CN114805117A - 抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 - Google Patents
抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 Download PDFInfo
- Publication number
- CN114805117A CN114805117A CN202210417140.3A CN202210417140A CN114805117A CN 114805117 A CN114805117 A CN 114805117A CN 202210417140 A CN202210417140 A CN 202210417140A CN 114805117 A CN114805117 A CN 114805117A
- Authority
- CN
- China
- Prior art keywords
- cells
- stem cells
- alkannin
- cell
- tumor
- 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.)
- Pending
Links
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 102
- UNNKKUDWEASWDN-UHFFFAOYSA-N alkannin Natural products CC(=CCC(O)c1cc(O)c2C(=O)C=CC(=O)c2c1O)C UNNKKUDWEASWDN-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 239000004229 Alkannin Substances 0.000 title claims abstract description 34
- 235000019232 alkannin Nutrition 0.000 title claims abstract description 34
- -1 alkannin oxime Chemical class 0.000 title claims abstract description 33
- NEZONWMXZKDMKF-JTQLQIEISA-N Alkannin Chemical class C1=CC(O)=C2C(=O)C([C@@H](O)CC=C(C)C)=CC(=O)C2=C1O NEZONWMXZKDMKF-JTQLQIEISA-N 0.000 title claims abstract description 32
- 230000000259 anti-tumor effect Effects 0.000 title description 30
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 96
- RQNVIKXOOKXAJQ-UHFFFAOYSA-N naphthazarin Chemical compound O=C1C=CC(=O)C2=C1C(O)=CC=C2O RQNVIKXOOKXAJQ-UHFFFAOYSA-N 0.000 claims abstract description 35
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 16
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 13
- 230000011987 methylation Effects 0.000 claims abstract description 12
- 238000007069 methylation reaction Methods 0.000 claims abstract description 12
- 206010006187 Breast cancer Diseases 0.000 claims description 41
- 208000026310 Breast neoplasm Diseases 0.000 claims description 41
- 239000003814 drug Substances 0.000 claims description 30
- 201000011510 cancer Diseases 0.000 claims description 23
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 2
- 101710160107 Outer membrane protein A Proteins 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims description 2
- 229960003328 benzoyl peroxide Drugs 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 229910052799 carbon Inorganic materials 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 120
- 150000001875 compounds Chemical class 0.000 abstract description 63
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 abstract description 20
- 229940041181 antineoplastic drug Drugs 0.000 abstract description 14
- 229930012538 Paclitaxel Natural products 0.000 abstract description 11
- 229960001592 paclitaxel Drugs 0.000 abstract description 11
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 abstract description 11
- 229940009456 adriamycin Drugs 0.000 abstract description 8
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 abstract description 6
- 229960002949 fluorouracil Drugs 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000011160 research Methods 0.000 abstract description 6
- 230000000857 drug effect Effects 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- KCLANYCVBBTKTO-UHFFFAOYSA-N Proparacaine Chemical compound CCCOC1=CC=C(C(=O)OCCN(CC)CC)C=C1N KCLANYCVBBTKTO-UHFFFAOYSA-N 0.000 abstract description 2
- 229940087458 alcaine Drugs 0.000 abstract description 2
- 150000002611 lead compounds Chemical class 0.000 abstract description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 2
- 230000001988 toxicity Effects 0.000 abstract description 2
- 210000005260 human cell Anatomy 0.000 abstract 1
- 239000002547 new drug Substances 0.000 abstract 1
- 229940079593 drug Drugs 0.000 description 20
- 230000000694 effects Effects 0.000 description 19
- 230000002829 reductive effect Effects 0.000 description 19
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- 210000004940 nucleus Anatomy 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 15
- 102000004169 proteins and genes Human genes 0.000 description 14
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 230000006907 apoptotic process Effects 0.000 description 12
- 230000005764 inhibitory process Effects 0.000 description 12
- 230000014509 gene expression Effects 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 241000699670 Mus sp. Species 0.000 description 9
- 239000006285 cell suspension Substances 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 230000012010 growth Effects 0.000 description 8
- 230000002401 inhibitory effect Effects 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 230000022131 cell cycle Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 6
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 6
- 230000001640 apoptogenic effect Effects 0.000 description 6
- 230000010261 cell growth Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 230000007246 mechanism Effects 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- JATZDQLMBLHKFK-AWEZNQCLSA-N (1S)-4-methyl-1-(1,4,5,8-tetramethoxynaphthalen-2-yl)pent-3-en-1-ol Chemical compound CC(C)=CC[C@H](O)C1=CC(OC)=C2C(OC)=CC=C(OC)C2=C1OC JATZDQLMBLHKFK-AWEZNQCLSA-N 0.000 description 5
