CN113577306A - Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment - Google Patents
Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment Download PDFInfo
- Publication number
- CN113577306A CN113577306A CN202110790475.5A CN202110790475A CN113577306A CN 113577306 A CN113577306 A CN 113577306A CN 202110790475 A CN202110790475 A CN 202110790475A CN 113577306 A CN113577306 A CN 113577306A
- Authority
- CN
- China
- Prior art keywords
- ucnp
- ucttd
- peg
- photo
- tpp
- 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
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 64
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 58
- 238000011282 treatment Methods 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 238000003745 diagnosis Methods 0.000 title abstract description 10
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 claims abstract description 79
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 claims abstract description 79
- 229940033123 tannic acid Drugs 0.000 claims abstract description 79
- 229920002258 tannic acid Polymers 0.000 claims abstract description 79
- 239000000463 material Substances 0.000 claims abstract description 37
- 239000001263 FEMA 3042 Substances 0.000 claims abstract description 34
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 claims abstract description 34
- 235000015523 tannic acid Nutrition 0.000 claims abstract description 34
- 238000006243 chemical reaction Methods 0.000 claims abstract description 29
- 239000002086 nanomaterial Substances 0.000 claims abstract description 25
- 239000002114 nanocomposite Substances 0.000 claims abstract description 21
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 claims abstract description 18
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims abstract description 10
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims abstract description 10
- 201000002528 pancreatic cancer Diseases 0.000 claims abstract description 10
- 208000008443 pancreatic carcinoma Diseases 0.000 claims abstract description 10
- 230000002195 synergetic effect Effects 0.000 claims abstract description 7
- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 claims abstract 4
- 239000003504 photosensitizing agent Substances 0.000 claims description 22
- 238000011068 loading method Methods 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 238000005406 washing Methods 0.000 claims description 12
- 239000003814 drug Substances 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 5
- 239000012190 activator Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 3
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 claims description 3
- 230000036571 hydration Effects 0.000 claims description 3
- 238000006703 hydration reaction Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 238000009210 therapy by ultrasound Methods 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 2
- 208000005016 Intestinal Neoplasms Diseases 0.000 claims description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 2
- 201000010881 cervical cancer Diseases 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 claims description 2
- 206010017758 gastric cancer Diseases 0.000 claims description 2
- 201000002313 intestinal cancer Diseases 0.000 claims description 2
- 201000007270 liver cancer Diseases 0.000 claims description 2
- 208000014018 liver neoplasm Diseases 0.000 claims description 2
- 201000005202 lung cancer Diseases 0.000 claims description 2
- 208000020816 lung neoplasm Diseases 0.000 claims description 2
- 201000011549 stomach cancer Diseases 0.000 claims description 2
- 229920001864 tannin Polymers 0.000 claims description 2
- 239000001648 tannin Substances 0.000 claims description 2
- 235000018553 tannin Nutrition 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 35
- 238000003384 imaging method Methods 0.000 abstract description 22
- 201000011510 cancer Diseases 0.000 abstract description 19
- 206010021143 Hypoxia Diseases 0.000 abstract description 9
- DULZJSBFYXKCJG-UHFFFAOYSA-M [OH-].[Si+4].CN(C)CCC[Si](C)(C)[O-].c1ccc2c3nc(nc4[n-]c(nc5nc(nc6[n-]c(n3)c3ccccc63)c3ccccc53)c3ccccc43)c2c1 Chemical compound [OH-].[Si+4].CN(C)CCC[Si](C)(C)[O-].c1ccc2c3nc(nc4[n-]c(nc5nc(nc6[n-]c(n3)c3ccccc63)c3ccccc53)c3ccccc43)c2c1 DULZJSBFYXKCJG-UHFFFAOYSA-M 0.000 abstract description 7
- 230000006870 function Effects 0.000 abstract description 6
- 230000001146 hypoxic effect Effects 0.000 abstract description 5
- 150000001875 compounds Chemical class 0.000 abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 3
- 210000003463 organelle Anatomy 0.000 abstract description 3
- 238000005481 NMR spectroscopy Methods 0.000 abstract description 2
- 238000000799 fluorescence microscopy Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 41
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 38
- 230000000694 effects Effects 0.000 description 27
- 238000002428 photodynamic therapy Methods 0.000 description 26
- 230000008685 targeting Effects 0.000 description 21
- 238000007626 photothermal therapy Methods 0.000 description 15
- 238000000338 in vitro Methods 0.000 description 14
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 13
- 239000001301 oxygen Substances 0.000 description 13
- 229910052760 oxygen Inorganic materials 0.000 description 13
- 239000003642 reactive oxygen metabolite Substances 0.000 description 12
- 210000001519 tissue Anatomy 0.000 description 12
- 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 11
- 230000008859 change Effects 0.000 description 11
- 238000010521 absorption reaction Methods 0.000 description 10
- 239000008367 deionised water Substances 0.000 description 8
- 229910021641 deionized water Inorganic materials 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 8
- 230000001225 therapeutic effect Effects 0.000 description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 6
- 229910052742 iron Inorganic materials 0.000 description 6
- 238000002595 magnetic resonance imaging Methods 0.000 description 6
- 210000003470 mitochondria Anatomy 0.000 description 6
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 239000002131 composite material Substances 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 231100000419 toxicity Toxicity 0.000 description 5
- 210000004881 tumor cell Anatomy 0.000 description 5
- 102000016938 Catalase Human genes 0.000 description 4
- 108010053835 Catalase Proteins 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 108010045325 cyclic arginine-glycine-aspartic acid peptide Proteins 0.000 description 4
- 230000007954 hypoxia Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- -1 rare earth ion Chemical class 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000001976 improved effect Effects 0.000 description 3
- 102000006495 integrins Human genes 0.000 description 3
- 108010044426 integrins Proteins 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- WNYJVAMZRBTOPE-YVSFHVDLSA-N 2-[(2s,5r,8r,11s)-5-benzyl-11-[3-(diaminomethylideneamino)propyl]-3,6,9,12,15-pentaoxo-8-(sulfanylmethyl)-1,4,7,10,13-pentazacyclopentadec-2-yl]acetic acid Chemical group N1C(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CS)NC(=O)[C@H]1CC1=CC=CC=C1 WNYJVAMZRBTOPE-YVSFHVDLSA-N 0.000 description 2
- MLOSJPZSZWUDSK-UHFFFAOYSA-N 4-carboxybutyl(triphenyl)phosphanium;bromide Chemical compound [Br-].C=1C=CC=CC=1[P+](C=1C=CC=CC=1)(CCCCC(=O)O)C1=CC=CC=C1 MLOSJPZSZWUDSK-UHFFFAOYSA-N 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 2
- 239000002616 MRI contrast agent Substances 0.000 description 2
- 238000006845 Michael addition reaction Methods 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- 238000003917 TEM image Methods 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 230000002789 catalaselike Effects 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000003759 clinical diagnosis Methods 0.000 description 2
- 238000011262 co‐therapy Methods 0.000 description 2
- 108010086416 cyclo(arginyl-glycyl-aspartyl-phenylalanyl-cysteinyl) Proteins 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 239000012737 fresh medium Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- 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 2
- 238000009097 single-agent therapy Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- 230000001960 triggered effect Effects 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- 208000020296 Abnormal vascular morphology Diseases 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102100022337 Integrin alpha-V Human genes 0.000 description 1
- 108010040765 Integrin alphaV Proteins 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 description 1
- 238000012084 abdominal surgery Methods 0.000 description 1
- 238000002679 ablation Methods 0.000 description 1
- 238000010317 ablation therapy Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 238000011203 antimicrobial therapy Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000002902 bimodal effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000005880 cancer cell killing Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 238000013170 computed tomography imaging Methods 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 108091008039 hormone receptors Proteins 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000000222 hyperoxic effect Effects 0.000 description 1
- 230000005917 in vivo anti-tumor Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 238000002324 minimally invasive surgery Methods 0.000 description 1
- 230000004065 mitochondrial dysfunction Effects 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000036457 multidrug resistance Effects 0.000 description 1
- 239000002120 nanofilm Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000011580 nude mouse model Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000007903 penetration ability Effects 0.000 description 1
- 238000006552 photochemical reaction Methods 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 230000005622 photoelectricity Effects 0.000 description 1
- 239000012221 photothermal agent Substances 0.000 description 1
- 230000002028 premature Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229910052761 rare earth metal Inorganic materials 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000002271 resection Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000007142 ring opening reaction Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000007669 thermal treatment Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 238000003325 tomography Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002723 toxicity assay Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 238000011277 treatment modality Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000005760 tumorsuppression 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
- A61K47/51—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 non-active ingredient being a modifying agent
- A61K47/56—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 non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—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 non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—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 non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7024—Esters of saccharides
-
- 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
- A61K33/26—Iron; 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
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- 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
- A61K41/0076—PDT with expanded (metallo)porphyrins, i.e. having more than 20 ring atoms, e.g. texaphyrins, sapphyrins, hexaphyrins, pentaphyrins, porphocyanines
-
- 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
- A61K47/51—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 non-active ingredient being a modifying agent
- A61K47/62—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 non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
-
- 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
- A61K47/6921—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 the form being a particulate, a powder, an adsorbate, a bead or a sphere
- A61K47/6925—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 the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a microcapsule, nanocapsule, microbubble or nanobubble
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
- A61K49/225—Microparticles, microcapsules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Abstract
The invention relates to the technical field of biomedicine, in particular to preparation of double-targeting pH stimulus-responsive nanoparticles and application thereof in tumor diagnosis and treatment. The invention uses tannic acid/Fe3+The complex compound is coated on UCNP and TPP and cRGD are modified, the constructed photo-thermal nano particle UCTTD has cancer cell and organelle dual-targeting capability, and simultaneously has good photo-thermal stability and photo-thermal performance, the photo-thermal conversion efficiency is up to 77.86%, and the photo-thermal nano particle UCTTD can be used as a multi-modal imaging diagnostic reagent to perform photo-acoustic, nuclear magnetic resonance, photo-thermal and up-conversion fluorescence imaging. The photo-thermal nano material has the characteristics of simple preparation method, low cost, good biocompatibility, good stability, multiple functions and the like. After a photosensitizer PC4 is loaded on UCTTD, the obtained nano composite material can realize the PTT and PDT synergistic treatment of hypoxic pancreatic cancer through laser irradiation of 808 nm.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to a double-targeting nanoparticle, a preparation method and application thereof, and a nanocomposite containing the nanoparticle.
Background
Pancreatic cancer is extremely high in incidence and malignancy and has become the cause of death in the seventh cancer world (abdominal surgery, 2020, Vol. 33, No. 6). Surgical resection is currently the most effective method for obtaining a cure opportunity for pancreatic cancer patients, however, most pancreatic cancer patients lose the opportunity for surgical treatment due to lack of timely diagnosis. With the progress of oncology research, research on the diagnosis and treatment of pancreatic cancer by people progresses rapidly, and a novel efficient cancer diagnosis and treatment means which has tumor targeting, reduces the toxicity to normal tissues and overcomes the multidrug resistance of tumor cells is widely developed and deeply researched.
Tumor Microenvironment (TME) is characterized by abnormal vascular morphology, dysregulated biological metabolic intermediates, hypoxia, microacidity and H2O2The characteristics of high content and the like (Chemical Society Reviews 2017,46,3830), provide internal environment for the growth and residence of cancer cells, play an important role in malignant growth, invasive metastasis and high drug resistance of tumors, and therefore, the search for new anti-cancer strategies aiming at TME is highly valued (Theransotics 2018,8, 1059). The unique properties of TME also provide new therapeutic targets for tumor therapy. To achieve accurate tumor treatment, the use of smart nanomaterials with TME stimulus responsive therapeutic capabilities is considered one of the most desirable approaches. In recent years, the design of a TME triggered minimally invasive surgery platform has attracted extensive attention and has been rapidly developed. It is composed ofAmong them, photothermal therapy (PTT), photodynamic therapy (PDT), etc. are potential anticancer strategies as emerging therapeutic approaches due to their advantages of negligible invasiveness, low toxicity, and high selectivity (ACS Nano 2017,11, 579). PTT can convert light energy into heat energy by using a near infrared light-induced photothermal conversion agent to be directly used for tumor ablation, and PDT generally needs to be carried out at O2In the presence of a light, the photosensitizer is activated by irradiation with light of a specific wavelength to generate Reactive Oxygen Species (ROS) having cytotoxicity to induce apoptosis and necrosis. PDT has received much attention as a novel tumor treatment modality due to its low invasiveness, high cure rate, and small damage to the body. However, due to the complexity of TME and the inherent drawbacks of each treatment approach, it is difficult for monotherapy to completely inhibit tumor growth (nanoscales 2018,10,4452). Therefore, co-therapy combining several therapeutic approaches is often used to overcome the deficiencies of monotherapy tumor suppression. However, there are certain challenges to developing therapeutic strategies that integrate two or more different anti-cancer approaches into one therapeutic nano-platform.
In general, PDT involves three basic conditions: light source, Photosensitizer (PS) and a hyperoxic environment in tissue. However, PS usually requires uv or visible light activation, but the penetration capability of these two wavelength light sources is limited, potential photodamage to the organism by uv light, and these problems greatly limit the practical application of PS in tumor therapy. Second, PS-mediated O during PDT due to the hypoxic microenvironment of a variety of solid tumors2Consumption further aggravates this anoxic condition and thereby impairs O dependence2PDT effect of (Nature 2012,491, 364-) (373). Furthermore, PDT produces ROS with a short lifetime and can only migrate short distances (R) ((R))<20nm) and thus the range of action of PDT is only in the vicinity of PS. Based on the shortcomings of conventional PDT, many new strategies have been developed. First, to solve the problem of laser penetration ability, Near Infrared (NIR) light (780-. Coincidently, rare earth ion based up-conversion nanoparticlesRice grains (UCNPs) can convert near-infrared light into ultraviolet light or visible light well, and the excitation wavelength of the UCNPs has higher tissue penetration depth, lower autofluorescence and good light stability, and can avoid direct damage of ultraviolet light to skin (Nature Communications 2018,9, 2415). After the photosensitizer is loaded, deep tissue PDT treatment stimulated by near infrared light can be realized.
In 2016, Yb was designed by He et al3+Enhancing synthesized core UCNPs (NaYbF)4:2%Er3+) And a mesoporous silica-coated upconversion/mesoporous silica nanocomposite (UCNP @ SiO)2) For continuous cellular imaging, photothermal drug delivery and cancer therapy, demonstrating its highly efficient photothermal performance and its great potential in simultaneous biomedical imaging and photothermal triggered cancer chemotherapy drug delivery (optical materials Express,2016,6, 1161). In addition, You et al successfully developed a Bi-based growth strategy in 20202Se3Novel hybrid nano-material of conjugated up-conversion nanoparticles (UCNPs), under 808nm near-infrared laser irradiation, UCNPs-Bi2Se3The visible light can emit bright visible light, the capability of the visible light in the aspects of efficient cell up-conversion luminescence (UCL), CT imaging, cancer cell ablation and the like is demonstrated, a treatment strategy designed under the guidance of bimodal imaging is realized, and the visible light has the characteristics of real-time dynamic monitoring, remote controllability, non-invasiveness and the like (Chemistry-AEuroplan Journal 2020,26 and 1127).
In 2017, Yan and the like utilize supramolecular coordination assembly to synthesize the nano composite material loaded with the liposoluble photosensitizer Ce 6. Due to Tannic Acid (TA) and Fe3+The hydrophobic Ce6 nano-particles have good stability in the complex assembled by the interface. Compared with the free Ce6 molecule, Ce6@ TA-Fe (III) NPs have longer blood circulation time, more tumor selective accumulation and better anti-tumor effect (Scientific reports 2017,7, 42978). In the same year, Wu is wrapped and synthesized into TA/Fe containing silver nano particles (AgNPs) by an ultrafast, green, simple and universal method3+A silver nanofilm. TA/Fe was investigated by in vitro and in vivo experiments3+Physical antimicrobial Activity and Photodynamic Antimicrobial Therapy (PAT) of AgNPsThe effect of (A) proves that it has strong antibacterial activity and good biocompatibility (ACS applied materials)&Interfaces,2017,9, 39657). The above studies have utilized only the biomedical imaging function of UCNP, TA/Fe3+The drug loading and the ability of prolonging blood circulation, promoting tumor selective accumulation of PSs and the like, and does not combine imaging and photothermal therapy/photodynamic therapy, particularly, the material has no tumor specific targeting effect, and the problem of tumor hypoxia is not considered to be solved in the photodynamic therapy.
Notably, as one of the main strategies to alleviate hypoxia in TME, catalase can catalyze high concentrations of H in TME2O2(100. mu.M-1 mM) in situ generation of O2(ACS Nano 2017,11, 7006). However, due to their sensitivity to complex biological environments, natural enzymes are easily inactivated and degraded. Moreover, their preparation is rather costly. Therefore, it is very important to design and develop a high-stability catalyst, nanoenzyme, similar in function to natural enzyme, as an oxygen generating agent to increase oxygen concentration in solid tumors.
In addition, the current preparation method of the nano material has the following problems: the preparation process is complex and time-consuming, most materials have larger nano size or are toxic, and can not be used for in vivo treatment, and especially the single up-conversion nanoparticles have no photodynamic and photothermal treatment capability; some materials only have single treatment function and have poor treatment effect; some materials have no active targeting effect, or only one targeting ability, and cannot specifically recognize cancer cells and organelles; some materials only have a therapeutic effect and do not have imaging diagnostic capability, so that diagnosis and treatment are difficult to combine for precise imaging-guided treatment; most materials do not have the ability to respond to stimuli, allowing for manageable treatment and reduced toxicity and damage to normal tissues.
Disclosure of Invention
In view of the above, the present invention aims to provide a preparation method of a dual-targeting pH stimulus-responsive nanoparticle and an application thereof in tumor diagnosis and treatment. The photo-thermal nano composite material not only has good photo-thermal stabilityAnd the film has high photo-thermal performance, and the photo-thermal conversion efficiency is up to 77.86%. After loading the photosensitizer PC4 on UCTTD, the obtained nano composite material has alpha v beta3The integrin receptor and mitochondria have double-targeting capability, and PTT and PDT can be used for synergistically treating hypoxic pancreatic cancer through laser irradiation at 808 nm.
In order to achieve the above object, the present invention provides the following technical solutions:
a dual-targeted nanoparticle, comprising:
an up-converting particle core; and
tannic acid/Fe coated on the surface of the inner core from inside to outside in sequence3+Complex, TPP and cRGD.
The double-targeting nanoparticle takes the conversion nano material as an inner core, and the complex of tannic acid and iron ions is coated at room temperature at an ultra-fast speed. Due to the addition of iron ions, the tumor cell has catalase activity, consumes hydrogen peroxide in tumor tissues, generates oxygen, improves the tumor anoxic environment, improves the aerobic PDT efficiency, and has the photo-thermal treatment capacity and the capacity of improving the tumor anoxic environment.
And (3) coating a complex of tannic acid and iron ions, and then modifying TPP and cRGD to obtain the post-nanoparticle UCTTD. The TPP and cRGD modified nano-particle can specifically recognize surface high-expression alpha v beta3Integrates the cancer cells of the hormone receptor and distributes in the mitochondria in a concentrated way. In the present invention, the sequence of the cRGD peptide is cyclo (Arg-Gly-Asp-d-Phe-Cys). The connection of cRGD peptide enables the nano material to express alpha v beta with high affinity3The integrin receptor cancer cell is easy to enter ROS sensitive organelle-mitochondria due to the connection of the mitochondrial targeting molecule TPP, thereby improving the concentration of Reactive Oxygen Species (ROS) in the mitochondria, accelerating the mitochondria dysfunction and inducing the cancer cell apoptosis. The double targeting has stronger targeting selectivity, improves the efficiency of the material entering tumor tissues, and simultaneously because the double targeting nanoparticles contain Fe3+Can catalyze H2O2React to generate oxygen to provide sufficient O for PDT2Improving PDT efficiency and obtaining more effective cancer cell killing effect。
Wherein, the hydration kinetic diameter of the dual-targeting nanoparticle is 150-200nm, and specifically can be 150nm, 174nm or 200 nm.
The invention also provides a preparation method of the double-targeting nano particle, which comprises the following steps:
step 1: mixing UCNP nano particles with tannic acid solution, performing ultrasonic treatment, and adding FeCl3Reacting the solution, centrifuging, and cleaning to obtain the product coated with tannin/Fe3+The nanoparticle of the complex UCNP @ TA/Fe;
and step 3: TPP and PEG-modified UCNP @ TA/Fe are mixed and react to obtain nano-particle UCNP @ TA/Fe-TPP;
and 4, step 4: and mixing the UCNP @ TA/Fe-TPP with the cRGD solution, and reacting to obtain the photo-thermal nano material UCNP @ TA/Fe-TPP-cRGD (UCTTD).
In some embodiments, in step 1, the concentration of the tannic acid solution is 1.6 to 6.4mg/mL, specifically 1.6mg/mL, 3.2mg/mL, or 6.4 mg/mL; the mass-to-volume ratio of the UCNP nano particles to the tannic acid solution is (2.5-10) in mg/mL: (0.5-2), specifically 5: 1.
In some embodiments, in step 2, the concentration of the PEG solution is 0.5-2mg/mL, and specifically may be 1mg/mL, wherein the PEG is DSPE-PEG2000、NH2-PEG2000-NH2And DSPE-PEG2000-a Maleimide composition; the DSPE-PEG2000、NH2-PEG2000-NH2And DSPE-PEG2000-Maleimide in a mass ratio of 90: 5: 5.
in some embodiments, in step 3, the reaction is carried out under rotational shaking conditions; the TPP is activated by an activator, and the activator is EDC and/or NHS.
In some embodiments, in step 4, the reaction is performed under conditions of 2-10 ℃ with rotary shaking for 6-24 h. In some embodiments, the reaction conditions are rotation shaking at 4 ℃ for 12 h.
In some embodiments, the method for preparing the dual-targeting nanoparticle comprises the following steps:
(1) preparation of Tannic Acid (TA) and Fe3+Complex coated upconversion nanoparticles of (UCNP @ TA/Fe)
Adding 2.5-10mg UCNP nano particles into 0.5-2mL TA solution (1.6-6.4mg/mL), performing ultrasonic treatment for 10min, and adding 0.25-1mL FeCl3Shaking the solution (0.8-3.2mg/mL) with a large force or stirring rapidly for 20-120s, centrifuging, washing with 50-75% ethanol twice, and washing with deionized water once to obtain UCNP @ TA/Fe.
(2) Preparation of UCNP @ TA/Fe-TPP
Firstly, adding 0.5-2mg/mL DSPE-PEG into UCNP @ TA/Fe2000In solution (wherein NH)2-PEG2000-NH2And DSPE-PEG2000-Maleimide in Total DSPE-PEG20005-20% of the amount), rotating and shaking for 6-24h, washing with deionized water for 2 times for later use. During this time, 4-carboxybutyltriphenylphosphonium bromide (TPP) was activated. TPP (0.2-0.5mmoL), EDC (0.5-1mmoL) and NHS (1-2mmoL) were weighed and added to 50-200. mu.L of anhydrous DMSO for activation for 0.5-6h, and added to 1mL of PEG-ylated UCNP @ TA/Fe and vortexed for 6-24 h. Washing with deionized water for three times to obtain UCNP @ TA/Fe-TPP, and storing at 4 ℃ for later use.
(3) Preparation of UCNP @ TA/Fe-TPP-cRGD
Adding UCNP @ TA/Fe-TPP to DSPE-PEG2000Rotating and shaking the solution of cRGD with the same molar amount of Maleimide at the temperature of 2-10 ℃ for 6-24h, washing with deionized water for 3 times to obtain UCNP @ TA/Fe-TPP-cRGD (UCTTD), and storing at the temperature of 4 ℃ for later use.
The invention also provides a nano composite material which comprises the double-targeting nano particle or the double-targeting nano particle prepared by the preparation method and a photosensitizer loaded on the double-targeting nano particle.
In some embodiments, a method of making a nanocomposite of the invention comprises:
under the condition of keeping out of the sun, adding the double-targeting nanoparticles (UCTTD) into a photosensitizer solution of 25-200 mu g/mL, and carrying out rotation oscillation reaction for 6-36 h in the absence of the sun to obtain the photosensitizer-loaded nanocomposite.
In some embodiments, the photosensitizer is PC 4.
In some embodiments, the photosensitizer is PC4, and the preparation method of the PC 4-loaded nanocomposite material comprises the following steps: under the condition of keeping out of the sun, adding the double-targeting nanoparticles (UCTTD) into a 25-200 mu g/mL PC4 solution, and carrying out a light-proof rotary oscillation reaction for 6-36 h to obtain the photosensitizer-loaded nanocomposite.
In one specific example, the present invention examined the loading efficiency of UCTTD photothermal nanoparticles in PC4 solutions of different concentrations, and found that the loading of UCTTD to PC4 was 80 μ g/mg.
The invention also provides application of the double-targeting nano particles or the nano composite material in preparation of antitumor drugs.
In some embodiments, the tumor comprises pancreatic cancer, liver cancer, lung cancer, stomach cancer, intestinal cancer, breast cancer, and cervical cancer.
In some embodiments, pancreatic cancer is specified, and more specifically hypoxic pancreatic cancer is specified.
The invention provides a dual-targeting nanoparticle, comprising: an up-converting particle core; and tannic acid/Fe coated on the surface of the inner core from inside to outside in sequence3+Complexes, TPP and RGD. The invention uses tannic acid/Fe3+The complex compound is coated on UCNP and TPP and cRGD are modified, the constructed photo-thermal nano particle UCTTD has cancer cell and organelle dual-targeting capability, and simultaneously has good photo-thermal stability and photo-thermal performance, and the photo-thermal conversion efficiency is up to 77.86%.
After the photosensitizer PC4 is loaded on the material, the synergistic treatment of PTT and PDT can be realized by only using laser with one wavelength of 808 nm. And the doping of iron makes the material possess catalase-like property, and can be used for treating H in cancer cells2O2Shows excellent catalytic performance, thereby overcoming the tumor hypoxia and improving the curative effect of PDT. At the same time, tannic acid/Fe3+The complex is sensitive to pH change, has the pH stimulation response capability, and can release more photosensitizer in a slightly acidic tumor microenvironment through the pH stimulation response performance(PC4) enabling a steerable PDT treatment. The UCTTD-PC4 nano-composite has the stability of nano-materials, and is beneficial to overcoming the instability and volatile activity of natural enzymes. In addition, due to the connection of RGD peptide, the nano material is highly expressed by the affinity of alpha v beta3The cancer cells of the integrin receptor can enter the ROS-sensitive organelle mitochondria more easily due to the mitochondrion-targeted connection, and the concentration of the ROS in the mitochondria is improved, so that the mitochondrial dysfunction and the apoptosis are accelerated. The dual-targeting nanoparticle UCTTD provided by the invention has higher targeting selectivity and can obtain better treatment effect. In addition, UCTTD can be used as a multi-modality imaging diagnostic reagent, and can perform Photoacoustic (PA), nuclear Magnetic Resonance (MR), Photothermal (PT) and up-conversion fluorescence (UCL) imaging. In a word, the material has the characteristics of simple manufacture, low cost, good biocompatibility, good stability, multiple functions and the like.
Drawings
FIG. 1 is a photograph of a UCNP @ TA/Fe nanomaterial before and after reaction, (a) UCNP in a TA solution, and (b) a photograph of a UCNP @ TA/Fe solution;
FIG. 2 characterization of nanomaterials; (a) transmission electron microscopy images of UCTTD. (b) Fourier transform infrared spectra of UCNP @ TA/Fe and UCTTD; (c) fluorescence emission spectra of UCNP, UCNP @ TA/Fe, UCTTD and UCTTD-PC 4. Hydrated particle size distribution (d) and zeta potential (e) of UCNP, UCNP @ TA/Fe, UCNP @ TA/Fe-TPP, UCTTD and UCTTD-PC 4; (f) ultraviolet absorption spectrograms of UCNP, PC4, UCNP @ TA/Fe, UCTTD and UCTTD-PC 4;
FIG. 3 measurement of photothermal performance and photothermal stability of UCTTD; (a)808nm laser at 0.5W/cm2The UCTTD is irradiated by the laser power, and the temperature is reduced for 20min after 6 times of photo-thermal stability measurement of temperature rise and temperature reduction, wherein the laser irradiation is carried out for 10min in each period; (b) measurement of photothermal conversion efficiency, 0.5W/cm2After respectively irradiating water and the UCTTD solution for 10min, naturally cooling to a temperature change curve at room temperature and a linear curve of the time of the cooling process and-ln (theta);
figure 4UCTTD versus photosensitizer PC4 loading and pH responsive release studies; (a) the load ratio of UCTTD to PC4(25, 50, 100, 150, 200 μ g/mL) at different time points; (b) the release efficiency of UCTTD-PC4 at different pH for PC 4;
FIG. 5 cell-targeted delivery study; (a) flow cytometry was used to quantify the flow images of the delivery efficiency of UCTTD-PC4 and UCNP @ TA/Fe-TPP-PC 4; (b) a statistical graph of delivery efficiency with or without cRGD targeting molecule connecting material;
FIG. 6 in vitro oxygen production study; UCTTD in H2O2O in solution (1mM)2Formation of (2), H of the same concentration2O2The solution is used as a control;
FIG. 7 in vitro ROS production study; (a) UCTTD-PC4+ H2O2+ DPBF solution and (b) DPBF solution at 808nm laser (0.3W/cm)2) Irradiating ultraviolet absorption spectra at different times;
FIG. 8. cell viability and toxicity; (a) cell viability of Capan-1 cells after 24h and 48h incubation with 0, 150, 300, 600, 900, 1200, 1500 and 2000. mu.g/mL UCTTD. (b) Through PBS, PBS + L, PC4, PC4+ L470Relative survival of Capan-1 cells after 24h and 48h treatment with different groups of UCTTD, UCTTD + L, UCTTD-PC4, UCTTD-PC4+ L. Data mean ± SD;
fig. 9 photoacoustic imaging intensity values and photoacoustic imaging pictures for different concentrations of UCTTD in vitro;
FIG. 10 MRI intensity values and MRI pictures weighted by T1(a) and T2(b) for different concentrations of UCTTD in vitro;
fig. 11 shows a study of the therapeutic effect of a living body (n-5); (a) representative photographs of female BALB/c nude mice bearing the Capan-1 tumor on day 14 of treatment and photographs of different groups of tumors (empty blue dotted circles indicate that the tumor was completely ablated); (b) changes in tumor volume during treatment for each group; l is470Represents a 470nm laser (laser density: 0.1W/cm)2) L represents a 808nm laser (laser density: 1.2W/cm2) And the irradiation time is 10 min.
Detailed Description
The invention provides a preparation method of double-targeting pH stimulus-responsive nanoparticles and application of the nanoparticles in tumor diagnosis and treatment. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
Photothermal therapy (PTT): photothermal therapy is a therapeutic method in which a material having a high photothermal conversion efficiency is injected into the inside of a human body, is concentrated near tumor tissue by using a targeting recognition technology, and converts light energy into heat energy under the irradiation of an external light source (generally, near infrared light) to kill cancer cells.
Photodynamic therapy (PDT): is a new method for treating tumor diseases by using photosensitive drugs and laser activation. The tumor part is irradiated by specific wavelength to activate the photosensitive medicine gathered selectively in tumor tissue, the photosensitive medicine transfers energy to surrounding oxygen to initiate photochemical reaction to generate singlet oxygen with strong activity, and the singlet oxygen can generate oxidation reaction with nearby biological macromolecules to generate cytotoxicity and further kill tumor cells.
pH responsive nanomaterials: refers to a kind of functional nano material that can produce chemical or physical change in or between molecules with the change of pH.
Catalase (Catalase, CAT): is an enzyme that catalyzes the decomposition of hydrogen peroxide into oxygen and water.
Nano-enzyme: is a mimic enzyme which not only has the unique performance of nano materials, but also has the catalytic function. The nano enzyme has the characteristics of high catalytic efficiency, stability, economy and large-scale preparation, and is widely applied to the fields of medicine, chemical industry, food, agriculture, environment and the like.
The reagents and materials adopted by the invention are all common commercial products and can be purchased in the market.
The invention is further illustrated by the following examples:
EXAMPLE 1 preparation of composite Material
(1) Preparation of Tannic Acid (TA) and Fe3+Complex coated upconversion nanoparticles of (UCNP @ TA/Fe)
5mg of UCNP nanoparticles were added to 1mL of TA solution (3.2mg/mL), sonicated for 10min, and 0.5mL of FeCl was added3Shaking the solution (1.6mg/mL) with a large force or stirring rapidly for 20-120s, centrifuging, washing with 50-75% ethanol twice, and washing with deionized water once to obtain UCNP @ TA/Fe.
(2) Preparation of UCNP @ TA/Fe-TPP
Firstly, adding 1mg/mL DSPE-PEG into UCNP @ TA/Fe2000In solution (wherein NH)2-PEG2000-NH2And DSPE-PEG2000-Maleimid accounts for the total DSPE-PEG 20005 percent of the amount), rotating and shaking for 12 hours, washing for 2 times by deionized water for later use. During this time, 4-carboxybutyltriphenylphosphonium bromide (TPP) was activated. TPP (0.2mmoL), EDC (0.5mmoL) and NHS (1mmoL) were weighed out and added to 100. mu.L of anhydrous DMSO for activation for 1h, and added to 1 mLPEG-treated UCNP @ TA/Fe and vortexed for 12 h. Washing with deionized water for three times to obtain UCNP @ TA/Fe-TPP, and storing at 4 ℃ for later use.
(3) Preparation of UCNP @ TA/Fe-TPP-cRGD
Adding UCNP @ TA/Fe-TPP to DSPE-PEG2000Rotating and shaking the cRGD solution with the same molar amount of Maleimid at 4 ℃ for 12h, washing with deionized water for 3 times to obtain UCNP @ TA/Fe-TPP-cRGD (UCTTD), and storing at 4 ℃ for later use.
FIG. 1 shows Tannic Acid (TA) and Fe3+Color change of solution before and after complex coating of upconverter, FIG. 1a shows UCNP dispersed in TA solution, which is TA yellowish, when Fe3+When added, TA and Fe3+The complex is formed instantly, coated on the surface of UCNP, and the color of the solution is instantly changed into black (figure 1b), so that the UCNP @ TA/Fe material has good photo-thermal property.
FIG. 2a is a Transmission Electron Micrograph (TEM) of UCNP @ TA/Fe-TPP-RGD (UCTTD), and it can be seen from FIG. 2 that TA and Fe are successfully coated on the outer surface of UCNP3+A complex of (a). FIG. 2b is a Fourier transform infrared spectrum of UCNP @ TA/Fe and UCNP @ TA/Fe-TPP-RGD (UCTTD), the UCTTD contains characteristic absorption peaks of benzene rings compared with UCNP @ TA/Fe: the C-H telescopic vibration of the benzene ring is 3068cm-1;1479cm-1And 1569cm-1The skeleton of the benzene ring vibrates. 692cm-1And 750cm-1The appearance indicates that the benzene ring is monosubstituted. These functional groups are consistent with the molecular structure of TPP, furthermore, at 1678cm-1A characteristic absorption peak at which an amide bond (-NH-C ═ O) appears, which is presumed to be a maleimide group and cRGDfc [ cyclo (Arg-Gly-Asp-d-Phe-Cys)]After the mercapto group in the compound is subjected to Michael addition condensation reaction, the compound is subjected to ring opening hydrolysis to generate a stable amido bond, which indicates that the cRGD is successfully connected. As shown in FIG. 2d, the hydration kinetic diameters of UCNP, UCTTD, and UCTTD-PC4 were 80nm, 174nm, and 262nm, respectively. FIG. 2e shows the change in zeta potential of the various particles. The potential of UCNP was seen to be +36.7mV, in TA vs Fe3+The potential of UCNP @ TA/Fe after coating with the complex of (1) was changed to-13.4 mV. After attachment of the targeting molecule, the potential changes were modest, with the potentials UCNP @ TA/Fe-TPP (UCTT) and UCTTD being-6.4 mV and-7.0 mV, respectively, but after loading with positively charged PC4, the potential of UCTTD-PC4 became +2.7mV, indicating that PC4 was successfully loaded onto the material. On the other hand, a small positive charge makes UCTTD-PC4 more accessible to cells and has little charge toxicity. In summary, the above characterization results demonstrate that UCTTD with dual targeting was successfully synthesized. FIG. 2f is a graph of the UV absorption spectra of UCNP, PC4, UCNP @ TA/Fe, UCTTD, and UCTTD-PC4, from which it can be seen that UCNP almost coincides with the baseline at the near infrared, and UCNP @ TA/Fe has significant absorption at the near infrared, especially after the attachment of targeting molecules, the near infrared absorption of UCTTD is further improved, which means that UCTTD has better photothermal properties. After UCTTD is loaded with PC4, the UV absorption peak of PC4 appears at 450-500 nm in UCTTD-PC4, which also proves the successful loading of PC 4. In addition, the maximum absorption peak of PC4 in the UV is about 470nm, and FIG. 2c shows the fluorescence emission spectra of UCNP, UCNP @ TA/Fe, UCTTD and UCTTD-PC4, although the fluorescence intensity is reduced after encapsulation, the emission spectrum still has strong emission at 470nm, and just provides a light source for the photosensitizer PC 4.
To measure the photothermal power and light stability of UCTTD, 1mL of UCTTD was placed in a colorless transparent cuvette and irradiated with a near-infrared laser (LOS 2-BLD-0808) -005W-C/P, high-tech photoelectricity, Inc., China) for 600s (0.5W/cm)2),And then cooling to room temperature, repeating the cycle for 6 periods, and detecting the photo-thermal stability of the material. The real-time temperature of the solution was recorded every 30s with a thermocouple (TES K thermometer 1319A, Taiwan, China). In addition, the solution temperature was read every 10s with a FLIR C2 thermographic camera (usa) during near infrared laser irradiation, in comparison with water as a cathode, and finally the photothermal conversion efficiency (η) of the material was calculated.
As shown in FIG. 3a, the UCTTD solution was passed through a 808nm laser (0.5W/cm)2) The irradiation is carried out for 10min, the interval is 20min, and after 6 times of heating and cooling cycle, the solution still can reach 46.8 ℃, which shows that the UCTTD nano material not only has higher photo-thermal performance, but also has better photo-thermal stability. As shown in FIG. 3b, the light-heat conversion efficiency of UCTTD is as high as 77.86%
The results show that the photothermal nanomaterial UCTTD provided by the invention has good photothermal stability and high photothermal conversion efficiency, and is a photothermal therapeutic agent with great potential.
Example 2 nanocomposite Loading of photosensitizer PC4
And adding 1mg of the photothermal nano material UCTTD prepared in the example 1 into 1mL of PC4 solution with different concentrations (25, 50, 100, 150 and 200 mu g/mL), and carrying out light-shielding rotary oscillation reaction for 36h to obtain the photosensitizer PC 4-loaded nano composite material UCTTD-PC 4.
Example 3 Loading and Release of photosensitizer (PC4)
Nanocomposites were prepared according to the method of example 2, and the concentration of PC4 (C) in the supernatant was measured at 6, 12, 24 and 36h, respectively, during the reactionx) Calculating the loading rate of PC 4:
(C) Loading Rate0-Cx)/C0×100%
Wherein, C0Is the initial concentration of PC 4. For PC4 release, UCTTD-PC4 was placed in PBS at pH 5.0, 6.5, 7.4, respectively, vortexed for 48h at 1,3, 6, 12, 24, 48h, and the concentration C of PC4 released from the supernatant was determinedxThe release rate is Cx/C0X 100% where C0Is the amount of material loading. The results are shown in FIG. 4.
As shown in FIG. 4a, the loading efficiency of 1mg of UCTTD nano-material in PC4 solutions with different concentrations is shown, and after loading for 36h, the loading of UCTTD on PC4 is calculated to be about 80 mug/mg. Since complexes of tannic acid and iron were pH sensitive, we examined the release behavior of UCTTD-PC4 in PBS at different pH (7.4, 6.5, 5.0), and we can see that UCTTD-PC4 has a release rate of only 2.6% at pH 7.4 and a release efficiency of 1.8 times at pH 5.0 (fig. 4 b). The results prove that the UCTTD-PC nano particles have good stability and can effectively reduce the premature release of the photosensitizer before reaching the lesion area. While the lower pH of the tumor tissue may stimulate the release of PC4 upon entry into the tumor lesion.
A great deal of research has found that integrin alpha v beta3Plays an important role in angiogenesis, and has high expression in tumor blood vessels and various invasive tumor cells, and low or even no expression in normal endothelial cells and tissues. cRGD peptide and alpha v beta3The receptor has high affinity and has been studied as a targeting molecule to specifically recognize cancer cells. The cRGD peptide is connected to the nano material through Michael addition reaction, so that the amount of the material entering tumor cells can be increased to improve the anti-tumor effect, and the side effect on normal cells can be reduced. Before in vitro treatment, we first determined the targeting ability of the composite of example 4 of the material
Capan-1 cells were seeded in a 96-well plate at 10000 cells/well, incubated at 37 ℃ for 12h, cultured with UCNP @ TA/Fe-TPP-PC4(-cRGD) and UCTTD-PC4(+ cRGD) for 6h, and then digested for quantitative delivery analysis using a flow cytometer.
As shown in fig. 5a and 5b, the delivery of UCTTD-PC4 and UCNP @ TA/Fe-TPP-PC4(UCTT-PC4) with RGD targeting was analyzed separately using flow cytometry. The delivery efficiencies of UCTTD-PC4 and UCTT-PC4 were 73.8% and 49.8%, respectively. This fully demonstrates the affinity of RGD targeting molecules to Capan-1 cells, and also demonstrates the reliability of UCTTD nanoparticles to target cancer cells.
Detection of O production from UCTTD Using YSI5000 dissolved oxygen-BOD determinator (USA)2The ability of the cell to perform. Typically, a dispersion of UCTTD nanoparticles (30 mL)) Put into a 50mL centrifuge tube, and H is added2O2Exposing the material to 1mM H2O2At the concentration. Under the condition of continuous stirring, a dissolved oxygen meter probe is adopted to detect the change of oxygen concentration, and H without UCTTD nano particles is added2O2(1mM) solution was used as negative control.
As shown in FIG. 6, H2O2The oxygen content of the solution is not obviously changed, but the oxygen concentration is increased along with the change of time when the solution is added into the solution of the nano material, which shows that the UCTTD nano particles have better catalase-like activity, namely can catalyze H2O2Production of O2. We speculate that it produces O2The reason is that the material contains Fe3+In H2O2The Fenton reaction is generated in the presence of the following reaction mechanism:
Fe3++H2O2→Fe2++O2+2H+
for the detection of ROS production in vitro, chemical oxidation using the chemical probe DPBF, DPBF and1O2an irreversible reaction occurs, and the absorbance of DPBF decreases. Confirmation of the change in absorption intensity of the probe DPBF at 410nm by UV-visible spectroscopy1O2Is generated. Generally, to UCTTD-PC4 solution containing 20. mu.g/mL DPBF, an appropriate amount of H was added2O2The final concentration was 1mM, and the mixture was protected from light by a 808nm laser (0.3W/cm)2) The irradiation was carried out for 5min, during which the UV was measured once per minute. DPBF (20. mu.g/mL) without UCTTD-PC4 was irradiated with laser light of the same power density as a control.
Fe in UCTTD is estimated according to the result of oxygen generation3+The synergistic effect of the catalytic center and PC4 may probably increase the production of ROS. To verify the ability of UCTTD-PC4 to generate ROS, UCTTD-PC4 was first generated in vitro by DPBF1O2The ability of (c) was studied. As shown in FIG. 7a, UCTTD-PC4 was at 808nm (0.3W/cm)2) The absorption peak of DPBF at 420 nm was decreasing during 5min of laser irradiation. The control group showed little change in the absorption intensity of DPBF under the same conditions without the addition of UCTTD-PC4 (FIG. 7 b). The above results indicate that UCTTD-PC4 has good ROS-generating ability in vitro.
EXAMPLE 5 evaluation of the Effect of the composite in vitro treatment
Due to the application of nanomaterials in photothermal therapy (PTT) and photodynamic therapy (PDT) in living beings, the active toxicity of UCTTD-PC4 in cells under laser irradiation and dark conditions is an important parameter.
Cell activity and toxicity assays were performed according to the standard procedures of the CCK-8 kit.
First, Capan-1 cells were seeded in a 96-well plate at a density of 10000 cells/well, cultured at 37 ℃ for 12 hours, and then fresh media (0, 150, 300, 600, 900, 1200, 1500, 2000. mu.g/mL) containing UCTTD at various concentrations were added. After further culturing for 24h and 48h, cell activity assay was performed. Cells plated for 12h were divided into 8 groups: PBS, PBS + L, UCTTD, UCTTD + L, PC4, PC4+ L470UCTTD-PC4 and UCTTD-PC4+ L. The laser irradiation group was irradiated with 470nm or 808nm laser, respectively, after 6h incubation. After 24h and 48h, the cells were washed with PBS and cultured for about 30min by adding 100. mu.L of fresh medium (containing 10. mu.L of CCK-8). Absorbance at 450nm was recorded using a microplate reader (Spark control Tecan, USA). Each set of experiments was repeated three times.
As shown in fig. 8a, after the cells are incubated with the UCTTD nanomaterial with the concentration of less than 1200 μ g/mL for 24 hours or 48 hours, the survival rate is still higher, which indicates that the biocompatibility and biosafety of UCTTD are good. However, after treatment with photodynamic agent, photothermal agent or photodynamic photothermal co-therapy agent, the cells showed significantly different survival rates (fig. 8b), with little effect on the survival rate of the cells with respect to the blank control, laser (PBS + L), material (UCTTD, UCTTD-PC4), photosensitizer (PC4), and PC4+ L470After 48h of treatment, the cell activity is only reduced to 68.6%, UCTTD + L808After treatment, the cell activity is reduced to 26.8%, and particularly after photo-thermal and photodynamic synergistic treatment for 48 hours, the cell survival rate is remarkably reduced to 8.1%. This is illustrated by the presence of Fe3+Under the catalysis of the PTT and the RGD targeting effect, the sensitivity of cells to photodynamic therapy is greatly enhanced, and finally the PTT synergistic PDT treatment effect is obvious. These results indicate that UCTTD-PC4 has high biocompatibility under dark conditions, is safe and nontoxic, is an effective PTT/PDT photodynamic co-therapeutic agent for hypoxic cancer cells under specific laser irradiation, and has excellent treatment effect.
Example 6 composite materials for evaluation of feasibility of tumor imaging
In order to more clearly guide the diagnosis and treatment of materials, this example studies the imaging and contrast effects of the nanocomposite material of the present invention and the enrichment thereof in tumors.
First, photoacoustic imaging was performed in vitro to confirm the photoacoustic imaging ability of the material itself, a mouse prosthesis was prepared according to the protocol, UCTTD (0.125, 0.25, 0.5, 1, 2, 4mg/mL) was injected into the prosthesis at different concentrations, and photoacoustic imaging was performed in a multispectral photoacoustic tomography system (MSOT inVision 128, i thermomedical, germany). The photoacoustic imaging picture and the photoacoustic signal intensity are processed using software provided by the manufacturer.
As can be seen from the experimental results of the in vivo photoacoustic imaging simulated by the in vitro prosthesis (FIG. 9), the photoacoustic intensity was increased and its linearity (R) was improved as the UCTTD concentration in the prosthesis was increased20.9939) exhibits concentration dependence, and the inset is an in vitro photoacoustic imaging picture, illustrating that a photoacoustic signal can be generated and that the signal intensity has a good linear relationship with concentration.
Magnetic Resonance Imaging (MRI) is one of the most common diagnostic tools that are indispensable and common in clinical diagnosis. The imaging technology has high safety factor and higher soft tissue resolution, can perform multi-azimuth and multi-parameter imaging under the condition of no trauma, and has the effect not influenced by the tissue depth. Therefore, in clinical diagnosis, MRI can safely, quickly, and accurately provide a high-resolution three-dimensional structural image to a patient, and is commonly used for detection of various diseases. To investigate the potential of UCTTD as an MRI contrast agent, T1 and T2 weighted MRIs at different concentrations of UCTTD (0, 0.5, 1, 2, 4mg/mL) as well as longitudinal and transverse MR relaxation rates were first measured by a 3.0T clinical MRI scanner.
MR images of aqueous solutions of UCTTD showed in vitro concentration-dependent imaging effects (fig. 10) with relatively high longitudinal (r1) and transverse (r2) relaxations, indicating that UCTTD is an effective MRI contrast agent.
Example 7 evaluation of composite materials for in vivo antitumor
This example treats female nude mice bearing the Capan-1 tumor and evaluates their anti-tumor capabilities in vivo.
For example, the figure shows the photographs of the mice and tumors (FIG. 11a) after 14 days of treatment in PBS group and UCTTD-PC4+ L group mice, and the change of tumor volume during the treatment period in 8 groups (FIG. 11 b). PC4+ L compared to PBS group470The tumors of the group are slightly reduced, the UCTTD + L group effectively inhibits the tumor growth, some tumors in the UCTTD-PC4+ L group are almost completely killed without any regeneration, while the tumors of the mice of the PBS + L, PC4, UCTTD and UCTTD-PC4 groups still keep higher growth speed, which shows that PDT per se has poor penetrability and no targeting due to laser (470nm), the in vivo circulation time is short, the treatment effect is not ideal, but under the photo-thermal synergistic treatment, the nano material has the targeting effect and Fe3+Capable of catalyzing H2O2Oxygen is generated to further improve the photodynamic sensitivity, so that the excellent treatment effect is shown in the photothermal and photodynamic therapy group.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (12)
1. A dual-targeted nanoparticle, comprising:
an up-converting particle core; and
tannic acid/Fe coated on the surface of the inner core from inside to outside in sequence3+Complex, TPP and cRGD.
2. The dual-targeted nanoparticle of claim 1, wherein the photo-thermal nanomaterial has a hydration kinetic diameter of 150-200 nm.
3. The method for preparing dual-targeted nanoparticles of claim 1 or 2, comprising the steps of:
step 1: mixing UCNP nano particles with tannic acid solution, performing ultrasonic treatment, and adding FeCl3Reacting the solution, centrifuging, and cleaning to obtain the product coated with tannin/Fe3+The nanoparticle of the complex UCNP @ TA/Fe;
step 2: adding a PEG solution into the UCNP @ TA/Fe solution, rotating and shaking for 6-24h, and washing with water to obtain PEG-ylated UCNP @ TA/Fe;
and step 3: TPP and PEG-modified UCNP @ TA/Fe are mixed and react to obtain nano-particle UCNP @ TA/Fe-TPP;
and 4, step 4: and mixing the UCNP @ TA/Fe-TPP with the cRGD solution, and reacting to obtain the photo-thermal nano material UCNP @ TA/Fe-TPP-cRGD.
4. The method according to claim 3, wherein in the step 1, the concentration of the tannic acid solution is 1.6 to 6.4 mg/mL; the mass-to-volume ratio of the UCNP nano particles to the tannic acid solution is (2.5-10) in mg/mL: (0.5-2).
5. The method of claim 3, wherein the concentration of the PEG solution in step 2 is 0.5-2mg/mL, wherein the PEG is DSPE-PEG2000、NH2-PEG2000-NH2And DSPE-PEG2000-a Maleimide composition; the DSPE-PEG2000、NH2-PEG2000-NH2And DSPE-PEG2000-Maleimide in a mass ratio of 90: 5: 5.
6. the method according to claim 3, wherein in step 3, the reaction is performed under a rotating shaking condition; the TPP is activated by an activator, and the activator is EDC and/or NHS.
7. The method according to claim 3, wherein in step 4, the reaction is carried out under conditions of 2-10 ℃ and 6-24h of rotary shaking.
8. A nano composite material, which comprises the dual-targeting nanoparticle of any one of claims 1 to 2 or the dual-targeting nanoparticle prepared by the preparation method of any one of claims 3 to 7, and a photosensitizer loaded on the dual-targeting nanoparticle.
9. The nanocomposite as claimed in claim 8, wherein the loading of the photosensitizer is 76-80 μ g/mg.
10. Nanocomposite as claimed in claim 8, wherein the photosensitizer is PC 4.
11. Use of the dual-targeted nanoparticles of any one of claims 1 to 2, the dual-targeted nanoparticles prepared by the preparation method of any one of claims 3 to 7, or the nanocomposite of any one of claims 8 to 10 for preparing a medicament for the PTT/PDT synergistic treatment of tumors.
12. The use of claim 11, wherein the tumor is pancreatic cancer, liver cancer, lung cancer, stomach cancer, intestinal cancer, breast cancer or cervical cancer.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110790475.5A CN113577306B (en) | 2021-07-13 | 2021-07-13 | Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110790475.5A CN113577306B (en) | 2021-07-13 | 2021-07-13 | Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113577306A true CN113577306A (en) | 2021-11-02 |
CN113577306B CN113577306B (en) | 2023-02-28 |
Family
ID=78247232
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110790475.5A Active CN113577306B (en) | 2021-07-13 | 2021-07-13 | Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113577306B (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114404429A (en) * | 2021-11-30 | 2022-04-29 | 重庆医科大学附属第二医院 | Nano-silver modified tannin-iron network drug-loaded nano-composite, preparation method thereof and application thereof in reversing tumor drug resistance |
CN114748420A (en) * | 2022-04-12 | 2022-07-15 | 武汉科技大学 | Preparation method of amphiphilic polymer micelle with charge self-reversal activation mitochondrial targeting effect |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014200441A1 (en) * | 2013-06-14 | 2014-12-18 | National University Of Singapore | Core-shell fluorescent upconversion nanoparticles for photoactivation of multiple biomolecules |
CN110478483A (en) * | 2019-08-22 | 2019-11-22 | 青岛大学 | Conversion nano probe and preparation method and application on a kind of polychrome |
CN112603999A (en) * | 2020-12-30 | 2021-04-06 | 重庆医科大学 | Tumor microenvironment response type nanoparticles based on bionic engineering and preparation method and application thereof |
-
2021
- 2021-07-13 CN CN202110790475.5A patent/CN113577306B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014200441A1 (en) * | 2013-06-14 | 2014-12-18 | National University Of Singapore | Core-shell fluorescent upconversion nanoparticles for photoactivation of multiple biomolecules |
CN110478483A (en) * | 2019-08-22 | 2019-11-22 | 青岛大学 | Conversion nano probe and preparation method and application on a kind of polychrome |
CN112603999A (en) * | 2020-12-30 | 2021-04-06 | 重庆医科大学 | Tumor microenvironment response type nanoparticles based on bionic engineering and preparation method and application thereof |
Non-Patent Citations (3)
Title |
---|
GUORUI JIN,ET AL.: "Near-infrared light-regulated cancer theranostic nanoplatform based on aggregation-induced emission luminogen encapsulated upconversion nanoparticles", 《THERANOSTICS》 * |
JUNGHAN LEE,ET AL.: "RGD Peptide–Conjugated Multimodal NaGdF4:Yb3+/Er3+ Nanophosphors for Upconversion Luminescence, MR, and PET Imaging of Tumor Angiogenesis", 《THE JOURNAL OF NUCLEAR MEDICINE》 * |
闫涛等: "肿瘤微环境刺激响应型上转换光动力诊疗体系的构建和发展", 《化学学报》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114404429A (en) * | 2021-11-30 | 2022-04-29 | 重庆医科大学附属第二医院 | Nano-silver modified tannin-iron network drug-loaded nano-composite, preparation method thereof and application thereof in reversing tumor drug resistance |
CN114748420A (en) * | 2022-04-12 | 2022-07-15 | 武汉科技大学 | Preparation method of amphiphilic polymer micelle with charge self-reversal activation mitochondrial targeting effect |
CN114748420B (en) * | 2022-04-12 | 2024-03-08 | 武汉科技大学 | Preparation method of amphiphilic polymer micelle with charge self-overturning activation mitochondrial targeting effect |
Also Published As
Publication number | Publication date |
---|---|
CN113577306B (en) | 2023-02-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chen et al. | NIR-II light activated photodynamic therapy with protein-capped gold nanoclusters | |
Jia et al. | Synthesis of carbon dots from Hypocrella bambusae for bimodel fluorescence/photoacoustic imaging-guided synergistic photodynamic/photothermal therapy of cancer | |
Jia et al. | Recent advances and prospects of carbon dots in cancer nanotheranostics | |
You et al. | Persistent regulation of tumor hypoxia microenvironment via a bioinspired Pt‐based oxygen nanogenerator for multimodal imaging‐guided synergistic phototherapy | |
He et al. | Tumor pH-responsive metastable-phase manganese sulfide nanotheranostics for traceable hydrogen sulfide gas therapy primed chemodynamic therapy | |
Yu et al. | Oxygen self-sufficient NIR-activatable liposomes for tumor hypoxia regulation and photodynamic therapy | |
Mo et al. | Multifunctional phototheranostic nanoplatform based on polydopamine-manganese dioxide-IR780 iodide for effective magnetic resonance imaging-guided synergistic photodynamic/photothermal therapy | |
Zhang et al. | Intelligent gold nanostars for in vivo CT imaging and catalase-enhanced synergistic photodynamic & photothermal tumor therapy | |
Zheng et al. | Biodegradable hypocrellin derivative nanovesicle as a near-infrared light-driven theranostic for dually photoactive cancer imaging and therapy | |
Wang et al. | Ag@ Fe3O4@ C nanoparticles for multi-modal imaging-guided chemo-photothermal synergistic targeting for cancer therapy | |
Han et al. | A magnetic polypyrrole/iron oxide core/gold shell nanocomposite for multimodal imaging and photothermal cancer therapy | |
Wang et al. | Photo-enhanced singlet oxygen generation of prussian blue-based nanocatalyst for augmented photodynamic therapy | |
Liu et al. | Carbon nanocage-based nanozyme as an endogenous H 2 O 2-activated oxygenerator for real-time bimodal imaging and enhanced phototherapy of esophageal cancer | |
CN113577306B (en) | Preparation of double-targeting pH stimulus-responsive nano particles and application of nano particles in tumor diagnosis and treatment | |
Soratijahromi et al. | Photothermal/sonodynamic therapy of melanoma tumor by a gold/manganese dioxide nanocomposite: In vitro and in vivo studies | |
CN108888767A (en) | A kind of nanocomposite, preparation method and application | |
Guo et al. | Emerging biocompatible nanoplatforms for the potential application in diagnosis and therapy of deep tumors | |
Bian et al. | Cu-based MOFs decorated dendritic mesoporous silica as tumor microenvironment responsive nanoreactor for enhanced tumor multimodal therapy | |
CN107469079B (en) | Preparation method of photodynamic therapeutic agent under guidance of T1-MRI imaging | |
CN109289050B (en) | Ferroferric oxide/polypyrrole/glucose oxidase composite multifunctional nano diagnosis and treatment agent and preparation method and application thereof | |
Chen et al. | Dual-source powered nanomotor with integrated functions for cancer photo-theranostics | |
Cao et al. | A multimodal imaging-guided nanoreactor for cooperative combination of tumor starvation and multiple mechanism-enhanced mild temperature phototherapy | |
Chen et al. | A Cu 2+ doped mesoporous polydopamine Fenton nanoplatform for low-temperature photothermal therapy | |
Wang et al. | Recent progress toward imaging application of multifunction sonosensitizers in sonodynamic therapy | |
Su et al. | Polydopamine nanoparticles coated with a metal-polyphenol network for enhanced photothermal/chemodynamic cancer combination therapy |
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 |