CN110152021A - A kind of medicament carrier system and preparation method thereof having target administration ability in cancer cell - Google Patents
A kind of medicament carrier system and preparation method thereof having target administration ability in cancer cell Download PDFInfo
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- CN110152021A CN110152021A CN201910563157.8A CN201910563157A CN110152021A CN 110152021 A CN110152021 A CN 110152021A CN 201910563157 A CN201910563157 A CN 201910563157A CN 110152021 A CN110152021 A CN 110152021A
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- prussian blue
- carrier system
- cancer cell
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- 239000003814 drug Substances 0.000 title claims abstract description 35
- 239000000969 carrier Substances 0.000 title claims abstract description 29
- 201000011510 cancer Diseases 0.000 title claims description 19
- 238000002360 preparation method Methods 0.000 title claims description 17
- 229960003351 Prussian blue Drugs 0.000 claims abstract description 53
- 108010039918 Polylysine Proteins 0.000 claims abstract description 31
- 229920000656 polylysine Polymers 0.000 claims abstract description 31
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 24
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 24
- 239000002105 nanoparticle Substances 0.000 claims abstract description 23
- 239000002096 quantum dot Substances 0.000 claims abstract description 23
- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims abstract description 22
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- -1 phenyl trifluoromethanesulfonate methane Chemical compound 0.000 claims abstract description 16
- 229960000304 Folic Acid Drugs 0.000 claims abstract description 14
- 235000019152 folic acid Nutrition 0.000 claims abstract description 14
- 239000011724 folic acid Substances 0.000 claims abstract description 14
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 10
- WHGYBXFWUBPSRW-FOUAGVGXSA-N β-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims abstract description 9
- 239000001116 FEMA 4028 Substances 0.000 claims abstract description 8
- 239000002840 nitric oxide donor Substances 0.000 claims abstract description 8
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims abstract description 6
- 229960004853 betadex Drugs 0.000 claims abstract description 6
- 239000003937 drug carrier Substances 0.000 claims abstract description 6
- 125000003929 folic acid group Chemical group 0.000 claims abstract description 6
- 239000011159 matrix material Substances 0.000 claims abstract description 4
- DNMNDNSFJMUUFM-UHFFFAOYSA-N Prussian blue Chemical group [Fe+3].[Fe+3].[Fe+3].[Fe+3].N#C[Fe-4](C#N)(C#N)(C#N)(C#N)C#N.N#C[Fe-4](C#N)(C#N)(C#N)(C#N)C#N.N#C[Fe-4](C#N)(C#N)(C#N)(C#N)C#N DNMNDNSFJMUUFM-UHFFFAOYSA-N 0.000 claims abstract 12
- 229920000729 poly(L-lysine) polymer Polymers 0.000 claims description 47
- 230000002194 synthesizing Effects 0.000 claims description 24
- 239000007853 buffer solution Substances 0.000 claims description 20
- 239000012265 solid product Substances 0.000 claims description 19
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-hydroxy-Succinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 17
- 239000005457 ice water Substances 0.000 claims description 17
- 238000004140 cleaning Methods 0.000 claims description 16
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 15
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 14
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 11
- 230000015572 biosynthetic process Effects 0.000 claims description 10
- 238000006011 modification reaction Methods 0.000 claims description 10
- 238000003786 synthesis reaction Methods 0.000 claims description 10
- YUFRRMZSSPQMOS-UHFFFAOYSA-N 2-(2-aminoethyldisulfanyl)ethanamine;hydron;dichloride Chemical compound Cl.Cl.NCCSSCCN YUFRRMZSSPQMOS-UHFFFAOYSA-N 0.000 claims description 9
- 230000004048 modification Effects 0.000 claims description 8
- 239000001301 oxygen Substances 0.000 claims description 7
- 229910052760 oxygen Inorganic materials 0.000 claims description 7
- 239000001184 potassium carbonate Substances 0.000 claims description 7
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 7
- MYMOFIZGZYHOMD-UHFFFAOYSA-N oxygen Chemical group O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 2
- 230000005284 excitation Effects 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims 7
- KZNICNPSHKQLFF-UHFFFAOYSA-N Succinimide Chemical compound O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 claims 1
- 150000001412 amines Chemical class 0.000 claims 1
- 150000002466 imines Chemical class 0.000 claims 1
- 229960002317 succinimide Drugs 0.000 claims 1
- 210000004027 cells Anatomy 0.000 abstract description 33
- 206010028980 Neoplasm Diseases 0.000 abstract description 15
- 230000000694 effects Effects 0.000 abstract description 15
- MWUXSHHQAYIFBG-UHFFFAOYSA-N nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 abstract description 6
- 229920000181 Ethylene propylene rubber Polymers 0.000 abstract description 5
- 238000005286 illumination Methods 0.000 abstract description 5
- 230000036457 multidrug resistance Effects 0.000 abstract description 4
- 210000004881 tumor cells Anatomy 0.000 abstract description 4
- 206010059512 Apoptosis Diseases 0.000 abstract description 3
- 230000006907 apoptotic process Effects 0.000 abstract description 3
- 230000002708 enhancing Effects 0.000 abstract description 3
- 230000001939 inductive effect Effects 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- DCYOBGZUOMKFPA-UHFFFAOYSA-N iron(2+);iron(3+);octadecacyanide Chemical compound [Fe+2].[Fe+2].[Fe+2].[Fe+3].[Fe+3].[Fe+3].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] DCYOBGZUOMKFPA-UHFFFAOYSA-N 0.000 description 42
- 239000000243 solution Substances 0.000 description 14
- BXGTVNLGPMZLAZ-UHFFFAOYSA-N N'-ethylmethanediimine;hydrochloride Chemical compound Cl.CCN=C=N BXGTVNLGPMZLAZ-UHFFFAOYSA-N 0.000 description 12
- 229940079593 drugs Drugs 0.000 description 11
- RWSXRVCMGQZWBV-WDSKDSINSA-N Glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 9
- 229960003180 Glutathione Drugs 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 238000006062 fragmentation reaction Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 231100000224 toxic side effect Toxicity 0.000 description 6
- 238000002512 chemotherapy Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 238000007626 photothermal therapy Methods 0.000 description 5
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 231100000614 Poison Toxicity 0.000 description 4
- 239000002131 composite material Substances 0.000 description 4
- 239000001963 growth media Substances 0.000 description 4
- 239000002539 nanocarrier Substances 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000002574 poison Substances 0.000 description 4
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 description 4
- 230000001225 therapeutic Effects 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-Toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 3
- 229940106305 Folic Acid 20 MG Drugs 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N acetic acid ethyl ester Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 238000011068 load Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002428 photodynamic therapy Methods 0.000 description 3
- 238000010791 quenching Methods 0.000 description 3
- 230000000171 quenching Effects 0.000 description 3
- 210000001519 tissues Anatomy 0.000 description 3
- 230000000007 visual effect Effects 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 230000001404 mediated Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 2
- 239000004810 polytetrafluoroethylene Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 231100000486 side effect Toxicity 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000004083 survival Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000024883 vasodilation Effects 0.000 description 2
- PIICEJLVQHRZGT-UHFFFAOYSA-N 1,2-ethanediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- TVFDJXOCXUVLDH-UHFFFAOYSA-N Cesium Chemical compound [Cs] TVFDJXOCXUVLDH-UHFFFAOYSA-N 0.000 description 1
- 206010010254 Concussion Diseases 0.000 description 1
- 229960003067 Cystine Drugs 0.000 description 1
- NBVXSUQYWXRMNV-UHFFFAOYSA-N Fluoromethane Chemical compound FC NBVXSUQYWXRMNV-UHFFFAOYSA-N 0.000 description 1
- 210000002216 Heart Anatomy 0.000 description 1
- 240000007871 Indigofera tinctoria Species 0.000 description 1
- 210000003734 Kidney Anatomy 0.000 description 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine zwitterion Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 description 1
- 210000004185 Liver Anatomy 0.000 description 1
- 238000010928 TGA analysis Methods 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000011358 absorbing material Substances 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 229920002892 amber Polymers 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002605 anti-dotal Effects 0.000 description 1
- 230000000259 anti-tumor Effects 0.000 description 1
- 239000000729 antidote Substances 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000003115 biocidal Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 229910052792 caesium Inorganic materials 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 230000022534 cell killing Effects 0.000 description 1
- 230000009087 cell motility Effects 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 229910052955 covellite Inorganic materials 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003247 decreasing Effects 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000029578 entry into host Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 235000003969 glutathione Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 1
- DCPMPXBYPZGNDC-UHFFFAOYSA-N hydron;methanediimine;chloride Chemical compound Cl.N=C=N DCPMPXBYPZGNDC-UHFFFAOYSA-N 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 230000003287 optical Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 230000000149 penetrating Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 229920000128 polypyrrole Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910052716 thallium Inorganic materials 0.000 description 1
- BKVIYDNLLOSFOA-UHFFFAOYSA-N thallium Chemical compound [Tl] BKVIYDNLLOSFOA-UHFFFAOYSA-N 0.000 description 1
- 238000002411 thermogravimetry Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0042—Photocleavage of drugs in vivo, e.g. cleavage of photolabile linkers in vivo by UV radiation for releasing the pharmacologically-active agent from the administered agent; photothrombosis or photoocclusion
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET 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 TOILET 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
- A61K47/54—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 compound
- A61K47/545—Heterocyclic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET 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
- A61K47/61—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 the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET 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
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
- A61K47/645—Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
- A61K49/0065—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle
- A61K49/0067—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the luminescent/fluorescent agent having itself a special physical form, e.g. gold nanoparticle quantum dots, fluorescent nanocrystals
-
- 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 discloses a kind of Drug Carrier Systems, be modified with carbon quantum dot and beta cyclodextrin it is Prussian blue for matrix, after being grafted in conjunction with polylysine with the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of NO donor 5-, then obtained from modified with folic acid.Medicament carrier system provided by the invention can realize effective enrichment in tumor locus by folate-targeted and tumor locus EPR effect, and under near-infrared laser irradiation, prussian blue nano particle can pass through excellent photothermal conversion efficiency ablated tumor cell.Simultaneously, the medicament carrier system can control nitric oxide production release under the illumination of 400nm wavelength, enhance nanoparticle in the enrichment of tumor locus so as to improve EPR effect, NO can inhibit tumour growth with inducing apoptosis of tumour cell, reverse multidrug resistance etc. simultaneously.
Description
Technical field
The invention belongs to pharmaceutical carrier fields, and in particular to a kind of pharmaceutical carrier for having target administration ability in cancer cell
System and preparation method thereof.
Background technique
In recent years, with the deterioration of natural environment, the disease incidence of cancer is higher and higher.Traditional anti-cancer therapies packet
It includes: operative treatment, chemotherapy and radiation.Although can extend the life of patient to a certain extent, these therapeutic modalities are general
The disadvantages of store-through is larger in toxic side effect, damages normal tissue and larger wound, to limit its therapeutic effect to tumour.
Therefore, new therapeutic modality is developed to get growing concern for.
Currently, chemotherapy is antitumor primary treatment scheme, but challenge of the chemotherapy by multidrug resistance (MDR), this is very
The curative effect of chemotherapy is limited in big degree.And often using high dose and increase administration frequency.But high dose and increase administration
Frequency will not significantly improve therapeutic effect, often bring instead to vitals (heart, liver and kidney) serious bad
Side effect, it is possible to further deteriorate drug resistance.Compared with traditional small-molecule drug chemotherapy of tumors, since near-infrared irradiation is set
It sets controllably, such as irradiation time, light source position, power output, so photo-thermal therapy (PTT) photodynamic therapy (PDT) is with low
The characteristics of poison, high specific.
Photo-thermal therapy (PTT) is a kind of new tumor therapeuticing method, has Noninvasive and space-time controllability.Photo-thermal therapy
Basic principle be: optothermal material by EPR effect tumor locus assemble, then under near infrared light, make tumour office
Portion's temperature is increased more than 42 DEG C, so that tumor cell damage be made even to be eliminated tumour cell, achievees the purpose that treating cancer.Pu Lu
Scholar's indigo plant (PB) due to special ion exchange, absorption and mechanical trapping characteristic, food and drug administration (FDA) in
Antidote of the approval lead in 2003 as thallium and caesium inner radiation contact scar, shows its good biological safety and biofacies
Capacitive.The nm near-infrared absorbing material (such as polypyrrole nanoparticle, gold nanoparticle, CuS nanoparticle) emerging with other
It compares, PB nanoparticle has good biocompatibility, higher photothermal conversion efficiency, photostability and easily-controllable size etc. excellent
Point.
Photodynamic therapy (PDT) is as a kind of emerging treatment means, since its invasion is small, Small side effects, drug resistance
The advantages that small, receives more and more attention in oncotherapy.Current optical dynamic therapy multi-pass crosses active oxygen (ROS) induction
Apoptosis, necrosis and disorganization.Unfortunately, angiogenesis abnormal in solid tumor and thrombosis will lead to oxygen supply with
Oxygen consumption is unbalance, causes high-caliber anoxic.The low-oxygen environment of tumor tissues may be decreased the concentration of effective oxygen in PDT, limit ROS
Generation.The generation of NO then needs not rely on the oxygen in tumor tissues, and the vasodilation that can be mediated by NO, mitigates swollen
The anoxic at tumor position.
Summary of the invention
The technical problem to be solved by the present invention is to provide a kind of drug load in view of the deficiency of the prior art
System system, compared to traditional load drug carrier, which has target administration ability in more efficient cancer cell, can effectively mention
High utilization ratio of drug simultaneously reduces its toxic side effect.
The present invention be solve the problems, such as it is set forth above used by technical solution are as follows:
Medicament carrier system provided by the present invention, be modified with carbon quantum dot and beta cyclodextrin it is Prussian blue for matrix,
After being grafted in conjunction with polylysine with the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of NO donor 5-, then obtained from modified with folic acid.
Said medicine carrier system, preparation method mainly include the following steps:
Step 1 synthesizes the Prussian blue (PB-CD-NH of beta cyclodextrin and 2-aminoethyl disulfide dihydrochloride modification2)
Prussian blue (PB) is dispersed in the buffer solution that pH is 5-6, then under conditions of ice-water bath, 1- is added
(3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDC) and n-hydroxysuccinimide react 20-30 hours；So
Afterwards, it is subsequently added into beta cyclodextrin (EDA- β-CD) and 2-aminoethyl disulfide dihydrochloride reacts at room temperature 20-30 hours, separate and clean solid production
Object obtains PB-CD-NH2；
Step 2, Prussian blue (PB-C-dots-CD) of synthesis carbon quantum dot modification
Carbon quantum dot (C-dots) is dispersed in the buffer solution that pH is 5-6, then under conditions of ice-water bath, is added
EDC and n-hydroxysuccinimide react 20-30 hours；Then, it is subsequently added into PB-CD-NH2, it is small that 20-30 is reacted at 25 DEG C
When, solid product is separated and cleaned, PB-C-dots-CD is obtained；
Step 3, chloro- 2- nitre phenyl trifluoromethanesulfonate methane grafting polylysine (PLL (NF)) of synthesis NO donor 5-
Polylysine (PLL), potassium carbonate and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of 5- are dissolved in solvent DMF, 25 DEG C to 35
It flows back at DEG C back flow reaction three days 72-96 hours, sufficiently after dialysis, freeze-drying obtains PLL (NF)；
Step 4, the Prussian blue (PB- of the chloro- 2- nitre phenyl trifluoromethanesulfonate methane grafting Mercapto-group modification of synthesis NO donor 5-
C-dots-CD-PLL(NF))
PB-C-dots-CD and PLL (NF) are dispersed in the buffer solution that pH is 7.2~7.4, are stirred to react under room temperature
45-55 hours, solid product is separated, obtains PB-C-dots-CD-PLL (NF) after washing is dry；
Step 5 synthesizes Prussian blue (PB-C-dots-CD-PLL (the NF)-FA) of modified with folic acid
Folic acid is dissolved in the buffer solution that pH is 5-6, then under conditions of ice-water bath, 1- (3- diformazan ammonia is added
Base propyl) after -3- ethyl-carbodiimide hydrochloride (EDC) and n-hydroxysuccinimide react 20-30 hours, add PB-
C-dots-CD-PLL (NF) isolates solid product, as modified with folic acid is Prussian blue in room temperature reaction 20-30 hours
(PB-C-dots-CD-PLL(NF)-FA)。
According to the above scheme, the Prussian blue prussian blue nano particle for citric acid surface modification, partial size is in 30nm-
Within the scope of 200nm.
According to the above scheme, the excitation wavelength of carbon quantum dot is in 400nm-660nm.
According to the above scheme, in step 1, the Prussian blue concentration in buffer solution is within the scope of 1-2.5mg/mL.
According to the above scheme, in step 1,1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride, N- hydroxyl amber
Concentration of the amber acid imide in buffer solution is 4-6mg/mL；EDA- β-CD, 2-aminoethyl disulfide dihydrochloride and Prussian blue mass ratio
Respectively (6-9): (1-4): 10.
According to the above scheme, in step 2, concentration of the carbon quantum dot in buffer solution is within the scope of 1-20mg/ml.
According to the above scheme, in step 2, the concentration of EDC, n-hydroxysuccinimide in buffer solution is 15-30mg/
ml；PB-CD-NH2Mass ratio with carbon quantum dot is (1-10): 1.
According to the above scheme, in step 3, the mass ratio of potassium carbonate and PLL are 1:(1-3)；PLL and the chloro- 2- nitre phenyl three of 5-
The mass ratio of fluoromethane is 1:(0.5-4).
According to the above scheme, in step 4, the concentration range difference of PB-C-dots-CD and PLL (NF) in buffer solution is equal
For 1-2mg/mL.
According to the above scheme, in step 5, folic acid is dissolved in the buffer solution that pH is 5-6, concentration 0.1-0.7mg/ml.
According to the above scheme, in step 5, the concentration of EDC, n-hydroxysuccinimide in buffer solution is 5-15mg/
ml；The mass ratio of PB-C-dots-CD-PLL (NF) and folic acid is 10:(0.5-3).
Compared with prior art, the beneficial effects of the present invention are:
Firstly, medicament carrier system provided by the invention, can be realized by folate-targeted and tumor locus EPR effect
Effective enrichment of tumor locus, under near-infrared laser irradiation, prussian blue nano particle can pass through excellent photothermal conversion
Efficiency ablated tumor cell.Meanwhile the medicament carrier system can control nitric oxide production release under the illumination of 400nm wavelength
It puts, enhances nanoparticle in the enrichment of tumor locus so as to improve EPR effect, while NO can be inverse with inducing apoptosis of tumour cell
Turn multidrug resistance etc. and inhibits tumour growth.
Second, NO donor is discharged by 400nm laser controlling in the medicament carrier system, will not be discharged, be can control in advance
Its toxic side effect to normal cell；And the vasodilation that NO can be mediated, so that carrier is more enriched in tumor locus, simultaneously
Mitigate the anoxic of tumor locus.
Third, it is Prussian blue can with carbon quantum dot formed fluorescent quenching, the carbon quantum that the present invention is grafted by cystine linkage
Point is modified Prussian blue, and the GSH response in tumour, which may be implemented, causes fluorescent quenching to disappear, and realizes the fixed point prison of drug
Depending on tracking carrier well in vivo, there is visual navigation.Since glutathion inside cell (GSH) concentration is extracellular
1000 times of GSH concentration in matrix, fluorescent quenching disappears in tumour cell, realizes fluorescent visual.
4th, polylysine has good biocompatibility, also has a large amount of unique film characters, can enhance cell pair
High molecular absorption；Folic acid has good targeting, and absorption of the tumour cell to carrier can be enhanced.
By the design of above series of, the present invention can construct one kind and have " tumour causes targeting " ability, based on PLL
(NF) then-FA/ β-CD/PB composite Nano medicament carrier system, the fluorescent visual in tumour cell apply exogenous stimulation
Discharge nitric oxide and photo-thermal ablated tumor cell, it can be ensured that normal cell will not be hurt, compared to traditional load drug carrier,
The system has target administration ability in more efficient cancer cell, can effectively improve utilization ratio of drug and reduces the secondary work of its poison
With.
Detailed description of the invention
The TEM figure that Fig. 1 is prussian blue nano particle PB used by embodiment；
The TEM figure that Fig. 2 is carbon quantum dot C-dots used by embodiment；
Fig. 3 is used by embodiment in 1W/cm2, under 808nm laser irradiation various concentration prussian blue nano particle
The photo-thermal figure of PB；
Fig. 4 is used by embodiment in 1W/cm2, 0.25mg/ml prussian blue nano particle under 808nm laser irradiation
The solar thermochemical cycle figure of PB；
Fig. 5 is the nuclear-magnetism figure of PLL (NF) used by embodiment；
The NO release that Fig. 6 is PLL (NF) used by embodiment；
Fig. 7 is the infrared figure of Fourier of each step products in embodiment 1；
Fig. 8 is the thermogravimetric analysis figure of each step products in embodiment 1；
Fig. 9 is the fluorescence of 1 products therefrom GSH of embodiment before and after the processing；
Figure 10 is that the cell of nano-carrier used by embodiment 1 absorbs the relationship with the time；
Figure 11 is photo-thermal fragmentation effect of nano-carrier when 50ug/mL and 100ug/mL used by embodiment 1；
Figure 12 is the MTT toxicity detection of nano-carrier used by embodiment 1.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but the present invention is not
It is limited only to the following examples.
PB, β-CD-OTs, EDA- β-CD, the C-dots being related in following embodiments can be prepared with the following method,
It can also be prepared using other methods.
1, the synthesis (PB) of prussian blue nano particle
It is prepared for the prussian blue nano particle (PB NPs) of citric acid surface modification in the following way, it is specific to prepare
Steps are as follows: by the FeCl of a certain amount3And K4[Fe(CN)6] it is added separately to the 20mL deionization containing 0.5mmol citric acid
In water, make FeCl3And K4[Fe(CN)6] final concentration be 1.0mmol/L；Then by above-mentioned FeCl3Solution and K4[Fe(CN)6]
Solution is separately heated to 60 DEG C, and by K4[Fe(CN)6] solution in 60 DEG C stirring under be added dropwise to FeCl3In solution；With dropwise addition
It carries out, solution can gradually become blue, be cooled to room temperature after which is persistently continued stirring 30min.
The acetone isometric with above-mentioned blue solution is added thereto, 30min is placed at room temperature, then in 12500r/m
Nanoparticle is collected after centrifugation 60min, and is washed repeatedly 3 times；The prussian blue nano particle of collection is finally set into vacuum drying
In case, dry 12h, spare under the conditions of 50 DEG C.
2. (1) preparation (β-CD-OTs) of tolysulfonyl oxygen group-beta-cyclodextrin
It weighs 25g beta-cyclodextrin (β-CD) and is dissolved in the NaOH solution of 300mL 0.4M, stirred in ice-water bath
It mixes until cyclodextrin is completely dissolved；The paratoluensulfonyl chloride (TsCl) of 18g is slowly added dropwise into aforementioned β-CD solution, in ice water
After being stirred to react 90min in bath, then filter；Taken filtrate adjusts its pH to 8.5 with HCl, then continues to be stirred to react at room temperature
2 hours, products therefrom was placed in refrigerator (4 DEG C) one nights of placement, is filtered, filter residue is washed with deionized three times, at 60 DEG C
After drying, β-CD-OTs is obtained.
(2) EDA- β-CD is synthesized
It weighs the β-CD-OTs 2.5g prepared and is added to ethylenediamine 15ml steams again after, dissolve it at normal temperature
And logical nitrogen 30min or so, it places into and continues back flow reaction 48h under nitrogen protection in 80 DEG C of water-baths, then simultaneously with acetone precipitation
It is filtered with organic filter membrane of 450nm, finally continues with acetone precipitation twice, to obtain yellowish white powder air drying with water dissolution
50h obtains the product EDA- β-CD of about 2.2g, and yield is about 95.7%
3. the synthesis (C-dots) of carbon quantum dot
It weighs 1.6g citric acid and 0.6g urea and is dissolved in the DMF of 10ml, then pour into the mixed solution
It into 20mL polytetrafluoroethylene (PTFE) autoclave, is reacted 12 hours at 200 DEG C, is centrifuged off solid after reaction, on gained
Clear liquid is cleaned three times with (petroleum ether: ethyl acetate=4:1) mixed liquor, and 12h is dried in vacuo at 70 DEG C can be obtained carbon quantum dot.
Prussian blue nano particle and carbon quantum dot are diluted to certain multiple, with its form of transmission electron microscope observing, such as Fig. 1
With shown in Fig. 2, carrier is the cube structure that partial size is about 45nm, and carbon quantum dot partial size is 3~4nm.
Fig. 3 is photo-thermal test of the prussian blue nano particle under various concentration, from the figure 3, it may be seen that prussian blue nano grain
Son has good light thermal property.
Fig. 4 is that the photo and thermal stability of prussian blue nano particle is tested, and as shown in Figure 4, prussian blue nano particle has good
Good photo and thermal stability, can be with Repeat-heating.
Fig. 5 is the nuclear-magnetism figure of PLL (NF), and as shown in Figure 7, the integral of PLL (NF), which compares, meets expection with chemical shift, shows
The synthesis success of PLL (NF).
Fig. 6 is the release experiment of NO, and under 400nm illumination, NO largely discharges, and also demonstrates the synthesis of PLL (NF) and connects
Branch success.
Embodiment 1
A kind of preparation method of composite Nano medicament carrier system, specifically comprises the following steps:
1. synthesizing PB-CD-NH2
It weighs PB 200mg to be dispersed in the PBS that the pH of 100ml is 5.5, then under the conditions of 4 DEG C of ice-water bath, EDC is added
(1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride 2.87g and n-hydroxysuccinimide 1.7262g, reaction 24
Hour；It is subsequently added into 140mgEDA- β-CD and 60mg 2-aminoethyl disulfide dihydrochloride to react 24 hours, eccentric cleaning obtained solid product,
As PB-CD-NH2。
2. synthesizing PB-C-dots-CD
It weighs carbon quantum dot 60mg to be dispersed in the PBS that the pH of 100ml is 5.5, EDC then is added at 4 DEG C of ice-water bath
1.7262g and n-hydroxysuccinimide 2.87g simultaneously reacts 24 hours, adds 200mg PB-CD-NH2 room temperature reaction for 24 hours,
Eccentric cleaning obtained solid product, as PB-C-dots-CD.
3. synthesizing PLL (NF)
200mg PLL, 100mg potassium carbonate and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane 200mg of 5- is weighed to flow back in 25mL DMF
It three days, dialyses three days at room temperature, changes within every 6 hours a deionized water, freeze-drying obtains PLL (NF).
4. synthesizing PB-C-dots-CD-PLL (NF)
100mg PB-C-dots-CD and 100mgPLL (NF) are weighed, is dispersed in the PBS buffer solution of 60ml
(pH7.4), it is stirred to react under room temperature two days, solid product is centrifuged out, cleaned for several times with deionized water, vacuum drying obtains
Product PB-C-dots-CD-PLL (NF).
5. synthesizing PB-C-dots-CD-PLL (NF)-FA
It weighs folic acid 20mg to be dissolved in the PBS that the pH of 200ml is 5.5, then under the conditions of 4 DEG C of ice-water bath, EDC is added
1.7262g and n-hydroxysuccinimide 2.87g reacts 24 hours, adds 200mg PB-C-dots-CD-PLL (NF), room
Temperature is reacted for 24 hours, eccentric cleaning obtained solid product, as PB-C-dots-CD-PLL (NF)-FA.
As shown in Figure 7, in above-mentioned steps, the stretching vibration peak of the specific groups of each step grafting it is observed that, show
Every single step reaction is successful.In conjunction with Fig. 8 it is found that the TG of grafting carbon quantum dot rises, the TG after other each step grafting declines,
Show the grafting success of nanoparticle.
Using test
1, influence of the GSH presence or absence to PB-C-dots-CD-PLL (NF)-FA: suitable PB-CD-C-dots-PLL is taken
(NF) the PBS buffer solution that-FA is dispersed in pH=7.4 respectively neutralizes the PBS buffer solution of the pH=7.4 containing 10mM GSH
In.It is placed in 37 degrees Celsius of shaking table 24 hours, is centrifuged, washing detects fluorescence.
As shown in Figure 9, PB-C-dots-CD-PLL (NF)-FA does not have fluorescence in the presence of no GSH, in the presence of GSH
Fluorescence occurs, and shows the grafting success of nanoparticle.
2, it inquires into cell to absorb and co-culture the relationship between the time: by HeLa cell with 1.0 × 104Cells/well's
Concentration kind enters in 6 orifice plates.After culture for 24 hours, the culture solution in original each orifice plate is substituted for the DMEM packet of fresh 1mL respectively
Containing PB-CD-C-dots-PLL (NF)-FA, PB-CD-C-dots-PLL (NF) and PB-CD-C-dots-PLL-FA, with its institute
Concentration containing PB is counted, and PB concentration is 100 μ g/mL-1.After cell incubation different time, PBS is washed three times, trypsinized,
5min is collected by centrifugation in 1100rpm, finally, being handled with concentrated nitric acid solution.Contained with ICP-MS (PerkinElmer company, the U.S.) test
Iron.
As shown in Figure 10, when the 8th hour, cell is to the uptake highest of nano-carrier, and PB-CD-C-
Dots-PLL (NF)-FA's and PB-CD-C-dots-PLL-FA is above PB-CD-C-dots-PLL (NF), can effectively demonstrate,prove
The targeting of bright folic acid.
3, photo-thermal cytotoxicity experiment carries out photo-thermal effect evaluation using HeLa cell.By HeLa cell with 1.0 ×
104The concentration kind of cells/well enters in 6 orifice plates, and 1mL is added in each orifice plate and contains 10%FBS and 1% antibiotic
DMEM culture medium.It include PBS by the DMEM that the culture solution in original each orifice plate is substituted for fresh 1mL respectively after culture for 24 hours
(control group), PB-CD-C-dots-PLL (NF)-FA, PB-CD-C-dots-PLL (NF) and PB-CD-C-dots-PLL-FA,
In terms of the concentration of PB contained by it is come, net PB concentration maintains always 100 μ g/mL.After cultivating 24 hours, it is with power density
1w/cm-2808nm laser irradiation 5min after, continue culture for 24 hours.Cell is washed three times with PBS, with depositing for mtt assay measurement cell
Motility rate.
As shown in Figure 11, control group is using 1W/cm2, under 808nm laser irradiation without obvious killing, it was demonstrated that laser photograph
Biggish harm will not be had to cell by penetrating.And the fragmentation effect of PB-CD-C-dots-PLL (NF)-FA is best, PB-CD-C-
The fragmentation effect of dots-PLL (NF) is secondly, the fragmentation effect of PB-CD-C-dots-PLL-FA is worst, the fragmentation effect of high concentration
Higher than low concentration.Drug has good targeting according to Figure 10 and Figure 12, and drug itself is without the secondary work of biggish poison
With.But drug NO can effectively discharge under light illumination, kill cell, and have good photo-thermal fragmentation effect.It demonstrates
The system has target administration ability in more efficient cancer cell, can effectively improve utilization ratio of drug and reduces the secondary work of its poison
With.
4, HeLa cell is inoculated in 96 orifice plates of 96 orifice plates with the density of 8000 cells/wells, culture for 24 hours, is added
PB-CD-C-dots-PLL (NF)-FA, PB-CD-C-dots-PLL (NF) and PB-CD-C-dots-PLL-FA of various concentration with
After cell co-cultures 48h, after culture, the fresh DMEM culture medium of 200 μ L and 20 μ L MTT solution will be changed in culture medium
(5mg/mL PBS).Continue after cultivating 4h, MTT solution is carefully removed, adds 150 μ L DMSO into hole, culture medium pallet
Concussion is rocked at room temperature, is uniformly mixed.The each hole (Model 550, Bio-Rad, USA) is recorded in microplate reader in 570nm
Absorbance, cell survival rate calculation formula is as follows:
Wherein OD570(control)It is the light absorption value measured when material not being added, OD570(treated)It is measured after material is added
Light absorption value.
As shown in Figure 12, the cell survival rate of the carrier system is above 90%, has good biocompatibility and nothing
Obvious toxic-side effects.
Embodiment 2
A kind of preparation method of composite Nano medicament carrier system, specifically comprises the following steps:
1. synthesizing PB-CD-NH2
PB 200mg is weighed to be dispersed in 100ml deionized water, with salt acid for adjusting pH to 5-6 (if adjust be lower than 5, can
A small amount of sodium hydroxide readjustment is added), then under the conditions of 4 DEG C of ice-water bath, EDC (1- (3- dimethylamino-propyl) -3- ethyl is added
Carbodiimide hydrochloride 2.87g and n-hydroxysuccinimide 1.7262g reacts 24 hours；It is subsequently added into 140mgEDA- β-CD
It is reacted 24 hours with 60mg 2-aminoethyl disulfide dihydrochloride, eccentric cleaning obtained solid product, as PB-CD-NH2.
2. synthesizing PB-C-dots-CD
It weighs carbon quantum dot 60mg to be dispersed in water, with salt acid for adjusting pH to 5-6, then be added at 4 DEG C of ice-water bath
EDC1.7262g and n-hydroxysuccinimide 2.87g simultaneously reacts 24 hours, adds 200mg PB-CD-NH2 room temperature reaction
For 24 hours, eccentric cleaning obtained solid product, as PB-C-dots-CD.
3. synthesizing PLL (NF)
200mg PLL, 100mg potassium carbonate and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane 200mg of 5- is weighed to flow back in 25mL DMF
It three days, dialyses three days at room temperature, freeze-drying obtains PLL (NF).
4. synthesizing PB-C-dots-CD-PLL (NF)
100mg PB-C-dots-CD and 100mgPLL (NF) are weighed, is dispersed in the PBS buffer solution of 60ml
(pH7.4), it is stirred to react under room temperature two days, solid product is centrifuged out, cleaned for several times with deionized water, vacuum drying obtains
Product PB-C-dots-CD-PLL (NF).
5. synthesizing PB-C-dots-CD-PLL (NF)-FA
It weighs folic acid 20mg to be dissolved in 200ml water, adjusts PH to 5-6 with hydrochloric acid, then under the conditions of 4 DEG C of ice-water bath, add
Enter EDC 1.7262g and n-hydroxysuccinimide 2.87g to react 24 hours, adds 200mg PB-C-dots-CD-PLL
(NF), 48h, eccentric cleaning obtained solid product, as PB-C-dots-CD-PLL (NF)-FA are reacted at room temperature.
Embodiment 3
A kind of preparation method of composite Nano medicament carrier system, specifically comprises the following steps:
1. synthesizing PB-CD-NH2
It weighs PB 200mg to be dispersed in the PBS that the pH of 100ml is 5.5, then under the conditions of 4 DEG C of ice-water bath, EDC is added
(1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride 2.87g and n-hydroxysuccinimide 1.7262g, reaction 24
Hour；It is subsequently added into 180mgEDA- β-CD and 20mg 2-aminoethyl disulfide dihydrochloride to react 24 hours, eccentric cleaning obtained solid product,
As PB-CD-NH2。
2. synthesizing PB-C-dots-CD
It weighs carbon quantum dot 60mg to be dispersed in the PBS that the pH of 100ml is 5.5, EDC then is added at 4 DEG C of ice-water bath
1.7262g and n-hydroxysuccinimide 2.87g simultaneously reacts 24 hours, adds 200mg PB-CD-NH2 room temperature reaction for 24 hours,
Eccentric cleaning obtained solid product, as PB-C-dots-CD.
3. synthesizing PLL (NF)
200mg PLL, 100mg potassium carbonate and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane 600mg of 5- is weighed to flow back in 25mL DMF
It three days, dialyses three days at room temperature, freeze-drying obtains PLL (NF).
4.PB-C-dots-CD-PLL(NF)
100mg PB-C-dots-CD and 100mgPLL (NF) are weighed, is dispersed in the PBS buffer solution of 60ml
(pH7.4), it is stirred to react under room temperature two days, solid product is centrifuged out, cleaned for several times with deionized water, vacuum drying obtains
Product PB-C-dots-CD-PLL (NF).
5. synthesizing PB-C-dots-CD-PLL (NF)-FA
It weighs folic acid 20mg to be dissolved in the PBS that the pH of 200ml is 5.5, then under the conditions of 4 DEG C of ice-water bath, EDC is added
1.7262g and n-hydroxysuccinimide 2.87g reacts 24 hours, adds 200mg PB-C-dots-CD-PLL (NF), room
Temperature is reacted for 24 hours, eccentric cleaning obtained solid product, as PB-C-dots-CD-PLL (NF)-FA.
The detection method of each intermediate product and final product obtained by embodiment 2-3 is same as Example 1, it is verified at
Medicament carrier system is made in function, has good targeting, and itself without biggish toxic side effect, NO can be effective under light illumination
Release, kill cell, and have good photo-thermal fragmentation effect.The medicament carrier system of the present invention is demonstrated to have more
Add target administration ability in efficient cancer cell, can effectively improve utilization ratio of drug and reduces its toxic side effect.
The above is only a preferred embodiment of the present invention, it is noted that come for those of ordinary skill in the art
It says, without departing from the concept of the premise of the invention, several modifications and variations can also be made, these belong to of the invention
Protection scope.
Claims (10)
1. a kind of medicament carrier system for having target administration ability in cancer cell, it is characterised in that it is with carbon quantum dot and β
It is cyclodextrin modified it is Prussian blue be matrix, by being grafted in conjunction with polylysine with the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of NO donor 5-
Afterwards, then obtained from modified with folic acid.
2. the preparation method described in claim 1 for having the medicament carrier system of target administration ability in cancer cell, feature
It is mainly to include the following steps:
Step 1 synthesizes the Prussian blue PB-CD-NH of beta cyclodextrin and 2-aminoethyl disulfide dihydrochloride modification2
Prussian blue PB is dispersed in the aqueous solution that pH is 5-6, then under conditions of ice-water bath, EDC and N- hydroxyl is added
Succinimide reacts 20-30 hours；Then, tolysulfonyl oxygen group-beta-cyclodextrin and 2-aminoethyl disulfide dihydrochloride room are subsequently added into
Temperature reaction 20-30 hours, separates and cleans solid product, obtain PB-CD-NH2；
Step 2, the Prussian blue PB-C-dots-CD of synthesis carbon quantum dot modification
Carbon quantum dot C-dots is dispersed in the aqueous solution that pH is 5-6, then under conditions of ice-water bath, EDC and N- is added
HOSu NHS reacts 20-30 hours；Then, it is subsequently added into PB-CD-NH2, react 20-30 hours, separate at 25 DEG C
And solid product is cleaned, obtain PB-C-dots-CD；
Step 3, chloro- 2- nitre phenyl trifluoromethanesulfonate methane grafting polylysine PLL (NF) of synthesis NO donor 5-
Polylysine PLL, potassium carbonate and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of 5- are dissolved in solvent DMF, flowed back at 25-35 DEG C
Reaction 72-96 hours, sufficiently after dialysis, freeze-drying obtains PLL (NF)；
Step 4, the Prussian blue PB-C- of the chloro- 2- nitre phenyl trifluoromethanesulfonate methane grafting Mercapto-group modification of synthesis NO donor 5-
dots-CD-PLL(NF)
PB-C-dots-CD and PLL (NF) are dispersed in the buffer solution that pH is 7.2-7.4, are stirred to react 45-55 under room temperature
Hour, solid product is separated, obtains PB-C-dots-CD-PLL (NF) after washing is dry；
Step 5 synthesizes Prussian blue PB-C-dots-CD-PLL (NF)-FA of modified with folic acid
Folic acid is dissolved in the aqueous solution that pH is 5-6, then under conditions of ice-water bath, EDC and N- hydroxysuccinimidyl acyl is added
After imine reaction 20-30 hours, PB-C-dots-CD-PLL (NF) is added in room temperature reaction 20-30 hours, isolates solid
Product, as PB-C-dots-CD-PLL (NF)-FA.
3. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 1, the Prussian blue prussian blue nano particle for citric acid surface modification, partial size is in 30nm-200nm
In range；In step 2, the excitation wavelength of carbon quantum dot is in 400nm-660nm.
4. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 1, Prussian blue concentration in aqueous solution is within the scope of 1-2.5mg/mL；1- (3- dimethylamino third
Base) -3- ethyl-carbodiimide hydrochloride, the concentration of n-hydroxysuccinimide in aqueous solution is 15-30mg/mL；EDA-
β-CD, 2-aminoethyl disulfide dihydrochloride and Prussian blue mass ratio are respectively (6-9): (1-4): 10.
5. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 2, the concentration of carbon quantum dot in aqueous solution is within the scope of 1-20mg/ml；EDC, N- hydroxysuccinimidyl acyl are sub-
Concentration of the amine in buffer solution is 15-30mg/ml；PB-CD-NH2Mass ratio with carbon quantum dot is (1-10): 1.
6. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 3, the mass ratio of potassium carbonate and PLL are 1:(1-3)；The matter of PLL and the chloro- 2- nitre phenyl trifluoromethanesulfonate methane of 5-
Amount is than being 1:(0.5-4).
7. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 4, the concentration range of PB-C-dots-CD and PLL (NF) in buffer solution is 1-2mg/mL respectively.
8. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 5, folic acid is dissolved in the aqueous solution that pH is 5-6, concentration 0.1-0.7mg/ml.
9. the preparation method according to claim 2 for having the medicament carrier system of target administration ability in cancer cell,
It is characterized in that in step 5, the concentration of EDC, n-hydroxysuccinimide in buffer solution is 5-15g/ml；PB-C-
The mass ratio of dots-CD-PLL (NF) and folic acid is 10:(0.5-3).
10. the pharmaceutical carrier for having target administration ability in cancer cell of any one of claim 2-9 the method preparation
System.
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