(8)
Schizantherin D 2.6±0.5(15) 2.1±0.5(10) 7.9±2.8(5)
Schizantherin E 3.8±1.0(10) 3.5±0.9(6) 8.2±3.1(5)
Wuweizi alcohol A 12±3(3) 8±3(3) 19±5(2)
Wuweizi alcohol B 10±2(4) 9±2(2) 21±3(2)
Numerical value in the bracket is represented drug-fast reverse multiple.
Conclusion: various couplet benzene ring octadiene lignanoid all can suppress the multi-drug resistance of the tumor of P-gp mediation, though the activity of every kind of couplet benzene ring octadiene lignanoid reverse multiple drug resistance of tumor has the branch of power.
Embodiment 6: couplet benzene ring octadiene lignanoid reverses the multi-drug resistance of the tumor of MRP1 mediation
Cell: the HL60/adr multidrug resistance cell strain is main resistance mechanism with MRP1 albumen high expressed.
Reagent: various couplet benzene ring octadiene lignanoid is dissolved in DMSO respectively, is made into 10mg/ml as mother solution, is made into working solution with RPMI-1640 again; The RPMI-1640 culture medium.
Instrument: culture bottle, culture plate, FCM.
Experimental technique:
Cell inoculation is to 96 porocyte culture plates, and 8000 cells in every hole after the overnight incubation, add various couplet benzene ring octadiene lignanoids and anticarcinogen (paclitaxel, amycin or vincristine etc.), continues to cultivate after 72 hours, does the MTT experiment.Cell inhibitory rate (IR)=[1-(experimental port A average/control wells A average)] * 100%.Calculating IR adopts Origin 7.0 data processing softwares to obtain half-inhibition concentration (IC with the match of Sigmodel function 50), every experiment triplicate.It is that IC50 (add a kind of couplet benzene ring octadiene lignanoid) is divided by IC50 (not adding couplet benzene ring octadiene lignanoid) that various couplet benzene ring octadiene lignanoid reverses multiple to the drug resistance of MDR cell.
Experimental result:
Various couplet benzene ring octadiene lignanoid can reverse the Drug resistance of multidrug resistance tumor cells HL60/adr.The results are shown in Table 6.
Table 6. couplet benzene ring octadiene lignanoid reverses the drug resistance of multidrug resistance tumor cells
Couplet benzene ring octadiene lignanoid (10 μ g/ml) IC50(ng/ml)
Amycin Vincristine Paclitaxel
Contrast 160±21 89±12 32±4
Wuweizisu A 52±11(3) 3.2±0.7(28) 4.2±0.8(8)
Wuweizisu B 48±11(3) 2.3±0.5(39) 4.1±1.2(8)
Schisandrin C 42±13(4) 3.1±0.8(29) 4.2±0.7(8)
Schizantherin A 56±11(3) 3.9±1.2(23) 4.3±1.4(8)
Schizantherin B 43±12(4) 2.6±0.8(34) 3.2±1.2(10)
Schizantherin C 61±20(3) 3.9±0.6(30) 5.2±2.1(6)
Schizantherin D 73±23(2) 2.8±1.3(32) 3.8±1.2(8)
Schizantherin E 35±11(5) 5.1±2.1(17) 2.9±0.6(11)
Wuweizi alcohol A 73±21(2) 21±3(4) 5.2±1.8(6)
Wuweizi alcohol B 81±20(2) 19±2(5) 8.3±2.1(4)
Numerical value in the bracket is represented drug-fast reverse multiple.
Conclusion: couplet benzene ring octadiene lignanoid can suppress the MRP1 function, reverses the multi-drug resistance of the tumor of MRP1 mediation.
Embodiment 7: the apoptotic effect of the short tumor cell of couplet benzene ring octadiene lignanoid.
(1) experiment material:
Cell strain: HMMC7721 is a hepatoma cell line, is provided by Chinese Academy of Sciences's Shanghai cell.
Reagent: various couplet benzene ring octadiene lignanoid is dissolved in DMSO respectively, is made into 10mg/ml, be made into working solution with RPMI-1640 again as mother solution; Amycin.
Instrument: culture bottle, culture plate, FCM.
(2) experimental technique:
The FACS method is analyzed the apoptotic effect of the short tumor cell of couplet benzene ring octadiene lignanoid: the HMMC7721 cell culture is in the fresh medium of no medicine.Experiment is divided into 4 groups, and contrast 1 does not have amycin for cell and lignanoid handles; Contrast 2 is handled for only using lignanoid (10 μ g/ml); The amycin group is handled for the amycin with 0.5 μ g/ml; Amycin+lignanoid's group is after handling 24 hours with lignanoid (10 μ g/ml) earlier, to add amycin (0.5 μ g/ml) again and handle.Above cell culture is after 24 hours, and (Propidium iodide, PI) staining and AnnexinV-FITC staining are measured the apoptosis number with FACS to use propidium iodide respectively.PI and AnnexinV-FITC test kit are all available from U.S. Sigma company.