- JATZDQLMBLHKFK-CQSZACIVSA-N (1r)-4-methyl-1-(1,4,5,8-tetramethoxynaphthalen-2-yl)pent-3-en-1-ol Chemical compound CC(C)=CC[C@@H](O)C1=CC(OC)=C2C(OC)=CC=C(OC)C2=C1OC JATZDQLMBLHKFK-CQSZACIVSA-N 0.000 description 5
- ZYNAFIJFZMCIPS-QFIPXVFZSA-N 8-[(1S)-4-methyl-1-(1,4,5,8-tetramethoxynaphthalen-2-yl)pent-3-enoxy]octan-1-ol Chemical compound CC(=CC[C@@H](C1=C(C2=C(C=CC(=C2C(=C1)OC)OC)OC)OC)OCCCCCCCCO)C ZYNAFIJFZMCIPS-QFIPXVFZSA-N 0.000 description 5
- 230000004543 DNA replication Effects 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 230000018199 S phase Effects 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 210000000170 cell membrane Anatomy 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 230000009036 growth inhibition Effects 0.000 description 5
- 229910052757 nitrogen Inorganic materials 0.000 description 5
- 239000003921 oil Substances 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 4
- HTSGKJQDMSTCGS-UHFFFAOYSA-N 1,4-bis(4-chlorophenyl)-2-(4-methylphenyl)sulfonylbutane-1,4-dione Chemical compound C1=CC(C)=CC=C1S(=O)(=O)C(C(=O)C=1C=CC(Cl)=CC=1)CC(=O)C1=CC=C(Cl)C=C1 HTSGKJQDMSTCGS-UHFFFAOYSA-N 0.000 description 4
- 108090000672 Annexin A5 Proteins 0.000 description 4
- 102000004121 Annexin A5 Human genes 0.000 description 4
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 102000004142 Trypsin Human genes 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000002601 intratumoral effect Effects 0.000 description 4
- 208000032839 leukemia Diseases 0.000 description 4
- 108010082117 matrigel Proteins 0.000 description 4
- 239000012044 organic layer Substances 0.000 description 4
- 230000037361 pathway Effects 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 238000010898 silica gel chromatography Methods 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 230000035519 G0 Phase Effects 0.000 description 3
- 230000010190 G1 phase Effects 0.000 description 3
- 241001071917 Lithospermum Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229910002651 NO3 Inorganic materials 0.000 description 3
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 3
- RAFKCLFWELPONH-UHFFFAOYSA-N acetonitrile;dichloromethane Chemical compound CC#N.ClCCl RAFKCLFWELPONH-UHFFFAOYSA-N 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 3
- 229940044683 chemotherapy drug Drugs 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 239000012065 filter cake Substances 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000004563 mammosphere formation Effects 0.000 description 3
- 239000012046 mixed solvent Substances 0.000 description 3
- 229930014626 natural product Natural products 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 238000006146 oximation reaction Methods 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 3
- 239000012679 serum free medium Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 2
- 102000000412 Annexin Human genes 0.000 description 2
- 108050008874 Annexin Proteins 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 230000027311 M phase Effects 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 238000011579 SCID mouse model Methods 0.000 description 2
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000021164 cell adhesion Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 231100000433 cytotoxic Toxicity 0.000 description 2
- 230000001472 cytotoxic effect Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229960003668 docetaxel Drugs 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 108091008039 hormone receptors Proteins 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 230000005918 in vitro anti-tumor Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000000394 mitotic effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000009456 molecular mechanism Effects 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 238000001959 radiotherapy Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 229910000104 sodium hydride Inorganic materials 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 208000022679 triple-negative breast carcinoma Diseases 0.000 description 2
- VIJSPAIQWVPKQZ-BLECARSGSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-acetamido-5-(diaminomethylideneamino)pentanoyl]amino]-4-methylpentanoyl]amino]-4,4-dimethylpentanoyl]amino]-4-methylpentanoyl]amino]propanoyl]amino]-5-(diaminomethylideneamino)pentanoic acid Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(C)=O VIJSPAIQWVPKQZ-BLECARSGSA-N 0.000 description 1
- FBYIJIHPWCXRHH-UHFFFAOYSA-N 1,4,5,8-tetramethoxynaphthalene-2-carbaldehyde Chemical compound O=CC1=CC(OC)=C2C(OC)=CC=C(OC)C2=C1OC FBYIJIHPWCXRHH-UHFFFAOYSA-N 0.000 description 1
- BUZZUHJODKQYTF-UHFFFAOYSA-N 1-iodo-3-methylbutane Chemical compound CC(C)CCI BUZZUHJODKQYTF-UHFFFAOYSA-N 0.000 description 1
- HAWSQZCWOQZXHI-FQEVSTJZSA-N 10-Hydroxycamptothecin Chemical compound C1=C(O)C=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 HAWSQZCWOQZXHI-FQEVSTJZSA-N 0.000 description 1
