CN115109052A - AIE compound with mitochondrion targeting function and synthesis method and application thereof - Google Patents
AIE compound with mitochondrion targeting function and synthesis method and application thereof Download PDFInfo
- Publication number
- CN115109052A CN115109052A CN202210819688.0A CN202210819688A CN115109052A CN 115109052 A CN115109052 A CN 115109052A CN 202210819688 A CN202210819688 A CN 202210819688A CN 115109052 A CN115109052 A CN 115109052A
- Authority
- CN
- China
- Prior art keywords
- compound
- aie
- triphenylamine
- tpa
- parent structure
- 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.)
- Granted
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 110
- 210000003470 mitochondria Anatomy 0.000 title claims abstract description 30
- 230000008685 targeting Effects 0.000 title claims abstract description 23
- 238000001308 synthesis method Methods 0.000 title abstract description 7
- 230000002438 mitochondrial effect Effects 0.000 claims abstract description 18
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 13
- 230000000694 effects Effects 0.000 claims abstract description 11
- 239000002246 antineoplastic agent Substances 0.000 claims abstract description 9
- 229940041181 antineoplastic drug Drugs 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- ODHXBMXNKOYIBV-UHFFFAOYSA-N triphenylamine Chemical compound C1=CC=CC=C1N(C=1C=CC=CC=1)C1=CC=CC=C1 ODHXBMXNKOYIBV-UHFFFAOYSA-N 0.000 claims description 24
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 23
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 17
- 125000003172 aldehyde group Chemical group 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 11
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 10
- 238000010992 reflux Methods 0.000 claims description 10
- 230000002194 synthesizing effect Effects 0.000 claims description 8
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 6
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 6
- 201000010881 cervical cancer Diseases 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 6
- PDQRQJVPEFGVRK-UHFFFAOYSA-N 2,1,3-benzothiadiazole Chemical compound C1=CC=CC2=NSN=C21 PDQRQJVPEFGVRK-UHFFFAOYSA-N 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000012043 crude product Substances 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- 125000000217 alkyl group Chemical group 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 2
- 125000004433 nitrogen atom Chemical group N* 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 63
- 230000006870 function Effects 0.000 abstract description 12
- 210000004881 tumor cell Anatomy 0.000 abstract description 11
- 201000011510 cancer Diseases 0.000 abstract description 10
- 210000001700 mitochondrial membrane Anatomy 0.000 abstract description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 5
- 230000004900 autophagic degradation Effects 0.000 abstract description 5
- 230000001939 inductive effect Effects 0.000 abstract description 5
- 239000001301 oxygen Substances 0.000 abstract description 5
- 229910052760 oxygen Inorganic materials 0.000 abstract description 5
- 230000002776 aggregation Effects 0.000 abstract description 4
- 238000004220 aggregation Methods 0.000 abstract description 4
- 231100000419 toxicity Toxicity 0.000 abstract description 3
- 230000001988 toxicity Effects 0.000 abstract description 3
- 206010034972 Photosensitivity reaction Diseases 0.000 abstract description 2
- 238000003745 diagnosis Methods 0.000 abstract description 2
- 238000000799 fluorescence microscopy Methods 0.000 abstract description 2
- 230000010354 integration Effects 0.000 abstract description 2
- 230000003287 optical effect Effects 0.000 abstract description 2
- 208000007578 phototoxic dermatitis Diseases 0.000 abstract description 2
- 231100000018 phototoxicity Toxicity 0.000 abstract description 2
- 229920000642 polymer Polymers 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 14
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 12
- 239000000523 sample Substances 0.000 description 11
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 10
- 239000003642 reactive oxygen metabolite Substances 0.000 description 9
- 238000011534 incubation Methods 0.000 description 8
- 239000012528 membrane Substances 0.000 description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- KBPLFHHGFOOTCA-UHFFFAOYSA-N caprylic alcohol Natural products CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 238000012512 characterization method Methods 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 4
- 230000005284 excitation Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 230000005311 nuclear magnetism Effects 0.000 description 4
- 239000012074 organic phase Substances 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 3
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 3
- 230000001464 adherent effect Effects 0.000 description 3
- 230000003698 anagen phase Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 3
- 229960004316 cisplatin Drugs 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 210000003463 organelle Anatomy 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 206010021143 Hypoxia Diseases 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000007810 chemical reaction solvent Substances 0.000 description 2
- 230000008045 co-localization Effects 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000001146 hypoxic effect Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 150000002634 lipophilic molecules Chemical class 0.000 description 2
- 230000002132 lysosomal effect Effects 0.000 description 2
- TVMXDCGIABBOFY-UHFFFAOYSA-N n-Octanol Natural products CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 description 2
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 238000002390 rotary evaporation Methods 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000004017 serum-free culture medium Substances 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 102000003954 Autophagy-Related Proteins Human genes 0.000 description 1
- 108010082399 Autophagy-Related Proteins Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 101000605835 Homo sapiens Serine/threonine-protein kinase PINK1, mitochondrial Proteins 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102100038376 Serine/threonine-protein kinase PINK1, mitochondrial Human genes 0.000 description 1
- 102000019259 Succinate Dehydrogenase Human genes 0.000 description 1
- 108010012901 Succinate Dehydrogenase Proteins 0.000 description 1
- COQLPRJCUIATTQ-UHFFFAOYSA-N Uranyl acetate Chemical compound O.O.O=[U]=O.CC(O)=O.CC(O)=O COQLPRJCUIATTQ-UHFFFAOYSA-N 0.000 description 1