The PI method:
Collecting cell is washed cell twice with cold PBS, cell fixed with 70% ethanol, and mixing, fixing, press the dyeing of test kit description pair cell, detect excitation wavelength 488nm, emission wavelength 630nm with FACS.Apoptotic cell is a hypodiploid.
The AnnexinV-FITC method
Collecting cell is washed cell twice with cold PBS, presses test kit and presses the dyeing of test kit description pair cell, detects excitation wavelength 488nm, emission wavelength 525nm (AnnexinV-FITC) with FACS.Apoptotic cell is by the cell of Annexin V-FITC labelling.
Experimental result:
After table 7 explanation, HMMC7721 hepatoma carcinoma cell are handled 24 hours through couplet benzene ring octadiene lignanoid (10 μ g/ml), significantly promote the apoptosis of the inductive tumor cell of antineoplastic agent.
(Propidium iodide, PI) staining detects the apoptosis (hypodiploid cell) that couplet benzene ring octadiene lignanoid promotion amycin is induced the HMMC7721 hepatoma carcinoma cell to table 7. propidium iodide.
Apoptotic cell (hypodiploid cell) %
Contrast 1 Contrast 2 The amycin group Amycin+lignanoid's group
Wuweizisu A 1.8±1.7 2.4±1.5 15±7 38±11(2.5)
Wuweizisu B 1.8±1.7 2.3±1.7 15±7 35±9(2.3)
Schizantherin A 1.8±1.7 2.1±1.4 15±7 32±7(2.1)
Schizantherin B 1.8±1.7 1.9±1.5 15±7 30±6(2.0)
In the table: contrast 1 does not have amycin for cell and lignanoid handles; Contrast 2 is handled for only using lignanoid (10 μ g/ml); The amycin group is handled for the amycin with 0.5 μ g/ml; Amycin+lignanoid's group is to handle 24 hours with lignanoid (10 μ g/ml) earlier, adds amycin (0.5 μ g/ml) again and handles.Data in the table bracket are the enhanced sensitivity multiple, and promptly the apoptotic cell of amycin group+lignanoid's group is divided by the apoptotic cell of amycin group.
Detect the apoptosis that couplet benzene ring octadiene lignanoid promotes the amycin inducing tumor cell with the AnnexinV-FITC method, obtain the result similar to table 7.
Table 8:AnnexinV-FITC method detects couplet benzene ring octadiene lignanoid and promotes amycin to induce the apoptosis of HMMC7721 hepatoma carcinoma cell.
Apoptotic cell (hypodiploid cell) %
Contrast 1 Contrast 2 The amycin group Amycin+lignanoid's group
Wuweizisu A 1.6±1.3 2.1±1.2 14±8 31±8(2.2)
Wuweizisu B 1.6±1.3 2.5±0.9 14±8 29±7(2.1)
Schizantherin A 1.6±1.3 2.3±1.1 14±8 30±5(2.1)
Schizantherin B 1.6±1.3 2.2±1.6 14±8 35±8(2.5)
In the table: contrast 1 does not have amycin for cell and lignanoid handles; Contrast 2 is handled for only using lignanoid (10 μ g/ml); The amycin group is handled for the amycin with 0.5 μ g/ml; Amycin+lignanoid's group is to handle 24 hours with lignanoid (10 μ g/ml) earlier, adds amycin (0.5 μ g/ml) again and handles.Data in the table bracket are the enhanced sensitivity multiple, and promptly the apoptotic cell of amycin group+lignanoid's group is divided by the apoptotic cell of amycin group.
HMMC7721 is not a drug resistance cell strain, is the hepatoma cell line of a pair of anticarcinogen sensitivity.Above result shows that couplet benzene ring octadiene lignanoid can significantly promote amycin to induce the apoptosis of HMMC hepatoma carcinoma cell, and therefore, couplet benzene ring octadiene lignanoid can be applicable to the preparation of the sensitizer of chemotherapy of tumors.
Embodiment 1 to 6 explanation couplet benzene ring octadiene lignanoid can effectively suppress P-gp and MRP1, and the resistance that this two ' Teat pipette ' can make tumor cell produce multiple anticarcinogen (sees Gottesman MM et al:Nature Medicine for details, 2:48-58,2001; Krishna R etal:European Journal of Pharmaceutical Science11:265-283,2000).In the research of MDR reversing drug, generally acknowledge as long as determine the target spot of MDR reversing drug effect, and energy reversion MDR cell is to the Drug resistance of typical anticarcinogen, but just can think Drug resistance (Teodori E etal:Il Parmaco 57:385-415,2002 of this medicine reversion MDR tumor to other drug; Seelig A etal:European Journal of Pharmaceutical Sceinces 12:31-40,2000; United StatesPatent Serial No 003215; United States Patent Serial No 714506; United StatesPatent Serial No 354443).Confirmed the action target spot (P-gp of couplet benzene ring octadiene lignanoid in an embodiment, MRP1), and clear and definite couplet benzene ring octadiene lignanoid effectively the Drug resistance of the right typical medicaments of reversion MDR tumor cell reverse, but so inference couplet benzene ring octadiene lignanoid also the reversion MDR tumor to the Drug resistance of other antitumor drug.
10 kinds of lignanoids that have the couplet benzene ring octadiene mother nucleus structure have been used in an embodiment, these chemical compounds all have the function of the multi-drug resistance of the tumor that reverses P-glycoprotein and MRP1 mediation, therefore, having the couplet benzene ring octadiene mother nucleus structure is the key structure that this compounds reverses the multi-drug resistance of the tumor of P-glycoprotein and MRP1 mediation.Therefore also can on the chemical compound that has the couplet benzene ring octadiene mother nucleus structure, do chemical modification and obtain new multidrug resistance reversing agent.
In sum: couplet benzene ring octadiene lignanoid can effectively reverse the multi-drug resistance of the tumor by mediations such as P-gp, MRP1, BCRP, and therefore, couplet benzene ring octadiene lignanoid can be made into the medicine of reverse multiple drug resistance of tumor.Its safety is good, therefore has bigger potential applicability in clinical practice.

Claims (12)

1. couplet benzene ring octadiene lignanoid is applied to prepare the medicine for treating tumor thing.
2. the application of couplet benzene ring octadiene as claimed in claim 1 lignanoid in preparation medicine for treating tumor thing is characterized in that described couplet benzene ring octadiene lignanoid structure is suc as formula shown in the I:
Figure A2005100493820002C1
In the formula, R 1, R 2, R 5, R 6Be respectively hydroxyl or methoxyl group, perhaps, R 1With R 2, R 5With R 6Not ring formation or R separately 1With R 2, R 5With R 6Form the alcoxyl ring separately:
R 3For one of following: 1.-O-CH 32.-OH;
Figure A2005100493820002C2
R 4For one of following: 1.-O-CH 32.-OH;
Figure A2005100493820002C3
R 7For one of following: 1.-H; 2.-OH;
Figure A2005100493820003C1
R 8, R 9One of it is following respectively to do for oneself: 1.-and H; 2.-OH;
R 10For one of following: 1.-H; 2.-OH;
Figure A2005100493820003C2
Figure A2005100493820003C3
Perhaps, R 7With R 10Be linked to be oxo bridge, R 1～R 6, R 8, R 9Definition as mentioned above;
Perhaps, R 3With R 7Form the acyl-oxygen ring, R 1, R 2, R 4～R 6, R 8～R 10Definition as mentioned above.
3. the application of couplet benzene ring octadiene as claimed in claim 2 lignanoid in preparation medicine for treating tumor thing is characterized in that described couplet benzene ring octadiene lignanoid structural formula is as follows:
In the formula, R 1, R 2, R 5, R 6Be respectively hydroxyl or methoxyl group, perhaps, R 1With R 2, R 5With R 6Not ring formation or R separately 1With R 2, R 5With R 6Form the alcoxyl ring separately;
R 7For one of following: 1.-H; 2.-OH;
Figure A2005100493820004C1
R 8, R 9One of it is following respectively to do for oneself: 1.-and H; 2.-OH;
R 10For one of following: 1.-H; 2.-OH;
Figure A2005100493820004C3
Figure A2005100493820004C4
4. the application of couplet benzene ring octadiene as claimed in claim 3 lignanoid in preparation medicine for treating tumor thing is characterized in that described couplet benzene ring octadiene lignanoid is one of following formula:
1. 2. 3. schisandrin C of wuweizisu B of wuweizisu A
4. 5. 6. schizantherin C of schizantherin B of schizantherin A
7. 8. 9. 10. wuweizi alcohol B of wuweizi alcohol A of schizantherin E of schizantherin D.
5. as the application of the described couplet benzene ring octadiene of one of claim 1～4 lignanoid in preparation medicine for treating tumor thing, it is characterized in that described couplet benzene ring octadiene lignanoid is applied to prepare tumor multidrug-resistance inversion agent medicine.
6. the application of couplet benzene ring octadiene lignanoid as claimed in claim 5 in preparation medicine for treating tumor thing, it is characterized in that described inversion agent medicine also can contain the antitumor drug as tumor multidrug-resistance inversion agent sensitivity to couplet benzene ring octadiene lignanoid, and drug excipient or carrier.
7. the application of couplet benzene ring octadiene as claimed in claim 6 lignanoid in preparation medicine for treating tumor thing is characterized in that described antitumor drug to tumor multidrug-resistance inversion agent sensitivity is one of following or the mixing of its arbitrary proportion:
1. the 2. 3. substrate of breast carcinoma sorcin of substrate of multidrug-associated protein of the substrate of P-glycoprotein.
8. the application of couplet benzene ring octadiene as claimed in claim 7 lignanoid in preparation medicine for treating tumor thing is characterized in that described antitumor drug to tumor multidrug-resistance inversion agent sensitivity is one of following or the mixing of its arbitrary proportion:
The Doxorubicin amycin; The Actinomycin D actinomycin D; Altreatamine; The Bleomycin bleomycin; The Busulphan busulfan; The Capecitabine capecitabine; The Carboplatin carboplatin; The Carmustine carmustine; The Chlorambucil chlorambucil; The Cisplatin cisplatin; The cyclophosphamide cyclophosphamide; The cytarbine cytosine arabinoside; Dacarabazine, the daunorubicin daunorubicin; The epirubicin epirubicin; The etoposide etoposide; Etoposide; Etoposide; The acid of fludarbine fluorine vidarabine; The fluorouracil fluorouracil; The gemcitabine gemcitabine; The herceptin Trastuzumab; The hydroxyurea hydroxyurea; The idarubicin idarubicin; The ifosfamide ifosfamide; The irinotecan Irinotecan; The lomustine lomustine; Lomustine; The melphalan melphalan; Alkeran; The mercaptopurine purinethol; The methotrexate methotrexate; The mitomycin mitomycin; The mitozantrone mitoxantrone; Dithranol; The oxaliplatin oxaliplatin; The procarbazine procarbazine; First (base) benzyl hydrazine; The rituxan Mabthera; The steroids steroid; The streptozocin streptozocin; Streptozotocin; The taxol paclitaxel, the taxotere taxotere; Tamozolomide, the thioguanine thioguanine; The thiotepa thio-tepa; Thiotef; Tespamin; Tomudex Raltitrexed (raltitrexed); The topotecan topotecan; The treosulfan treosulfan; The uracil-tegufur uracil; The vinblastine vinblastine; Vincaleucoblastine; The vindesine vindesine; The vinorelbine vinorelbine.
9. the application of couplet benzene ring octadiene as claimed in claim 5 lignanoid in preparation medicine for treating tumor thing is characterized in that described medicine for treating tumor thing can be made into one of following dosage form:
1. 2. 3. 4. 5. slow releasing agent of granule of capsule of tablet of injection.
10. as the application of the described couplet benzene ring octadiene of one of claim 1～4 lignanoid in preparation medicine for treating tumor thing, it is characterized in that described couplet benzene ring octadiene lignanoid is applied to prepare improves the sensitizer medicine of tumor to chemotherapeutics.
11., it is characterized in that described couplet benzene ring octadiene lignanoid is applied to prepare the medicine that promotes apoptosis of tumor cells as the application of the described couplet benzene ring octadiene of one of claim 1～4 lignanoid in preparation medicine for treating tumor thing.
12., it is characterized in that described medicine for treating tumor thing is the mixture of described couplet benzene ring octadiene lignanoid as the application of the described couplet benzene ring octadiene of one of claim 1～4 lignanoid in preparation medicine for treating tumor thing.
CN 200510049382 2005-03-16 2005-03-16 Application of biphenyl cyclooctadiene lignin in treatment of tumour medicine Pending CN1833636A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102048827A (en) * 2010-01-25 2011-05-11 浙江大学 Application of shiandra for preparing medicaments for resisting tumor invasion and metastasis
CN103601621A (en) * 2013-10-18 2014-02-26 河北大学 Schisandrin and pharmaceutical application thereof
CN111410645A (en) * 2020-02-16 2020-07-14 浙江工业大学 Lignans compound and preparation and application thereof
CN114533724A (en) * 2022-04-08 2022-05-27 温州医科大学 Application of schisantherin D in preparing antineoplastic medicine

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102048827A (en) * 2010-01-25 2011-05-11 浙江大学 Application of shiandra for preparing medicaments for resisting tumor invasion and metastasis
CN103601621A (en) * 2013-10-18 2014-02-26 河北大学 Schisandrin and pharmaceutical application thereof
CN111410645A (en) * 2020-02-16 2020-07-14 浙江工业大学 Lignans compound and preparation and application thereof
CN114533724A (en) * 2022-04-08 2022-05-27 温州医科大学 Application of schisantherin D in preparing antineoplastic medicine

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