- JCXZKUZXVQKENT-UHFFFAOYSA-N 2-(4-methylpiperazin-1-ium-1-yl)acetate Chemical compound CN1CCN(CC(O)=O)CC1 JCXZKUZXVQKENT-UHFFFAOYSA-N 0.000 description 1
- JCRBYQZIJFWGOO-UHFFFAOYSA-N 2-(8-bromooctoxy)oxane Chemical compound BrCCCCCCCCOC1CCCCO1 JCRBYQZIJFWGOO-UHFFFAOYSA-N 0.000 description 1
- JSBBMMHTNIUVTC-UHFFFAOYSA-N 2-hydroxyethanone Chemical class OC[C]=O JSBBMMHTNIUVTC-UHFFFAOYSA-N 0.000 description 1
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 1
- ZYNAFIJFZMCIPS-JOCHJYFZSA-N 8-[(1R)-4-methyl-1-(1,4,5,8-tetramethoxynaphthalen-2-yl)pent-3-enoxy]octan-1-ol Chemical compound CC(=CC[C@H](C1=C(C2=C(C=CC(=C2C(=C1)OC)OC)OC)OC)OCCCCCCCCO)C ZYNAFIJFZMCIPS-JOCHJYFZSA-N 0.000 description 1
- 102000004000 Aurora Kinase A Human genes 0.000 description 1
- 108090000461 Aurora Kinase A Proteins 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 102000000905 Cadherin Human genes 0.000 description 1
- 108050007957 Cadherin Proteins 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- 108010067225 Cell Adhesion Molecules Proteins 0.000 description 1
- 102000016289 Cell Adhesion Molecules Human genes 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000012746 DNA damage checkpoint Effects 0.000 description 1
- 229940123780 DNA topoisomerase I inhibitor Drugs 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 230000010337 G2 phase Effects 0.000 description 1
- 230000004668 G2/M phase Effects 0.000 description 1
- 102100032606 Heat shock factor protein 1 Human genes 0.000 description 1
- 101710190344 Heat shock factor protein 1 Proteins 0.000 description 1
- 101000740519 Homo sapiens Syndecan-4 Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 1
- 238000011789 NOD SCID mouse Methods 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 108010017324 STAT3 Transcription Factor Proteins 0.000 description 1
- 101100243565 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) PAC10 gene Proteins 0.000 description 1
- 102100024040 Signal transducer and activator of transcription 3 Human genes 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 102100037220 Syndecan-4 Human genes 0.000 description 1
- 208000035199 Tetraploidy Diseases 0.000 description 1
- 108010046075 Thymosin Proteins 0.000 description 1
- 102000007501 Thymosin Human genes 0.000 description 1
- 239000000365 Topoisomerase I Inhibitor Substances 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000002280 anti-androgenic effect Effects 0.000 description 1
- 229940046836 anti-estrogen Drugs 0.000 description 1
- 230000001833 anti-estrogenic effect Effects 0.000 description 1
- 239000000051 antiandrogen Substances 0.000 description 1
- 229940030495 antiandrogen sex hormone and modulator of the genital system Drugs 0.000 description 1
- 230000005775 apoptotic pathway Effects 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 239000003886 aromatase inhibitor Substances 0.000 description 1
- 229940046844 aromatase inhibitors Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000010009 beating Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000003766 bioinformatics method Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 210000005056 cell body Anatomy 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000025084 cell cycle arrest Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000010428 chromatin condensation Effects 0.000 description 1
- 229940121657 clinical drug Drugs 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 238000010520 demethylation reaction Methods 0.000 description 1
- 230000023077 detection of light stimulus Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000328 estrogen antagonist Substances 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 201000007741 female breast cancer Diseases 0.000 description 1
- 201000002276 female breast carcinoma Diseases 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 239000003163 gonadal steroid hormone Substances 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005917 in vivo anti-tumor Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 210000004216 mammary stem cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003990 molecular pathway Effects 0.000 description 1
- 229940125645 monoclonal antibody drug Drugs 0.000 description 1
- FRASJONUBLZVQX-UHFFFAOYSA-N naphthoquinone group Chemical group C1(C=CC(C2=CC=CC=C12)=O)=O FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 1
- 229930082768 natural naphthoquinone Natural products 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000002135 phase contrast microscopy Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000583 progesterone congener Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- BJDYCCHRZIFCGN-UHFFFAOYSA-N pyridin-1-ium;iodide Chemical compound I.C1=CC=NC=C1 BJDYCCHRZIFCGN-UHFFFAOYSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000005932 reductive alkylation reaction Methods 0.000 description 1
- 208000016691 refractory malignant neoplasm Diseases 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000004017 serum-free culture medium Substances 0.000 description 1
