CN116239584B - 一种单体m1、二聚体d1及其制备方法和应用 - Google Patents
一种单体m1、二聚体d1及其制备方法和应用 Download PDFInfo
- Publication number
- CN116239584B CN116239584B CN202310114965.2A CN202310114965A CN116239584B CN 116239584 B CN116239584 B CN 116239584B CN 202310114965 A CN202310114965 A CN 202310114965A CN 116239584 B CN116239584 B CN 116239584B
- Authority
- CN
- China
- Prior art keywords
- tumor
- dimer
- membrane
- monomer
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000539 dimer Substances 0.000 title claims abstract description 64
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 239000000178 monomer Substances 0.000 title abstract description 27
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 127
- 239000003504 photosensitizing agent Substances 0.000 claims abstract description 27
- 230000006907 apoptotic process Effects 0.000 claims abstract description 21
- 238000011282 treatment Methods 0.000 claims abstract description 15
- 239000003814 drug Substances 0.000 claims abstract description 5
- 230000004614 tumor growth Effects 0.000 claims abstract description 5
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 22
- MUQFSLMELVMDSR-UHFFFAOYSA-N 5-[4-(4-methyl-n-(4-methylphenyl)anilino)phenyl]thiophene-2-carbaldehyde Chemical compound C1=CC(C)=CC=C1N(C=1C=CC(=CC=1)C=1SC(C=O)=CC=1)C1=CC=C(C)C=C1 MUQFSLMELVMDSR-UHFFFAOYSA-N 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 11
- -1 octyl-1, 8-diyl Chemical group 0.000 claims description 11
- 239000003054 catalyst Substances 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 6
- FKNQCJSGGFJEIZ-UHFFFAOYSA-O 4-methylpyridin-1-ium Chemical compound CC1=CC=[NH+]C=C1 FKNQCJSGGFJEIZ-UHFFFAOYSA-O 0.000 claims description 4
- 239000011261 inert gas Substances 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 238000010992 reflux Methods 0.000 claims description 2
- 230000005764 inhibitory process Effects 0.000 claims 1
- 239000012528 membrane Substances 0.000 abstract description 64
- 238000009169 immunotherapy Methods 0.000 abstract description 13
- 230000002776 aggregation Effects 0.000 abstract description 6
- 238000004220 aggregation Methods 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 6
- 230000012010 growth Effects 0.000 abstract description 5
- 206010027476 Metastases Diseases 0.000 abstract description 4
- 239000012620 biological material Substances 0.000 abstract description 4
- 238000009826 distribution Methods 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract description 4
- 238000003384 imaging method Methods 0.000 abstract description 4
- 230000009401 metastasis Effects 0.000 abstract description 4
- 230000017188 evasion or tolerance of host immune response Effects 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 238000001338 self-assembly Methods 0.000 abstract description 2
- 230000008685 targeting Effects 0.000 description 45
- 210000004027 cell Anatomy 0.000 description 40
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 20
- 210000000170 cell membrane Anatomy 0.000 description 15
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 10
- 150000001875 compounds Chemical class 0.000 description 10
- 239000003642 reactive oxygen metabolite Substances 0.000 description 10
- 201000011510 cancer Diseases 0.000 description 9
- 239000000243 solution Substances 0.000 description 7
- 102100037387 Gasdermin-A Human genes 0.000 description 6
- 101001026276 Homo sapiens Gasdermin-A Proteins 0.000 description 6
- 230000037449 immunogenic cell death Effects 0.000 description 6
- 238000001126 phototherapy Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000002757 inflammatory effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000002428 photodynamic therapy Methods 0.000 description 5
- 230000005778 DNA damage Effects 0.000 description 4
- 231100000277 DNA damage Toxicity 0.000 description 4
- 230000005975 antitumor immune response Effects 0.000 description 4
- 238000013459 approach Methods 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- 201000006549 dyspepsia Diseases 0.000 description 4
- 208000024798 heartburn Diseases 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 230000036961 partial effect Effects 0.000 description 4
- 238000007626 photothermal therapy Methods 0.000 description 4
- 208000024981 pyrosis Diseases 0.000 description 4
- 239000012265 solid product Substances 0.000 description 4
- 230000009885 systemic effect Effects 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- XDFNWJDGWJVGGN-UHFFFAOYSA-N 2-(2,7-dichloro-3,6-dihydroxy-9h-xanthen-9-yl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC(Cl)=C(O)C=C2OC2=CC(O)=C(Cl)C=C21 XDFNWJDGWJVGGN-UHFFFAOYSA-N 0.000 description 3
- 102000011727 Caspases Human genes 0.000 description 3
- 108010076667 Caspases Proteins 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 230000003213 activating effect Effects 0.000 description 3
- 230000005809 anti-tumor immunity Effects 0.000 description 3
- 238000002619 cancer immunotherapy Methods 0.000 description 3
- 239000003086 colorant Substances 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- AZKSAVLVSZKNRD-UHFFFAOYSA-M 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide Chemical compound [Br-].S1C(C)=C(C)N=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=CC=C1 AZKSAVLVSZKNRD-UHFFFAOYSA-M 0.000 description 2
- FKNQCJSGGFJEIZ-UHFFFAOYSA-N 4-methylpyridine Chemical compound CC1=CC=NC=C1 FKNQCJSGGFJEIZ-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 2
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical compound C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000012223 aqueous fraction Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000005779 cell damage Effects 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 2
- 230000002601 intratumoral effect Effects 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- 210000003463 organelle Anatomy 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
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- ODHXBMXNKOYIBV-UHFFFAOYSA-N triphenylamine Chemical compound C1=CC=CC=C1N(C=1C=CC=CC=1)C1=CC=CC=C1 ODHXBMXNKOYIBV-UHFFFAOYSA-N 0.000 description 2
- DEQOVKFWRPOPQP-UHFFFAOYSA-N (5-formylthiophen-2-yl)boronic acid Chemical compound OB(O)C1=CC=C(C=O)S1 DEQOVKFWRPOPQP-UHFFFAOYSA-N 0.000 description 1
- DKEGCUDAFWNSSO-UHFFFAOYSA-N 1,8-dibromooctane Chemical compound BrCCCCCCCCBr DKEGCUDAFWNSSO-UHFFFAOYSA-N 0.000 description 1
- VFNKZQNIXUFLBC-UHFFFAOYSA-N 2',7'-dichlorofluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(O)C=C1OC1=C2C=C(Cl)C(O)=C1 VFNKZQNIXUFLBC-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 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 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 102000004082 Calreticulin Human genes 0.000 description 1
- 108090000549 Calreticulin Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 230000005971 DNA damage repair Effects 0.000 description 1
- 101710185235 High mobility group protein 1 Proteins 0.000 description 1
- 102100037907 High mobility group protein B1 Human genes 0.000 description 1
- 101710168537 High mobility group protein B1 Proteins 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000003782 apoptosis assay Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000012300 argon atmosphere Substances 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 208000002352 blister Diseases 0.000 description 1
- 230000005880 cancer cell killing Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008045 co-localization Effects 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000004624 confocal microscopy Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000004041 dendritic cell maturation Effects 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 239000001046 green dye Substances 0.000 description 1
- 239000001307 helium Substances 0.000 description 1
- 229910052734 helium Inorganic materials 0.000 description 1
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 1
- 230000008073 immune recognition Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000003859 lipid peroxidation Effects 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000030589 organelle localization Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 239000012221 photothermal agent Substances 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000005522 programmed cell death Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000002829 reductive 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
- 150000003839 salts Chemical class 0.000 description 1
- 102000034285 signal transducing proteins Human genes 0.000 description 1
- 108091006024 signal transducing proteins Proteins 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 125000003698 tetramethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- JLZUZNKTTIRERF-UHFFFAOYSA-N tetraphenylethylene Chemical group C1=CC=CC=C1C(C=1C=CC=CC=1)=C(C=1C=CC=CC=1)C1=CC=CC=C1 JLZUZNKTTIRERF-UHFFFAOYSA-N 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000001544 thienyl group Chemical group 0.000 description 1
- 229930192474 thiophene Natural products 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms 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
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
-
- 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
- C09K2211/1007—Non-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
- C09K2211/1092—Heterocyclic compounds characterised by ligands containing sulfur as the only heteroatom
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种单体M1及其制备方法、一种二聚体D1及其制备方法和一种光敏剂。本发明还公开了所述单体M1和/或二聚体D1和/或光敏剂在制备治疗肿瘤诱导细胞焦亡和/或治疗肿瘤和/或抑制肿瘤生长的药物中的应用。所述单体M1、二聚体D1具有更好的自组装和聚集诱导发射，其在成像和光免疫治疗方面具有更大的潜力；具有近红外聚集诱导的发射和良好的亚细胞分布，是一种很有前途的光诱导细胞焦亡的候选材料。本发明开创性地将肿瘤膜靶向聚集诱导发射光敏二聚体生物材料，并将其用于细胞焦亡介导的光免疫协同治疗。本发明公开的应用可以通过防止免疫逃逸来消除肿瘤和防止肿瘤转移，在抑制原发肿瘤和远端肿瘤的生长方面都非常有效。
Description
技术领域
本发明涉及一种靶向光敏剂及其制备方法和应用，尤其涉及一种单体M1、二聚体D1及其制备方法和应用，属于肿瘤光疗领域。
背景技术
免疫治疗是一种很有前途的肿瘤治疗方法，它通过增强免疫反应来消除肿瘤细胞。它避免了癌细胞的转移和残留，因此优于传统的放疗和化疗，直接杀死癌细胞。实现高效的免疫原性细胞死亡(ICD)作为癌症免疫治疗的关键步骤，已经成为一个重要的问题。在ICD期间，癌细胞释放各种损伤相关的分子模式(DAMP)，包括钙网蛋白、高迁移率族蛋白1和三磷酸腺苷。这些DAMP随后作为天然佐剂触发免疫识别，促进树突状细胞成熟，协助肿瘤相关抗原运送到淋巴结中的T细胞，最终启动适应性抗肿瘤免疫，作为癌症免疫治疗的完整过程。然而，目前诱导ICD的有效途径仅限于对癌细胞具有抵抗力的凋亡或坏死。迫切需要开发一种有效的途径来诱导ICD用于癌症免疫治疗，同时也可以克服癌细胞的耐药性。
细胞焦亡是由炎症小体引发的程序性细胞死亡，死亡的细胞释放抗原并有效地触发抗原特异性免疫反应。细胞焦亡不仅可以绕过细胞凋亡抵抗，而且还可以启动肿瘤特异性免疫，使细胞焦亡介导的免疫治疗成为一种有前景的策略，可以诱导ICD，增强免疫反应，并最终抑制肿瘤生长。细胞焦亡主要依靠炎症小体激活caspase家族的部分蛋白，使其切割gasdermin蛋白，激活gasdermin蛋白，活化的gasdermin蛋白转位到膜上，形成孔洞，细胞肿胀，胞质外流，最终导致细胞膜破裂，细胞焦亡。释放的细胞内容物包括各种肿瘤相关抗原，如乳酸脱氢酶(LDH)和炎性细胞因子(IL-1β和IL-18)，以启动抗肿瘤免疫反应。细胞焦亡可以进一步诱导释放足够的损伤相关的分子模式，以触发强大和持续的抗肿瘤免疫反应。细胞焦亡介导的免疫疗法已被用于癌症化疗；然而，开发一种有效的、非侵入性的、有针对性的癌症光免疫疗法是非常重要的。
光疗具有光损伤可调、无侵袭性、副作用小等优点，是一种很有前途的肿瘤介入治疗方法。光动力疗法和光热疗法是实现光疗的两种合理途径。在光动力学过程中，光敏剂在光照射下产生细胞毒性活性氧物种(ROS)，而在光热过程中，热能通过光热试剂有效地转化为光能。在这种情况下，光动力治疗和光热治疗相结合的协同策略为加速细胞死亡和提高治疗效率提供了一种最佳途径。光动力疗法可以有效地触发caspase的激活，通过产生ROS来启动ICD，而光热疗法可以加速细胞肿胀和膜破裂，这可以提高光免疫疗法。由于ROS的寿命短，有效半径有限，光敏剂的精确亚细胞器定位对于在细胞器中原位产生ROS以避免快速衰减和降解至关重要。在各种亚细胞器靶向光敏剂中，细胞膜靶向光敏剂被认为可以增加膜的通透性，破坏膜的完整性，诱导脂质过氧化，使膜锚定的信号蛋白失活，最终导致细胞膜破裂，细胞内容物迅速释放，这是用于细胞焦亡介导的光免疫治疗的光敏剂的首选方案。尽管基于各种光敏剂的光免疫治疗已经有了广泛的研究，但用于细胞焦亡介导的光免疫治疗的膜靶向光敏剂还很少。
发明内容
发明目的：本发明所要解决的技术问题是提供了一种单体M1及其制备方法。
本发明还要解决的技术问题是提供一种二聚体D1及其制备方法。
本发明还要解决的技术问题是提供一种光敏剂，包括所述单体M1、所述二聚体D1或中的一种或两种。
本发明还要解决的技术问题是提供所述单体M1和/或所述二聚体D1和/或所述光敏剂在制备治疗肿瘤诱导细胞焦亡和/或治疗肿瘤和/或抑制肿瘤生长的药物中的应用。
技术方案：为解决上述技术问题，本发明提供了一种单体M1，化学结构式如下：
本发明还提供了所述单体M1的制备方法，包括以下步骤：将4-甲基-1-(3-(三甲基氨基)丙基)吡啶-1-鎓和5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛加到无水乙醇中，加入哌啶作为催化剂，并在惰性气体中回流过夜，即得单体M1。
本发明还提供了一种二聚体D1，化学结构式如下：
在成像和光免疫治疗方面具有更大的潜力，具有近红外聚集诱导的发射和良好的亚细胞分布，是一种很有前途的光诱导细胞焦亡的候选材料。
本发明还提供了所述二聚体D1的制备方法，包括以下步骤：将5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)加至无水乙醇中，在惰性气体中70～95℃回流过夜，反应得到二聚体D1。
其中，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)的摩尔比为2:0.60～1.2。
其中，还包括催化剂，所述催化剂包括哌啶，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和哌啶的摩尔比为2:0.05～0.3。
本发明还提供了一种光敏剂，所述光敏剂的结构式为D-π-A，D为三苯胺或四苯乙烯的衍生物，π为苯环，噻酚类似的平面结构以及双键三键，A为季铵盐。
其中，包括权利要求1所述单体M1、权利要求2所述二聚体D1中的一种或两种。
本发明还提供了所述单体M1和/或所述二聚体D1和/或所述光敏剂在制备治疗肿瘤诱导细胞焦亡和/或治疗肿瘤和/或抑制肿瘤生长的药物中的应用，可以通过防止免疫逃逸来消除肿瘤和防止肿瘤转移，在抑制原发肿瘤和远端肿瘤的生长方面都非常有效。
发明原理：本发明利用三苯胺、噻吩基和吡啶基形成分子内电荷转移强的π偶联光敏发色团，并用长辛基将两个发色团连接起来形成肿瘤膜靶向D-π-A共轭结构光敏二聚体D1。在光照射下，光动力学和光热过程可以协同进行，在此过程中，产生的I型ROS和热能分别用于启动和增强肿瘤的细胞焦亡。此外，光疗诱导的细胞焦亡促进细胞内容物和炎性细胞因子的释放，从而激活抗肿瘤免疫反应，促进肿瘤特异性抗原的产生和树突状细胞的成熟，导致活化的T细胞增殖，提供全身抗肿瘤免疫。
有益效果：与现有技术相比，本发明具有如下显著优点：
1、肿瘤膜靶向光敏二聚体D1和肿瘤膜靶向光敏单体M1具有更好的自组装和聚集诱导发射，表明其在成像和光免疫治疗方面具有更大的潜力；
2、肿瘤膜靶向光敏二聚体D1和肿瘤膜靶向光敏单体M1具有近红外聚集诱导的发射和良好的亚细胞分布，是一种很有前途的光诱导细胞焦亡的候选材料；
3、开创性地将肿瘤膜靶向聚集诱导发射光敏二聚体生物材料，并将其用于细胞焦亡介导的光免疫协同治疗；
4、本发明提供的全身性抗肿瘤免疫可以通过防止免疫逃逸来消除肿瘤和防止肿瘤转移，在抑制原发肿瘤和远端肿瘤的生长方面都非常有效。
附图说明
图1A为光敏剂单体M1的合成路线，图1B为肿瘤膜靶向光敏二聚体D1的合成路线；
图2为在激发波长为500nm时，肿瘤膜靶向光敏二聚体D1(10μM)在DMSO溶液中的紫外光谱和荧光光谱；
图3A为肿瘤膜靶向光敏二聚体D1在不同水分数(fw)的DMSO/H2O混合物中的荧光光谱，图3B为肿瘤膜靶向光敏二聚体D1在不同水分数(fW)的H2O/DMSO混合物中肿瘤膜靶向光敏二聚体D1在669nm处的荧光强度；
图4为肿瘤膜靶向光敏二聚体D1在水溶液中下产生活性氧ROS的情况；
图5为肿瘤膜靶向光敏二聚体D1在激光(0.5W cm-2)照射下的升温情况；
图6为细胞膜着色剂DiO和肿瘤膜靶向光敏二聚体D1染色的共聚焦图像；
图7A为肿瘤膜靶向光敏二聚体D1分别在光照(520nm，0.3W cm-2，1min)和黑暗条件下，经不同浓度的肿瘤膜靶向光敏二聚体D1处理后的4T1细胞的细胞毒性情况(D1为黑暗条件，D1+L为光照条件)；7B为光敏剂单体M1分别在光照(520nm，0.3W cm-2，1min)和黑暗条件下，经不同浓度的光敏剂单体M1处理后的4T1细胞的细胞毒性情况(D1为黑暗条件，M1+L为光照条件)；
图8A为经肿瘤膜靶向光敏二聚体D1治疗前后的细胞膜扩张和内容物释放情况；图8B为经光敏剂单体M1治疗前后的细胞膜扩张和内容物释放情况；
图9为经肿瘤膜靶向光敏二聚体D1处理4T1细胞损伤后γH2AX蛋白的表达水平；
图10为瘤内注射光敏剂单体M1和肿瘤膜靶向光敏二聚体D1后4T1荷瘤小鼠的体内荧光图像；
图11为激光照射后，4T1荷瘤小鼠的原发肿瘤(上)和远端肿瘤(下)的情况(n＝3)。
具体实施方式
下面结合附图对本发明的技术方案作进一步说明。
实施例1
如图1B所示，一种肿瘤膜靶向光敏二聚体D1的制备，包括以下步骤：
(1)制备化合物5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛：将4-溴-4',4”-二甲基三苯胺(3.38g，9.6mmol)，5-醛基-2-噻吩硼酸(1g，6.4mmol)，碳酸钾(35.3g，25.6mmol)，四氢呋喃(90mL)/水(30mL)和四(三苯基膦)钯(0.37g，0.32mmol)脱气并充入氮气。将反应混合物在70℃下搅拌24h，冷却至室温后，用二氯甲烷和水提取，用无水Na2SO4干燥。待溶剂挥发后，用硅胶柱层析，以正己烷/乙酸乙酯(25/1，v/v)为洗脱剂，得到黄色固体5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛(1.53g，62.5％)。
(2)制备1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)：将4-甲基吡啶(0.93g，10mmol)和1，8-二溴辛烷(2.72g，10mmol)溶于无水N,N-二甲基甲酰胺(DMF，20mL)，装入50mL的圆底烧瓶中。再将混合物在100℃回流过夜，冷却后，用乙醚洗涤，过滤沉淀物，得到白色固体化合物d 1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)(2.7g，90.5％)。该d 1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)的核磁表征如下：1H NMR(600MHz,DMSO-d6)δ8.99(d,J＝6.4Hz,4H),8.01(d,J＝6.2Hz,4H),4.55(t,J＝7.3Hz,4H),2.61(s,6H),1.92-1.82(m,4H),1.26(dd,J＝20.4,5.0Hz,8H).
(3)制备肿瘤膜靶向光敏二聚体D1：将步骤(1)制备的化合物5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛(0.77g,2mmol)和步骤(2)制备的化合物1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)(0.46g，1mmol)加至无水乙醇(30mL)中，加入哌啶(4.26mg，0.05mmo1)作为催化剂，在氮气气氛中(95℃)回流过夜。反应完成后，冷却至室温，过滤，乙醇洗涤三次，干燥得到红色固体产品肿瘤膜靶向光敏二聚体D1(0.73g，61.4％)。所得肿瘤膜靶向光敏二聚体D1具有两个D-π-A结构。
该肿瘤膜靶向光敏二聚体D1的核磁表征数据如下：1H NMR(600MHz,DMSO-d6)δ8.90(d,J＝6.6Hz,4H),8.23(d,J＝15.9Hz,2H),8.19(d,J＝6.7Hz,4H),7.57(d,J＝8.7Hz,4H),7.49(dd,J＝7.4,3.9Hz,4H),7.16(d,J＝8.2Hz,8H),7.12(d,J＝15.8Hz,2H),6.98(d,J＝8.3Hz,8H),6.90(d,J＝8.7Hz,4H),4.46(t,J＝7.2Hz,4H),2.29(s,12H),1.91-1.84(m,4H),1.28(dd,J＝10.6,6.8Hz,8H).13C NMR(151MHz,DMSO-d6)δ153.10,148.73,148.08,144.48,144.39,138.84,134.58,134.39,133.74,130.71,127.17,125.71,125.57,124.34,123.65,121.45,121.21,59.91,30.91,28.69,25.83,20.92.HR-ESI-MS:calcd.forC70H68Br2N4S2:m/z:[M-2Br]2+/2:514.2437,found:m/z 514.2434.
如图1A所示，一种肿瘤膜靶向光敏剂单体M1的制备，包括以下步骤：
将化合物b 4-甲基-1-(3-(三甲基氨基)丙基)吡啶-1-鎓(0.35g，1mmol)和化合物c 5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛(0.38g，1mol)添加到无水乙醇(30mL)中，加入哌啶(4.26mg，0.05mmol)作为催化剂，并在氮气气氛中回流(95℃)过夜。然后，将其冷却至室温，通过减压蒸发除去溶剂。再使用DCM/MeOH(2/1，v/v)作为洗脱剂，通过硅胶柱色谱纯化粗产物，获得紫色固体产物肿瘤膜靶向光敏剂单体M1(0.4g，55.6％)。所得肿瘤膜靶向光敏剂单体M1具有单个D-π-A结构。
该肿瘤膜靶向光敏剂单体M1的核磁表征数据如下：1H NMR(600MHz,DMSO-d6)δ8.96(d,J＝6.8Hz,2H),8.29(d,J＝15.9Hz,1H),8.25(d,J＝6.8Hz,2H),7.58(d,J＝8.7Hz,2H),7.52(d,J＝3.9Hz,1H),7.49(d,J＝3.9Hz,1H),7.16(dd,J＝12.4,6.5Hz,5H),6.99(d,J＝8.3Hz,4H),6.90(d,J＝8.7Hz,2H),4.57(t,J＝7.3Hz,2H),3.45-3.41(m,2H),3.11(s,9H),2.48-2.42(m,2H),2.29(s,6H).13C NMR(151MHz,DMSO-d6)δ153.54,148.79,148.26,144.61,144.49,138.86,134.91,134.55,133.76,130.73,127.21,125.72,125.60,124.40,123.70,121.44,121.20,62.27,56.88,52.96,24.55,20.92.HR-ESI-MS:calcd.forC37H41Br2N3S:m/z:[M-2Br]2+/2:279.6505,found:m/z 279.6504.
实施例2
取实施例1中的化合物5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛(0.77g,2mmol)和化合物1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)加至无水乙醇(30mL)中，加入哌啶作为催化剂，在氩气气氛中(70℃)回流过夜。反应完成后，冷却至室温，过滤，乙醇洗涤三次，干燥得到红色固体产品肿瘤膜靶向光敏二聚体D1(0.73g，61.4％)。所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)摩尔比为2：1.2，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛与哌啶摩尔比为2:0.3。
实施例3
取实施例1中的化合物5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛(0.77g,2mmol)和化合物1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)加至无水乙醇(30mL)中，加入哌啶作为催化剂，在氦气气氛中(85℃)回流过夜。反应完成后，冷却至室温，过滤，乙醇洗涤三次，干燥得到红色固体产品肿瘤膜靶向光敏二聚体D1(0.73g，61.4％)。所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-(辛-1,8-二基)双(4-甲基吡啶-1-鎓)摩尔比为2：0.6，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛与哌啶摩尔比为2:0.2。
实施例4肿瘤膜靶向光敏二聚体D1光物理性质的研究
如图2所示，肿瘤膜靶向光敏二聚体D1的DMSO溶液(10μM)在497nm处显示出主吸收峰，在675nm处显示了荧光发射峰。此外，如图3所示，肿瘤膜靶向光敏二聚体D1在DMSO和水中显示出显著的聚集诱导发射性质。如图4所示，通过用二氯荧光素(DCFH)作ROS指示剂，用20mW cm-2的白光(500-600nm)照射光敏剂肿瘤膜靶向光敏二聚体D1(10μM)和DCFH(50μM)的混合物不同时间后，立即记录光谱。DCFH的荧光在488nm处激发，这显示了肿瘤膜靶向光敏二聚体D1有优异的活性氧生成能力。如图5所示，使用520nm激光评估肿瘤膜靶向光敏二聚体D1的光热行为。用520nm,0.5W cm-2的光照射肿瘤膜靶向光敏二聚体D1溶液(1mM)，温度从25℃逐渐升温至138.4℃。当照射180秒时，肿瘤膜靶向光敏二聚体D1的DMSO溶液显示出强烈的光热效应，并达到138.4℃的最高温度。肿瘤膜靶向光敏二聚体D1的光热转换效率(η)计算为64.19％，说明了肿瘤膜靶向光敏二聚体D1在体外具有光热效应。
实施例5肿瘤膜靶向光敏二聚体D1的膜靶向能力的研究
通过共定位分析，研究肿瘤膜靶向光敏二聚体D1的细胞成像和亚细胞器分布。将HeLa细胞与肿瘤膜靶向光敏二聚体D1(10μM)在共聚焦培养皿中培养30分钟，然后去除培养基并用PBS洗涤细胞。再加入商业细胞膜绿色染料DiO并培养8分钟。最后，用PBS洗涤细胞3次，并在共聚焦激光扫描显微镜(CLSM)下分析。如图6所示，当肿瘤膜靶向光敏二聚体D1与细胞膜着色剂DiO共染色时，肿瘤膜靶向光敏二聚体D1的红色荧光与细胞膜着色剂DiO的绿色荧光有很好的重叠，说明了肿瘤膜靶向光敏二聚体D1具有靶向细胞膜的能力；肿瘤膜靶向光敏二聚体D1和细胞膜着色剂DiO之间的Pearson相关系数为0.90，表明肿瘤膜靶向光敏二聚体D1具有良好的膜靶向能力。由于带正电荷的肿瘤膜靶向光敏二聚体D1和带负电荷的细胞膜之间的静电相互作用，它们往往以细胞膜为靶标，由于其良好的亲水性而不能快速通过细胞膜的磷脂双层。
实施例6在有无光照以及不同浓度的肿瘤膜靶向光敏二聚体D1处理癌细胞的研究
用四甲基偶氮唑盐比色法(MTT法)检测肿瘤膜靶向光敏二聚体D1对4T1肿瘤细胞的杀伤作用。分别在光照(520nm，0.3W cm-2，1min)(D1+L组)和黑暗条件(D1组)下，用不同浓度(0、0.5、1、2、5、10、20、30、40、50μm)的肿瘤膜靶向光敏二聚体D1处理4T1细胞，并对其细胞毒性进行测定(图7A)。同理，分别在光照(520nm，0.3W cm-2，1min)(M1+L组)和黑暗条件(M1组)下，用不同浓度(0、0.5、1、2、5、10、20、30、40、50μm)的肿瘤膜靶向光敏单体M1处理4T1细胞，并对其细胞毒性进行测定(图7B)。MTT法测定细胞活力。在加入不同浓度的肿瘤膜靶向光敏二聚体D1或肿瘤膜靶向光敏单体M1后，用520nm激光(0.3W cm-2)对每个孔进行暗处理或照射1min。培养24小时后，用标准3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑鎓溴化物(MTT)溶液(1mg/ml，100μL)替换每个孔中的培养基。4小时后，向每个孔中加入100μL二甲基亚砜以溶解甲醛，并使用微孔板读取器(VarioskanTM LUX)分析490nm处的吸光度。每个实验应至少进行3次。
如图7所示，在黑暗条件下，4T1细胞在0-50μM的肿瘤膜靶向光敏二聚体D1和肿瘤膜靶向光敏单体M1溶液中的细胞存活率达90％以上，说明肿瘤膜靶向光敏二聚体D1和肿瘤膜靶向光敏单体M1对黑暗中细胞的毒性可忽略不计。在光照条件下，肿瘤膜靶向光敏二聚体D1和肿瘤膜靶向光敏单体M1组细胞死亡率显著增加，达95％以上，表现出极高的癌细胞杀伤能力。
实施例7肿瘤膜靶向光敏二聚体D1处理后的细胞焦亡的研究
激光共聚焦显微镜对肿瘤膜靶向光敏二聚体D1处理的细胞焦亡的形态分析。
细胞焦亡主要依靠炎症小体激活caspase家族的部分蛋白，使其切割gasdermin蛋白，激活gasdermin蛋白，活化的gasdermin蛋白转位到膜上，形成孔洞，细胞肿胀，胞质外流，最终导致细胞膜破裂，细胞焦亡。为了直接研究细胞膜靶向的二聚体D1诱导的细胞焦亡，用共聚焦显微镜观察经肿瘤膜靶向光敏二聚体D1(10μM)处理前后的细胞的形态变化(光照：520nm，0.3W cm-2)。如图8A所示，当肿瘤膜靶向光敏二聚体D1染色的癌细胞被光照射8分钟时，细胞肿胀(有细胞焦亡的迹象)而不是皱缩(有细胞凋亡的迹象)，泡泡从质膜上移除并逐渐扩张，表现出明显的细胞焦亡的过程。如图8B所示，肿瘤膜靶向光敏单体M1，也会产生细胞焦亡的现象。
实施例8
测定肿瘤膜靶向光敏二聚体D1处理4T1细胞损伤后γ-H2AX蛋白的表达水平。分别在光照L1(520nm,0.1W cm–2，15min)、光照L2(520nm，0.3W cm–2，5min)和黑暗条件下，用肿瘤膜靶向光敏二聚体D1(10μM)处理4T1细胞，收集不同组的4T1细胞。加入Triton X-100处理20分钟，然后离心除去Triton X-00。最后，将4T1细胞与兔源组蛋白γ-H2AX多克隆抗体按1:500的比例孵育，然后与山羊抗兔二级抗体(1:500)孵育，然后通过流式细胞术分析细胞。如图9所示，光动力疗法诱导肿瘤DNA损伤，导致DNA损伤标记物γ-H2AX的表达显著增加，而光热疗法的引入进一步加强了DNA损伤。与光动力疗法造成的DNA损伤相比，光热处理组所造成的更强的损伤触发了DNA损伤修复的发生。
实施例9
采用瘤内注射法建立4T1荷瘤小鼠模型(小鼠乳腺癌肿瘤)，评价光敏剂肿瘤膜靶向光敏二聚体D1的治疗作用。将肿瘤膜靶向光敏二聚体D1溶液(5mg/kg；每kg小鼠，5mg肿瘤膜靶向光敏二聚体D1)注射到4T1荷瘤BALB/c小鼠体内，肿瘤膜靶向光敏二聚体D1的荧光在注射后24小时仍然聚集在肿瘤部位(图10)。为进一步证明协同光免疫治疗对全身抗肿瘤免疫反应的影响，建立了小鼠双侧腋窝4T1肿瘤模型，以评价细胞焦亡介导的光免疫治疗是否能有效地诱导特异性全身抗肿瘤效应。治疗14天后，监测原发肿瘤(已治疗)和远处肿瘤(未治疗)的生长情况，以评估治疗效果。如图11所示(1,2,3,4,5,6依次分别代表(PBS,Light,M1,D1,M1+L,D1+L)，光免疫疗法后7天内，原发肿瘤完全消退，远处肿瘤的生长也完全受到抑制。因此，肿瘤膜靶向光敏二聚体D1通过光免疫疗法，通过全身免疫有效地抑制远处肿瘤。
Claims (6)
1.一种二聚体D1，其特征在于，化学结构式如下：
。
2.权利要求1所述二聚体D1的制备方法，其特征在于，包括以下步骤：将5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-（辛-1,8-二基）双（4-甲基吡啶-1-鎓）加至无水乙醇中，在惰性气体中70~95℃回流过夜，反应得到二聚体D1。
3.根据权利要求2所述的方法，其特征在于，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和1,1'-（辛-1,8-二基）双（4-甲基吡啶-1-鎓）的摩尔比为2:0.60~1.2。
4.根据权利要求2所述的方法，其特征在于，还包括催化剂，所述催化剂包括哌啶，所述5-(4-(二对甲苯基氨基)苯基)噻吩-2-甲醛和哌啶的摩尔比为2:0.05~0.3。
5.一种光敏剂，其特征在于，包括权利要求1所述二聚体D1。
6.权利要求1所述二聚体D1和/或权利要求5所述光敏剂在制备治疗肿瘤诱导细胞焦亡和/或治疗肿瘤和/或抑制肿瘤生长的药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310114965.2A CN116239584B (zh) | 2023-02-15 | 2023-02-15 | 一种单体m1、二聚体d1及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310114965.2A CN116239584B (zh) | 2023-02-15 | 2023-02-15 | 一种单体m1、二聚体d1及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116239584A CN116239584A (zh) | 2023-06-09 |
| CN116239584B true CN116239584B (zh) | 2024-06-11 |
Family
ID=86629161
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310114965.2A Active CN116239584B (zh) | 2023-02-15 | 2023-02-15 | 一种单体m1、二聚体d1及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116239584B (zh) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107849441A (zh) * | 2015-06-24 | 2018-03-27 | 香港科技大学 | 用于癌症的可视化和治疗的aie发光体 |
| CN111263751A (zh) * | 2017-10-24 | 2020-06-09 | 香港科技大学 | 具有聚集诱导发光特性的水溶性化合物 |
| CN113234065A (zh) * | 2020-01-22 | 2021-08-10 | 香港科技大学 | 用于光动力学疗法的聚集诱导的发射发光体 |
| CN113582917A (zh) * | 2021-08-12 | 2021-11-02 | 广东工业大学 | 一种三苯胺-季铵盐型聚合物及其制备方法和应用 |
| CN113683605A (zh) * | 2021-07-21 | 2021-11-23 | 南京大学 | 一种基于聚集诱导发光的高效靶向高尔基体的化合物及生物应用 |
| CN113956190A (zh) * | 2021-10-29 | 2022-01-21 | 大连理工大学 | 可激活肿瘤细胞焦亡的细胞器靶向光敏剂及其制备方法和应用 |
| CN114040962A (zh) * | 2019-06-18 | 2022-02-11 | 香港科技大学 | 用于细胞器成像的发射红色荧光的化合物 |
| CN114105967A (zh) * | 2021-11-01 | 2022-03-01 | 浙江大学 | 一种能够诱发肿瘤细胞焦亡的靶向线粒体aie荧光探针及其制备方法和应用 |
| CN114163427A (zh) * | 2021-12-16 | 2022-03-11 | 深圳大学 | 两亲性聚集诱导发光材料、近红外聚集诱导发光有机硅纳米粒子及其制备方法、应用 |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB201910239D0 (en) * | 2019-07-17 | 2019-08-28 | Lightox Ltd | Fluorescent systems for biological imaging and uses thereof |
-
2023
- 2023-02-15 CN CN202310114965.2A patent/CN116239584B/zh active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107849441A (zh) * | 2015-06-24 | 2018-03-27 | 香港科技大学 | 用于癌症的可视化和治疗的aie发光体 |
| CN111263751A (zh) * | 2017-10-24 | 2020-06-09 | 香港科技大学 | 具有聚集诱导发光特性的水溶性化合物 |
| CN114040962A (zh) * | 2019-06-18 | 2022-02-11 | 香港科技大学 | 用于细胞器成像的发射红色荧光的化合物 |
| CN113234065A (zh) * | 2020-01-22 | 2021-08-10 | 香港科技大学 | 用于光动力学疗法的聚集诱导的发射发光体 |
| CN113683605A (zh) * | 2021-07-21 | 2021-11-23 | 南京大学 | 一种基于聚集诱导发光的高效靶向高尔基体的化合物及生物应用 |
| CN113582917A (zh) * | 2021-08-12 | 2021-11-02 | 广东工业大学 | 一种三苯胺-季铵盐型聚合物及其制备方法和应用 |
| CN113956190A (zh) * | 2021-10-29 | 2022-01-21 | 大连理工大学 | 可激活肿瘤细胞焦亡的细胞器靶向光敏剂及其制备方法和应用 |
| CN114105967A (zh) * | 2021-11-01 | 2022-03-01 | 浙江大学 | 一种能够诱发肿瘤细胞焦亡的靶向线粒体aie荧光探针及其制备方法和应用 |
| CN114163427A (zh) * | 2021-12-16 | 2022-03-11 | 深圳大学 | 两亲性聚集诱导发光材料、近红外聚集诱导发光有机硅纳米粒子及其制备方法、应用 |
Non-Patent Citations (3)
| Title |
|---|
| "A tailored and red-emissive type I photosensitizer to potentiate photodynamic immunotherapy";Sirong Zhou，et al.;《J. Mater. Chem. B》;20220829;第10卷;第8003-8012页 * |
| "Pyroptosis-Mediated Synergistic Photodynamic and Photothermal Immunotherapy Enabled by a Tumor-Membrane-Targeted Photosensitive Dimer";Yuqi Tang，et al.;《Adv. Mater.》;20230425;第35卷;第1-12页 * |
| "聚集诱导发光型光敏剂用于线粒体靶向光动力治疗";郭玲香 等;《化学进展》;20220623;第34卷(第11期);第2489-2502页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116239584A (zh) | 2023-06-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Niu et al. | Highly photostable two-photon NIR AIEgens with tunable organelle specificity and deep tissue penetration | |
| Zhu et al. | Anti-Stokes shift luminescent materials for bio-applications | |
| CN110312708B (zh) | 用于生物应用的发光材料 | |
| Che et al. | Ultrafast and noninvasive long-term bioimaging with highly stable red aggregation-induced emission nanoparticles | |
| Patel et al. | Highly effective dual-function near-infrared (NIR) photosensitizer for fluorescence imaging and photodynamic therapy (PDT) of cancer | |
| Xu et al. | Deep-red fluorescent organic nanoparticles with high brightness and photostability for super-resolution in vitro and in vivo imaging using STED nanoscopy | |
| Sourdon et al. | Dendrimeric nanoparticles for two‐photon photodynamic therapy and imaging: Synthesis, Photophysical properties, innocuousness in daylight and cytotoxicity under two‐photon irradiation in the NIR | |
| Yang et al. | Organic small molecular nanoparticles based on self-assembly of amphiphilic fluoroporphyrins for photodynamic and photothermal synergistic cancer therapy | |
| CN114040962B (zh) | 用于细胞器成像的发射红色荧光的化合物 | |
| CN108727256B (zh) | 一种基于三苯胺多吡啶盐的光敏剂及其制备方法与应用 | |
| CN111689955A (zh) | 一类萘并噻二唑自由基型光敏剂及其制备方法与应用 | |
| CN113831331B (zh) | 用于多模态成像及诊疗的近红外二区聚集诱导发光分子及其应用 | |
| Zhang et al. | Diketopyrrolopyrrole derivatives-based NIR-II fluorophores for theranostics | |
| Eçik et al. | Novel Bodipy-triazine conjugates: Synthesis and the generation of singlet oxygen | |
| Ma et al. | Enhanced singlet oxygen generation of a soft salt through efficient energy transfer between two ionic metal complexes | |
| CN114045045B (zh) | 一类单光子上转换五甲川菁类光敏染料、其制备方法和应用 | |
| CN115385851A (zh) | 具有不对称二乙腈基结构的近红外聚集诱导发光型超高效光敏剂、制备方法及应用 | |
| Liang et al. | Esterase‐Activated, pH‐Responsive, and Genetically Targetable Nano‐Prodrug for Cancer Cell Photo‐Ablation | |
| CN113321644B (zh) | 一种具有刺激响应性的超分子光热剂化合物及其组合物和应用 | |
| CN116239584B (zh) | 一种单体m1、二聚体d1及其制备方法和应用 | |
| Peng et al. | Red-to-near-infrared self-reporting photosensitizers with high photostability for photodynamic therapy | |
| WO2008119950A1 (en) | Porphyrin compounds | |
| Li et al. | Novel D-π-AD Type aggregation induced emission luminogens based nanoparticles as efficient 1O2 photosensitizer and bright NIR imaging agent | |
| CN108358972B (zh) | 邻菲罗啉钌配合物类光敏染料及其制备方法和用途 | |
| Chen et al. | Anti-stokes luminescent organic nanoparticles for frequency upconversion biomedical imaging |
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 |