CN115779101B - Preparation and application of nanoparticle for resisting tumor metastasis - Google Patents
Preparation and application of nanoparticle for resisting tumor metastasis Download PDFInfo
- Publication number
- CN115779101B CN115779101B CN202111069967.1A CN202111069967A CN115779101B CN 115779101 B CN115779101 B CN 115779101B CN 202111069967 A CN202111069967 A CN 202111069967A CN 115779101 B CN115779101 B CN 115779101B
- Authority
- CN
- China
- Prior art keywords
- tumor
- nanoparticles
- psn
- metastasis
- nanoparticle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 153
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 76
- 206010027476 Metastases Diseases 0.000 title claims abstract description 62
- 230000009401 metastasis Effects 0.000 title claims abstract description 61
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000003814 drug Substances 0.000 claims abstract description 72
- 229960000590 celecoxib Drugs 0.000 claims abstract description 50
- 229960002528 ticagrelor Drugs 0.000 claims abstract description 49
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 41
- 239000003446 ligand Substances 0.000 claims abstract description 25
- RZEKVGVHFLEQIL-UHFFFAOYSA-N celecoxib Chemical compound C1=CC(C)=CC=C1C1=CC(C(F)(F)F)=NN1C1=CC=C(S(N)(=O)=O)C=C1 RZEKVGVHFLEQIL-UHFFFAOYSA-N 0.000 claims abstract description 23
- 230000000694 effects Effects 0.000 claims abstract description 23
- OEKWJQXRCDYSHL-FNOIDJSQSA-N ticagrelor Chemical compound C1([C@@H]2C[C@H]2NC=2N=C(N=C3N([C@H]4[C@@H]([C@H](O)[C@@H](OCCO)C4)O)N=NC3=2)SCCC)=CC=C(F)C(F)=C1 OEKWJQXRCDYSHL-FNOIDJSQSA-N 0.000 claims abstract description 20
- 230000008685 targeting Effects 0.000 claims abstract description 18
- 239000003112 inhibitor Substances 0.000 claims abstract description 16
- 239000003255 cyclooxygenase 2 inhibitor Substances 0.000 claims abstract description 13
- 230000006870 function Effects 0.000 claims description 15
- 206010006187 Breast cancer Diseases 0.000 claims description 12
- 208000026310 Breast neoplasm Diseases 0.000 claims description 12
- 239000002539 nanocarrier Substances 0.000 claims description 11
- 230000000259 anti-tumor effect Effects 0.000 claims description 8
- 229920001577 copolymer Polymers 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 230000001105 regulatory effect Effects 0.000 claims description 6
- 238000011282 treatment Methods 0.000 claims description 6
- 230000001276 controlling effect Effects 0.000 claims description 5
- 230000002708 enhancing effect Effects 0.000 claims description 2
- 229940093444 Cyclooxygenase 2 inhibitor Drugs 0.000 claims 2
- 229940122204 Cyclooxygenase inhibitor Drugs 0.000 claims 1
- 239000004480 active ingredient Substances 0.000 claims 1
- 239000002599 prostaglandin synthase inhibitor Substances 0.000 claims 1
- 229940079593 drug Drugs 0.000 abstract description 63
- 238000012546 transfer Methods 0.000 abstract description 16
- 108010035766 P-Selectin Proteins 0.000 abstract description 11
- 229940111134 coxibs Drugs 0.000 abstract description 11
- 108010037462 Cyclooxygenase 2 Proteins 0.000 abstract description 10
- 102100023472 P-selectin Human genes 0.000 abstract description 10
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 abstract description 10
- 238000009825 accumulation Methods 0.000 abstract description 8
- 230000009545 invasion Effects 0.000 abstract description 7
- 238000002271 resection Methods 0.000 abstract description 5
- ZRVUJXDFFKFLMG-UHFFFAOYSA-N Meloxicam Chemical compound OC=1C2=CC=CC=C2S(=O)(=O)N(C)C=1C(=O)NC1=NC=C(C)S1 ZRVUJXDFFKFLMG-UHFFFAOYSA-N 0.000 abstract description 3
- 230000003993 interaction Effects 0.000 abstract description 3
- 229960001929 meloxicam Drugs 0.000 abstract description 3
- 230000005012 migration Effects 0.000 abstract description 3
- 238000013508 migration Methods 0.000 abstract description 3
- HYWYRSMBCFDLJT-UHFFFAOYSA-N nimesulide Chemical compound CS(=O)(=O)NC1=CC=C([N+]([O-])=O)C=C1OC1=CC=CC=C1 HYWYRSMBCFDLJT-UHFFFAOYSA-N 0.000 abstract description 3
- 229960000965 nimesulide Drugs 0.000 abstract description 3
- 229960000371 rofecoxib Drugs 0.000 abstract description 3
- RZJQGNCSTQAWON-UHFFFAOYSA-N rofecoxib Chemical compound C1=CC(S(=O)(=O)C)=CC=C1C1=C(C=2C=CC=CC=2)C(=O)OC1 RZJQGNCSTQAWON-UHFFFAOYSA-N 0.000 abstract description 3
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 abstract description 2
- 239000005552 B01AC04 - Clopidogrel Substances 0.000 abstract description 2
- 239000005465 B01AC22 - Prasugrel Substances 0.000 abstract description 2
- 229960001138 acetylsalicylic acid Drugs 0.000 abstract description 2
- 230000004087 circulation Effects 0.000 abstract description 2
- 229960003009 clopidogrel Drugs 0.000 abstract description 2
- GKTWGGQPFAXNFI-HNNXBMFYSA-N clopidogrel Chemical compound C1([C@H](N2CC=3C=CSC=3CC2)C(=O)OC)=CC=CC=C1Cl GKTWGGQPFAXNFI-HNNXBMFYSA-N 0.000 abstract description 2
- 229960004197 prasugrel Drugs 0.000 abstract description 2
- DTGLZDAWLRGWQN-UHFFFAOYSA-N prasugrel Chemical compound C1CC=2SC(OC(=O)C)=CC=2CN1C(C=1C(=CC=CC=1)F)C(=O)C1CC1 DTGLZDAWLRGWQN-UHFFFAOYSA-N 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 36
- 241000699670 Mus sp. Species 0.000 description 22
- 210000001519 tissue Anatomy 0.000 description 21
- 210000004072 lung Anatomy 0.000 description 20
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- 238000000034 method Methods 0.000 description 16
- 229920001223 polyethylene glycol Polymers 0.000 description 12
- 108090000765 processed proteins & peptides Proteins 0.000 description 11
- 201000011510 cancer Diseases 0.000 description 10
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 9
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 9
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 9
- 210000003462 vein Anatomy 0.000 description 9
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 8
- 239000002245 particle Substances 0.000 description 8
- 230000001225 therapeutic effect Effects 0.000 description 8
- 238000011725 BALB/c mouse Methods 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- COXVTLYNGOIATD-HVMBLDELSA-N CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O Chemical compound CC1=C(C=CC(=C1)C1=CC(C)=C(C=C1)\N=N\C1=C(O)C2=C(N)C(=CC(=C2C=C1)S(O)(=O)=O)S(O)(=O)=O)\N=N\C1=CC=C2C(=CC(=C(N)C2=C1O)S(O)(=O)=O)S(O)(=O)=O COXVTLYNGOIATD-HVMBLDELSA-N 0.000 description 6
- 201000005389 breast carcinoma in situ Diseases 0.000 description 6
- 229960003699 evans blue Drugs 0.000 description 6
- 238000000338 in vitro Methods 0.000 description 6
- 238000011534 incubation Methods 0.000 description 6
- 230000006907 apoptotic process Effects 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000012292 cell migration Effects 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 238000010172 mouse model Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 230000008728 vascular permeability Effects 0.000 description 5
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 230000002001 anti-metastasis Effects 0.000 description 4
- 230000003915 cell function Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 150000002632 lipids Chemical class 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 3
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 3
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 3
- 208000007536 Thrombosis Diseases 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000033115 angiogenesis Effects 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 230000021164 cell adhesion Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000002296 dynamic light scattering Methods 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 238000010253 intravenous injection Methods 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000001394 metastastic effect Effects 0.000 description 3
- 206010061289 metastatic neoplasm Diseases 0.000 description 3
- 239000000106 platelet aggregation inhibitor Substances 0.000 description 3
- 230000002980 postoperative effect Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 230000004614 tumor growth Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- IPJDHSYCSQAODE-UHFFFAOYSA-N 5-chloromethylfluorescein diacetate Chemical compound O1C(=O)C2=CC(CCl)=CC=C2C21C1=CC=C(OC(C)=O)C=C1OC1=CC(OC(=O)C)=CC=C21 IPJDHSYCSQAODE-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 206010006189 Breast cancer in situ Diseases 0.000 description 2
- 102000016289 Cell Adhesion Molecules Human genes 0.000 description 2
- 108010067225 Cell Adhesion Molecules Proteins 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 229930040373 Paraformaldehyde Natural products 0.000 description 2
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 2
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000009435 amidation Effects 0.000 description 2
- 238000007112 amidation reaction Methods 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 238000005415 bioluminescence Methods 0.000 description 2
- 230000029918 bioluminescence Effects 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 210000002865 immune cell Anatomy 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 108010082117 matrigel Proteins 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229920002866 paraformaldehyde Polymers 0.000 description 2
- 150000003904 phospholipids Chemical class 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 239000012679 serum free medium Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 102100036150 C-X-C motif chemokine 5 Human genes 0.000 description 1
- 238000007808 Cell invasion assay Methods 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010053567 Coagulopathies Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 108010040476 FITC-annexin A5 Proteins 0.000 description 1
- 229920000855 Fucoidan Polymers 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 101000947186 Homo sapiens C-X-C motif chemokine 5 Proteins 0.000 description 1
- 101000947178 Homo sapiens Platelet basic protein Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- KTDZCOWXCWUPEO-UHFFFAOYSA-N NS-398 Chemical compound CS(=O)(=O)NC1=CC=C([N+]([O-])=O)C=C1OC1CCCCC1 KTDZCOWXCWUPEO-UHFFFAOYSA-N 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 102100036154 Platelet basic protein Human genes 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920002873 Polyethylenimine Polymers 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 101710180316 Protease 2 Proteins 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- 208000003721 Triple Negative Breast Neoplasms Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000702 anti-platelet effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 229940127218 antiplatelet drug Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 description 1
- 208000034158 bleeding Diseases 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- BQRGNLJZBFXNCZ-UHFFFAOYSA-N calcein am Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=C(OC(C)=O)C=C1OC1=C2C=C(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(=O)C)C(OC(C)=O)=C1 BQRGNLJZBFXNCZ-UHFFFAOYSA-N 0.000 description 1
- 230000004709 cell invasion Effects 0.000 description 1
- 238000007806 cell migration and invasion assay Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- VBVAVBCYMYWNOU-UHFFFAOYSA-N coumarin 6 Chemical compound C1=CC=C2SC(C3=CC4=CC=C(C=C4OC3=O)N(CC)CC)=NC2=C1 VBVAVBCYMYWNOU-UHFFFAOYSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000012137 double-staining Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000007705 epithelial mesenchymal transition Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 description 1
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920001610 polycaprolactone Polymers 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920000166 polytrimethylene carbonate Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 150000003180 prostaglandins Chemical class 0.000 description 1
- 208000005333 pulmonary edema Diseases 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 239000013074 reference sample Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 208000022679 triple-negative breast carcinoma Diseases 0.000 description 1
- 230000005747 tumor angiogenesis Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method of active targeting ligand modified nano-particles for entrapping a platelet inhibitor and a cyclooxygenase-2 (COX-2) inhibitor and application thereof in treating tumor metastasis. The platelet inhibitor is at least one of ticagrelor, aspirin, clopidogrel and prasugrel, and the COX-2 inhibitor is at least one of nimesulide, meloxicam, celecoxib and rofecoxib; wherein the platelet inhibitor can inhibit interaction of platelets with tumor cells, and the COX-2 inhibitor can inhibit expression of COX-2 by tumor cells, so as to weaken invasion and migration capacity. The P-selectin targeting ligand is modified on the surface of the nanoparticle, and accumulation of the nanoparticle at the primary tumor position and the transfer position is realized through targeting activated platelets. The ligand modified drug-loaded nanoparticle prepared by the invention has obvious effects in the aspects of treating diffuse circulation tumor cell metastasis, primary tumor metastasis and metastasis after tumor resection.
Description
Technical Field
The invention relates to preparation of active targeting ligand modified nano particles for regulating platelet and tumor cell functions and application thereof in treating tumor metastasis. In particular to a P-selectin targeting ligand modified nanoparticle for entrapping a platelet inhibitor and a COX-2 inhibitor and application thereof, belonging to the technical field of medicines.
Background
Cancer is a major disease threatening human health. Tumor metastasis is the leading cause of high mortality in cancer, with 90% of tumor patients dying from metastasis. Conventional treatments such as radiation and chemotherapy can remove localized cancer cells, but it is difficult to achieve the desired metastasis inhibiting effect. Surgical resection is an effective clinical strategy for treating solid tumors, and although surgery can ablate more than 90% of tumors, residual micro-tumors that are difficult to eliminate intraoperatively are prone to tumor recurrence. In addition, residual micro-tumors invade surrounding tissues, leading to distant metastasis of tumor cells. Therefore, metastasis inhibition is a critical and challenging task in cancer treatment.
Studies have shown that a suitable microenvironment is critical for the transplantation and growth of tumor cells. Primary tumors induce the establishment of a pre-transfer microenvironment (PMN) at secondary organ and tissue sites, facilitating the spread and survival of Circulating Tumor Cells (CTCs) prior to reaching these sites. The pre-transfer microenvironment is called as "soil" of metastasis, is composed of Circulating Tumor Cells (CTCs), endothelial cells, activated platelets, intercellular adhesion molecules (ICAMs) and the like, has the characteristics of immunosuppression, inflammation, angiogenesis and the like, and is favorable for tumor cell colonization. Each stage of metastasis requires a tight interaction between the tumor cells and the immune cells, stroma, inflammation, etc. components of the tumor microenvironment. Therefore, designing a combined administration strategy to regulate the transfer process from different mechanisms will help to achieve better transfer therapeutic effect.
Platelets play an important role in tumor metastasis. At the tumor site, tumor cells express Adenosine Diphosphate (ADP) to activate platelets, activate platelets to secrete transforming growth factor beta (TGF- β), vascular Endothelial Growth Factor (VEGF) and Platelet Derived Growth Factor (PDGF), promote epithelial-mesenchymal transition of cancer cells and induce angiogenesis; during the transfer process, platelets form aggregates with tumor cells entering the blood circulation, helping CTCs to undergo shear stress and avoid immune cell clearance, promoting their invasion into surrounding tissues; at the secondary site of metastasis, activated platelets secrete pro-inflammatory factors CXCL5 and CXCL7, which aid granulocyte recruitment to create a pre-metastatic microenvironment. The role of platelets in cancer-related inflammation and thrombosis suggests that platelets are potential therapeutic targets. P-selectin is an adhesion molecule stored in platelet alpha-particles. When platelets are activated, P-selectin rapidly migrates from the alpha-particles to the platelet surface and plays an important role in cancer metastasis by interacting with P-selectin glycoprotein ligands that are overexpressed on cancer cells. The P-selectin targeting ligand can specifically bind to P-selectin to bind to activated platelets, and has excellent targeting ability to primary tumors and metastatic sites.
Cyclooxygenase-2 (COX-2) is overexpressed in various malignant tumors, playing an important role in tumor growth and metastasis. COX-2 expression is increased under inflammatory stimulus conditions, inducing vascular permeability of primary breast tumors and distant organs, and extravasating CTC. In addition, COX-2 at the tumor site promotes angiogenesis by catalyzing prostaglandin production, up-regulating Vascular Endothelial Growth Factor (VEGF) expression, induces invasion and increases metastasis. Specific inhibitors of COX-2 such as celecoxib, nimesulide, meloxicam and rofecoxib can produce a strong inhibition effect on tumor angiogenesis by inhibiting the activity of COX-2, down regulate the expression of Matrix Metalloproteinases (MMP) and Vascular Endothelial Growth Factor (VEGF), weaken the migration, invasion and adhesion ability of cancer cells, and effectively inhibit tumor metastasis. Celecoxib is used for research on anti-tumor drug delivery systems at present, but an anti-tumor metastasis drug delivery system prepared by combining COX-2 inhibitor and platelet inhibitor and loading nanoparticles is not reported yet.
Disclosure of Invention
In order to solve the problem that the tumor metastasis is difficult to treat clinically at present. The inventor creatively uses nano carrier materials to encapsulate platelet inhibitor drugs and COX-2 inhibitor drugs, and realizes the regulation and control of the transfer process by the combined use and the regulation and control of the functions of platelets and tumor cells. On the basis, the P-selectin ligand is used for modifying the nanoparticle, so that the targeting ability of the nanoparticle to activate platelets is provided. By tracking platelets and tumor cells in the transfer process, the accumulation of the medicine at the primary tumor part and the transfer part is increased, the interaction between platelets and tumor cells is inhibited, and the curative effect of resisting tumor transfer is further improved.
One of the purposes of the invention is to overcome the defects of the existing treatment scheme, provide an active targeting anti-tumor metastasis nanoparticle which can regulate and control the functions of platelets and tumor cells at the same time, block the metastasis promotion capability of the platelets and the invasion capability of the tumor cells, inhibit the formation of microenvironment before the metastasis, and obviously enhance the inhibition effect on tumor metastasis.
The invention also aims to overcome the safety problem of systemic application of platelet inhibitors and the problem of less accumulation of free drugs in focus, and prepare ligand modified nanoparticles capable of simultaneously targeting primary tumors and metastasis and inhibiting tumor metastasis, thereby realizing targeted delivery of the existing antiplatelet drugs and COX-2 inhibitors.
It is another object of the present invention to provide the use of nanoparticles capable of simultaneously modulating platelet and tumor cell function in anti-tumor metastasis.
The invention is realized by the following technical scheme: provides a nanoparticle for simultaneously regulating and controlling the functions of platelets and tumor cells and enhancing the anti-tumor metastasis effect and application thereof in the aspects of treating diffuse circulation tumor cell metastasis, primary tumor metastasis and metastasis after tumor resection, and comprises a platelet inhibitor for regulating and controlling the functions of the platelets, a COX-2 inhibitor for regulating and controlling the functions of the tumor cells, a nano carrier material and a targeting ligand.
The platelet inhibitor is at least one of ticagrelor, aspirin, clopidogrel and prasugrel; preferably ticagrelor.
Specifically, ticagrelor Lei Zhiliang accounts for 30-40% of the total mass of the nano particles, and the balance is nano carrier material.
The COX-2 inhibitor is at least one of celecoxib, nimesulide, meloxicam, rofecoxib and NS 398; celecoxib is preferred.
Specifically, the mass of celecoxib accounts for 5-15% of the total mass of the nano particles, and the balance is nano carrier material.
The nano carrier material is a polymer capable of encapsulating a hydrophobic drug, and is preferably one or more of polylactic acid-glycolic acid copolymer, poly (epsilon-caprolactone) and copolymer, polylactic acid, poly (trimethylene carbonate) and copolymer thereof and polyethyleneimine; more preferably a polylactic acid-glycolic acid copolymer.
The targeting ligand can bind to P-selectin on the surface of activated platelets, and is preferably one or more of PSN peptide (DAEWVDVS), fucoidan sulfate, sialic acid Lewis X and GPIba. More preferably PSN peptide (DAEWVDVS).
The ligand modified drug-loaded nanoparticle of the entrapped ticagrelor or celecoxib is preferably prepared by the following method: (1) preparing a ligand-modified nanocarrier: PSN peptides are conjugated to NHS ester modified lipids by reductive amidation to form polypeptide modified NHS esters with stable amide linkages. Namely: the NHS-modified lipid was dissolved in DMSO, and DMSO in which PSN peptide was dissolved was added dropwise with stirring, and the reaction was stirred at room temperature for 24 hours. And after the reaction is finished, removing free PSN peptide by deionized water dialysis, and freeze-drying to obtain the ligand modified nano-carrier.
Preferably, the lipid of step (1) is distearoyl phosphatidylethanolamine-polyethylene glycol.
(2) The drug is packed in a ligand-modified nanocarrier: and preparing the singly-loaded ligand modified drug-loaded nanoparticles by using a nano precipitation method. Dissolving a ligand-modified nano-carrier, a polylactic acid-glycolic acid copolymer and phospholipid in a certain mass ratio (10:4:2) into DMSO to be used as an oil phase, adding ticagrelor or celecoxib, and slowly dripping the oil phase into deionized water under stirring to prepare ligand-modified ticagrelor celecoxib-carrying nanoparticles.
The ligand modified nanoparticle can also be loaded with platelet inhibitor drugs and COX-2 inhibitor drugs simultaneously. The preparation method of the double-carrier nano-particle comprises the steps of simultaneously adding two medicines in a certain proportion into an oil phase, and the rest methods are the same as those of the single-carrier nano-particle. The dual-drug nanoparticle can achieve the equivalent effect of respectively releasing two drugs when the single-carrier nanoparticle is combined by simultaneously releasing the two drugs.
The tumor comprises breast cancer, gastric cancer, colon cancer, pancreatic cancer, melanoma, liver cancer, lung cancer, bladder cancer, cervical cancer and prostate cancer; lung metastasis of breast cancer is preferred.
The invention is to add platelet inhibitor drugs and COX-2 inhibitor drugs into tumor cells after physical mixing or to inject the drugs into animals intravenously for combined medication.
In the combined drug, the P-selectin targeting ligand modified carrier-substituted kagrelor nanoparticles and celecoxib nanoparticles can inhibit the tumor cell metastasis effect of platelets through accumulation in tumor tissues and metastasis sites, inhibit COX-2 expression, induce tumor cell apoptosis, down regulate VEGF and MMP, reduce vascular permeability, destroy microenvironment soil before transfer, and inhibit breast cancer metastasis.
The beneficial effects of the invention are as follows:
the invention provides a method for realizing regulation and control of a metastasis process by considering the functions of regulating and controlling platelets and tumor cells at the same time, and relates to the use of antiplatelet inhibitor drugs and non-steroidal anti-inflammatory drug COX-2 inhibitor drugs. The P-selectin targeting ligand modified drug-loaded nanoparticle provided by the invention can effectively improve the accumulation of drugs at primary tumor positions and metastasis positions by 'tracking' platelets and tumor cells in the metastasis process, compared with free drugs, thereby realizing the selective inhibition of platelet functions and tumor cell functions, increasing the therapeutic effect of the drugs and reducing the bleeding risk of the platelet inhibitors applied to the whole body.
Drawings
FIG. 1 is a schematic illustration of the preparation of ligand-modified drug-loaded nanoparticles.
FIG. 2 is a representation of ligand-modified drug-loaded nanoparticles.
FIG. 3 is a graph showing the effect of ligand-modified drug-loaded nanoparticles on the in vitro migration and invasion capacity of triple negative breast cancer 4T1 cells.
FIG. 4 is a graph showing the effect of ligand-modified drug-loaded nanoparticles on platelet function.
FIG. 5 is a graph showing the effect of ligand-modified drug-loaded nanoparticles on tumor cell function.
FIG. 6 is a graph showing the effect of ligand-modified drug-loaded nanoparticles on platelet-tumor cell adhesion.
Figure 7 is a graph showing the in vitro binding activation platelet effect of ligand-modified nanoparticles.
FIG. 8 is a tissue profile of ligand-modified nanoparticles in 4T1-Luc breast cancer metastasis and 4T1 breast cancer in situ tumor model mice.
FIG. 9 shows the anti-metastatic therapeutic effect of ligand-modified drug-loaded nanoparticles in 4T1-Luc breast cancer metastasis model mice.
FIG. 10 is a graph showing the effect of ligand-modified drug-loaded nanoparticles on inhibiting microenvironment formation prior to transfer.
FIG. 11 shows the anti-metastatic therapeutic effect of ligand-modified drug-loaded nanoparticles in 4T1 in situ breast cancer model mice.
FIG. 12 is a graph showing the effect of in vivo safety inspection of ligand-modified drug-loaded nanoparticles.
FIG. 13 shows the anti-metastatic therapeutic effect of ligand-modified drug-loaded nanoparticles in 4T1 in situ breast cancer surgical resection model mice.
Detailed Description
The technical scheme of the present invention is described in further detail below in conjunction with specific embodiments. It will be appreciated by those skilled in the art that the embodiments of the invention are not limited to the examples described.
Example 1: synthesis and characterization of ligand-modified DSPE-PEG
Conjugation of PSN peptides to NHS ester modified lipids (DSPE-PEG) by reductive amidation 2000 -NHS) to form DSPE-PEG with stable amide bond 2000 -PSN. 10mg DSPE-PEG 2000 NHS was dissolved in 1ml DMSO, 200. Mu.l DMSO in which 3mg PSN peptide was dissolved was added dropwise with stirring, and the reaction was stirred at room temperature for 24 hours. After the reaction is finished, using a dialysis bag with the molecular weight cut-off of 3500, dialyzing for 24 hours by using deionized water as a medium to remove unreacted free peptide, and freeze-drying to obtain DSPE-PEG 2000 -PSN。
Taking the freeze-dried DSPE-PEG 2000 PSN sample 4mg, dissolved in 1mL DMSO-d 6. And weighing 4mg DSPE-PEG 2000 NHS was dissolved in 1mL of DMSO-d6 as a reference sample. Nuclear magnetic resonance hydrogen spectrum at 400MHz 1 H-NMR) analysis.
The results are shown in FIG. 2A, DSPE-PEG 2000 NHS shows a characteristic peak of NHS at 2.8ppm, DSPE-PEG after reaction with PSN peptide 2000 The disappearance of the NHS peak in PSN demonstrated successful attachment of PSN peptide to DSPE-PEG 2000 -NHS.
Example 2: preparation and characterization of ligand modified drug-loaded nanoparticles
The ligand modified drug-loaded nanoparticle is prepared by adopting a nano precipitation method. Specifically, DSPE-PEG is used 2000 PSN, polylactic acid-glycolic acid copolymer (PLGA) and phospholipid are dissolved in DMSO according to a certain mass ratio (10:4:2) to be used as oil phases, and ticagrelor (100%, w/w) or celecoxib (12) with a certain PLGA mass fraction is respectively addedAnd (3) in percent, w/w), uniformly mixing, and slowly dropwise adding the mixture into deionized water under stirring at 1000rpm to prepare ligand-modified supported ticagrelor nanoparticles (PSN-supported ticagrelor nanoparticles) and ligand-modified supported celecoxib nanoparticles (PSN-supported celecoxib nanoparticles). The prepared nanoparticles were concentrated with an ultrafiltration tube (10 kDa) and washed twice with low temperature deionized water to remove the organic solvent, followed by dispersing the nanoparticles in PBS (ph=7.4). The particle size distribution and zeta potential of freshly prepared nanoparticles were measured by Dynamic Light Scattering (DLS) using a malvern laser particle sizer.
As shown in FIGS. 2B-C, the particle diameters of the two ligand-modified drug-loaded nanoparticles are about 85nm, the electric potential is about-20 mV, and PDI is less than 0.3, which indicates that the nanoparticle distribution is uniform.
Example 3: stability test of ligand-modified drug-loaded nanoparticles
1mL of the ligand-modified drug-loaded nanoparticle prepared in example 2 was dispersed in phosphate buffer (PBS, pH=7.4) or PBS containing 10% Fetal Bovine Serum (FBS). The particle size change was determined by dynamic light scattering at predetermined time points (0,2,4,8,12 and 24 h) by incubation in a shaker at 37℃for 24h at 75 rpm.
As shown in fig. 2D, the particle size distribution of the nanoparticle after 24h incubation in PBS or 10% fbs did not significantly change, indicating that the ligand-modified drug-loaded nanoparticle has better in vitro stability and serum stability.
Example 4: in vitro Transwell cell migration and invasion assay
In the cell migration test, 4T1 cells in the logarithmic phase were taken, the cells were digested, washed 2 times with serum-free medium and resuspended, counted, and the cell suspension concentration was adjusted to 8X 10 5 cell/ml. Adding 700 μl of 1640 culture medium containing 10% serum into the bottom of a 24-well plate, adding 100 μl of cell suspension into the upper chamber, culturing for 4 hr, adding different drugs (containing no drug culture medium, platelet, platelet+ticagrelor+celecoxib, platelet+PSN-ticagrelor nanoparticle, platelet+PSN-celecoxib nanoparticle+PSN-celecoxib nanoparticle, and platelet+PSN-ticagrelor nanoparticle+PSN-celecoxib nanoparticle)Nanoparticles) were incubated for 24h. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2. The liquid in the upper chamber of the Transwell cell was aspirated, the lower chamber was fixed with 4% paraformaldehyde at room temperature and stained with 0.1% crystal violet stain for 10min, the upper non-migrated cells were wiped off with a cotton swab, washed 3-4 times with PBS, and cells migrated to the lower chamber were observed under a microscope. Finally, the purple crystals were dissolved using a 33% acetic acid solution and absorbance was measured at 570nm on a microplate reader. Mobility was calculated as follows: cell mobility (%) = absorbance of experimental group/absorbance of control group x 100%. In the cell invasion assay, 3mg/ml matrigel was previously spread on the upper chamber of a Transwell cell having a pore size of 8. Mu.m, and left at 37℃for 2 hours to solidify the matrigel into a gel. The remaining steps were the same as in the cell migration test.
The results are shown in fig. 3, where the mobility of 4T1 cells is significantly increased after incubation with platelets, indicating that platelet stimulation can promote tumor cell migration. PNS-celecoxib nanoparticles or PSN-ticagrelor nanoparticles can inhibit the increase of cell migration caused by platelets to a certain extent. Further, after the two nano-particles are combined, the mobility and invasion rate of the 4T1 cells are reduced to the minimum, and the optimal anti-tumor cell migration and invasion capacity is shown.
Example 5: influence of ligand-modified drug-loaded nanoparticles on platelet function
To study the effect of ticagrelor on platelet function blockade, platelet aggregation function was first evaluated. After incubating platelets labeled with the fluorescent dye DiI with PBS, free ticagrelor, and ligand-modified drug-loaded nanoparticles, thrombin was added to induce platelet aggregation. The light transmittance of the treated platelet suspension was measured at 650nm using the PBS-treated platelet aggregation rate as a positive control to quantitatively evaluate platelet aggregation. Wherein the ligand-modified drug-loaded nanoparticle is from example 2.
Results as shown in fig. 4A, platelets treated with PBS and PSN-celecoxib nanoparticles aggregated into large particles, while platelets treated with ticagrelor or PSN-ticagrelor nanoparticles retained their original size. From fig. 4B, it can be seen that the aggregation rate of platelets in the PSN-celecoxib nanoparticle treated group was close to that in the control group. In contrast, PSN-ticagrelor nanoparticles can significantly inhibit platelet aggregation, and the platelet aggregation rate is reduced to 18.97%.
The effect of the nanoparticles on the platelet clotting function was further investigated by clot retraction assay.
As shown in fig. 4C-E, PSN-ticagrelor nanoparticles significantly inhibited the contraction of blood clots, resulting in an increase in the weight of blood clots and a decrease in serum release, suggesting that platelet coagulation function was inhibited.
The above results indicate that PSN-ticagrelor nanoparticles can block platelet functions, including platelet aggregation and platelet coagulation.
Example 6: ligand modified drug-loaded nanoparticle induced tumor cell apoptosis
Inoculating 4T1 cells into a 12-well plate, culturing for 24 hours in a cell culture box, sucking liquid in the well plate, adding 1mL of a drug-containing culture medium for continuous culture for 24 hours, digesting and collecting the cells with pancreatin, washing the cells twice with PBS, operating according to the instruction of an Annexin V-FITC/APC double-staining kit, and detecting by using a flow cytometer after staining. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.
As shown in fig. 5A-B, celecoxib can obviously induce apoptosis of 4T1 tumor cells, and the PSN-celecoxib nanoparticles have stronger apoptosis induction capacity than free celecoxib.
The ability of the nanoparticles to inhibit tumor metastasis related proteins was further examined by Western-blot immunoblotting. 4T1 cells were inoculated into 6-well plates, cultured in a cell culture tank for 12 hours, the culture solution was aspirated off, and 2mL of the medicated medium was added for further culture for 24 hours. The medium was discarded, washed 2 times with PBS and the cells lysed using a complete lysate ice bath. Centrifuging to obtain supernatant, adding SDS loading buffer solution, and boiling for 10min to obtain loading sample. The samples were subjected to protein separation by SDS-PAGE gel electrophoresis, transferred to membrane, blocked with 5% milk powder, incubated overnight with primary antibody, washed with TBS/T and incubated with secondary antibody for 2h. The procedure was performed with reference to the chemiluminescent kit instructions and imaging was performed using a gel imager. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.
As shown in FIG. 5B, PSN-celecoxib nanoparticles can obviously reduce the expression of Vascular Endothelial Growth Factor (VEGF), metal matrix protease-2 (MMP-2) and metal matrix protease-9 (MMP-9) in 4T1 tumor cells.
Example 7: in vitro and in vivo assay quantitative analysis of ligand-modified drug-loaded nanoparticles to inhibit platelet-tumor cell adhesion: platelets were obtained from BALB/c mice by extraction, washed with PBS and labeled with 5. Mu.M calcein AM. 4T1 cells were seeded in 96-well plates (black, transparent at the bottom) and after growing to about 80% confluence, fluorescent-labeled platelets were added to each well after 1h of pretreatment of the cells with different drugs, incubation was continued for 45min at 37 ℃, and the fluorescent signal was detected for each well using an enzyme-labeled instrument at ex=490 nm and em=515 nm. The cells were washed 3 times with PBS to remove non-adherent platelets, 100uL of serum-free medium was added, and the remaining fluorescence intensity per well was determined, leaving the wells free of tumor cells as blanks. Platelet adhesion rate was calculated as: (residual fluorescence-blank)/(total fluorescence-blank) ×100%. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.
Qualitative analysis: 4T1 cells in the logarithmic growth phase are inoculated on 18mm borosilicate cover slips and incubated for 24 hours to enable the cells to adhere to the wall and grow adaptively. After washing the medium with PBS, 300. Mu.L of CMFDA staining solution at a concentration of 5. Mu.M was added to each well, and incubated at 37℃for 30min in the absence of light to prepare 4T1-CFSE cells. 1mL of the liquid medicine was added to each well and incubated at 37℃for 1 hour, respectively, to control the culture medium without the medicine. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2. Freshly extracted platelets were stained with DiI dye for 30min and collected by centrifugation. To each well 1mL of fluorescence labelled platelet suspension (containing 2X 10) 6 Platelets) and incubated at 37 ℃ for 45min to allow for adequate adhesion to tumor cells. Washed three times with PBS and fixed with 4% paraformaldehyde. DAPI stained nuclei for 5min, washed five times with PBS, and confocal photographed.
As shown in fig. 6A-B, the blank group had 75.53% of platelets adhered to 4T1 tumor cells, and the PSN-ticagrelor nanoparticle + PSN-celecoxib nanoparticle group had the best inhibitory effect on platelet adhesion to 4T1 cells, significantly lower than the PSN-ticagrelor nanoparticle alone or the PSN-celecoxib nanoparticle treated group.
Platelet-tumor cell adhesion in vivo was further studied. Platelets were extracted, washed and stained with DiI to prepare red fluorescent-labeled platelets. The 4T1 tumor cells in the logarithmic growth phase were also digested and incubated with CMFDA to prepare green fluorescent-labeled 4T1 cells. BALB/c small tail vein injection of drug from the tail vein, respectively, and 4T1 cell suspension (5×10) was injected into each mouse after 1h 5 cell/alone) followed immediately by injection of platelets via the contralateral tail vein (1X 10) 9 In mice, mice were sacrificed by cervical dislocation after 24h, and lung tissues were obtained. Cutting a small amount of lung tissue, embedding with tissue embedding agent on an objective table, quick-freezing at-25deg.C for 15min, cutting into frozen slices with thickness of 10 μm on a manual frozen slicer, washing with PBS for 2 times to remove water-soluble embedding agent, sucking water with filter paper, and dripping a small amount of anti-fluorescence quenching agent on the slices. The sections were observed under a confocal microscope and photographed.
As a result, as shown in FIG. 6C, platelets in the lung tissue sections of the control mice were in an aggregated state, and red fluorescence was mostly aggregated around the green fluorescence of tumor cells. And the combined use of the PSN-ticagrelor nanoparticles and the PSN-celecoxib nanoparticles can well inhibit the adhesion of platelets to tumor cells.
In summary, the combined administration of PSN-ticagrelor nanoparticles and PSN-celecoxib nanoparticles can effectively inhibit platelet adhesion induced by tumor cells by simultaneously inhibiting platelets and tumor cells.
Example 8: investigation of in vitro binding and activation platelet ability of ligand-modified nanoparticles
Platelets were extracted, washed and labeled with DiI as described above. Platelets were activated by incubation with ADP (20. Mu.M) for 30min at 37 ℃. The activated platelets were then incubated with coumarin 6-labeled PSN modified or unmodified nanoparticles for 30 minutes at 37 ℃ and then centrifuged at 3500rpm for 10 minutes to remove unbound nanoparticles. Finally, the platelet pellet was washed and resuspended in PBS (ph=7.4) and fluorescent images were captured under CLSM.
As shown in FIG. 7, the PSN modified nanoparticles have significantly higher fluorescence intensity than unmodified nanoparticles, and the green fluorescence of the nanoparticles is co-localized with the red fluorescence of activated platelets, indicating that the ligand-modified nanoparticles provided by the invention can effectively bind activated platelets.
Example 9: in-vivo distribution investigation of ligand modified nanoparticles in 4T1-Luc metastasis and 4T1 breast cancer in-situ tumor model mice
BALB/c mice metastasis model was established by intravenous injection of 4T1-Luc cells, and tumor metastasis progression was monitored by bioluminescence imaging. Mice successfully modeled for lung metastasis were selected, randomly divided into 3 groups (3 per group), and free DiR and DiR-labeled PSN modified or unmodified nanoparticles (DiR equivalent 1 mg/kg) were injected by tail vein. Mice were sacrificed 24h after injection, tissues and organs were rapidly removed, fluorescence signals were photographed ex vivo, and the fluorescence signals of the tissues were semi-quantitatively analyzed using ROI tools. The results are shown in FIGS. 8A-B.
The fluorescence imaging and semi-quantitative results of 24h isolated organ tissue show that the DiR-labeled PSN modified nanoparticles in the isolated lung showed the highest accumulation, 2.13 times and 3.21 times the fluorescence intensity of the unmodified nanoparticles and free DiR, respectively.
Further establishes a primary 4T1 breast cancer animal model to investigate the targeting of the ligand-modified nanoparticles to the primary tumor. By inoculating 3X 10 to the right side of female BALB/c mice with a third pair of breast pads 5 The 4T1 cells establish a 4T1 mouse orthotopic tumor model. At 14 days after tumor inoculation, 9 tumor-bearing mice with similar tumor sizes were randomly divided into 3 groups of 3 mice each. Each group was injected with free DiR and DiR-labeled PSN modified or unmodified nanoparticles (DiR equivalent 1 mg/kg) from the tail vein, respectively. Mice were sacrificed 24h after dosing, tumor tissue and lung tissue were dissected, fluorescent signals were taken ex vivo, and fluorescent signals of the tissue were semi-quantitatively analyzed using ROI tools.
As shown in fig. 8C-D, PSN modified nanoparticles have stronger fluorescent signals in tumor tissue, 1.59 and 9.60 times that of unmodified nanoparticles and free DiR, respectively, indicating that ligand modification can enhance the targeting of nanoparticles to in situ tumors. In addition, the accumulation of the ligand modified nanoparticle in the lung before the metastasis of the primary tumor model mouse is improved by 2.2 times compared with that of the unmodified nanoparticle.
The above results demonstrate that the ligand-modified nanoparticles provided by the present invention can effectively enhance accumulation at the primary tumor site and the metastatic site, as compared to the free drug.
Example 10: therapeutic effect of ligand-modified drug-loaded nanoparticles on metastasis of diffuse breast cancer cells in order to verify whether the nanoparticles can block the spreading process of diffuse circulating tumor cells and eliminate tumor metastasis, 1×10 is inoculated through tail vein 5 The individual 4T1-Luc cells established a BALB/c mouse breast cancer metastasis model. Days of intravenous injection of tumor cells were defined as day 0, and the drug was administered by intravenous injection 1 time every 3 days 6 times from day 3. On day 20 after tail vein injection of tumor cells, each mouse was intraperitoneally injected with 200 μ L D-potassium fluorescein salt (15 mg/ml) and the luminescence signal of the cells was detected by an IVIS Spectrum in vivo imaging system to monitor the formation of metastases. Mice were sacrificed and lungs were collected for weighing. The number of transfers was counted using Bouin's dye for fixation.
The administration method comprises the following steps:
control group: saline was injected intravenously through the tail of the mice on day 0,3,6,9,12,15, respectively;
free drug group: physiological saline (containing ticagrelor Lei Dangliang mg/kg, celecoxib equivalent 2 mg/kg) was injected intravenously via the tail of the mice on day 0,3,5,7,9,11, respectively;
PSN-ticagrelor Lei Nami granule group: physiological saline containing PSN-ticagrelor nanoparticles (containing ticagrelor Lei Dangliang mg/kg) was injected intravenously through the tail of the mice on day 0,3,5,7,9,11, respectively;
PSN-celecoxib nanoparticle group: physiological saline containing PSN-ticagrelor nanoparticles (containing celecoxib equivalent 2 mg/kg) was injected intravenously via the tail of the mice on day 0,3,5,7,9,11, respectively;
PSN-ticagrelor nanoparticle combination PSN-celecoxib nanoparticle group: on day 0,3,5,7,9,11, physiological saline containing PSN-ticagrelor nanoparticles and PSN-celecoxib nanoparticles (containing ticagrelor Lei Dangliang 10mg/kg, celecoxib equivalent 2 mg/kg) was injected intravenously through the tail of the mice, respectively;
wherein the PSN-ticagrelor nanoparticles and PSN-celecoxib nanoparticles are from example 2.
As shown in fig. 9, the bioluminescence intensity, lung weight index (lung weight/body weight x 100%) results and metastasis nodule count results of the in vitro lung of the 20 th day mice all showed that the combination of PSN-ticagrelor nanoparticles and PSN-celecoxib nanoparticles had the best effect of resisting metastasis of circulating tumor cells.
Example 11: investigation of the ability of ligand-modified drug-loaded nanoparticles to inhibit the formation of the pre-transfer microenvironment the influence of leakage of the co-suspension nanoparticles from Evan Blue (EB) on the lung vascular permeability of the pre-transfer microenvironment was carried out. BALB/c mice carrying 4T1 in situ breast cancer were randomly grouped, 4 per group (grouping method same as example 10). The drug solution was injected by tail vein (once every 3 days, 3 total) on day 5 after tumor inoculation. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.2 days after the last treatment, mice were injected intravenously with 100. Mu.L (20 mg/kg) of EB solution. 2. After hours, mice were sacrificed and bronchoalveolar lavage fluid (BALF) was extracted. Lung tissue was harvested, weighed and immersed in formamide to extract EB (2 mL formamide was added per 100 mg wet lung weight). After 48h incubation at 37℃the supernatant was centrifuged at 5000rpm for 20min and the fluorescence intensity of the supernatant was measured using an enzyme-labeled instrument (Ex: 620nm, em:683 nm). The EB content (mass of EB per gram of lung, μg/g) in the lung tissue was then calculated. The total protein content in BALF was determined by BCA kit.
As shown in fig. 10, the primary tumor significantly induced vascular permeability of the lung, resulting in increased EB leakage and total protein content in BALF, compared to healthy mice. And the combination of the PSN-ticagrelor nanoparticles and the PSN-celecoxib nanoparticles has the optimal effect of reversing the high vascular permeability of the microenvironment before pulmonary transfer.
Example 12: therapeutic effects of ligand-modified drug-loaded nanoparticles on primary breast cancer metastasis by inoculating 3×10 to the right third pair of breast pads of female BALB/c mice 5 The 4T1 cells establish a 4T1 mouse orthotopic tumor model. To the length of tumor to 50mm 3 At time (day 0), mice were randomized and 5 animals per group were dosed. The animals were grouped and dosed as in example 10. Mice were sacrificed on day 24, the number of tumor nodules on the lungs of mice was used as an indicator to examine the anti-metastatic capacity in vivo in each group, and the lung tissues of mice were collected for H&E, pathological analysis. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.
As shown in fig. 11, the tumor growth curve and the tumor weight result show that the combination of the PSN-celecoxib nanoparticles and the PSN-ticagrelor nanoparticles can inhibit the growth of in-situ tumors, and tumor slices show that the tumor tissues undergo apoptosis. From the results of the metastasis nodule counts and H & E stained sections, PSN-celecoxib nanoparticle + PSN-ticagrelor nanoparticle treated mouse lung tissue had little metastasis. In addition, the ligand-modified drug-loaded nanoparticle has good biocompatibility (fig. 12).
Example 13: the treatment effect of the ligand-modified drug-loaded nanoparticle on breast cancer postoperative metastasis is further examined, and a 4T1 mouse in-situ breast cancer model is established. Tumor-bearing BALB/c mice were operated to ablate more than 90% of tumors and 4T1 cells were injected via tail vein to mimic circulating tumor cells that invaded blood vessels during the procedure. The animals were grouped and dosed as in example 10. Wherein the PSN-ticagrelor nanoparticle and the PSN-celecoxib nanoparticle are from example 2.
As shown in fig. 13, the combined use of free celecoxib and ticagrelor has a certain effect of inhibiting postoperative tumor recurrence, and the volume and weight of the tumor recurrence 30 days after tumor resection are obviously lower than those of a normal saline group. In addition, the combination of PSN-celecoxib nanoparticles with PSN-ticagrelor nanoparticles has an optimal resistance to postoperative tumor recurrence and metastasis compared to free drug.
Claims (3)
1. A medicament for simultaneously regulating and controlling functions of platelets and tumor cells and enhancing anti-tumor metastasis effect is characterized in that: the medicine is applied to anti-tumor metastasis treatment; the medicament is prepared from nanoparticles simultaneously loaded with a platelet inhibitor and a cyclooxygenase inhibitor or respectively loaded with the platelet inhibitor and the cyclooxygenase-2 inhibitor for later combined use, wherein the nanoparticles comprise an active ingredient, a nano-carrier material and a targeting ligand, the platelet inhibitor is ticagrelor, and the cyclooxygenase-2 inhibitor is celecoxib; the carried ticagrelor Lei Zhiliang accounts for 30-40% of the total mass of the nano particles, the celecoxib accounts for 5-15% of the total mass of the nano particles, and the nano carrier material is polylactic acid-glycolic acid copolymer; the targeting ligand is PSN peptide-DAEWVDVS.
2. A medicament as claimed in claim 1, wherein: the tumor is breast cancer.
3. Use of a medicament according to any one of claims 1-2 for the preparation of a medicament having an anti-tumour metastasis effect, characterized in that: the tumor is breast cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111069967.1A CN115779101B (en) | 2021-09-13 | 2021-09-13 | Preparation and application of nanoparticle for resisting tumor metastasis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111069967.1A CN115779101B (en) | 2021-09-13 | 2021-09-13 | Preparation and application of nanoparticle for resisting tumor metastasis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115779101A CN115779101A (en) | 2023-03-14 |
| CN115779101B true CN115779101B (en) | 2024-03-15 |
Family
ID=85473377
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111069967.1A Active CN115779101B (en) | 2021-09-13 | 2021-09-13 | Preparation and application of nanoparticle for resisting tumor metastasis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115779101B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107441492A (en) * | 2016-05-30 | 2017-12-08 | 复旦大学 | The medical composition and its use of Cyclooxygenase-2 Inhibitor and Nano medication delivery system |
| WO2018141810A1 (en) * | 2017-01-31 | 2018-08-09 | Alf Lamprecht | Use of nanoparticles for immunotherapy |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10426820B2 (en) * | 2011-04-13 | 2019-10-01 | Case Western Reserve University | Synthetic platelets |
| US9107963B2 (en) * | 2012-04-13 | 2015-08-18 | Case Western Reserve University | Heteromultivalent nanoparticle compositions |
-
2021
- 2021-09-13 CN CN202111069967.1A patent/CN115779101B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107441492A (en) * | 2016-05-30 | 2017-12-08 | 复旦大学 | The medical composition and its use of Cyclooxygenase-2 Inhibitor and Nano medication delivery system |
| WO2018141810A1 (en) * | 2017-01-31 | 2018-08-09 | Alf Lamprecht | Use of nanoparticles for immunotherapy |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115779101A (en) | 2023-03-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Nanophotosensitizer-engineered Salmonella bacteria with hypoxia targeting and photothermal-assisted mutual bioaccumulation for solid tumor therapy | |
| Yin et al. | Dual receptor recognizing liposomes containing paclitaxel and hydroxychloroquine for primary and metastatic melanoma treatment via autophagy-dependent and independent pathways | |
| Cai et al. | Hybrid cell membrane-functionalized biomimetic nanoparticles for targeted therapy of osteosarcoma | |
| Yao et al. | TMZ magnetic temperature-sensitive liposomes-mediated magnetothermal chemotherapy induces pyroptosis in glioblastoma | |
| Wang et al. | Photo-responsive hydrogel facilitates nutrition deprivation by an ambidextrous approach for preventing cancer recurrence and metastasis | |
| Akbarian et al. | Biological aspects in controlling angiogenesis: current progress | |
| Long et al. | A hybrid membrane coating nanodrug system against gastric cancer via the VEGFR2/STAT3 signaling pathway | |
| Zhang et al. | Near-infrared-light induced nanoparticles with enhanced tumor tissue penetration and intelligent drug release | |
| CN111568924B (en) | Application of prussian blue in preparation of medicine for treating vascular restenosis | |
| Wang et al. | Tumor-selective lipopolyplex encapsulated small active RNA hampers colorectal cancer growth in vitro and in orthotopic murine | |
| Deng et al. | Biodegradable polymeric micelle-encapsulated doxorubicin suppresses tumor metastasis by killing circulating tumor cells | |
| CN107157950A (en) | A kind of albumin nano granular and its production and use | |
| NL2033447B1 (en) | Brain-targeting erythrocyte membrane-enveloped salvianolic acid b nanoparticles as well as preparation method and application thereof | |
| Chen et al. | Inflammatory responsive neutrophil-like membrane-based drug delivery system for post-surgical glioblastoma therapy | |
| Wang et al. | Biomimetic camouflaged nanoparticle-based folfirinox platform for optimizing clinical pancreatic cancer treatment | |
| Huo et al. | A pHe sensitive nanodrug for collaborative penetration and inhibition of metastatic tumors | |
| Ma et al. | Enhanced EPR effects by tumour stromal cell mimicking nanoplatform on invasive pituitary adenoma | |
| CN115779101B (en) | Preparation and application of nanoparticle for resisting tumor metastasis | |
| Yao et al. | Targeted long-term noninvasive treatment of choroidal neovascularization by biodegradable nanoparticles | |
| CN110339232A (en) | A kind of Chinese medicine composition prevented and/or treat ischemical reperfusion injury | |
| Zhang et al. | Dual-Targeting Biomimetic Nanomaterials for Photo-/Chemo-/Antiangiogenic Synergistic Therapy | |
| Wang et al. | Sensitizing chemotherapy for glioma with fisetin mediated by a microenvironment-responsive nano-drug delivery system | |
| Li et al. | Enhanced antitumour efficiency of R8GD-modified epirubicin plus tetrandrine liposomes in treatment of gastric cancer via inhibiting tumour metastasis | |
| CN115845059A (en) | Small molecule inhibitor nanoparticle based on polyamino acid and preparation method and application thereof | |
| Zhang et al. | Comparative study on the antitumor effects of gemcitabine polybutylcyanoacrylate nanoparticles coupled with anti-human MUC1 and CA199 monoclonal antibodies on pancreatic cancer in vitro and in vivo |
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 |