- 208000002491 severe combined immunodeficiency Diseases 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 210000001626 skin fibroblast Anatomy 0.000 description 1
- 239000012312 sodium hydride Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C251/00—Compounds containing nitrogen atoms doubly-bound to a carbon skeleton
- C07C251/32—Oximes
- C07C251/34—Oximes with oxygen atoms of oxyimino groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals
- C07C251/44—Oximes with oxygen atoms of oxyimino groups bound to hydrogen atoms or to carbon atoms of unsubstituted hydrocarbon radicals with the carbon atom of at least one of the oxyimino groups being part of a ring other than a six-membered aromatic ring
- C07C251/46—Quinone oximes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
- C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
- C07D295/14—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D295/145—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with the ring nitrogen atoms and the carbon atoms with three bonds to hetero atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
- C07D295/15—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals with the ring nitrogen atoms and the carbon atoms with three bonds to hetero atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/04—One of the condensed rings being a six-membered aromatic ring
- C07C2602/10—One of the condensed rings being a six-membered aromatic ring the other ring being six-membered, e.g. tetraline
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
技术领域
本发明属于医药领域,涉及一种抗肿瘤干细胞的紫草素及阿卡宁肟衍生物,具体涉及一种紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物及其医药用途。
背景技术
肿瘤是严重危害人类健康的一种疾病,近年来其发病率逐年上升,发病情况也呈现明显的年轻化趋势(S.Gupta,et al.International trends in the incidence ofcancer among adolescents and young adults,J.Natl.Cancer Inst.,2020,112(11),1105-1117)。全球癌症观察站(GLOBOCAN)近期发布的统计报告表明,2020年全球新增癌症病例约1929万,当年死亡病例995万(H.Sung,et al.Global cancer statistics 2020:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185countries.CA:a cancer journal for clinicians,2021,71(3),209-249)。当年新发病例中,女性乳腺癌的发病率最高,占到全球新增癌症病例的11.7%;其次为肺癌 (11.4%)和结肠直肠癌(10.0%)。随着新诊疗方法的出现,近年来癌症患者总体生存率显著提高,但在治疗中出现的肿瘤的迁移、复发及耐药现象,是疾病难以治愈的关键原因。
肿瘤是由异质性细胞群体构成,这些细胞群具有不同的表观形态、功能、表面抗原类型及基因的表达水平。其中的一小部分细胞(0.2-3%)具有与干细胞相似的自我更新能力并可以产生肿瘤内所有异质性细胞,这部分细胞被称为肿瘤干细胞(E.Batlle,etal.Cancer stem cells revisited,Nat.Med.,2017,23,1124-1134.)。肿瘤干细胞具有自我更新及无限增殖的特性,可以产生构成实体瘤内所有异质性的癌细胞群。肿瘤干细胞的存在首次于1997年发表的关于急性髓性白血病(AML) 的研究中得到证实(D.Bonnet,etal.Human acute myeloid leukemia is organized as a hierarchy that originatesfrom a primitive hematopoietic cell.Nat.Med.,1997,3, 730-737)。少量的原代AML细胞在转移到非肥胖型糖尿病/严重联合免疫缺陷 (NOD/SCID)小鼠时可以重现白血病并维持白血病生长。2003年,Al-Hajj等人首先证实了乳腺癌中肿瘤干细胞的存在,该类细胞的表面抗原类型为 CD44+CD24-/low。乳腺癌患者的CD44+CD24-/low肿瘤细胞在NOD/SCID小鼠中比相应的 CD44+CD24+细胞具有更高的致瘤性。此外,由CD44+CD24-/low乳腺癌细胞所形成的肿瘤可以连续传代,这表明该细胞具有自我更新能力,且可以连续复制并产生具有细胞异质性的肿瘤。
肿瘤干细胞具有高度的耐药性,对临床常用的化疗药物及放射性疗法均不敏感(C.T.Jordan,et al.Cancer stem cells,New Engl.J.Med.,2006,355,1253-1261)。在化疗及放疗过程中,处于快速有丝分裂期的普通癌细胞被杀伤,而具有耐药性的癌症干细胞在特定条件下持续分裂并维持肿瘤的生长,导致肿瘤复发。由肿瘤干细胞分化的癌细胞也会获得干细胞的特性,增加化疗耐药性。此外,研究表明乳腺癌中的肿瘤干细胞也与肿瘤的侵袭性相关。因此,寻找选择性杀伤肿瘤干细胞的药物,对于解决目前肿瘤治疗中出现的迁移、复发及耐药现象,具有非常重要的意义。
目前临床使用的抗肿瘤药物可以分为细胞毒类药物、激素类药物、生物反应调节剂、单克隆抗体药物等(孙燕。关于抗肿瘤药物分类的共识建议。循证医学, 2004,3,190-191)。细胞毒类药物主要通过抑制肿瘤细胞的复制或生长来发挥作用,分为作用于DNA化学结构的药物、影响核酸合成的药物、作用于核酸转录的药物、作用于DNA复制的拓扑异构酶Ⅰ抑制剂、主要作用于有丝分裂M期干扰微管蛋白合成的药物等。激素类药物通过特异性与激素受体结合而发挥作用,形成激素受体复合物,被活化并进入细胞体内,调节DNA的复制与细胞分裂。激素类药物分为抗雌激素、芳香化酶抑制剂、孕激素、性激素、抗雄激素、RH-LH激动剂/拮抗剂等。生物反应调节剂为通过免疫系统增强机体抗肿瘤效应并对肿瘤有治疗效果的药剂,分为干扰素、白细胞介素2、胸腺肽类等。单克隆抗体主要通过抗体作用于细胞膜表面的特定抗原,调节细胞的生长,提高肿瘤细胞的对于化疗药物的敏感性。上述临床抗肿瘤药物作用于快速生长的细胞,由于肿瘤干细胞一般处于静止期,因此上述靶向快速分裂细胞的临床药物,对肿瘤干细胞不敏感。治疗乳腺癌的一线临床抗肿瘤药物紫杉醇,对于三阴性乳腺癌细胞系 MDA-MB-231具有非常强的生长抑制活性,其IC50值仅为8nM;而对于具有肿瘤干细胞性质的三阴性乳腺癌细胞系MDA-MB-231PAC10,紫杉醇的IC50大于1 μM,IC50值的差异大于100倍(Liu,P.,et al.Disulfiram targets cancer stem-like cells andreverses resistance and cross-resistance in acquired paclitaxel-resistanttriple-negative breast cancer cells.Br.J.Cancer,2013,109,1876–1885)。对于分选所得的CD44+/CD24-/lowMCF-7乳腺癌干细胞,紫杉醇的IC50大于100μM(J.Cui, etal.,Naturalproducts targeting cancer stem cells:a revisit,Curr.Med.Chem.,2021, 28,6773-6804)。因此,不能简单的根据药物具有抗肿瘤活性的特点,即推测其具有抗肿瘤干细胞活性。
从天然产物中寻找预防和治疗恶性肿瘤的药物始终是药学领域的研究热点之一。目前临床中使用的抗肿瘤药物紫杉醇、多西紫杉醇、博来霉素、阿霉素、多柔比星、喜树碱、羟基喜树碱等均源于天然产物。作为传统化疗药物,这些抗肿瘤药对体内快速生长的细胞均有较强的生长抑制活性,但对于肿瘤干细胞几乎无影响。同时,这些抗肿瘤药物缺乏选择性,即在杀伤肿瘤细胞的同时,也对正常细胞及免疫细胞表现出极大的杀伤作用。天然萘醌类化合物紫草素有广谱抗肿瘤活性,其萘醌结构产生活性氧及侧链的生物还原烷基化作用是该化合物具有抗肿瘤活性的分子机制(X.Zhang,et al.Advance in Anti-tumorMechanisms of Shikonin,Alkannin and Their Derivatives.Mini-Rev.Med.Chem.,2018,18(2), 164-172)。近期研究报道表明,该化合物对于肿瘤干细胞具有生长抑制作用(R. Thakur,et al.Inhibition of STAT3,FAK and Src mediated signaling reducescancer stem cell load,tumorigenic potential and metastasis in breastcancer.Sci.Rep.,2015, 5(1),1-16),然而紫草素的广泛细胞毒性,限制了该化合物作为抗肿瘤干细胞药物的临床应用。
发明内容
本发明的目的在于克服现有技术的不足,提供一种抗肿瘤干细胞的紫草素及阿卡宁肟衍生物。具体而言,本发明以(S)-2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8- 四甲氧基萘(附图1,Ⅹ-S)及其对映体(R)-2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8- 四甲氧基萘(附图1,Ⅹ-R)为原料,在侧链羟基上引入烷基链或者末端具有含氮杂环的烷基链,再通过氧化脱甲基化羰基肟化方法,制备了一种具有抗肿瘤干细胞活性的紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物。药理结果显示,本发明所述紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物具有很强的抗肿瘤干细胞活性,抗肿瘤干细胞作用的机理为靶向肿瘤干细胞的黏附分子通路,杀伤肿瘤干细胞,这一抗肿瘤干细胞的机理完全不同于现有技术(Z L201310044877.6)中所公开化合物的抗肿瘤作用机理。本专利所公开的化合物其抗肿瘤干细胞作用机理新颖,药效显著高于阿霉素、紫杉醇、氟尿嘧啶等临床常用抗肿瘤药物,而且于正常细胞几乎无毒性,有良好的开发前景。
本发明的目的是通过以下技术方案实现的:
第一方面,本发明涉及一种抗肿瘤干细胞的紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物,所述衍生物的结构式如式(Ⅰ)所示:
其中,R代表含3,3-二甲基正丙基、8-羟基正辛基、8-(1-甲基-4-哌嗪乙酰氧基)正辛基;手性碳(*)的构型为R型或S型。
第二方面,本发明涉及一种所述紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物在制备抗肿瘤干细胞药物中的用途。
所述抗肿瘤干细胞药物中所述衍生物为主要活性成分。
优选的,所述肿瘤干细胞为细胞表面抗原类型为CD44+CD24-/low的乳腺癌干细胞。
第三方面,本发明涉及一种所述紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物与其它抗肿瘤药物联用在制备用于恶性肿瘤的治疗的药物中的用途。
优选的,所述药物可制成下列之一的剂型:①注射剂;②片剂;③胶囊剂;④颗粒剂。
与现有技术相比,本发明具有如下的有益效果:
1)本发明所述紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物结构明确,制备方法简便,收率较高且原料易得。
2)体外抗肿瘤活性实验研究表明,该类化合物抗肿瘤干细胞活性较强,显著优于临床常用的抗肿瘤药如阿霉素、紫杉醇及氟尿嘧啶。
3)本发明所公开的紫草素及阿卡宁萘茜母核羟基甲基化羰基肟衍生物抗肿瘤干细胞作用的机理新颖,完全不同于现有技术(ZL201310044877.6)中所公开化合物的抗肿瘤作用机理。
4)体外细胞生长抑制活性实验研究表明,本发明所述化合物对人体正常细胞几乎无生长抑制活性,表现出良好的选择性。
附图说明
通过阅读以下附图对非限制性实施例所作的详细描述,本发明的其它特征、目的和优点将会变得更明显:
图1为紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物的制备方法示意图。
图2为分选前后乳腺癌干细胞MCF-7 CD44+CD24-/low在细胞群落中的比例变化。
图3为分选所得乳腺癌干细胞MCF-7 CD44+CD24-/low的成瘤性实验结果。
图4为化合物Ⅱ-2对MCF-7 CD44+CD24-/low乳腺癌干细胞球形态的影响实验结果。
图5为化合物Ⅱ-2对MCF-7 CD44+CD24-/low乳腺癌干细胞的细胞周期阻滞效果。
图6为化合物Ⅱ-2诱导MCF-7 CD44+CD24-/low乳腺癌干细胞凋亡。
图7为化合物Ⅱ-2的体内抗肿瘤干细胞活性测试结果。
具体实施方式
下面结合附图和具体实施例对本发明进行详细说明。以下实施例将有助于本领域的技术人员进一步理解本发明,但不以任何形式限制本发明。应当指出的是,对本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干调整和改进。这些都属于本发明的保护范围。下列实施例中未注明具体条件的实验方法,通常按照常规条件,或按照制造厂商所建议的条件。
实施例1
本实施例涉及一种具有结构式(Ⅱ-1)的(S)-2-(1-异戊氧基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘的制备方法,如图1所示,包括以下步骤:
采用文献(Liming Zhao et al,Letters in Organic Chemistry,2008,5,234-236;徐彦等,紫草素与阿卡宁的不对称全合成。沈阳药科大学学报,2014,3 1(6),440-443,482)报道的方法,以1,4,5,8-四甲氧基-2-萘甲醛为原料,制备(S) -2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅹ-S)。取(S)-2-(1-羟基-4-甲基 -3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅹ-S,1.7g,5.0mmol)溶于无水DMF(30.0 mL),冰浴以及氮气保护条件下加入60%NaH(1.0g,25.0mmol),30min 后,将碘代异戊烷(5.0g,25.0mmol)缓慢滴加到反应液中,20min后升温至60℃搅拌过夜,反应完毕加入二氯甲烷和水稀释反应液,萃取,无水Na2SO4干燥,减压浓缩,粗产品用硅胶柱层析得到无色油状物(1.90g,91.3%)。1H NMR(400MHz,CDCl3):δ6.96(s,1H),6.81(s,2H),5.25–5.21(m, 1H),4.88–4.84(m,1H),3.94–3.90(m,9H),3.72(s,3H),3.34–3.28 (m,2H),2.48–2.44(m,2H),1.68(s,3H),1.52(s,3H),1.44–1.40(m,1 H),1.27–1.23(m,2H),0.85(s,3H),0.80(s,3H)。
实施例2
本实施例涉及一种具有结构式(Ⅱ-2)的(S)-6-(1’-异戊氧基-4’-甲基-3’-戊烯基)-5,8-二甲氧基-1,4-萘二酮二肟的制备方法,如图1所示,包括以下步骤:
将(S)-2-(1-异戊氧基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅱ-1,1.25g,3.0mmol)溶于二氯甲烷-乙腈混合溶剂(3:1,V/V,45mL)中,冰浴下加入硝酸铈铵(4.24g,7.7mmol)的水溶液(4.5mL),反应完成后用二氯甲烷萃取,有机层经无水Na2SO4干燥,减压浓缩,粗产品用硅胶柱层析得到黄色油状物(0. 31g)。将该黄色溶于无水乙醇(24.0mL)和吡啶(5.50mL),氮气保护下加入盐酸羟胺(0.39g,5.6mmol),53℃搅拌过夜,减压蒸除溶剂,加水抽滤,滤饼干燥后用无水乙醇重结晶得化合物Ⅱ-2,呈淡黄色固体(0.25g,收率75.1%)。1H NMR(400MHz,CDCl3):δ=11.9(s,2H,=N-OH),7.68(d,2H, HQuin),7.23(s,1H,HAr),5.24(t,1H,-CH=),4.75(t,1H,-CHOH(CH2)), 3.98(S,9H,ArOCH3),3.78(s,3H,ArOCH3),3.37(t,2H,-CH2-O-),2.4 2(m,2H,-CH2-CH=),1.67(s,3H,CH3)1.54(s,3H,CH3),1.47(m,1H), 1.24(m,2H),0.88(2s,3H,CH3),0.84(2s,3H,CH3)。
实施例3
本实施例涉及一种具有结构式(Ⅱ-2)的(R)-6-(1’-异戊氧基-4’-甲基-3’-戊烯基)-5,8-二甲氧基-1,4-萘二酮二肟的制备方法,如图1所示,包括以下步骤:
采用文献(Liming Zhao et al,Letters in Organic Chemistry,2008,5,234-236;徐彦等,紫草素与阿卡宁的不对称全合成。沈阳药科大学学报,2014,31(6), 440-443,482)报道的方法,以1,4,5,8-四甲氧基-2-萘甲醛为原料,制备(R)-2-(1- 羟基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅹ-R)。后以(R)-2-(1-羟基-4-甲基-3- 戊烯基)-1,4,5,8-四甲氧基萘(Ⅹ-R)为原料,采用与实施例1及实施例2相同的实验步骤,制备化合物Ⅱ-3。化合物呈淡黄色粉末,收率63.6%。1H NMR(400 MHz,DMSO-d6)δ12.02(s,2H),7.42–7.28(m,2H),7.05(s,1H),5.23– 5.13(m,1H),4.68–4.55(m,1H),3.74(s,3H),3.55(s,3H),3.30–3.19 (m,2H),2.32–2.26(m,2H),1.74–1.65(m,1H),1.60(s,3H),1.47(s,3 H),1.40–1.28(m,2H),0.88–0.65(m,6H)
实施例4
本实施例涉及一种具有结构式(Ⅲ-1)的(S)-2-(1-(8-羟基辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘的制备方法,如图1所示,包括以下步骤:
将(S)-2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(0.5g,1.4mmol)溶于无水DMF(15mL)中,N2保护,冰浴条件下加入的氢化钠(0.45g,60%, 11.2mmol),继续搅拌30min。然后滴加2-(8-溴辛氧基)四氢-2H-吡喃(2.8m mol)和碘化钾(46mg,0.28mmol),60℃反应过夜。反应完毕后,用乙酸乙酯萃取,合并有机层;有机相减压浓缩,柱层析分离得淡黄色油状物。将该黄色油状物溶于甲醇(10mL)中,室温下滴加浓盐酸(1mL),继续搅拌30min。待反应完毕后,向反应液中加入CH2Cl2(10mL)稀释,用饱和碳酸氢钠溶液中和盐酸,分出有机相;有机相用饱和NaCl洗,无水硫酸钠干燥,减压浓缩,柱层析分离,得(S)-2-(1-(8-羟基辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅲ -1),呈淡黄色油状液体(收率94.7%)。1H NMR(400MHz,CDCl3):δ6.89 (s,1H),6.72–6.66(m,2H),5.21–5.15(m,1H),4.82–4.74(m,1H),3.82 (s,3H),3.79(s,3H),3.75(s,3H),3.63(s,3H),3.46–3.40(m,2H),3.25 –3.16(m,2H),2.80–2.71(m,1H),2.44–2.35(m,2H),1.55(s,3H),1.43 (s,3H),1.40–1.34(m,2H),1.31–1.08(m,10H)。
实施例5
本实施例涉及一种具有结构式(Ⅲ-2)的(S)-6-(1-(8-羟基辛氧基)-4-甲基戊-3-烯-1-基)-5,8-二甲氧基-1,4-萘二酮二肟的制备方法,如图1所示,包括以下步骤:
将(S)-2-(1-(8-羟基辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(1.42g,3. 0mmol)溶于二氯甲烷-乙腈混合溶剂(3:1,V/V,45mL)中,冰浴下加入硝酸铈铵(4.24g,7.7mmol)的水溶液(4.5mL),反应完成后用二氯甲烷萃取,有机层经无水Na2SO4干燥,减压浓缩,粗产品用硅胶柱层析得到黄色油状物(0. 31g)。将该黄色溶于无水乙醇(24.0mL)和吡啶(5.50mL),氮气保护下加入盐酸羟胺(0.39g,5.6mmol),53℃搅拌过夜,减压蒸除溶剂,加水抽滤,滤饼干燥后用无水乙醇重结晶得目标化合物,呈淡黄色固体(0.45g,31.6%)。1 HNMR(400MHz,DMSO-d6)δ11.98(s,2H),7.38–7.32(m,2H),7.04(s, 1H),5.18(t,J=6.3Hz,1H),4.60(t,J=6.3Hz,1H),4.24–4.16(m,1 H),3.74(s,3H),3.55(s,3H),3.34–3.28(m,2H),3.25–3.18(m,2H),2.3 2–2.24(m,2H),1.59(s,3H),1.46(s,3H),1.48–1.38(m,12H)。