- 235000005811 Viola adunca Nutrition 0.000 description 1
- 240000009038 Viola odorata Species 0.000 description 1
- 235000013487 Viola odorata Nutrition 0.000 description 1
- 235000002254 Viola papilionacea Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 230000005775 apoptotic pathway Effects 0.000 description 1
- 230000002886 autophagic effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- HOQPTLCRWVZIQZ-UHFFFAOYSA-H bis[[2-(5-hydroxy-4,7-dioxo-1,3,2$l^{2}-dioxaplumbepan-5-yl)acetyl]oxy]lead Chemical compound [Pb+2].[Pb+2].[Pb+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HOQPTLCRWVZIQZ-UHFFFAOYSA-H 0.000 description 1
- 230000003913 calcium metabolism Effects 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 230000004700 cellular uptake Effects 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000004110 gluconeogenesis Effects 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000003712 lysosome Anatomy 0.000 description 1
- 230000001868 lysosomic effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000008965 mitochondrial swelling Effects 0.000 description 1
- 230000021125 mitochondrion degradation Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 230000030589 organelle localization Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- -1 phenylimino Chemical group 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000035806 respiratory chain Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- VLLMWSRANPNYQX-UHFFFAOYSA-N thiadiazole Chemical compound C1=CSN=N1.C1=CSN=N1 VLLMWSRANPNYQX-UHFFFAOYSA-N 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 230000004143 urea cycle Effects 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- 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
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1011—Condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1014—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
- C09K2211/1051—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms with sulfur
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Materials Engineering (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an AIE compound with mitochondrion targeting, which enhances the charge intensity of the compound by combining molecules with an aggregation-induced emission structure and long-chain molecules with positive charges, and the obtained compound can target the mitochondrion of tumor cells and realize the treatment effect of generating dark toxicity and phototoxicity on the tumor cells; the compound has the functions of inducing the generation of active oxygen in cancer cells, reducing mitochondrial membrane potential, changing the form of mitochondria, damaging the functions of mitochondria and inducing autophagy of tumor cells (mitochondria); in addition, due to the inherent optical property of the AIE molecules, cancer cells and normal cells can be distinguished through fluorescence imaging, so that diagnosis and treatment integration can be realized, and the AIE molecular structure has a wide application prospect. The invention also discloses a synthesis method of the AIE compound with the mitochondrial targeting function and application of the AIE compound in preparation of antitumor drugs.
Description
Technical Field
The invention relates to an AIE compound with mitochondrion targeting, a synthesis method of the compound and application of the compound in preparation of antitumor drugs.
Technical Field
Mitochondria are important organelles in human cells, with the outer membrane having a negative potential and the inner membrane being rich in proteins involved in protein import mechanisms, ATP synthase and respiratory chain complexes. As a cellular motive force, mitochondria synthesize energy-rich molecular Adenosine Triphosphate (ATP) to support various cellular processes, including metabolism, cell division, and biomolecule synthesis. In addition to its role as an energy factory, mitochondria are also responsible for many other important functions, including the initiation of apoptotic pathways, calcium metabolism, electron transport, lipid metabolism, reactive oxygen species production, urea cycle, citrate cycle, and gluconeogenesis and steroid hormone synthesis.
The concept of aggregation-induced emission (AIE) was proposed in 2001 by Tang-loyalty courtyard et al. In this phenomenon, the molecules fluoresce strongly when aggregated or in the solid state, but weakly when dispersed in solution. In view of their advantages, such as high light stability, large stokes shift, and ease of preparation. A number of AIE luminescent substances (AIEgens) have been widely explored, particularly in relation to the fields of bioimaging, sensing and therapy. Side chain engineering refers to the modification of the side chain structure of a molecule, thereby modulating the properties of the molecule. Side chain engineering can be an effective method in the aspect of constructing an AIE active multifunctional photochemical system.
Disclosure of Invention
The purpose of the invention is as follows: based on the characteristics that the mitochondria of tumor cells are different from normal cells in structure and function, one of the purposes of the invention is to provide an AIE compound capable of targeting the mitochondria of tumor cells, wherein the AIE compound can still keep the fluorescence emission performance related to radiation transition by controlling the length of an introduced side chain, and simultaneously can improve the D-A intensity of the compound by introducing the side chain, so as to realize the generation of active oxygen in induced cancer cells; the invention also aims to provide a synthesis method of the compound and application of the compound in preparing antitumor drugs.
The technical scheme is as follows: the AIE compound with mitochondrion targeting has a chemical structural formula shown in a formula (I):
the synthesis method of the AIE compound with the mitochondria targeting function specifically comprises the following steps: firstly synthesizing a triphenylamine parent structure compound with aldehyde group, and then reacting the triphenylamine parent structure compound with aldehyde group with pyridine with a long chain to obtain an AIE compound with mitochondrion targeting;
wherein, the chemical structural formula of the triphenylamine parent structure compound with aldehyde group is as follows:
wherein in the pyridine with the long chain, one side of the N atom of the pyridine is grafted with a C4-12 straight-chain alkyl group.
Further preferably, one side of the pyridine atom is grafted with C8 straight-chain alkyl, and the chemical structural formula of the pyridine with the long chain is as follows:
the method specifically comprises the following steps: heating and refluxing a triphenylamine parent structure compound with aldehyde group and pyridine with a long chain in absolute ethyl alcohol for reaction, extracting and separating a crude product after the reaction is finished, and purifying by column chromatography to obtain the AIE compound with mitochondrion targeting.
Wherein, the triphenylamine parent structure molecule with aldehyde group is prepared by the following method: heating a compound (4,4 '- [ [4- (4, 5-tetramethyl-1, 3,2-dioxaborolan 2-ethyl) phenylimino di, 1,1' -dimethyl ester) with a triphenylamine parent structure and 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole in a mixed solution of tetrahydrofuran and water for reflux reaction in an inert atmosphere, extracting and separating a crude product after the reaction is finished, and purifying by column chromatography to obtain a triphenylamine parent structure compound with aldehyde group;
the chemical structural formula of the compound with the triphenylamine parent structure is as follows:
7-bromo-4-aldehyde benzo [ C][1,2,5]The chemical structural formula of thiadiazole is as follows:
wherein, the reaction molar ratio of the triphenylamine parent structure compound with aldehyde group to pyridine with long chain is 1: 1.2; the reflux reaction time is 4-4.5 h; the reaction temperature is 80-85 ℃.
Wherein the reaction molar ratio of the compound with the triphenylamine parent structure to the 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole is 1: 1.2; the reflux reaction time is 18-19 h; the reaction temperature is 70-75 ℃.
The AIE compound with the mitochondrion targeting function is applied to the preparation of antitumor drugs and antitumor drug components.
Wherein the tumor is cervical cancer.
The chemical reaction formula of the compound of the invention is as follows:
has the advantages that: the invention firstly prepares a molecule with an aggregation-induced emission structure (a triphenylamine rotor structure), one end of the molecule structure is an electron-withdrawing end, the other end is an electron-donating end, and meanwhile, the molecule with the aggregation-induced emission structure is combined with a long-chain molecule with positive charges, so that the charge intensity of the compound is further enhanced, and the obtained compound can target mitochondria of tumor cells and realize the therapeutic action of generating dark toxicity and phototoxicity on the tumor cells; the compound has the functions of inducing the generation of active oxygen in cancer cells, reducing mitochondrial membrane potential, changing the form of mitochondria, damaging the functions of mitochondria and inducing autophagy of tumor cells (mitochondria); in addition, due to the inherent optical property of the AIE molecules, cancer cells and normal cells can be distinguished through fluorescence imaging, so that diagnosis and treatment integration can be realized, and the AIE molecular structure has a wide application prospect.
Drawings
FIG. 1 is a cellular uptake map of the compound TPA-N-8 of example 1;
FIG. 2 is a subcellular organelle localization map of compound TPA-N-8 of example 1;
FIG. 3 is a schematic diagram of the compound TPA-N-8 of example 1 inducing the production of reactive oxygen species by HeLa cells;
FIG. 4 is a graph showing that compound TPA-N-8 of example 1 induces a decrease in mitochondrial membrane potential in HeLa cells;
FIG. 5 is a schematic representation of the effect of the compound TPA-N-8 of example 1 on mitochondrial morphology and function;
FIG. 6 is a graph showing that the compound TPA-N-8 of example 1 induces the change of the content of related proteins in HeLa cells.
Detailed Description
The technical solution of the present invention is further described with reference to the following specific embodiments.
Example 1
The invention relates to a method for synthesizing a long-chain AIE compound (TPA-N-8) with mitochondrion targeting, which comprises the following steps:
(1) preparation of triphenylamine precursor structure compound (TPA-CHO) having aldehyde group: weighing compound 1 (a compound with a triphenylamine parent structure) (1.461g, 3 mol), compound 2 (7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole) (0.7533g, 3.1mol) and anhydrous potassium carbonate (4.14g, 30mol) in a three-neck flask, isolating air, using argon as a protective gas, adding tetrakis (triphenylphosphine) palladium (0.1155g, 0.1mol), using tetrahydrofuran/water (the volume ratio of tetrahydrofuran/water is 3:1) as a reaction solvent, and heating and refluxing for 18h at 70 ℃. And (3) post-treatment: stopping stirring, and removing tetrahydrofuran by rotary evaporation; adding more water, extracting for several times, adding anhydrous magnesium sulfate, and drying. The mixture was filtered with suction, the resulting filtrate was freed of the solvent by means of a rotary evaporator, an orange solid appeared, and the solid was dissolved with methylene chloride and then stirred with silica gel. Performing column chromatography separation by using a developing solvent (the volume ratio of petroleum ether to ethyl acetate is 15: 1-petroleum ether to ethyl acetate is 5:1) to obtain a product TPA-CHO (orange solid, the yield is 65%), wherein the chemical structural formula of the TPA-CHO is as follows:
the chemical reaction formula of the triphenylamine parent structure compound (TPA-CHO) with aldehyde group is as follows:
by the nuclear magnetism and the mass spectrum characterization, 1 H NMR(400MHz,DMSO-d 6 )δ(ppm):8.34(d,J＝7.4Hz,1H),8.09(dd,J＝12.0,7.9Hz,3H),7.91(d,J＝8.5Hz,4H),7.30(d,J＝8.4Hz,2H),7.17(d,J＝8.6Hz,4H),3.83(s,6H). 13 C NMR(101MHz,DMSO-d 6 )δ(ppm):189.50,166.09,153.56,153.40,150.68,147.06,138.34,133.55,132.84,131.71,131.46,127.44,126.34,125.94,124.62,123.54,52.45.ESI-MS(CH 3 OH):calcd for[TPA-CHO]m/z＝523.1，found m/z＝523.2。
(2) weighing TPA-CHO (0.1046g, 0.2mol) in a reaction device, vacuumizing, and taking argon as protective gas; adding long chain (8-N) + ) (0.027g, 0.22mol) in absolute ethanol, heated at 80 ℃ and stirred for 10min to dissolve completely, then a small amount of piperidine (50. mu.L) was added, and heated under reflux at 80 ℃ for 4 h. And (3) post-treatment: stopping stirring, and removing the reaction solvent by rotary evaporation; the solid was dissolved with dichloromethane and stirred with silica gel. Performing column chromatography separation by using a developing solvent (the volume ratio is that dichloromethane is 20:1 to dichloromethane is 10:1) to obtain a product TPA-N-8 (a red brown solid, the yield is 40 percent), wherein the chemical structural formula of the TPA-N-8 is as follows:
the chemical reaction formula for a long chain AIE compound with mitochondrial targeting (TPA-N-8) is:
by the nuclear magnetism and the mass spectrum characterization, 1 H NMR(400MHz，DMSO-d 6 )δ(ppm):9.04(d,J＝6.6Hz,2H),8.45–8.31(m,4H),8.19–8.10(m,3H),8.05(d,J＝7.5Hz,1H),7.93(d,J＝8.8Hz,4H),7.34–7.27(m,2H),7.21–7.15(m,4H),4.54(t,J＝7.4Hz,2H),3.84(s,6H),1.97–1.89(m,2H),1.31–1.20(m,12H),0.88–0.83(m,3H). 13 C NMR(101MHz,DMSO-d 6 )δ166.13,153.12,150.76,146.53,144.86,136.97,134.23,132.89,131.47,131.43,128.36,127.88,127.25,126.14,124.70,124.53,123.40,60.29,52.46,31.61,31.09,29.47,28.93,28.85,25.93,22.51,14.41.ESI-MS(CH 3 OH):calcd for[TPA-N-8]m/z＝711.3，found m/z＝711.4。
comparative example 1: synthesis of TPA-N-4
Comparative example 1TPA-N-4 Synthesis method was the same as in example 1 except that long chain (4-N) was added in step (2) + ) The structural formula of the obtained compound TPA-N-4 is as follows:
by the nuclear magnetism and the mass spectrum characterization, 1 H NMR(400MHz,Chloroform-d)δ(ppm):9.13(d,J＝6.5Hz,2H),8.34–8.25(m,3H),8.17(d,J＝16.2Hz,1H),8.07(d,J＝7.5Hz,1H),7.99–7.91(m,6H),7.75(d,J＝7.4Hz,1H),7.27(d,J＝10.6Hz,2H),7.20–7.16(m,4H),4.83(t,J＝7.4Hz,2H),3.92(s,6H),2.04(dd,J＝8.7,6.5Hz,2H),1.50–1.44(m,2H),1.00(t,J＝7.4Hz,3H). 13 C NMR(101MHz,Chloroform-d)δ(ppm):166.52,153.80,150.60,146.91,144.18,137.93,135.27,133.60,132.59,131.17,130.68,127.47,126.74,126.34,125.29,124.92,124.61,123.31,61.01,52.08,33.53,29.70,19.43,13.62,1.03.ESI-MS(CH 3 OH):calcd for[TPA-N-4]m/z＝655.2，found m/z＝655.3。
comparative example 2: synthesis of TPA-N-12
Comparative example 2TPA-N-12 Synthesis Process is the same as in example 1, with the only difference that in step (2) is added long chain (12-N) + ) The structural formula of the obtained compound TPA-N-12 is as follows:
by the nuclear magnetism and the mass spectrum characterization, 1 H NMR(400MHz,DMSO-d6)δ(ppm):9.04(d,J＝6.6Hz,2H),8.45–8.29(m,4H),8.14(dd,J＝18.6,8.1Hz,3H),8.05(d,J＝7.5Hz,1H),7.92(d,J＝8.8Hz,4H),7.34–7.26(m,2H),7.19–7.14(m,4H),4.54(t,J＝7.3Hz,2H),3.83(s,6H),1.92(s,2H),1.30–1.20(m,20H),0.85–0.80(m,3H). 13 C NMR(101MHz,DMSO-d6)δ(ppm):166.11,153.10,150.74,146.51,144.86,136.97,133.33,132.93,131.46,131.41,128.35,127.84,126.11,124.68,124.51,123.39,60.28,52.46,31.77,31.07,29.49,29.38,29.26,29.19,28.87,25.90,22.56,14.42.ESI-MS(CH 3 OH):calcd for[TPA-N-12]m/z＝767.3，found m/z＝767.4。
testing the lipid-water partition coefficients logP of the compounds TPA-N-4, TPA-N-8 and TPA-N-12 o/w ：
Liposolubility of compounds by classical shake flask method (using lipid-water partition coefficient logP) o/w Indicated) and analyzed using a uv-vis spectrometer. Mixing the water phase with deionized water and the organic phase with n-octanol, oscillating for 24h to reach mutual saturation, dissolving compound (100 μ M) in the water phase saturated with n-octanol, adding equal volume of water-saturated organic phase, oscillating for 4h at 37 deg.C on a 500rpm shaking table, centrifuging at 3000rpm for 5min, detecting absorbance values of the compounds in the water phase and the organic phase respectively with an ultraviolet-visible spectrometer, and determining logarithm of the concentration ratio of the compound in the organic phase and the water phase to obtain a distribution coefficient log P of lipid-water o/w The value is obtained.
Compound TPA-N-4, TPAThe distribution coefficients of lipid and water logP of-N-8 and TPA-N-12 o/w As shown in table 1.
TABLE 1
As can be seen from Table 1, the lipid solubility of the compound TPA-N-8 is slightly higher than that of the compounds TPA-N-4 and TPA-N-12, and more lipophilic molecules enter cells more easily.
The application of the compounds TPA-N-4, TPA-N-8 and TPA-N-12 in the antitumor activity of the cervical cancer cell HeLa:
the anti-proliferative effect of the compounds TPA-N (N-4/8/12) and cisplatin CDDP on cervical cancer cells HeLa was analyzed using MTT colorimetry. In living cells, succinate dehydrogenase within the mitochondria can reduce MTT to produce a blue-violet product: formazan (soluble in DMSO), and the product has an absorption peak at 570nm, so A can be used 570nm To analyze cell proliferation.
The specific experimental steps are as follows:
(1) resuscitating a tube of tumor cells, culturing with fresh culture solution (wherein, the HeLa cells use DMEM culture medium + 10% fetal calf serum + 1% penicillin and streptomycin), and using after passage for 2 times;
(2) when the cells reached the logarithmic growth phase, the cells were seeded at a cell density of 5000 cells/well into a 96-well plate (100. mu.L of culture medium per well), followed by being placed in a thermostat (37 ℃, 5% CO) 2 ) Medium culture;
(3) after the cells adhere to the wall, 100 mu L of fresh culture solution containing TPA-N-N (N is 4/8/12) and cisplatin CDDP with different concentration gradients is respectively added into each hole, and then the cells are placed in a thermostat for continuous incubation;
(4) after 24 hours of incubation, 20. mu.L of MTT (5mg/mL) was added to each well, after incubation for 4 hours at 37 ℃ in an incubator, the supernatant was aspirated, 150. mu.L of dimethyl sulfoxide (DMSO) was added to each well, and A was detected using an enzyme-linked immunosorbent assay 570nm Calculating the inhibition rate of cell proliferation to obtain IC 50 Value (drug concentration corresponding to an inhibition equal to 50%). MTT assay of compound TPA-N-N (N-4/8/12) and cisplatin CDDPThe test results are shown in table 2:
TABLE 2
The results show that: the proliferation inhibition activity of the compound TPA-N-8 on tumor cells is higher than that of the compound TPA-N-4/12 and CDDP, which shows that the compound TPA-N-8 with the best lipophilic effect has good inhibition effect on the cervical cancer cells HeLa; in addition, the TPA-N-8 compound not only shows better toxicity, but also can improve the treatment effect under the condition of illumination; the compound TPA-N-8 still shows good treatment effect on the cervical cancer cell HeLa in the hypoxic environment, and solves the condition limitation of tumor hypoxic environment.
Example 1 use of the compound TPA-N-8 for HeLa cell uptake:
and (3) detecting the uptake of the compound TPA-N-8 in HeLa cells. Plating 5 x 10 onto coverslips in 12-well plates 4 HeLa cells in logarithmic growth phase are placed in an incubator to grow in an adherent way overnight, and a cell culture solution which is prepared in advance and contains a compound TPA-N-8 is added to incubate for different times (0.5, 1, 1.5, 2, 3 and 4 hours). The culture broth was aspirated and washed three times with PBS. The photographs were then observed under a confocal microscope. The compound TPA-N-8: lambda [ alpha ] ex ＝488nm，λ em ＝700nm。
The uptake of TPA-N-8 prepared in example 1 into HeLa cells is shown in FIG. 1. The results show that: the lipophilic compound TPA-N-8 can completely enter cells after being incubated with HeLa for 2 hours, and lays a foundation for improving the anti-tumor curative effect.
The application of the compounds TPA-N-4, TPA-N-8 and TPA-N-12 to the localization of intracellular subcellular organelles (mitochondria):
the distribution of the compounds TPA-N-4, TPA-N-8 and TPA-N-12 in mitochondria and lysosomes in HeLa cells was detected by using commercial probes MitoGreen and LysoGreen. Plating 5 x 10 onto coverslips in 12-well plates 4 Placing HeLa cells in logarithmic growth phase in incubator for adherent growth overnight, respectively adding pre-prepared TPA-N-4 containing 5 μ L compound and TPA-N-8 and TPA-N-12 cell culture medium, and further incubation for 2 hours. The culture broth was aspirated and washed three times with PBS. Subcellular organelle staining was done by adding culture medium containing commercial probes at a certain concentration and incubating for a certain time, which was 30min, before testing, and then observing and photographing under a confocal microscope. The compound TPA-N-8: lambda [ alpha ] ex ＝488nm，λ em 700 nm; excitation and emission wavelength ranges for MitoGreen commercial probes: lambda [ alpha ] ex ＝488nm，λ em 525 nm; excitation and emission wavelength ranges for LysoGreen commercial probes: lambda [ alpha ] ex ＝405nm，λ em 525 nm. Co-localization coefficient analysis was analyzed with ImageJ software.
The confocal image of TPA-N-8 prepared in example 1 after co-incubation with mitochondrial and lysosomal probes is shown in FIG. 2. The results show that: the co-localization coefficient of TPA-N-8 with mitochondrial probes was 0.805 and with lysosomal probes was 0.259, indicating that TPA-N-8 was able to localize efficiently in mitochondria within HeLa cells.
Use of TPA-N-8 prepared in example 1 to induce intracellular reactive oxygen species production:
the method comprises the following steps: confocal microscopy detects ROS in cancer cells. HeLa cells were seeded on 35mm Corning confocal laser culture dishes, and when the cell density reached 70%, the compound TPA-N-84. mu.L was added for treatment for 2h, and then the cells were stained with 10. mu.M DCF-DA in a serum-free medium at 37 ℃ in the dark for 30min, followed by immediate observation with a confocal microscope at 488nm for excitation wavelength and 530. + -. 20nm for emission wavelength.
The method 2 comprises the following steps: flow cytometry detects ROS in cancer cells. After the HeLa cells are respectively treated by a compound TPA-N-8(1/2/4/6/8 mu L) for 2h, the HeLa cells are dyed for 30min at 37 ℃ in a dark place by using a serum-free culture medium containing 10 mu M DCF-DA, the supernatant is removed by centrifugation, and the HeLa cells are washed three times by using the serum-free culture medium to remove the DCF-DA which does not enter the cells; measuring the green fluorescence intensity by using a flow cytometer within half an hour after the cells are collected; the excitation wavelength is 488nm, and the emission wavelength is 530 +/-20 nm. The mean fluorescence intensity of the green light was analyzed with the FlowJo7.6 (Tree Star, OR, USA) software.
The results of the compound TPA-N-8 on induction of intracellular reactive oxygen species are shown in FIG. 3. The results show that: compared with a control group, the content of active oxygen in cells can be effectively induced to increase after the compound TPA-N-8 is treated, after 6 mu L of the compound TPA-N-8 is added into HeLa cells, the content of active oxygen in the cells is increased by about 10 times, and meanwhile, green fluorescence in a confocal picture is obviously enhanced. The control group (only DMSO) has no obvious change in the intracellular reactive oxygen species, which indicates that the compound TPA-N-8 induces the cancer cells to generate Reactive Oxygen Species (ROS) after entering the cancer cells, thereby causing the cancer cells to die.
Use of TPA-N-8 prepared in example 1 to induce intracellular mitochondrial membrane potential changes:
the flow cytometry detects the change of mitochondrial membrane potential in tumor cells. Adding cell culture solution containing compound TPA-N-8(1/2/4/6/8 μ L) into 6-well plate inoculated with HeLa cell with good morphology and normal growth condition, treating with drug for 2 hr, collecting cell, washing with PBS, adding prepared JC-1 working solution, and staining for 25-30 min; the cells were washed and resuspended with buffer, the test samples were immediately examined using a BDFACStersersersersery flow cytometer, and the results were processed and analyzed with FlowJo7.6 software. Detecting the fluorescence channel as lambda ex ＝488nm，λ em ＝530nm；λ ex ＝488nm，λ em ＝590nm。
The results of the compound TPA-N-8 on the induction of intracellular mitochondrial membrane potential changes are shown in FIG. 4. The results show that: an increase in JC-1 monomer and a decrease in JC-1 aggregates were observed after treatment with compound TPA-N-8 compared to the control, indicating that compound TPA-N-8 effectively induces a decrease in mitochondrial membrane potential.
Effect of the Compound TPA-N-8 prepared in example 1 on mitochondrial morphology and function:
mitochondrial morphology of HeLa cells after treatment with the compound TPA-N-8 was observed by Transmission Electron Microscopy (TEM). To 100mm which had been seeded with HeLa cells 3 Adding pre-prepared cell culture solution containing 4 μ L of TPA-N-8 compound into culture dish, treating with drug, incubating for 24 hr, collecting cells, washing with PBS, fixing with 2.5% glutaraldehyde, performing gradient dehydration with alcohol, embedding with resin, and ultra-thin cuttingThe tablet is dyed by uranyl acetate and lead citrate, a sample to be observed is prepared on a copper net, and the mitochondrial morphology is observed by using a Hitachi H-7650 transmission microscope.
The results of the experiment on the effect of the compound TPA-N-8 on mitochondrial morphology are shown in FIG. 5. The results show that compared with the intact mitochondrial cristae structure of the Control group which is not treated by adding medicine, the mitochondrial swelling of the HeLa cells treated by the compound TPA-N-8(4 mu M) becomes round, and the cristae structure disappears. This morphologically demonstrated the characteristic of the compound TPA-N-8 damaging HeLa cells mitochondria.
Application of TPA-N-8 prepared in example 1 to the induction of HeLa cell autophagy:
changes in autophagic protein content were detected using western immunoblotting (WB). Adding a prepared cell culture solution containing a compound TPA-N-8(2/4/8 mu L) and a control group (containing DMSO) into a 100mL culture dish of HeLa cells which grow in an adherent manner, centrifugally collecting the cells after the drug treatment is carried out for 24h, washing by PBS to remove serum in the residual culture solution, adding a strong Biyuntian RIPA lysis solution containing PMSF to carry out whole cell lysis for 20min, and ensuring a low-temperature environment of 4 ℃ in the whole process so as to ensure protein invariance. Centrifuging at 13400rpm for 20min at low temperature, and sucking supernatant obtained by centrifugation, namely a cell whole protein sample required by the experiment; determining the protein concentration in the protein sample by using a BCA protein content detection kit; and detecting the expression contents of different proteins in the sample by SDS-PAGE gel electrophoresis. After the gel is prepared, adding protein samples with the same volume into each hole to perform a gel electrophoresis experiment, and stopping electrophoresis immediately after proper separation; the protein of interest was transferred to PVDF membrane using a wet process, and after completion, the membrane was blocked in a solution containing 5% skimmed milk powder for 2 h. Diluting the primary antibody with skimmed milk powder according to the corresponding ratio according to the use instruction of the antibody, and placing the sealed membrane in a primary antibody incubation solution for incubation at room temperature for a period of time to allow the membrane to be specifically combined with the target protein. After completion, the cells were washed with PBST (4X 8 min/time). The washed membrane was incubated in a pre-formulated secondary antibody incubation for a period of time to allow binding of the primary antibody, and washed with PBST as well. Preparing equal-volume ECL developing solution, covering the ECL developing solution on a PVDF membrane, and shooting by using a chemiluminescence imaging system after treating for 2 min.
The experimental results of the compound TPA-N-8 on HeLa cells to induce the expression of autophagy-related proteins are shown in FIG. 6. The results show that: compared with a control group which is not added with drugs, after the cells are treated by TPA-N-8, the expression of the mitophagy-related protein PINK1 is obviously up-regulated, and meanwhile, the content of the autophagy marker protein LC3-II is increased in a concentration-dependent manner, which indicates that the death mode of the HeLa cells induced by the compound TPA-N-8 is autophagy.
Claims (8)
1. An AIE compound having mitochondrial targeting, wherein the compound has the chemical structural formula shown in formula (I):
2. the method of synthesizing an AIE compound with mitochondrial targeting according to claim 1, wherein the method comprises: firstly synthesizing a triphenylamine parent structure compound with aldehyde group, and then reacting the triphenylamine parent structure compound with aldehyde group with pyridine with a long chain to obtain an AIE compound with mitochondrion targeting;
wherein, the chemical structural formula of the triphenylamine parent structure compound with aldehyde group is as follows:
wherein in the pyridine with the long chain, one side of the N atom of the pyridine is grafted with a C4-12 straight-chain alkyl group.
3. The method for synthesizing an AIE compound with mitochondrial targeting according to claim 2, specifically comprising: heating and refluxing a triphenylamine parent structure compound with aldehyde group and pyridine with a long chain in absolute ethyl alcohol for reaction, extracting and separating a crude product after the reaction is finished, and purifying by column chromatography to obtain the AIE compound with mitochondrion targeting.
4. The method for synthesizing an AIE compound having a mitochondrial targeting effect according to claim 2, wherein the triphenylamine parent structural molecule having an aldehyde group is prepared by the following method: heating a compound with a triphenylamine parent structure and 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole in a mixed solution of tetrahydrofuran and water for reflux reaction in an inert atmosphere, extracting, separating and extracting a crude product after the reaction is finished, and purifying by column chromatography to obtain the compound with the triphenylamine parent structure and aldehyde groups;
the chemical structural formula of the compound with the triphenylamine parent structure is as follows:
5. the method of synthesizing an AIE compound with mitochondrial targeting according to claim 3, wherein: the reaction molar ratio of the triphenylamine parent structure compound with aldehyde group to pyridine with long chain is 1: 1.2; the reflux reaction time is 4-4.5 h; the reaction temperature is 80-85 ℃.
6. The method of synthesizing an AIE compound with mitochondrial targeting according to claim 4, wherein: the reaction molar ratio of the compound with the triphenylamine parent structure to the 7-bromo-4-aldehyde benzo [ C ] [1,2,5] thiadiazole is 1: 1.2; the reflux reaction time is 18-19 h; the reaction temperature is 70-75 ℃.
7. The use of the AIE compound having mitochondrial targeting according to claim 1 for the preparation of antitumor drugs and antitumor drug components.
8. The use of the AIE compound having mitochondrial targeting according to claim 7 for the preparation of antitumor drugs and antitumor drug components, characterized in that: the tumor is cervical cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210819688.0A CN115109052B (en) | 2022-07-13 | 2022-07-13 | AIE compound with mitochondrial targeting and synthesis method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210819688.0A CN115109052B (en) | 2022-07-13 | 2022-07-13 | AIE compound with mitochondrial targeting and synthesis method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115109052A true CN115109052A (en) | 2022-09-27 |
| CN115109052B CN115109052B (en) | 2023-08-22 |
Family
ID=83331254
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210819688.0A Active CN115109052B (en) | 2022-07-13 | 2022-07-13 | AIE compound with mitochondrial targeting and synthesis method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115109052B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110642852A (en) * | 2019-10-25 | 2020-01-03 | 南开大学 | Organic AIE photosensitive probe with mitochondrion targeting function and preparation method and application thereof |
| CN111690005A (en) * | 2020-06-27 | 2020-09-22 | 北京化工大学 | Sensing array with mitochondrial targeting and aggregation-induced emission effects and application of sensing array in cell identification |
| CN113956292A (en) * | 2021-10-29 | 2022-01-21 | 南京师范大学 | Mitochondrial targeting compound with aggregation-induced emission property and synthesis method and application thereof |
| CN114195774A (en) * | 2021-11-04 | 2022-03-18 | 徐州医科大学 | Photosensitizer with hypochlorous acid activated fluorescence and mitochondrion targeting functions and preparation method and application thereof |
-
2022
- 2022-07-13 CN CN202210819688.0A patent/CN115109052B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110642852A (en) * | 2019-10-25 | 2020-01-03 | 南开大学 | Organic AIE photosensitive probe with mitochondrion targeting function and preparation method and application thereof |
| CN111690005A (en) * | 2020-06-27 | 2020-09-22 | 北京化工大学 | Sensing array with mitochondrial targeting and aggregation-induced emission effects and application of sensing array in cell identification |
| CN113956292A (en) * | 2021-10-29 | 2022-01-21 | 南京师范大学 | Mitochondrial targeting compound with aggregation-induced emission property and synthesis method and application thereof |
| CN114195774A (en) * | 2021-11-04 | 2022-03-18 | 徐州医科大学 | Photosensitizer with hypochlorous acid activated fluorescence and mitochondrion targeting functions and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115109052B (en) | 2023-08-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102924524B (en) | Manganese complex two-photon absorption material with living cell development function and synthesis method thereof | |
| CN112159522B (en) | Water-soluble rhodamine-based fluorescent/colorimetric dual-mode probe and preparation method and application thereof | |
| CN111848633B (en) | coumarin-Tr baby's base Fe3+ fluorescent probe and preparation method thereof | |
| CN106854210A (en) | The water-soluble porphyrin of phenolic ketone containing adjacent nitro and its Schiff copper porphyrin complex, its synthetic method and application | |
| CN105949222B (en) | A kind of water-soluble acylhydrazone class Schiff porphyrin metal Cu (II) complexs and its synthesis and application | |
| CN104744453A (en) | Hemicyanine compound for detecting polarity of mitochondria | |
| CN114853810A (en) | Curcumin derivative and preparation method and application thereof | |
| CN111793371B (en) | 3, 5-asymmetrically modified BODIPY near-infrared fluorescent dye and preparation method thereof | |
| CN107987085B (en) | Water-soluble nitro-copper-containing porphyrin, water-soluble Schiff base copper porphyrin complex thereof, and synthesis method and application thereof | |
| CN103012401A (en) | Preparation method and application of anthraquinone polypyridine ligand and ruthenium-anthraquinone complex | |
| CN108948093B (en) | Phosphorescent metal iridium complex with singlet oxygen detection effect and preparation method and application thereof | |
| CN115109052B (en) | AIE compound with mitochondrial targeting and synthesis method and application thereof | |
| CN113956292B (en) | Mitochondrial targeting compound with aggregation-induced emission property and synthesis method and application thereof | |
| CN102250149A (en) | Ionic iridium coordination compounds having weak coordinate bonds and preparation method and use thereof | |
| CN114524853B (en) | All-trans retinoic acid-aryl metal complex, preparation method and application | |
| CN112876414B (en) | Polyamine-modified naphthalimide conjugate, and preparation method and application thereof | |
| CN105601676A (en) | Ruthenium complex and application thereof | |
| CN110818739B (en) | Metal iridium complex with synergistic response to tumor microenvironment pH/hypoxic and application thereof | |
| CN112940059B (en) | Glycosyl modified naphthalimide-polyamine conjugate, preparation method and application thereof | |
| Hu et al. | A series of Zn/Cd coordination polymers constructed from 1, 4-naphthalenedicarboxylate and N-donor ligands: Syntheses, structures and luminescence sensing of Cr3+ in aqueous solutions | |
| CN108373487B (en) | Preparation method of two-photon absorption ruthenium complex and application of two-photon absorption ruthenium complex as tumor probe | |
| CN107857766B (en) | Synthetic method and application of spiroindole compound based on phenylalanine and polycarbonyl cyclic ketone compound | |
| CN109824735A (en) | A kind of naphthalimide-platinum (II) complex and its preparation method and application | |
| CN115160376B (en) | Cinnamic acid modified cyclometalated iridium (III) complex and synthetic method and application thereof | |
| CN107857765B (en) | Synthesis method and application of spiroindole compound based on sarcosine and multi-carbonyl cyclic ketone compound |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |