CN114748641A - 一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 - Google Patents
一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 Download PDFInfo
- Publication number
- CN114748641A CN114748641A CN202210425294.7A CN202210425294A CN114748641A CN 114748641 A CN114748641 A CN 114748641A CN 202210425294 A CN202210425294 A CN 202210425294A CN 114748641 A CN114748641 A CN 114748641A
- Authority
- CN
- China
- Prior art keywords
- silicon
- nanocomposite
- hollow mesoporous
- composite
- photosensitizer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 title claims abstract description 108
- 229910052710 silicon Inorganic materials 0.000 title claims abstract description 106
- 239000010703 silicon Substances 0.000 title claims abstract description 106
- 239000002114 nanocomposite Substances 0.000 title claims abstract description 93
- 208000005718 Stomach Neoplasms Diseases 0.000 title claims abstract description 42
- 206010017758 gastric cancer Diseases 0.000 title claims abstract description 42
- 201000011549 stomach cancer Diseases 0.000 title claims abstract description 42
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 230000028993 immune response Effects 0.000 title abstract description 12
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 44
- 238000000034 method Methods 0.000 claims abstract description 32
- 239000002539 nanocarrier Substances 0.000 claims abstract description 30
- 239000003504 photosensitizing agent Substances 0.000 claims abstract description 24
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 22
- 239000002105 nanoparticle Substances 0.000 claims abstract description 17
- 239000000243 solution Substances 0.000 claims description 40
- 238000003756 stirring Methods 0.000 claims description 21
- 238000006243 chemical reaction Methods 0.000 claims description 16
- 230000006378 damage Effects 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 239000003814 drug Substances 0.000 claims description 10
- 230000019491 signal transduction Effects 0.000 claims description 8
- 229920000642 polymer Polymers 0.000 claims description 7
- 230000001939 inductive effect Effects 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 5
- 239000012670 alkaline solution Substances 0.000 claims description 5
- 229910002621 H2PtCl6 Inorganic materials 0.000 claims description 4
- 239000003638 chemical reducing agent Substances 0.000 claims description 4
- 239000002131 composite material Substances 0.000 claims description 4
- 238000005457 optimization Methods 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 239000012279 sodium borohydride Substances 0.000 claims description 4
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 4
- 238000011398 antitumor immunotherapy Methods 0.000 claims description 3
- 150000003377 silicon compounds Chemical class 0.000 claims description 3
- 239000011258 core-shell material Substances 0.000 claims description 2
- 239000006185 dispersion Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 102100031256 Cyclic GMP-AMP synthase Human genes 0.000 claims 1
- 101710118064 Cyclic GMP-AMP synthase Proteins 0.000 claims 1
- 101710196623 Stimulator of interferon genes protein Proteins 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 21
- 238000011065 in-situ storage Methods 0.000 abstract description 6
- 230000009467 reduction Effects 0.000 abstract description 5
- 206010062016 Immunosuppression Diseases 0.000 abstract description 3
- 238000002619 cancer immunotherapy Methods 0.000 abstract description 3
- 230000001506 immunosuppresive effect Effects 0.000 abstract description 3
- 230000000973 chemotherapeutic effect Effects 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 30
- 210000004027 cell Anatomy 0.000 description 26
- 239000002244 precipitate Substances 0.000 description 16
- 238000012360 testing method Methods 0.000 description 15
- 238000005406 washing Methods 0.000 description 15
- WOWHHFRSBJGXCM-UHFFFAOYSA-M cetyltrimethylammonium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+](C)(C)C WOWHHFRSBJGXCM-UHFFFAOYSA-M 0.000 description 14
- 108020005196 Mitochondrial DNA Proteins 0.000 description 13
- 206010028980 Neoplasm Diseases 0.000 description 13
- 108091093105 Nuclear DNA Proteins 0.000 description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- 230000015572 biosynthetic process Effects 0.000 description 12
- 238000003786 synthesis reaction Methods 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 11
- 239000000203 mixture Substances 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 description 9
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 239000001301 oxygen Substances 0.000 description 9
- 229910052760 oxygen Inorganic materials 0.000 description 9
- 238000001514 detection method Methods 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 description 7
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 7
- 235000011114 ammonium hydroxide Nutrition 0.000 description 7
- 230000000259 anti-tumor effect Effects 0.000 description 7
- 239000002245 particle Substances 0.000 description 7
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 6
- 239000006285 cell suspension Substances 0.000 description 6
- 230000001276 controlling effect Effects 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 235000019441 ethanol Nutrition 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 6
- 238000010166 immunofluorescence Methods 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- 210000003855 cell nucleus Anatomy 0.000 description 5
- 210000004443 dendritic cell Anatomy 0.000 description 5
- 238000005530 etching Methods 0.000 description 5
- 238000009169 immunotherapy Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 230000005778 DNA damage Effects 0.000 description 4
- 231100000277 DNA damage Toxicity 0.000 description 4
- 108010067028 Mitochondrial Permeability Transition Pore Proteins 0.000 description 4
- 230000006907 apoptotic process Effects 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 239000012154 double-distilled water Substances 0.000 description 4
- 238000000684 flow cytometry Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 210000001700 mitochondrial membrane Anatomy 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 206010021143 Hypoxia Diseases 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- 210000000805 cytoplasm Anatomy 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 230000001472 cytotoxic effect Effects 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 239000012894 fetal calf serum Substances 0.000 description 3
- 239000012467 final product Substances 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 229920001427 mPEG Polymers 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 230000004792 oxidative damage Effects 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 229940002612 prodrug Drugs 0.000 description 3
- 239000000651 prodrug Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000013049 sediment Substances 0.000 description 3
- 238000000527 sonication Methods 0.000 description 3
- SLIOYUPLNYLSSR-UHFFFAOYSA-J tetrachloroplatinum;hydrate;dihydrochloride Chemical compound O.Cl.Cl.Cl[Pt](Cl)(Cl)Cl SLIOYUPLNYLSSR-UHFFFAOYSA-J 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- ZKSVYBRJSMBDMV-UHFFFAOYSA-N 1,3-diphenyl-2-benzofuran Chemical compound C1=CC=CC=C1C1=C2C=CC=CC2=C(C=2C=CC=CC=2)O1 ZKSVYBRJSMBDMV-UHFFFAOYSA-N 0.000 description 2
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 2
- 238000000116 DAPI staining Methods 0.000 description 2
- 102000053602 DNA Human genes 0.000 description 2
- 108010069941 DNA receptor Proteins 0.000 description 2
- 102000002227 Interferon Type I Human genes 0.000 description 2
- 108010014726 Interferon Type I Proteins 0.000 description 2
- 102100026720 Interferon beta Human genes 0.000 description 2
- 108090000467 Interferon-beta Proteins 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 108010050904 Interferons Proteins 0.000 description 2
- 102100040678 Programmed cell death protein 1 Human genes 0.000 description 2
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000000861 blow drying Methods 0.000 description 2
- 230000009702 cancer cell proliferation Effects 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229940044683 chemotherapy drug Drugs 0.000 description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 2
- 229960004316 cisplatin Drugs 0.000 description 2
- ZOOGRGPOEVQQDX-KHLHZJAASA-N cyclic guanosine monophosphate Chemical compound C([C@H]1O2)O[P@](O)(=O)O[C@@H]1[C@H](O)[C@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-KHLHZJAASA-N 0.000 description 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 230000001146 hypoxic effect Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 210000003470 mitochondria Anatomy 0.000 description 2
- 230000004898 mitochondrial function Effects 0.000 description 2
- 229960001756 oxaliplatin Drugs 0.000 description 2
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 238000002428 photodynamic therapy Methods 0.000 description 2
- 238000007626 photothermal therapy Methods 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- HRTZJTKYLWFENK-UHFFFAOYSA-N triethoxy-[3-(3-triethoxysilylpropylsilyl)propyl]silane Chemical compound CCO[Si](CCC[SiH2]CCC[Si](OCC)(OCC)OCC)(OCC)OCC HRTZJTKYLWFENK-UHFFFAOYSA-N 0.000 description 2
- 210000003171 tumor-infiltrating lymphocyte Anatomy 0.000 description 2
- 210000004366 CD4-positive T-lymphocyte Anatomy 0.000 description 1
- 210000001239 CD8-positive, alpha-beta cytotoxic T lymphocyte Anatomy 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102100034533 Histone H2AX Human genes 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101001067891 Homo sapiens Histone H2AX Proteins 0.000 description 1
- 102000003960 Ligases Human genes 0.000 description 1
- 108090000364 Ligases Proteins 0.000 description 1
- 101710122864 Major tegument protein Proteins 0.000 description 1
- 102000003832 Nucleotidyltransferases Human genes 0.000 description 1
- 108090000119 Nucleotidyltransferases Proteins 0.000 description 1
- 101710148592 PTS system fructose-like EIIA component Proteins 0.000 description 1
- 101710169713 PTS system fructose-specific EIIA component Proteins 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 101710199973 Tail tube protein Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000005773 cancer-related death Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000008102 immune modulation Effects 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000005918 in vitro anti-tumor Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000015788 innate immune response Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002357 laparoscopic surgery Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004065 mitochondrial dysfunction Effects 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000013379 physicochemical characterization Methods 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011470 radical surgery Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 210000004767 rumen Anatomy 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 230000003007 single stranded DNA break Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000004544 sputter deposition Methods 0.000 description 1
- 238000011272 standard treatment Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- -1 triethoxysilyl Chemical group 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/243—Platinum; Compounds thereof
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- 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
- A61P37/04—Immunostimulants
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Inorganic Chemistry (AREA)
- Organic Chemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明提供一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用。本发明以中空介孔硅为纳米载体,并采用原位还原法在包裹有光敏剂的中空介孔硅纳米复合物表面负载铂纳米颗粒,形成兼具光热、光动力、化疗疗效的一体化硅纳米复合物。该硅纳米复合物可应用于逆转胃癌免疫抑制微环境、扩大胃癌免疫治疗的适用人群。
Description
技术领域
本发明属于生物医用材料领域,尤其涉及一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用。
背景技术
胃癌是全球第五大常见的恶性肿瘤,也是癌症相关死亡的第三大原因。一般情况下,80%的胃癌患者在确诊时已处于进展期,预后差、死亡率高。目前,根治性手术联合术后综合治疗仍然是进展期胃癌的标准治疗方案,其中胃癌根治术已由传统开腹手术演进为更加微创、更加精准的腹腔镜手术。然而,进展期胃癌患者的术后综合治疗仍主要是以顺铂、奥沙利铂等为代表的系统性化疗药物,极易导致耐药及肿瘤复发。近年来,以PD-1抗体为代表的免疫疗法在肿瘤治疗领域中取得了突破性进展,它通过充分利用、调动肿瘤患者体内的杀伤性T细胞、杀伤肿瘤细胞。然而,由于胃癌病灶局部缺乏肿瘤浸润淋巴细胞,胃癌患者对免疫治疗的有效应答率低于20%。因此,提高肿瘤微环境中肿瘤浸润淋巴细胞水平,提高胃癌患者的免疫治疗应答率是亟需解决的临床难题。
近年来,研究发现cGAS-STING通路是启动抗肿瘤天然免疫应答的重要途径,有望成为使肿瘤由“冷”转“热”的新一代免疫治疗靶点。其中,环鸟苷酸-腺苷酸合成酶(cGAS)是一种核酸转移酶,具有DNA感受器的功能,能识别各种内外源性的胞质DNA并产生环鸟苷酸-腺苷酸(cGAMP)、激活干扰素刺激蛋白(STING),调控下游的I型干扰素的分泌,激活抗肿瘤天然免疫应答。线粒体DNA(mitoDNA)是真核细胞内中唯一的非核基因,由于mitoDNA缺乏组蛋白的保护、修复能力差,容易受到mitoROS的攻击氧化形成Ox-mitoDNA,甚至泄露在细胞浆内,因此Ox-mitoDNA的释放有望通过激活细胞内的DNA感受器(cGAS),启动抗肿瘤天然免疫应答。前期研究证实,光热和光动力治疗能诱导氧化应激损伤、催生释放的Ox-mitoDNA,这提示光热和光动力效应可激活c-GAS/STING信号通路启动天然免疫应答、协同PD-1单抗发挥增强的抗肿瘤效应。然而,胃癌微环境本身缺氧,在一定程度上限制了光热和光动力效应;此外,蕴含更多遗传信息的细胞核DNA(nDNA)在单纯光热和光动力激发下未能得到充分氧化释放。因此,如何增加肿瘤局部的氧气生成、双向诱导nDNA和mitoDNA损伤,将是强化胃癌天然免疫应答的关键。
目前,大量高级别临床研究与基础研究发现:基于铂类的化疗药物诱导的肿瘤细胞DNA损伤可通过非同步突变产生新抗原、增加肿瘤细胞的免疫原性、抗原呈递细胞向肿瘤微环境募集,进而扩大胃癌免疫治疗获益人群、增敏胃癌免疫治疗。进一步研究证实:临床中使用的铂类药物(包括顺铂和奥沙利铂等)是一类2价药物,其在细胞质中经过水解、羟化作用形成配位离子,并进入细胞核与DNA的两个鸟嘌呤碱基N-7位络合成一个封闭的五元螯合环,从而破坏了两条多核苷酸链上嘌呤和胞嘧啶之间的氢键,扰乱DNA的复制与转录,增加nDNA暴露于细胞质的机会,而暴露的细胞核DNA可进一步激活cGAS-STING信号通路,最终激活I型干扰素和炎症分子介导的天然免疫应答。然而,传统的铂类药物是一类泛细胞性的细胞毒药物,除肿瘤细胞外,其还会对机体生长代谢更新较快的正常细胞产生毒副反应,包括造血干细胞等,这将导致患者出现骨髓抑制、影响免疫细胞的增殖及功能维持。
发明内容
本发明旨在至少解决上述现有技术中存在的技术问题之一。基于此,本发明第一个方面提出一种硅纳米复合物,能够通过双损伤nDNA和mitoDNA,双重激活cGAS/STING信号依赖的天然免疫应答效应,发挥强效的免疫调控,从而增强抗肿瘤免疫治疗。
本发明的第二个方面提出了一种上述硅纳米复合物的制备方法。
本发明的第三个方面提出了一种上述硅纳米复合物在制备诱导mitoDNA和nDNA的双重损伤、激活cGAS/STING信号通路进而增强抗肿瘤免疫治疗的药物中的应用。
根据本发明的第一个方面,提出了一种硅纳米复合物,包括中空介孔硅、铂纳米颗粒、光敏剂,所述中空介孔硅具有核壳结构,所述中空介孔硅的内核负载有所述光敏剂,所述中空介孔硅表面负载有所述铂纳米颗粒。
本发明中,将光敏剂负载于中空介孔硅内核,并在中空介孔硅表面负载0价铂化疗前药(铂纳米颗粒,Pt-NPs),形成硅纳米复合物。其中,Pt-NPs具有类过氧化物酶活性,可以触发H2O2分解为O2,从而缓解胃癌局部缺氧微环境、增强光动力效果,诱导mitoDNA损伤;同时,增强的光动力效应能诱导胃癌细胞氧化应激,活化0价铂化疗前药为具有细胞毒效应的2价铂类药物,诱导nDNA损伤。
在本发明的一些实施方式中,所述中空介孔硅为球状或类球状,所述中空介孔硅的直径为90nm~110nm。
在本发明的一些实施方式中,所述硅纳米复合物中,1mg中空介孔硅负载有230μg~260μg的光敏剂。
在本发明的一些实施方式中,所述硅纳米复合物中,1mg中空介孔硅负载有180μg~210μg的铂纳米粒子。
在本发明的一些实施方式中,所述光敏剂选自IR820、IR783、IR806、IR825、IR808中的至少一种。
根据本发明的第二个方面,提出了一种上述硅纳米复合物的制备方法,包括以下步骤:
S1:将中空介孔硅纳米载体分散液和光敏剂溶液混合,避光反应,得到负载光敏剂的中空介孔硅复合物;
S2:将S1所述负载光敏剂的中空介孔硅复合物的碱性溶液与H2PtCl6反应后,加入还原剂反应,离心,得到所述硅纳米复合物。
在本发明的一些实施方式中,为了进一步提高所述硅纳米复合物的水溶性及生物相容性,还包括对所述硅纳米复合物进行优化处理的步骤,所述优化处理具体为:将S2所述硅纳米复合物溶解于有机溶剂,与两亲性聚合物混合搅拌,再去除有机溶剂,得到优化后的硅纳米复合物。
在本发明的一些实施方式中,所述两亲性聚合物选自C18PMH-mPEG、SH-mPEG-SH、NH2-mPEG-NH2中的至少一种。
在本发明的一些优选的实施方式中,所述两亲性聚合物为C18PMH-mPEG。
在本发明中,通过温和的选择性刻蚀工艺成功地合成中空介孔硅纳米载体,将光敏剂负载于中空介孔硅内核,并利用强还原剂在中空介孔硅表面原位还原成0价铂化疗前药(铂纳米颗粒,Pt-NPs),形成硅纳米复合物。
在本发明的一些优选的实施方式中,S1所述中空介孔硅纳米载体与所述光敏剂的用量比为1:0.1~0.5。
在本发明的一些优选的实施方式中,S2所述负载光敏剂的中空介孔硅复合物与所述H2PtCl6的用量比为1:0.4~0.85。
在本发明的一些更优选的实施方式中,S1所述中空介孔硅纳米载体与所述光敏剂的用量比为1:0.2~0.4。
在本发明的一些更优选的实施方式中,S2所述负载光敏剂的中空介孔硅复合物与所述H2PtCl6的用量比为1:0.5~0.8。
在本发明的一些优选的实施方式中,S1所述避光反应的时间为18h~26h。
在本发明的一些更优选的实施方式中,以十六烷基三甲基氯化铵(CTAC)作为模板剂、利用氨水通过选择性刻蚀方法来构建中空介孔硅(HMON)纳米载体。
根据本发明的第三个方面,提出了一种上述硅纳米复合物在制备诱导胃癌细胞mitoDNA和nDNA的双重损伤、激活cGAS/STING信号通路进而增强免疫治疗的药物中的应用。
本发明的有益效果为:
(1)本发明所述的硅纳米复合物性质稳定、制备条件温和、所需原料廉价、操作方便、工艺简单、高效实用。
(2)本发明创新性地利用“原位还原法”在中空介孔硅纳米载体表面形成铂纳米颗粒,首先,铂纳米颗粒能被光动力效应释放的活性氧活化为具有细胞毒效应的2价形式、引起细胞核DNA损伤;同时,铂纳米颗粒能催化肿瘤细胞内的过氧化氢生成氧气、增强光敏剂的光动力效应、诱导线粒体DNA的氧化损伤。
(3)本发明的硅纳米复合物能通过双向诱导损伤mitoDNA和nDNA、激活cGAS-STING信号通路,触发I型干扰素释放,启动抗肿瘤天然免疫应答并招募浸润性淋巴细胞,实现在制备免疫增敏治疗药物中的应用。
附图说明
下面结合附图和实施例对本发明做进一步的说明,其中:
图1为本发明实施例1制备的硅纳米复合物的理化表征图,其中A为透射电镜图(TEM);B为单线氧生成水平;C为氧气生成水平。
图2为本发明实施例1与对比例制备的硅纳米复合物抗肿瘤效果,其中A为胃癌细胞增殖效果图;B为细胞凋亡水平结果图,***表示P<0.001。
图3为本发明实施例1与对比例制备的硅纳米复合物对线粒体功能的影响,其中A为活性氧生成水平;B为线粒体膜电势(MTPs)图;C为线粒体膜通透性(mPTP)效果图;D为免疫荧光实验结果图,比例尺10μm,***表示P<0.001。
图4为本发明实施例1与对比例制备的硅纳米复合物对胃癌细胞核DNA的损伤检测结果,比例尺10μm。
图5为本发明实施例1与对比例制备的硅纳米复合物激活cGAS/STING信号图,其中A为胃癌细胞内cGAS相对表达水平图;B为胃癌细胞内STING相对表达水平图;C为IFN-β的分泌水平,***表示P<0.001。
图6为本发明实施例1与对比例制备的硅纳米复合物体内免疫招募探索,其中A为胃癌组织内CD8+T淋巴细胞的比例图;B为淋巴结组织中成熟DC细胞的比例图,其中***表示P<0.001。
具体实施方式
以下将结合实施例对本发明的构思及产生的技术效果进行清楚、完整地描述,以充分地理解本发明的目的、特征和效果。显然,所描述的实施例只是本发明的一部分实施例,而不是全部实施例,基于本发明的实施例,本领域的技术人员在不付出创造性劳动的前提下所获得的其他实施例,均属于本发明保护的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
实施例1
本实施例制备了一种硅纳米复合物HMON@IR820/Pt-NPs,具体过程为:
(1)中空介孔硅(HMON)纳米载体的合成:
以十六烷基三甲基氯化铵(CTAC)作为模板剂、利用氨水通过选择性刻蚀方法来构建中空介孔硅纳米载体。首先,将2.1mL的CTAC溶液以及50μL的三乙醇胺(TEA)溶液加入到20mL的去离子水(ddH2O)中,通过磁力加热搅拌装置加热至95℃、恒温搅拌5分钟(500r/min);然后,将1mL的四乙氧基硅烷(TEOS)逐滴滴入上述混合溶液中、溶液逐渐变成乳白色,继续在95℃中持续搅拌1小时(500r/min);随后,将双[3-(三乙氧基甲硅烷基)丙基]四硫化物(BTES,1mL)和TEOS(1mL)混匀,逐滴滴入后继续在95℃条件下反应4小时;反应结束、冷却至室温后,用无水乙醇洗涤3遍,离心(8000r/min,10min)后可获得MSN@MON沉淀。为了去除反应体系中游离的CTAC,用含有8.4mL HCl溶液(37wt%)的30mL ddH2O重悬MSN@MON沉淀,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀;再次加入含有8.4mLHCl溶液(37wt%)的30mL ddH2O,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀。最后,将20mL的ddH2O和13.5mL氨水溶液(25wt%)混匀后、重悬先前的沉淀,在60℃下搅拌反应3h,用无水乙醇和ddH2O离心洗涤3次即可获得HMON纳米载体,并进行干燥称重。
(2)HMON@IR820纳米复合物的合成:
首先,将先前合成的50mg HMON纳米载体溶解于25mL的乙醇溶液中;然后,利用5mL的二甲基亚砜(DMSO)溶液溶解25mg的光敏剂IR820粉末后,并迅速与含HMON纳米载体的乙醇溶液混匀。避光反应18小时后,离心(8000rpm/min,10min)、ddH2O洗涤纯化后,可获得蓝绿色沉淀(HMON@IR820)。
(3)HMON@IR820/Pt-NPs纳米复合物的合成:
通过“原位还原法”将Pt-NPs负载于HMON@IR820表面。具体合成步骤如下:首先,用ddH2O配制pH=10的NaOH溶液,并将20mg HMON@IR820纳米复合物溶解于10mL上述的碱性溶液中,加入氯铂酸水合物(H2PtCl6,15mg)后,在37℃的环境中间歇超声搅拌3小时;随后,将预先配置好的硼氢化钠(50mM,1mL)逐滴加入上述的溶液中,涡旋搅拌12小时后,经离心(8000rpm/min,10min)、ddH2O洗涤后,即可获得黑色产物(HMON@IR820/Pt-NPs)。此外,为了进一步提高HMON@IR820/Pt-NPs纳米复合物的水溶性及生物相容性,将HMON@IR820/Pt-NPs纳米复合物溶解于氯仿溶液中并加入两亲性聚合物C18PMH-mPEG(30mg)搅拌,在37℃的环境中搅拌1小时,然后通过吹干去除氯仿。最后,将最终产物(HMON@IR820/Pt-NPs纳米成分)通过超声处理重新分散在水中,并在25℃下保存。
实施例2
本实施例制备了一种硅纳米复合物HMON@IR820/Pt-NPs,具体过程为:
(1)中空介孔硅(HMON)纳米载体的合成:
以十六烷基三甲基氯化铵(CTAC)作为模板剂、利用氨水通过选择性刻蚀方法来构建中空介孔硅纳米载体。首先,将2.1mL的CTAC溶液以及50μL的三乙醇胺(TEA)溶液加入到20mL的去离子水(ddH2O)中,通过磁力加热搅拌装置加热至95℃、恒温搅拌5分钟(500r/min);然后,将1mL的四乙氧基硅烷(TEOS)逐滴滴入上述混合溶液中、溶液逐渐变成乳白色,继续在95℃中持续搅拌1小时(500r/min);随后,将双[3-(三乙氧基甲硅烷基)丙基]四硫化物(BTES,1mL)和TEOS(1mL)混匀,逐滴滴入后继续在95℃条件下反应4小时;反应结束、冷却至室温后,用无水乙醇洗涤3遍,离心(8000r/min,10min)后可获得MSN@MON沉淀。为了去除反应体系中游离的CTAC,用含有8.4mL HCl溶液(37wt%)的30mL ddH2O重悬MSN@MON沉淀,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀;再次加入含有8.4mLHCl溶液(37wt%)的30mL ddH2O,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀。最后,将20mL的ddH2O和13.5mL氨水溶液(25wt%)混匀后、重悬先前的沉淀,在60℃下搅拌反应3h,用无水乙醇和ddH2O离心洗涤3次即可获得HMON纳米载体,并进行干燥称重。
(2)HMON@IR820纳米复合物的合成:
首先,将先前合成的50mg HMON纳米载体溶解于25mL的乙醇溶液中;然后,利用5mL的二甲基亚砜(DMSO)溶液溶解10mg的光敏剂IR820粉末后,并迅速与含HMON纳米载体的乙醇溶液混匀。避光反应20小时后,离心(8000rpm/min,10min)、ddH2O洗涤纯化后,可获得蓝绿色沉淀(HMON@IR820)。
(3)HMON@IR820/Pt-NPs纳米复合物的合成:
通过“原位还原法”将Pt-NPs负载于HMON@IR820表面。具体合成步骤如下:首先,用ddH2O配制pH=10的NaOH溶液,并将20mg HMON@IR820纳米复合物溶解于10mL上述的碱性溶液中,加入氯铂酸水合物(H2PtCl6,10mg)后,在37℃的环境中间歇超声搅拌3小时;随后,将预先配置好的硼氢化钠(50mM,1mL)逐滴加入上述的溶液中,涡旋搅拌12小时后,经离心(8000rpm/min,10min)、ddH2O洗涤后,即可获得黑色产物(HMON@IR820/Pt-NPs)。此外,为了进一步提高HMON@IR820/Pt-NPs纳米复合物的水溶性及生物相容性,将HMON@IR820/Pt-NPs纳米复合物溶解于氯仿溶液中并加入两亲性聚合物SH-mPEG-SH(30mg)搅拌,在37℃的环境中搅拌1小时,然后通过吹干去除氯仿。最后,将最终产物(HMON@IR820/Pt-NPs纳米成分)通过超声处理重新分散在水中,并在25℃下保存。
实施例3
本实施例制备了一种硅纳米复合物HMON@IR820/Pt-NPs,具体过程为:
(1)中空介孔硅(HMON)纳米载体的合成:
以十六烷基三甲基氯化铵(CTAC)作为模板剂、利用氨水通过选择性刻蚀方法来构建中空介孔硅纳米载体。首先,将2.1mL的CTAC溶液以及50μL的三乙醇胺(TEA)溶液加入到20mL的去离子水(ddH2O)中,通过磁力加热搅拌装置加热至95℃、恒温搅拌5分钟(500r/min);然后,将1mL的四乙氧基硅烷(TEOS)逐滴滴入上述混合溶液中、溶液逐渐变成乳白色,继续在95℃中持续搅拌1小时(500r/min);随后,将双[3-(三乙氧基甲硅烷基)丙基]四硫化物(BTES,1mL)和TEOS(1mL)混匀,逐滴滴入后继续在95℃条件下反应4小时;反应结束、冷却至室温后,用无水乙醇洗涤3遍,离心(8000r/min,10min)后可获得MSN@MON沉淀。为了去除反应体系中游离的CTAC,用含有8.4mL HCl溶液(37wt%)的30mL ddH2O重悬MSN@MON沉淀,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀;再次加入含有8.4mLHCl溶液(37wt%)的30mL ddH2O,将温度控制在80℃持续搅拌12小时,用无水乙醇洗涤3遍后收集沉淀。最后,将20mL的ddH2O和13.5mL氨水溶液(25wt%)混匀后、重悬先前的沉淀,在60℃下搅拌反应3h,用无水乙醇和ddH2O离心洗涤3次即可获得HMON纳米载体,并进行干燥称重。
(2)HMON@IR820纳米复合物的合成:
首先,将先前合成的50mg HMON纳米载体溶解于25mL的乙醇溶液中;然后,利用5mL的二甲基亚砜(DMSO)溶液溶解7mg的光敏剂IR820粉末后,并迅速与含HMON纳米载体的乙醇溶液混匀。避光反应24小时后,离心(8000rpm/min,10min)、ddH2O洗涤纯化后,可获得蓝绿色沉淀(HMON@IR820)。
(3)HMON@IR820/Pt-NPs纳米复合物的合成:
通过“原位还原法”将Pt-NPs负载于HMON@IR820表面。具体合成步骤如下:首先,用ddH2O配制pH=10的NaOH溶液,并将20mg HMON@IR820纳米复合物溶解于10mL上述的碱性溶液中,加入氯铂酸水合物(H2PtCl6,12mg)后,在37℃的环境中间歇超声搅拌3小时;随后,将预先配置好的硼氢化钠(50mM,1mL)逐滴加入上述的溶液中,涡旋搅拌12小时后,经离心(8000rpm/min,10min)、ddH2O洗涤后,即可获得黑色产物(HMON@IR820/Pt-NPs)。此外,为了进一步提高HMON@IR820/Pt-NPs纳米复合物的水溶性及生物相容性,将HMON@IR820/Pt-NPs纳米复合物溶解于氯仿溶液中并加入两亲性聚合物NH2-mPEG-NH2(30mg)搅拌,在37℃的环境中搅拌1小时,然后通过吹干去除氯仿。最后,将最终产物(HMON@IR820/Pt-NPs纳米成分)通过超声处理重新分散在水中,并在25℃下保存。
对比例
本对比例制备了一种硅纳米复合物HMON@IR820,具体过程与实施例1中(1)和(2)相同,与实施例1的区别在于本对比例未负载铂纳米颗粒。
试验例1
本试验例将实施例1制备的硅纳米复合物HMON@IR820/Pt-NPs进行如下各理化表征:
(1)透射电镜检查:
将硅纳米复合物滴加在Cu网上,常温下,待溶液蒸发后,用离子溅射仪镀上导电金膜,在200kV的加速电压下,采用TEM观察纳米颗粒的原始粒径及形状,见图1A。
从图1A的透射电镜结果可见该硅纳米复合物被成功合成,其是一个直径约100nm的球形纳米颗粒、表面有很多黑色的小颗粒(即为铂纳米颗粒)。
(2)粒径及电势分析:
将实施例1制备的中空介孔硅纳米载体HMON和硅纳米复合物HMON@IR820/Pt-NPs分别复合物用足量的去离子水分散;然后,用马尔文动态光散射仪检测、分析其粒径与电位,见表1。
表1粒径与电位
实施例1制备的HMON | 实施例1制备的HMON@IR820/Pt-NPs | |
粒径(nm) | 91.23±2.76 | 93.98±3.12 |
电位(mV) | -24.56±1.45 | -29.54±1.23 |
从表1可看出,HMON纳米载体与HMON@IR820/Pt-NPs纳米复合物的粒径约90nm;HMON纳米载体的电势为-24.56±1.45mV,负载Pt-NPs的硅纳米复合物的电势为-29.54±1.23mV,表明负载铂纳米颗粒可以降低电势使HMON@IR820/Pt-NPs纳米复合物更容易在血液循环中运输。
(3)光动力特性检测:
为了检测硅纳米复合物的光动力效应,用1,3-Diphenylisobenzofuran(DPBF)探针检测经NIR分别照射1mM过氧化氢溶液与加入HMON@IR820/Pt-NPs纳米复合物的1mM过氧化氢溶液后,上述两种溶液中单线态氧的产生水平,结果见图1B。
从图1B可看出该硅纳米复合物具有良好的光动力效应,能诱导单线氧的生成。
(4)氧气生成水平检测:
将HMON@IR820/Pt-NPs硅纳米复合物与2mM的H2O2溶液进行共孵育,利用氧气检测装置监测溶液内氧气的生成水平,见图1C。
从图1C可看出该硅纳米复合物可以催化过氧化氢产生氧气,有望应用于缓解肿瘤局部的缺氧微环境。
试验例2
本试验例将实施例1和对比例制备的硅纳米复合物进行体外抗肿瘤测试,具体过程为:
(1)胃癌细胞增殖实验:
将MFC胃癌细胞分为4组分别接种于96孔板中;待细胞贴壁后,两组加入HMON@IR820,另外两组加入HMON@IR820/Pt-NPs纳米复合物,进行共孵育4小时后,一组加入HMON@IR820的和一组加入HMON@IR820/Pt-NPs的进行NIR照射(808nm,0.7W/cm2,5min),剩下两组则不进行照射,继续孵20小时后在各组孔中加入10wt%的CCK-8试剂,继续37℃孵育2小时后,在酶标仪测定490nm波长各孔的吸光度(A)值,结果见图2A。
图2A结果显示:在近红外光照射下,HMON@IR820和HMON@IR820/Pt-NPs均能显著抑制胃癌细胞的增殖活性,但HMON@IR820/Pt-NPs处理组的细胞活性最低。
(2)细胞凋亡水平检测:
将MFC胃癌细胞分3组接种于6孔板中,其中两组分别加入HMON@IR820和HMON@IR820/Pt-NPs纳米复合物,剩下一组用含10%胎牛血清的DMEM培养基处理作为对照组(Control),进行共孵育4小时后进行NIR照射(808nm,0.7W/cm2,5min),继续孵20小时后收集细胞沉淀,加入细胞凋亡检测试剂,避光孵育15分钟,最后在流式细胞仪上进行检测,结果见图2B。
图2B流式细胞学结果一致显示,在近红外光照射下,HMON@IR820/Pt-NPs纳米复合物能诱导更多的胃癌细胞发生凋亡。
试验例3
本试验例将实施例1和对比例制备的硅纳米复合物进行诱导线粒体功能障碍测试,具体过程为:
(1)线粒体功能检测:将MFC胃癌细胞分为三组,接种于6孔板中,分别为HMON@IR820合并NIR照射(808nm,0.7W/cm2,5min)组、HMON@IR820/Pt-NPs纳米复合物合并NIR照射(808nm,0.7W/cm2,5min)组和用含10%胎牛血清的DMEM培养基处理作为对照组(Control);收集细胞分别用JC-1检测试剂盒、ROS检测试剂盒、mPTP检测试剂盒测试对细胞内线粒体膜电势(MTP)、ROS的生成水平以及mPTP的开放程度。
(2)免疫荧光实验:8-羟脱氧鸟苷(8-OHdG)是mitoDNA损害的敏感标志物,利用根据8-OHdG的表达量来判断细胞内氧化型线粒体DNA(Ox-mitoDNA)的表达水平。收集处理后的MFC胃癌细胞进行免疫荧光实验,即经固定、破膜、封闭后,与泛素化后的8-OHdG抗体孵育后用Cy3标记的人抗鼠IgG二抗进行显色、与TOM20抗体孵育后用FITC标记的人抗兔IgG二抗进行显色、与DAPI染色液孵育进行细胞核染色,最后在荧光线显微镜下拍照分析。
结果显示,HMON@IR820/Pt-NPs纳米复合物的处理能下调胃癌细胞内的线粒体膜电势(图3A)、诱导ROS生成(图3B)、增加线粒体通透性转换孔(mPTP)的开放(图3C)。此外,在图3D中,8-OHdG抗体标记的是氧化损伤型DNA,而TOM20抗体标记的是线粒体;通过免疫荧光实验发现光热和光动力治疗后胃癌细胞内的8-OHdG水平显著上调;且8-OHdG的荧光与线粒体的荧光(TOM20)完全重合,这提示光热和光动力导致的DNA氧化损伤均为线粒体DNA。
试验例4
本试验例将实施例1和对比例制备的硅纳米复合物进行诱导细胞核DNA受损测试,具体过程为:
免疫荧光实验:γH2AX是双链DNA损伤的关键标记物,其表达上调常常提示细胞核DNA损伤,其简要的操作步骤如下所述:收集分别经HMON@IR820和HMON@IR820/Pt-NPs纳米复合物以及含10%胎牛血清的DMEM培养基处理后的MFC胃癌细胞进行免疫荧光实验,即经固定、破膜、封闭后,与γ-H2AX孵育后用FITC标记的人抗兔IgG二抗进行显色、与DAPI染色液孵育进行细胞核染色,最后在荧光线显微镜下拍照分析。
图4结果显示:γ-H2AX标记的是损伤断裂的双链DNA;在近红外光驱动下,HMON@IR820/Pt-NPs纳米复合物能引起细胞核内γ-H2AX的荧光增强,这提示HMON@IR820/Pt-NPs纳米复合物能引起细胞核内的双链DNA断裂。
试验例5
本试验例将实施例1制备的硅纳米复合物进行激活cGAS/STING信号通路测试,具体过程为:
(1)RT-qPCR实验:
将MFC胃癌细胞接种于六孔板中,分三组,分别加入HMON@IR820、HMON@IR820/Pt-NPs、含10%胎牛血清的DMEM培养基处理(Control对照组)后分别收集各组细胞进行总RNA提取、逆转录、荧光定量PCR等实验步骤后,收集数据进行分析。
(2)酶联免疫吸附实验(ELISA):
将胃癌细胞接种于六孔板中,分三组,分别加入HMON@IR820、HMON@IR820/Pt-NPs、含10%胎牛血清的DMEM培养基处理(Control对照组)后收集细胞上清液进行ELISA实验,检测IFN-β的分泌水平。
结果分析:RT-qPCR实验结果显示,HMON@IR820/Pt-NPs纳米复合物能引起cGAS及STING转录水平上调(图5A和图5B)、诱导肿瘤细胞分泌更多的IFN-β(图5C)。这提示,HMON@IR820/Pt-NPs纳米复合物能激活cGAS/STING信号通路,有望逆转胃癌免疫抑制微环境。
试验例6
本试验例将实施例1制备的硅纳米复合物进行免疫调控效应测试,具体过程为:
首先构建皮下荷瘤胃癌小鼠模型,并随机分为三组:Control组、HMON@IR820+NIR组、HMON@IR820/Pt-NPs+NIR组。模型构建成功后,分别在第1、3、5天时间点经皮下注射方式进行处理(Control组对应注射生理盐水处理),在药物注射24小时后进行近红外光(NIR)照射(808nm,0.7W/cm2,5min)。在处理后的第七天,将收集的肿瘤组织、淋巴结及脾脏组织充分剪碎、研磨成匀浆,加入PBS用移液枪反复吹打,使之尽量变成单细胞悬液,随后与特异性抗体孵育后进行流式分选:①根据说明书,将收集不同组织的单细胞悬液与anti-CD80-cy5.5、anti-CD86-FITC和anti-CD11c-PE流式抗体共孵育,用流式细胞术测定肿瘤组织中成熟树突状细胞(DC)的含量;②将收集不同组织的单细胞悬液与anti-CD3-cy5.5和anti-CD4-FITC流式抗体共孵育,用流式细胞术测定肿瘤组织中CD4+T细胞的含量;③将收集不同组织的单细胞悬液与anti-CD3-cy5.5和anti-CD8-PE流式抗体共孵育,用流式细胞术测定肿瘤组织中CD8+T细胞的含量。
结果分析:在近红外光的照射下,HMON@IR820/Pt-NPs能通过光热和光动力以及化疗效应双重激活cGAS/STING信号通路,进一步活化DC细胞为成熟的DC细胞、并招募更多的CD4和CD8阳性的T淋巴细胞聚集于胃癌病灶(图6A和图6B)。因此,体内预实验一致说明HMON@IR820/Pt-NPs纳米复合物能激活cGAS/STING信号通路、逆转胃癌免疫抑制微环境,启动抗肿瘤天然免疫应答。
上面对本发明实施例作了详细说明,但是本发明不限于上述实施例,在所属技术领域普通技术人员所具备的知识范围内,还可以在不脱离本发明宗旨的前提下作出各种变化。此外,在不冲突的情况下,本发明的实施例及实施例中的特征可以相互组合。
Claims (10)
1.一种硅纳米复合物,其特征在于,包括中空介孔硅、铂纳米颗粒、光敏剂,所述中空介孔硅具有核壳结构,所述中空介孔硅的内核负载所述光敏剂,所述中空介孔硅表面负载有所述铂纳米颗粒。
2.根据权利要求1所述的一种硅纳米复合物,其特征在于,所述中空介孔硅为球状或类球状,所述中空介孔硅的直径为90nm~110nm。
3.根据权利要求1所述的一种硅纳米复合物,其特征在于,所述硅纳米复合物中,1mg中空介孔硅负载有230μg~260μg的光敏剂和180μg~210μg的铂纳米粒子。
4.一种如权利要求1~3任一项所述的硅纳米复合物的制备方法,其特征在于,包括以下步骤:
S1:将中空介孔硅纳米载体分散液和光敏剂溶液混合,避光反应,得到负载光敏剂的中空介孔硅复合物;
S2:S1所述负载光敏剂的中空介孔硅复合物的碱性溶液与H2PtCl6反应后,加入还原剂反应,离心,得到所述硅纳米复合物。
5.根据权利要求4所述的硅纳米复合物的制备方法,其特征在于,还包括对所述硅纳米复合物进行优化处理的步骤,所述优化处理具体为:将S2所述硅纳米复合物溶解于有机溶剂,与两亲性聚合物混合搅拌,去除有机溶剂。
6.根据权利要求4所述的硅纳米复合物的制备方法,其特征在于,S1所述中空介孔硅纳米载体与所述光敏剂的用量比为1:0.1~0.5。
7.根据权利要求4所述的硅纳米复合物的制备方法,其特征在于,S2所述负载光敏剂的中空介孔硅复合物与所述H2PtCl6的用量比为1:0.4~0.85。
8.根据权利要求4所述的硅纳米复合物的制备方法,其特征在于,S2所述还原剂选自硼氢化钠。
9.根据权利要求4所述的硅纳米复合物的制备方法,其特征在于,S1所述避光反应的时间为18h~26h。
10.一种硅纳米复合物在制备诱导胃癌细胞mitoDNA和nDNA的双重损伤、激活cGAS/STING信号通路进而增强抗肿瘤免疫治疗的药物中的应用,其特征在于,所述硅纳米复合物为权利要求1~3任一项所述的硅纳米复合物或根据权利要求4~9任一项所述的制备方法制备的硅纳米复合物。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210425294.7A CN114748641A (zh) | 2022-04-22 | 2022-04-22 | 一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210425294.7A CN114748641A (zh) | 2022-04-22 | 2022-04-22 | 一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN114748641A true CN114748641A (zh) | 2022-07-15 |
Family
ID=82332129
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210425294.7A Pending CN114748641A (zh) | 2022-04-22 | 2022-04-22 | 一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114748641A (zh) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110196285A1 (en) * | 2008-10-10 | 2011-08-11 | Dong Chen | Hollow Mesoporous Silica Sphere Coated with Gold and Preparation Method Thereof and Use in Cancer Therapy |
CN103011067A (zh) * | 2012-12-28 | 2013-04-03 | 哈尔滨工业大学 | 介孔二氧化硅纳米马达及其制备方法和应用 |
US20160067354A1 (en) * | 2014-08-29 | 2016-03-10 | University Of South Carolina | Preparations of gold/mesoporous silica hybrid nanoparitcle and applications |
CN107684626A (zh) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | 介孔二氧化硅‑6‑巯基嘌呤‑顺铂纳米粒, 其制备, 活性和应用 |
US20180153796A1 (en) * | 2014-10-14 | 2018-06-07 | The University Of Chicago | Nanoparticles for photodynamic therapy, x-ray induced photodynamic therapy, radiotherapy, radiodynamic therapy, chemotherapy, immunotherapy, and any combination thereof |
CN108992419A (zh) * | 2018-06-27 | 2018-12-14 | 南京师范大学 | 一种介孔-大孔纳米马达及其制备方法和应用 |
US20200246179A1 (en) * | 2015-12-21 | 2020-08-06 | Gholam A. Peyman | Cancer Treatment Methods Using Thermotherapy And/Or Enhanced Immunotherapy |
-
2022
- 2022-04-22 CN CN202210425294.7A patent/CN114748641A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110196285A1 (en) * | 2008-10-10 | 2011-08-11 | Dong Chen | Hollow Mesoporous Silica Sphere Coated with Gold and Preparation Method Thereof and Use in Cancer Therapy |
CN103011067A (zh) * | 2012-12-28 | 2013-04-03 | 哈尔滨工业大学 | 介孔二氧化硅纳米马达及其制备方法和应用 |
US20160067354A1 (en) * | 2014-08-29 | 2016-03-10 | University Of South Carolina | Preparations of gold/mesoporous silica hybrid nanoparitcle and applications |
US20180153796A1 (en) * | 2014-10-14 | 2018-06-07 | The University Of Chicago | Nanoparticles for photodynamic therapy, x-ray induced photodynamic therapy, radiotherapy, radiodynamic therapy, chemotherapy, immunotherapy, and any combination thereof |
US20200246179A1 (en) * | 2015-12-21 | 2020-08-06 | Gholam A. Peyman | Cancer Treatment Methods Using Thermotherapy And/Or Enhanced Immunotherapy |
CN107684626A (zh) * | 2016-08-05 | 2018-02-13 | 首都医科大学 | 介孔二氧化硅‑6‑巯基嘌呤‑顺铂纳米粒, 其制备, 活性和应用 |
CN108992419A (zh) * | 2018-06-27 | 2018-12-14 | 南京师范大学 | 一种介孔-大孔纳米马达及其制备方法和应用 |
Non-Patent Citations (1)
Title |
---|
WEIHONG GUO,等: "Improved immunotherapy for gastric cancer by nanocomposites with capability of triggering Dual-Damage of Nuclear/Mitochondrial DNA and cGAS/STING-Mediated innate immunity", 《CHEMICAL ENGINEERING JOURNAL》 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113559084B (zh) | 一种基于微流控芯片的载药超小四氧化三铁纳米团簇及其制备方法和应用 | |
Wang et al. | NIR-II light triggered nitric oxide release nanoplatform combined chemo-photothermal therapy for overcoming multidrug resistant cancer | |
CN110201169B (zh) | 氧气自给型靶向纳米光动力治疗系统 | |
Yu et al. | Graphene quantum dots‐based targeted nanoprobes detecting drug delivery, imaging, and enhanced chemotherapy of nasopharyngeal carcinoma | |
Yang et al. | Type I macrophage activator photosensitizer against hypoxic tumors | |
Yang et al. | Construction of PEI‐EGFR‐PD‐L1‐siRNA dual functional nano‐vaccine and therapeutic efficacy evaluation for lung cancer | |
Yang et al. | Tirapazamine-loaded UiO-66/Cu for ultrasound-mediated promotion of chemodynamic therapy cascade hypoxia-activated anticancer therapy | |
Jia et al. | Preparation of responsive “dual-lock” nanoparticles and their application in collaborative therapy based on CuS coordination | |
Wu et al. | Ultrasound‐Driven Piezoelectrocatalytic Immunoactivation of Deep Tumor | |
Xu et al. | Development of fullerene nanospherical miRNA and application in overcoming resistant breast cancer | |
CN113230418A (zh) | 一种超小核壳结构铁纳米颗粒的制备方法及应用 | |
Wang et al. | Effect of Gambogic Acid–Loaded Porous-Lipid/PLGA Microbubbles in Combination With Ultrasound-Triggered Microbubble Destruction on Human Glioma | |
CN114748641A (zh) | 一种硅纳米复合物的制备方法及其在增强胃癌细胞天然免疫应答中的应用 | |
Xue et al. | The dependence of radio-sensitization efficiency on mitochondrial targeting with NaGdF4: Yb, Er nanoparticles | |
Faustova et al. | Polymer particles containing Fe-based metalloporphyrin as a highly efficient stimulator of reactive oxygen species formation in vitro and in vivo | |
CN114887061A (zh) | 靶向肿瘤的光热基因联合治疗纳米系统的制备方法及应用 | |
Hu et al. | Dendritic cells reprogrammed by CEA messenger RNA loaded multi-functional silica nanospheres for imaging-guided cancer immunotherapy | |
Hao et al. | The GSH responsive indocyanine green loaded PD-1 inhibitory polypeptide AUNP12 modified MOF nanoparticles for photothermal and immunotherapy of melanoma | |
Yao et al. | The application of drug loading and drug release characteristics of two-dimensional nanocarriers for targeted treatment of leukemia | |
Xu et al. | Cu2+-pyropheophorbide-a-cystine conjugate-mediated multifunctional mesoporous silica nanoparticles for photo-chemodynamic therapy/GSH depletion combined with immunotherapy cancer | |
Perota et al. | A Study of Sonodynamic Therapy of Melanoma C540 Cells in Vitro by Titania/Gold Nanoparticles | |
Fu et al. | Cu2WS4‐PEG Nanozyme as Multifunctional Sensitizers for Enhancing Immuno‐Radiotherapy by Inducing Ferroptosis | |
Bai et al. | Design of a Nanozyme− Based Magnetic Nanoplatform to Enhance Photodynamic Therapy and Immunotherapy | |
CN110628719B (zh) | 一种诱导细胞快速产生囊泡的方法及其应用 | |
Wang et al. | Dual‐Programmable Semiconducting Polymer NanoPROTACs for Deep‐Tissue Sonodynamic‐Ferroptosis Activatable Immunotherapy |
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 | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220715 |