实施例6
本实施例涉及一种具有结构式(Ⅲ-3)的(R)-2-(1-(8-羟基辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘的制备方法,如图1所示,包括以下步骤:
以(R)-2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘为原料,采用与实施例 4及实施例5相同的实验步骤,制备化合物Ⅲ-3。化合物呈淡黄色粉末,收率31. 2%。1HNMR(400MHz,DMSO-d6):δ11.98(s,2H),7.38–7.32(m,2H),7. 04(s,1H),5.18(t,J=6.3Hz,1H),4.61(t,J=6.3Hz,1H),4.24–4.16 (m,1H),3.74(s,3H),3.55(s,3H),3.34–3.28(m,2H),3.25–3.18(m,2 H),2.32–2.24(m,2H),1.60(s,3H),1.47(s,3H),1.46–1.38(m,12H)。
实施例7
本实施例涉及一种具有结构式(IV-1)的(S)-2-(1-(8-(1-甲基-4-哌嗪乙酰氧)辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘的制备方法,如图1所示,包括以下步骤:
将(S)-2-(1-(8-羟基辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(Ⅲ-1,1.42 g,3.0mmol)溶于无水二氯甲烷中,加入催化量的对二甲氨基吡啶及1.5倍当量的缩合剂N,N’-二环己基碳二亚胺。反应液室温搅拌10min后,加入1.5倍当量的4-甲基-1-哌嗪乙酸。反应液室温搅拌24小时后,用二氯甲烷稀释并抽滤。滤液经水、饱和食盐水清洗,减压浓缩至干后得目标化合物IV-1,呈淡黄色油状物(1.72g,产率93.4%)。1H NMR(400MHz,CDCl3)δ6.91(s,1H),6.76 (d,J=2.2Hz,2H),5.19(t,J=6.7Hz,1H),4.81(t,J=6.9Hz,1H),4.02(t,J=6.7Hz,2H),3.85(s,9H),3.67(s,3H),3.30–3.18(m,2H),3.14(s, 2H),2.69–2.48(m,8H),2.43–2.39(m,2H),2.25(s,3H),1.61(s,3H), 1.55–1.51(m,2H),1.47(s,3H),1.35–1.10(m,10H)。
实施例8
本实施例涉及一种具有结构式(IV-2)的(S)-(1E,4E)-6-(1-(8-(1-甲基-4-哌嗪乙酰氧)辛氧基)-4-甲基戊-3-烯-1-基)-5,8-二甲氧基萘-1,4-二酮二肟的制备方法,如图1所示,包括以下步骤:
将(S)-2-(1-(8-(1-甲基-4-哌嗪乙酰氧)辛氧基)-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘(1.84g,3.0mmol)溶于二氯甲烷-乙腈混合溶剂(3:1,V/V,45mL) 中,冰浴下加入硝酸铈铵(4.24g,7.7mmol)的水溶液(4.5mL),反应完成后用二氯甲烷萃取,有机层经无水Na2SO4干燥,减压浓缩,粗产品用硅胶柱层析得到黄色油状物(0.31g)。将该黄色溶于无水乙醇(24.0mL)和吡啶(5.50 mL),氮气保护下加入盐酸羟胺(0.39g,5.6mmol),53℃搅拌过夜,减压蒸除溶剂,加水抽滤,滤饼干燥后用无水乙醇重结晶得化合物IV-2,呈淡黄色固体(0.36g,产率19.7%)。1H NMR(400MHz,DMSO-d6)δ11.99(s,2H),7.3 5(d,J=2.6Hz,2H),7.05(s,1H),5.18(t,J=6.7Hz,1H),4.62(t,J=6. 6Hz,1H),3.99(d,J=7.1Hz,2H),3.75(s,3H),3.56(s,3H),3.25–3.21 (m,4H),2.82–2.73(s,4H),2.64(s,3H),2.48–2.44(m,4H),2.31(s,2H), 1.61(s,3H),1.53–1.50(m,2H),1.48(s,3H),1.35–1.10(m,10H)。
实施例9
本实施例涉及一种具有结构式(IV-3)的(R)-(1E,4E)-6-(1-(8-(1-甲基-4-哌嗪乙酰氧)辛氧基)-4-甲基戊-3-烯-1-基)-5,8-二甲氧基萘-1,4-二酮二肟的制备方法,如图1所示,包括以下步骤:
以(R)-2-(1-羟基-4-甲基-3-戊烯基)-1,4,5,8-四甲氧基萘为原料,采用与实施例 4、实施例7及实施例8相同的实验步骤,制备化合物IV-3。化合物呈淡黄色粉末,总收率16.2%。1H NMR(400MHz,DMSO-d6)δ12.03(s,1H),10.59(s, 1H),7.41–7.36(m,2H),7.08(s,1H),5.22(t,J=7.1Hz,1H),4.65(t,J =6.4Hz,1H),4.06(t,J=6.6Hz,2H),3.78(s,3H),3.59(s,3H),3.44(d, J=11.8Hz,2H),3.28(q,J=6.4,6.0Hz,2H),3.17(s,6H),2.93(s,2H), 2.77(s,3H),2.35(s,2H),1.64(s,3H),1.60–1.53(m,2H),1.51(s,5H),1. 25(s,8H)。
实施例10
本实施例涉及乳腺癌干细胞MCF-7 CD44+CD24-/low的分选及表征方法,包括以下步骤:
参考文献(Muhammad Al-Hajj et al,PNAS,100(2003),3983-3988)所报道的方法,我们采用荧光标记抗体CD44-APC及CD24-PE,通过流式细胞术分选了乳腺癌MCF-7 CD44+CD24-/low细胞亚群(乳腺癌干细胞)。将贴壁的MCF-7细胞用预冷的PBS清洗后,用稀释的胰蛋白酶-乙二胺四乙酸(EDTA)溶液处理以使细胞从培养皿表面脱离。为了尽量减少对细胞的损害,用相差显微镜监测脱离。目视确认脱离后,加入5ml含有血清的培养基终止消化。将细胞悬液离心后,用5mlPBS洗涤MCF-7细胞。离心5分钟后,将离心管底部的细胞用500μL 的PBS重悬,根据制造商的说明,使用荧光标记抗体CD44-APC及CD24-PE(BD Pharmingen,SanDiego,CA)将细胞染色。细胞染色结束后,在室温下将细胞用PBS冲洗3次,然后转移到5ml流式细胞管。BD FACS Calibur(Becton& Dickinson Biosciences,San Jose,CA)上对细胞进行分选,分选后的细胞于 Mammocult无血清培养基中进行悬浮培养。悬浮培养的细胞2周使用荧光标记抗体CD44-APC及CD24-PE重新分析细胞表面抗原(CD44及CD24)的表达,乳腺癌干细胞MCF-7 CD44+CD24-/low在所培养的细胞中,比例达到75%。分选前的 MCF-7贴壁细胞,分化抗原(CD)类型为CD44+CD24-/low的细胞亚群(乳腺癌干细胞)比例仅为1.89%(附图2)。
实施例11
本实施例涉及分选所得的乳腺癌干细胞MCF-7 CD44+CD24-/low的体内成瘤性实验,包括以下步骤:
将免疫缺陷小鼠NOD/SCID饲养于SPF级实验动物中心,温度、湿度和光照分别控制在21±2℃,50±10%和12h光照/12h黑暗循环。所有动物实验的操作均参照动物实验伦理委员会指导手册要求执行。将分选所得的乳腺癌干细胞 MCF-7 CD44+CD24-/low及分选前的MCF-7细胞正常传代,培养至所需数目。细胞经胰酶消化后,用PBS清洗并制备两种细胞的细胞悬液,调整细胞悬液中的细胞数至所需浓度。将该细胞悬液与基质胶(BD MatrigelTM)等体积混合后,接种于NOD/SCID小鼠皮下。接种2周后,将小鼠实施安乐死并取出肿瘤。结果如附图3所示,接种105个、2*104个及104个MCF-7 CD44+CD24-/low乳腺癌干细胞的小鼠,皮下均有移植瘤生成。而接种107个MCF-7细胞的小鼠,无移植瘤产生。实验结果表明,分选所得的MCF-7 CD44+CD24-/low乳腺癌干细胞的具有很强的成瘤性,与文献(Muhammad Al-Hajj etal,PNAS,100(2003),3983-3988)所报道的乳腺癌干细胞的成瘤性一致。
实施例12
本实施例涉及紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物对乳腺癌干细胞MCF-7 CD44+CD24-/low的生长抑制活性测试,包括以下步骤:
取对数生长期的MCF-7 CD44+CD24-/low肿瘤干细胞,用0.25%胰蛋白酶消化后,用磷酸盐缓冲液稀释后离心,加入Mammocult无血清培养基混悬,并调整细胞悬液至合适的浓度,在96孔板中每孔加入100μL细胞悬液,使得每孔中的细胞数为5000个。将96孔板静置于37℃,5%CO2恒温培养箱中培养12h,待细胞贴壁后每孔加入用完全培养基稀释的化合物100μL,各组均设三个复孔。每块96孔板上需设置调零孔(只加化合物与培养基,不含细胞)与空白对照孔 (只含有细胞与培养基,不加化合物)。继续培养48h后,加入之前配好的MTS溶液20μL/孔,避光继续培养4h,在多功能酶标仪上490nm波长处检测各孔的吸光度(OD值)。测试中选用临床常用抗肿瘤药阿霉素、紫杉醇及氟尿嘧啶为阳性对照药物。
细胞生长抑制率的计算公式:
生长抑制率(%)=(1–给药组OD值/对照组OD值)×100
根据计算得到的不同给药浓度下的细胞生长抑制率,采用Prism软件中的 Dose-response-Inhibition里面的log(inhibitor)vs.normalized response程序拟合曲线,计算IC50值。
表1目标化合物抑制乳腺癌干细胞MCF-7 CD44+CD24-/low的IC50值
化合物 | IC<sub>50</sub>(μM) |
Ⅱ-2 | 0.77 |
Ⅱ-3 | 1.08 |
Ⅲ-2 | 0.96 |
Ⅲ-3 | 1.76 |
IV-2 | 1.21 |
IV-3 | 2.09 |
阿霉素 | 38.6 |
紫杉醇 | >100 |
氟尿嘧啶 | >250 |
在上述肿瘤干细胞生长抑制活性测试中,所采用的阳性对照药物阿霉素、紫杉醇及氟尿嘧啶均是临床常用的抗肿瘤药物,文献(Sun W H.Synthesis and cytotoxicityevaluation of oleanolic acid derivatives.Bioorganic&Medicinal Ch emistryLetters,2013;)研究表明,这三种药物对于乳腺癌MCF-7细胞有非常强的生长抑制活性(阿霉素IC50=1.65μM;),然而几种化合物对于相应的乳腺癌肿瘤干细胞均不敏感,不能根据化合物具有抗肿瘤活性推测出其具有抗肿瘤干细胞活性。现有技术(ZL 201310044877.6)报道了化合物Ⅱ-2的抗肿瘤活性,其抑制乳腺癌MCF-7细胞的IC50值为17.6μM,具有中等活性的抗肿瘤活性;而在本发明中,该化合物抗乳腺癌肿瘤干细胞的IC50值仅为0.77μM,具有极强的抗肿瘤干细胞活性,不能根据其具有中等活性的抗肿瘤活性,来推测其具有强抗肿瘤干细胞的活性。
实施例13
乳腺细胞球形成分析法(Mammosphere formation assay)是检验乳腺干细胞、乳腺癌癌干细胞活性及自我更新能力的一种公认的方法(Eguiara A,et al.Ma mmosphereFormation in Breast Carcinoma Cell Lines Depends upon Expressio n of E-cadherin.PLoS ONE,2013,8(10),e77281)。本实施例涉及高光学纯度阿卡宁萘茜母核羟基甲基化羰基肟衍生物对乳腺癌干细胞乳腺细胞球形成能力的测试,包括以下步骤:
参考文献(Gupta P,et al.Identification of selective inhibitors ofcancer stem cells by high-throughput screening.Cell,2009,138(4),645-659.)的方法,取对数生长期的MCF-7 CD44+CD24-/low肿瘤干细胞,用胰酶分离细胞。调整细胞悬液的浓度为10,000个细胞/mL,并将细胞接种在6孔超低附着板中,使用Mammocult 无血清培养基培养。培养72h后,给药组用化合物Ⅱ-2(2μM in DMSO)处理,对照组用相同体积的DMSO处理。分别于8h及24小时观察肿瘤干细胞球的数量及形态。结果如附图4所示,经化合物Ⅱ-2处理后的乳腺癌干细胞,细胞球的数量随药物处理时间的增加显著下降,同时单细胞数量显著增加。实验结果说明化合物Ⅱ-2可显著降低肿瘤干细胞的自我更新能力及细胞活性。
实施例14
本实施例涉及高光学纯度阿卡宁萘茜母核羟基甲基化羰基肟衍生物对MCF-7 CD44 +CD24-/low肿瘤干细胞的生长周期抑制实验。
参考文献(Chefetz I.,et al.Inhibition of Aurora-A kinase induces cellcycle arrest in epithelial ovarian cancer stem cells by affecting NFκBpathway.Cell Cycle. 2011 Jul 1;10(13):2206–2214)方法进行细胞周期抑制实验。将5×106个MCF-7 CD44+CD24-/low肿瘤干细胞接种至6孔板内,培养24小时后,给药不同浓度的化合物Ⅱ-2(给药浓度依次为0,1.0,2.0μM),给药后继续孵育24小时。用胰酶消化细胞,加入完全培养基终止消化后,收集所有细胞至离心管内,1000g 离心5min,弃上清,用预冷的PBS重悬细胞沉淀,以漂洗离心管内剩余的培养基。1000g离心5min后弃上清,PBS重悬沉淀后计数,调节细胞悬液浓度为2 ×106cells/mL,转移细胞至10mL离心管内,再次离心沉淀细胞,小心吸除上清。后加入1mL预冷的70%乙醇,轻轻吹打混匀细胞沉淀,静置4℃固定12h。 4℃,1000g离心5min,沉淀细胞,小心吸除上清,加入预冷的PBS重悬细胞,再次离心沉淀细胞,小心吸除上清。每管样品内加入0.5mL已配制好的PI染色液,缓慢并充分重悬细胞沉淀,37℃避光温育30min,避光置于冰上待用。用流式细胞仪在激发波长488nm处检测红色荧光,同时检测光散射情况。染色完成后的样品宜在24h内使用流式细胞仪进行检测。用FlowJo7.0软件分析数据, Dean-Jett-Fox模型周期。
荧光染料碘化吡啶能在嵌入双链DNA后显示红色荧光,根据细胞周期内各阶段DNA含量的不同,能在直方图上直观地将细胞分为三类,如附图5所示, G0期和G1期细胞还未开始DNA的复制,都含有相同的二倍体DNA,在图中归为一个峰;而G2期与M期的细胞已完成DNA的复制但还未或正在分裂为两个细胞,因均含有四倍体DNA而显示为一个峰;分布在两个峰之间的则为正在进行DNA复制的S期细胞。从上图5中我们可以看到,当浓度达到1μM时能发现 S期细胞比例由对照组的15.97%明显减少为13.63%,而G0/G1期细胞从70.72%增加到75.92%,G2/M期细胞也由13.31%下降至10.45%。当化合物Ⅱ浓度升高至2μM时,处理组G0/G1期细胞明显增加,比例增加至83.23%;S期细胞显著减少,比例下降至6.70%。即在该浓度下细胞周期开始明显阻滞在G0/G1期,进入S期的细胞数目降低。这说明肿瘤干细胞的DNA细胞调控因素的影响下,可能受到损伤,不能通过G1期的DNA损伤检测点,细胞周期被阻断不能进入S期,细胞在试图修复DNA不成功后可能启动凋亡程序。
表2目标化合物Ⅱ抑制乳腺癌干细胞MCF-7 CD44+CD24-/low的细胞周期
实施例15
本实施例涉及高光学纯度阿卡宁萘茜母核羟基甲基化羰基肟衍生物诱导 MCF-7CD44+CD24-/low肿瘤干细胞凋亡的实验。给药不同浓度的化合物Ⅱ后,采用AnnexV-FITC/PI染色法,对细胞凋亡情况进行定量分析。
在正常细胞中,磷脂酰丝氨酸分布在细胞膜脂质双层的内侧,当细胞处于凋亡早期时,在细胞皱缩、染色质浓缩、细胞膜通透性增加等凋亡现象还未开始出现时,磷脂酰丝氨酸从膜的内侧翻向外侧。AnnexinV与磷脂酰丝氨酸有高度的亲和力,能在凋亡早期与暴露在细胞外侧的磷脂酰丝氨酸相结合,标记了FITC 的AnnexinV能做为荧光探针用于检测早期凋亡的细胞。前面已经提到,PI是一种不能透过完整细胞膜的核酸染料,但当细胞处于凋亡的中晚期或坏死时,细胞膜的通透性增加,PI即能透过细胞膜而使细胞核染色。因此,根据AnnexinV和 PI对细胞染色情况能将处于不同凋亡时期的细胞区分开来。
如图附图6所示,左下象限为正常细胞(Annexin V-/PI-),右下象限为凋亡早期细胞(Annexin V+/PI-),右上象限内的则为凋亡晚期或坏死细胞(Annexin V+/PI+)。当用化合物Ⅱ-2对MCF-7 CD44+CD24-/low肿瘤干细胞进行处理时,凋亡细胞的比例随着药物处理浓度的增加而增加,我们对比1μM化合物处理24h 后的细胞与正常生长的阴性对照细胞(control)可以发现,凋亡早期细胞的比例由接近于0增加到19.4%。当化合物Ⅱ-2浓度增加至2μM时,早期凋亡细胞的比例进一步增加至38.5%,晚期凋亡细胞的比例则增加至27.7%。这一研究结果提示,凋亡途径是化合物Ⅱ-2浓度导致肿瘤干细胞死亡的原因之一,且随化合物浓度的升高,早期凋亡及晚期凋亡细胞的比例均显著上升。
实施例16
本实施例涉及紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物抗MCF-7 CD44+CD24-/low干细胞瘤的分子机制研究。
实验方法:
将生长状态良好的MCF-7 CD44+CD24-/low乳腺癌干细胞分为药物处理组 (Treated)及空白对照组(Control)。处理组细胞培养基中加入化合物Ⅱ-2(终浓度为2.0μM),共孵育24h;对照组加入同等浓度的DMSO,孵育24h。孵育结束后,通过离心方法收集细胞,用预冷的PBS清洗三次后。药物处理组及空白对照组,分别进行三次生物重复。所收集的6份样品,经细胞裂解、总蛋白提取及蛋白定量后,分别进行还原烷基化处理及Trypsin酶解,用TMT试剂标记肽段。利用RP-HPLC技术对标记后的肽段进行预分离,所收集的各蛋白洗脱液浓缩后、用液相色谱串联质谱进行分析。分析所得的数据采用Proteome DiscovererTM Software2.2搜库,结果进行数据统计及生物信息学分析。
实验结果:
在TMT蛋白组学研究中,最终成功鉴定蛋白6848个,所鉴定的蛋白约60%位于细胞质中。通过软件分析,我们找到了在三次生物重复实验中均存在表达差异、且差异倍数超过1.2倍的蛋白共186个,其中经化合物Ⅱ处理后表达下调的蛋白135个,上调蛋白51个。在所鉴定的差异蛋白,5个表达下调的差异蛋白均归属于细胞黏附分子通路(表3);数据的统计分析表明,在三次生物重复中,这5个蛋白在化合物Ⅱ-2处理组的表达与在相应对照组的表达量相比,具有显著的差异(P<0.05)。
表3乳腺癌干细胞MCF-7 CD44+CD24-/low经化合物Ⅱ-2处理后,黏附分子通路表达下调的蛋白
表3所示的细胞黏附分子通路的5个差异蛋白(CD171,CD166,SDC4,CD111,CD138),均为肿瘤干细胞的标志物,与肿瘤干细胞的自我更新、细胞粘附及增殖、肿瘤耐药密切相关(Dana,H.,et al.CD166 as a Stem Cell Marker?A Potential Target forTherapy Colorectal Cancer?J Stem Cell Res Ther.2016,1(6),226-229; Shimada,etal.Syndecan-1(CD138)contributes to prostate cancer progression by stabilizingtumour-initiating cells.J Pathol,2013,231,495-504;Onyeisi,J.et al. Syndecan-4as a pathogenesis factor and therapeutic target in cancer.Biomolecules, 2021,11(4),503.)。化合物Ⅱ-2处理后,上述标志物的表达量下调,肿瘤干细胞的粘附性降低,同时细胞的增殖及其恶性程度降低,即化合物Ⅱ-2靶向肿瘤干细胞的标记物,产生抗肿瘤干细胞的活性。
结论:化合物Ⅱ-2处理后,肿瘤干细胞粘附分子通路上的蛋白表达发生变化,蛋白表达下调后影响肿瘤干细胞的生长。
实施例17
本实施例涉及紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物抗MCF-7 CD44+CD24-/low乳腺癌干细胞瘤的体内活性研究。参考文献(Celikoglu F, etal.Intratumoral administration of cisplatin through a bronchoscope followedby irradiation for treatment of inoperable non-small cell obstructive lungcancer.Lung Cancer,2006,51(2),225-236)方法,采用瘤内注射的方式进行化合物Ⅱ-2抗MCF-7 CD44+CD24-/low干细胞瘤的体内药效实验。
实验方法:
NOD-SCID小鼠(雌性)饲养在SPF级实验动物中心,温度、湿和光照分别控制21±2℃,50±10%及12h光照/12h黑暗循环。所有动物实验的操作均参照所在实验动物中心的动物实验伦理委员会指导手册。
称取所需重量的化合物Ⅱ-2,将其溶于聚氧乙烯蓖麻油与无水乙醇1:1的混合液中,制备浓度为6毫克/毫升的储备液,置于玻璃瓶中待用,给药前用生理盐水稀释6倍至1毫克/毫升浓度,一次性过滤器过滤后即可给药。
MCF-7 CD44+CD24-/low乳腺癌干细胞正常传代培养至所需数目,细胞用胰酶消化;离心收取细胞后,使用培养基与基质胶(BD MatrigelTM)等体积混合液稀释,调整细胞浓度至(2×10^6个/mL)后接种于小鼠右前肢腋窝皮下(0.2mL/ 只)。接种后,待移植瘤瘤体积大于50mm3时,将小鼠按5只/组随机分组,分为给药组(化合物Ⅱ-2)及空白对照组(control),并称重记录。给药组用已配好的化合物Ⅱ-2溶液对小鼠进行瘤内注射(0.2mL/只),空白对照组用相应的空白制剂进行瘤内注射,隔天给药,共给药10次。开始实验后每天记录肿瘤的体积,给药结束后再记录瘤重2天。
肿瘤体积计算公式:
体积(mm3)=长径(mm)×短径(mm)×短径(mm)/2
肿瘤体积变化率计算公式:
肿瘤体积变化率(%)=肿瘤体积/初次给药时肿瘤体积×100
实验结果:
实验结果如附图7所示,随化合物Ⅱ-2的瘤内注射,给药组的肿瘤体积变化不大,在第20天,给药组的平均肿瘤体积变化率仅为初始值的135%;空白对照组的平均肿瘤体积变化率已达到初始值的270%。这一结果提示,在所采用的给药方式下,化合物Ⅱ-2可显著抑制MCF-7 CD44+CD24-/low乳腺癌干细胞瘤的生长。
实施例18
本实施例涉及紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟衍生物对人皮肤成纤细胞HSF(人正常细胞)的生长抑制活性测试,包括以下步骤:
取对数生长期的HSF,用0.25%胰蛋白酶消化后,用磷酸盐缓冲液稀释后离心,加入无血清培养基混悬,并调整细胞悬液至合适的浓度,在96孔板中每孔加入100μL细胞悬液,使得每孔中的细胞数为5000个。将96孔板静置于37℃, 5%CO2恒温培养箱中培养12h,待细胞贴壁后每孔加入用完全培养基稀释的化合物100μL,各组均设三个复孔。每块96孔板上需设置调零孔与空白对照孔。继续培养48h后,加入之前配好的MTT溶液20μL/孔,避光继续培养4h。后将上清液吸出,每孔加100微升DMSO,并于多功能酶标仪上570nm波长处检测各孔的吸光度(OD值)。测试中选用紫草素为阳性对照药物。细胞生长抑制率及半数抑制浓度(IC50)计算方法同实施例12。
表2目标化合物抑制人正常细胞HSF的IC50值
化合物 | IC<sub>50</sub>(μM) |
Ⅱ-2 | >50 |
Ⅱ-3 | >50 |
Ⅲ-2 | >50 |
Ⅲ-3 | >50 |
IV-2 | >50 |
IV-3 | >50 |
紫草素 | 1.3 |
综上所述,本发明化合物的制备方法简便,收率较高且原料易得。体外抗肿瘤活性实验研究表明,所制备紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟化衍生物对乳腺癌干细胞有非常强的生长抑制作用,其去除肿瘤干细胞的药效显著高于临床应用的紫杉醇、多西紫杉醇及阿霉素。与紫草素相比,紫草素及其对映体阿卡宁萘茜母核羟基甲基化羰基肟化衍生物对于正常细胞HSF-1毒性显著降低。该类化合物可选择性抗肿瘤干细胞的特性,为后续抗肿瘤药物筛选提供了先导化合物,具有重大开发前景。
以上对本发明的具体实施例进行了描述。需要理解的是,本发明并不局限于上述特定实施方式,本领域技术人员可以在权利要求的范围内做出各种变形或修改,这并不影响本发明的实质内容。
Claims (5)
2.一种如权利要求1所述的衍生物在制备抗肿瘤干细胞药物中的用途,其特征在于,所述药物中所述衍生物为主要活性成分。
3.如权利要求2所述的用途,其特征在于,所述肿瘤干细胞为细胞表面抗原类型为CD44+CD24-/low的乳腺癌干细胞。
4.一种如权利要求1所述的衍生物与其它抗肿瘤药物联用在制备治疗恶性肿瘤药物中的用途。
5.如权利要求2或4所述的用途,其特征在于,所述药物制成下列之一的剂型:①注射剂、②片剂、③胶囊剂、④颗粒剂。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210417140.3A CN114805117A (zh) | 2022-04-20 | 2022-04-20 | 抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210417140.3A CN114805117A (zh) | 2022-04-20 | 2022-04-20 | 抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114805117A true CN114805117A (zh) | 2022-07-29 |
Family
ID=82504817
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210417140.3A Pending CN114805117A (zh) | 2022-04-20 | 2022-04-20 | 抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114805117A (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115991664A (zh) * | 2022-12-09 | 2023-04-21 | 上海交通大学 | 一种具有抗菌活性的萘醌肟衍生物及其医药用途 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101194920A (zh) * | 2006-09-29 | 2008-06-11 | 杭州贺博生物技术有限公司 | 紫草及其有效成分在制备治疗肿瘤干细胞的药物中的应用 |
CN102557914A (zh) * | 2012-01-31 | 2012-07-11 | 上海交通大学 | 萘茜母核氧烷基、酰基取代的阿卡宁衍生物及制备和用途 |
CN103130680A (zh) * | 2013-02-04 | 2013-06-05 | 上海交通大学 | 高光学纯度紫草素和阿卡宁萘茜母核羟基甲基化羰基肟衍生物及其制备和用途 |
CN103145583A (zh) * | 2013-02-04 | 2013-06-12 | 上海交通大学 | 外消旋体紫草素萘茜母核羟基甲基化羰基肟衍生物及其制备和用途 |
CN110041180A (zh) * | 2019-04-25 | 2019-07-23 | 上海交通大学 | 含氮杂侧链的紫草素肟衍生物及其制备方法和医药用途 |
-
2022
- 2022-04-20 CN CN202210417140.3A patent/CN114805117A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101194920A (zh) * | 2006-09-29 | 2008-06-11 | 杭州贺博生物技术有限公司 | 紫草及其有效成分在制备治疗肿瘤干细胞的药物中的应用 |
CN102557914A (zh) * | 2012-01-31 | 2012-07-11 | 上海交通大学 | 萘茜母核氧烷基、酰基取代的阿卡宁衍生物及制备和用途 |
CN103130680A (zh) * | 2013-02-04 | 2013-06-05 | 上海交通大学 | 高光学纯度紫草素和阿卡宁萘茜母核羟基甲基化羰基肟衍生物及其制备和用途 |
CN103145583A (zh) * | 2013-02-04 | 2013-06-12 | 上海交通大学 | 外消旋体紫草素萘茜母核羟基甲基化羰基肟衍生物及其制备和用途 |
CN110041180A (zh) * | 2019-04-25 | 2019-07-23 | 上海交通大学 | 含氮杂侧链的紫草素肟衍生物及其制备方法和医药用途 |
Non-Patent Citations (1)
Title |
---|
JIAHUA CUI: "DMAKO-20 as a New Multitarget Anticancer Prodrug Activated by the Tumor Specific CYP1B1 Enzyme", MOLECULAR PHARMACEUTICS, vol. 16, pages 409 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115991664A (zh) * | 2022-12-09 | 2023-04-21 | 上海交通大学 | 一种具有抗菌活性的萘醌肟衍生物及其医药用途 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20120071477A1 (en) | Cdki pathway inhibitors and uses thereof | |
Tan et al. | Purified vitexin compound 1 suppresses tumor growth and induces cell apoptosis in a mouse model of human choriocarcinoma | |
CN109734714B (zh) | 一种吴茱萸生物碱衍生物及其合成方法和应用 | |
CN103347520A (zh) | 用作癌症化学治疗的晚期sv40因子(lsf)的抑制剂 | |
KR20210027382A (ko) | 접히지 않은 단백질 반응의 활성화제 | |
CN113304151B (zh) | 一种硝基呋喃类小分子化合物在制备诱导铁死亡和/或减缓胃癌化疗耐药药物中的应用 | |
CN112656795A (zh) | 防己诺林碱在抗结膜黑色素瘤中的作用机制及应用 | |
CN111658644B (zh) | 一种小分子stat3抑制剂wz-2-033及其在制备治疗乳腺癌和胃癌药物中的应用 | |
CN114805117A (zh) | 抗肿瘤干细胞的紫草素及阿卡宁肟衍生物 | |
WO2008109717A1 (en) | Compositions and methods for treating cancer | |
EP1516620B1 (en) | Rifampicin for treating angiogenesis | |
CN117230132A (zh) | 一种口山酮类化合物Austocystin R的培养方法及制药用途 | |
Awasthi et al. | Antitumor activity of a pexidartinib bioisostere inhibiting CSF1 production and CSF1R kinase activity in human hepatocellular carcinoma | |
CN109328062A (zh) | 小分子活性氧簇诱导剂和线粒体活性抑制剂 | |
CN114195779A (zh) | 9-0-乙基乙醚小檗红碱的合成方法及其在制备抗肿瘤药物中的用途 | |
CN109172548B (zh) | 叶黄素及其衍生物在制备抗脑胶质瘤药物中的应用 | |
CN108727459B (zh) | 雷公藤红素适配子偶合物及其制备方法和应用 | |
CN113024400A (zh) | 一种秋水仙碱衍生物及其制备方法和应用 | |
TWI486341B (zh) | 抑制atr與fancd2激活之組成物與方法 | |
RU2454232C2 (ru) | Применение производных трииндолилметана в качестве противоопухолевых средств | |
CN1331469C (zh) | 一种苯酞类二聚体的用途 | |
CN114984007B (zh) | Pradx-ezh2小分子抑制剂及其在制备肿瘤治疗药物中的用途 | |
US11142530B2 (en) | Deep-sea fungus-derived anthraquinone compound and use thereof in preparing anti-allergic drugs | |
CN109503495B (zh) | 一种2-亚胺苯并咪唑类化合物及医药用途 | |
CN114437041B (zh) | 一类具有抗肿瘤活性的4-四唑基取代-3,4-二氢喹唑啉衍生物及其制备方法和应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination |