CN108042509A - Nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification and preparation method and application - Google Patents
Nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification and preparation method and application Download PDFInfo
- Publication number
- CN108042509A CN108042509A CN201810136097.7A CN201810136097A CN108042509A CN 108042509 A CN108042509 A CN 108042509A CN 201810136097 A CN201810136097 A CN 201810136097A CN 108042509 A CN108042509 A CN 108042509A
- Authority
- CN
- China
- Prior art keywords
- particle
- nano
- mesoporous silica
- spnc
- msn
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 title claims abstract description 176
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 156
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 115
- 239000000377 silicon dioxide Substances 0.000 title claims abstract description 81
- 230000004048 modification Effects 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 238000002715 modification method Methods 0.000 title 1
- 238000012986 modification Methods 0.000 claims abstract description 38
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 31
- 238000001338 self-assembly Methods 0.000 claims abstract description 25
- 229910052814 silicon oxide Inorganic materials 0.000 claims abstract description 14
- OJBXRIGCBKNSQC-UHFFFAOYSA-N 2-[[2-[[2-[(2-methylpropan-2-yl)oxycarbonylamino]-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]acetic acid Chemical group C=1C=CC=CC=1CC(C(=O)NCC(O)=O)NC(=O)C(NC(=O)OC(C)(C)C)CC1=CC=CC=C1 OJBXRIGCBKNSQC-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000003937 drug carrier Substances 0.000 claims abstract description 11
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 77
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 76
- 239000007788 liquid Substances 0.000 claims description 56
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 52
- 239000000243 solution Substances 0.000 claims description 51
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 42
- 229910052757 nitrogen Inorganic materials 0.000 claims description 38
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 36
- 229910017163 MnFe2O4 Inorganic materials 0.000 claims description 36
- 239000006185 dispersion Substances 0.000 claims description 34
- 235000019441 ethanol Nutrition 0.000 claims description 34
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 29
- 239000011572 manganese Substances 0.000 claims description 29
- 238000010438 heat treatment Methods 0.000 claims description 28
- 229910052748 manganese Inorganic materials 0.000 claims description 27
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 claims description 26
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N iron oxide Inorganic materials [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 claims description 23
- HEDRZPFGACZZDS-UHFFFAOYSA-N CHCl3 Substances ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 22
- 239000007795 chemical reaction product Substances 0.000 claims description 22
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 20
- 229910017052 cobalt Inorganic materials 0.000 claims description 20
- 239000010941 cobalt Substances 0.000 claims description 20
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 claims description 20
- 235000013339 cereals Nutrition 0.000 claims description 19
- 238000010992 reflux Methods 0.000 claims description 16
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 15
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 15
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 14
- 238000001704 evaporation Methods 0.000 claims description 14
- 230000008020 evaporation Effects 0.000 claims description 14
- 239000002245 particle Substances 0.000 claims description 12
- PZJJKWKADRNWSW-UHFFFAOYSA-N trimethoxysilicon Chemical compound CO[Si](OC)OC PZJJKWKADRNWSW-UHFFFAOYSA-N 0.000 claims description 12
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 11
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 11
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 11
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 11
- 239000005642 Oleic acid Substances 0.000 claims description 11
- ZITKDVFRMRXIJQ-UHFFFAOYSA-N dodecane-1,2-diol Chemical class CCCCCCCCCCC(O)CO ZITKDVFRMRXIJQ-UHFFFAOYSA-N 0.000 claims description 11
- WTFXARWRTYJXII-UHFFFAOYSA-N iron(2+);iron(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Fe+2].[Fe+3].[Fe+3] WTFXARWRTYJXII-UHFFFAOYSA-N 0.000 claims description 11
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 11
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 11
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 11
- -1 propyl trimethoxy silicon Alkane Chemical class 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 11
- CUJRVFIICFDLGR-UHFFFAOYSA-N acetylacetonate Chemical compound CC(=O)[CH-]C(C)=O CUJRVFIICFDLGR-UHFFFAOYSA-N 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- QGLWBTPVKHMVHM-KTKRTIGZSA-N (z)-octadec-9-en-1-amine Chemical compound CCCCCCCC\C=C/CCCCCCCCN QGLWBTPVKHMVHM-KTKRTIGZSA-N 0.000 claims description 9
- DDEHZCWFAUZBEH-UHFFFAOYSA-O azanium;ethanol;nitrate Chemical compound [NH4+].CCO.[O-][N+]([O-])=O DDEHZCWFAUZBEH-UHFFFAOYSA-O 0.000 claims description 9
- 125000001743 benzylic group Chemical group 0.000 claims description 9
- 229910052742 iron Inorganic materials 0.000 claims description 9
- LZKLAOYSENRNKR-LNTINUHCSA-N iron;(z)-4-oxoniumylidenepent-2-en-2-olate Chemical compound [Fe].C\C(O)=C\C(C)=O.C\C(O)=C\C(C)=O.C\C(O)=C\C(C)=O LZKLAOYSENRNKR-LNTINUHCSA-N 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 239000007987 MES buffer Substances 0.000 claims description 8
- 235000007164 Oryza sativa Nutrition 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 8
- 235000009566 rice Nutrition 0.000 claims description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 7
- 229910052708 sodium Inorganic materials 0.000 claims description 7
- 239000011734 sodium Substances 0.000 claims description 7
- 239000000758 substrate Substances 0.000 claims description 7
- VNNDVNZCGCCIPA-FDGPNNRMSA-N (z)-4-hydroxypent-3-en-2-one;manganese Chemical compound [Mn].C\C(O)=C\C(C)=O.C\C(O)=C\C(C)=O VNNDVNZCGCCIPA-FDGPNNRMSA-N 0.000 claims description 6
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 claims description 6
- 125000003277 amino group Chemical group 0.000 claims description 6
- BKFAZDGHFACXKY-UHFFFAOYSA-N cobalt(II) bis(acetylacetonate) Chemical compound [Co+2].CC(=O)[CH-]C(C)=O.CC(=O)[CH-]C(C)=O BKFAZDGHFACXKY-UHFFFAOYSA-N 0.000 claims description 6
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- RPNUMPOLZDHAAY-UHFFFAOYSA-N Diethylenetriamine Chemical compound NCCNCCN RPNUMPOLZDHAAY-UHFFFAOYSA-N 0.000 claims description 5
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 5
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims description 5
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 150000002171 ethylene diamines Chemical class 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims description 3
- 229910021529 ammonia Inorganic materials 0.000 claims description 3
- NDLPOXTZKUMGOV-UHFFFAOYSA-N oxo(oxoferriooxy)iron hydrate Chemical compound O.O=[Fe]O[Fe]=O NDLPOXTZKUMGOV-UHFFFAOYSA-N 0.000 claims description 3
- 239000004408 titanium dioxide Substances 0.000 claims description 3
- 241000790917 Dioxys <bee> Species 0.000 claims description 2
- 229910003978 SiClx Inorganic materials 0.000 claims description 2
- 239000005543 nano-size silicon particle Substances 0.000 claims description 2
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 125000005211 alkyl trimethyl ammonium group Chemical group 0.000 claims 1
- SWLVFNYSXGMGBS-UHFFFAOYSA-N ammonium bromide Chemical compound [NH4+].[Br-] SWLVFNYSXGMGBS-UHFFFAOYSA-N 0.000 claims 1
- PRORZGWHZXZQMV-UHFFFAOYSA-N azane;nitric acid Chemical compound N.O[N+]([O-])=O PRORZGWHZXZQMV-UHFFFAOYSA-N 0.000 claims 1
- 229910052710 silicon Inorganic materials 0.000 claims 1
- 239000010703 silicon Substances 0.000 claims 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 31
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 26
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 26
- 229960000975 daunorubicin Drugs 0.000 description 25
- 229940079593 drug Drugs 0.000 description 25
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 16
- 229940020947 fluorescein sodium Drugs 0.000 description 15
- 238000012360 testing method Methods 0.000 description 14
- 229910002518 CoFe2O4 Inorganic materials 0.000 description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 11
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 10
- 238000001228 spectrum Methods 0.000 description 10
- 239000007864 aqueous solution Substances 0.000 description 9
- 231100000135 cytotoxicity Toxicity 0.000 description 9
- 230000003013 cytotoxicity Effects 0.000 description 9
- 239000008367 deionised water Substances 0.000 description 9
- 229910021641 deionized water Inorganic materials 0.000 description 9
- 150000001413 amino acids Chemical group 0.000 description 8
- 239000007787 solid Substances 0.000 description 8
- 241000209094 Oryza Species 0.000 description 7
- 238000001142 circular dichroism spectrum Methods 0.000 description 7
- RPENMORRBUTCPR-UHFFFAOYSA-M sodium;1-hydroxy-2,5-dioxopyrrolidine-3-sulfonate Chemical compound [Na+].ON1C(=O)CC(S([O-])(=O)=O)C1=O RPENMORRBUTCPR-UHFFFAOYSA-M 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 229910052799 carbon Inorganic materials 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 238000002296 dynamic light scattering Methods 0.000 description 5
- 238000003746 solid phase reaction Methods 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- HYZQBNDRDQEWAN-LNTINUHCSA-N (z)-4-hydroxypent-3-en-2-one;manganese(3+) Chemical compound [Mn+3].C\C(O)=C\C(C)=O.C\C(O)=C\C(C)=O.C\C(O)=C\C(C)=O HYZQBNDRDQEWAN-LNTINUHCSA-N 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 229910018557 Si O Inorganic materials 0.000 description 4
- 150000001408 amides Chemical class 0.000 description 4
- 238000003795 desorption Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 235000012239 silicon dioxide Nutrition 0.000 description 4
- LIVNPJMFVYWSIS-UHFFFAOYSA-N silicon monoxide Inorganic materials [Si-]#[O+] LIVNPJMFVYWSIS-UHFFFAOYSA-N 0.000 description 4
- VSIVTUIKYVGDCX-UHFFFAOYSA-M sodium;4-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].COC1=CC([N+]([O-])=O)=CC=C1[N+]1=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=NN1C1=CC=C([N+]([O-])=O)C=C1 VSIVTUIKYVGDCX-UHFFFAOYSA-M 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 229910003321 CoFe Inorganic materials 0.000 description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 238000013270 controlled release Methods 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000009616 inductively coupled plasma Methods 0.000 description 3
- 230000005415 magnetization Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 238000001757 thermogravimetry curve Methods 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- SJECZPVISLOESU-UHFFFAOYSA-N 3-trimethoxysilylpropan-1-amine Chemical compound CO[Si](OC)(OC)CCCN SJECZPVISLOESU-UHFFFAOYSA-N 0.000 description 2
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- 229910004298 SiO 2 Inorganic materials 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 230000006399 behavior Effects 0.000 description 2
- 238000005452 bending Methods 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 238000002737 cell proliferation kit Methods 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 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 2
- 238000007654 immersion Methods 0.000 description 2
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 239000012299 nitrogen atmosphere Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000004886 process control Methods 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000002411 thermogravimetry Methods 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- 238000000733 zeta-potential measurement Methods 0.000 description 2
- GPAAEXYTRXIWHR-UHFFFAOYSA-N (1-methylpiperidin-1-ium-1-yl)methanesulfonate Chemical compound [O-]S(=O)(=O)C[N+]1(C)CCCCC1 GPAAEXYTRXIWHR-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 229910002483 Cu Ka Inorganic materials 0.000 description 1
- WEAHRLBPCANXCN-UHFFFAOYSA-N Daunomycin Natural products CCC1(O)CC(OC2CC(N)C(O)C(C)O2)c3cc4C(=O)c5c(OC)cccc5C(=O)c4c(O)c3C1 WEAHRLBPCANXCN-UHFFFAOYSA-N 0.000 description 1
- 238000003775 Density Functional Theory Methods 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 238000004639 Schlenk technique Methods 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- 229910002808 Si–O–Si Inorganic materials 0.000 description 1
- 108090001109 Thermolysin Proteins 0.000 description 1
- 241001193851 Zeta Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- HRHBQGBPZWNGHV-UHFFFAOYSA-N azane;bromomethane Chemical compound N.BrC HRHBQGBPZWNGHV-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000002983 circular dichroism Methods 0.000 description 1
- 230000001427 coherent effect Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- IDLFZVILOHSSID-OVLDLUHVSA-N corticotropin Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)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)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)CO)C1=CC=C(O)C=C1 IDLFZVILOHSSID-OVLDLUHVSA-N 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- 229910003460 diamond Inorganic materials 0.000 description 1
- MHDVGSVTJDSBDK-UHFFFAOYSA-N dibenzyl ether Chemical compound C=1C=CC=CC=1COCC1=CC=CC=C1 MHDVGSVTJDSBDK-UHFFFAOYSA-N 0.000 description 1
- OGQYPPBGSLZBEG-UHFFFAOYSA-N dimethyl(dioctadecyl)azanium Chemical compound CCCCCCCCCCCCCCCCCC[N+](C)(C)CCCCCCCCCCCCCCCCCC OGQYPPBGSLZBEG-UHFFFAOYSA-N 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 229960002143 fluorescein Drugs 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- SZVJSHCCFOBDDC-UHFFFAOYSA-N iron(II,III) oxide Inorganic materials O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical compound [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 description 1
- 239000002121 nanofiber Substances 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 238000006053 organic reaction Methods 0.000 description 1
- 230000003534 oscillatory effect Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000008729 phenylalanine Nutrition 0.000 description 1
- 150000002994 phenylalanines Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000000634 powder X-ray diffraction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- SCPYDCQAZCOKTP-UHFFFAOYSA-N silanol Chemical compound [SiH3]O SCPYDCQAZCOKTP-UHFFFAOYSA-N 0.000 description 1
- 238000002336 sorption--desorption measurement Methods 0.000 description 1
- 239000011029 spinel Substances 0.000 description 1
- 229910052596 spinel Inorganic materials 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000008542 thermal sensitivity Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
- A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5115—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5123—Organic compounds, e.g. fats, sugars
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Nanotechnology (AREA)
- Optics & Photonics (AREA)
- Inorganic Chemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention provides a kind of nucleocapsid mesoporous silica nano-particles of controllable temperature-sensitive peptide nanometer valve modification, the amphipathic small peptide that hud typed mesoporous silica nano-particle surface is grafted on by hud typed mesoporous silica nano-particle and by long chain amino forms, the hud typed mesoporous silica nano-particle is made of the mesoporous silicon oxide shell of superparamagnetic nano particle core and package superparamagnetic nano particle core, the amphipathic small peptide on hud typed mesoporous silica nano-particle surface is grafted on by long chain amino can respond the variation self assembly of temperature or unlocking self assembly makes duct on mesoporous silicon oxide shell be in closure or opening state, the amino acid sequence of the amphipathic small peptide is Boc Phe Phe Gly Gly COOH, long chain amino isOrApplication the present invention also provides the preparation method of the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification and its as pharmaceutical carrier.
Description
Technical field
The invention belongs to drug carrier material fields, are related to a kind of mesoporous dioxy of nucleocapsid of controllable temperature-sensitive peptide nanometer valve modification
SiClx nano particle and preparation method thereof and the nucleocapsid mesoporous silica nano-particle of the controllable temperature-sensitive peptide nanometer valve modification
Application as pharmaceutical carrier.
Background technology
Mesoporous silicon dioxide nano particle (MSN) since its surface area is big, porous structure and good biocompatibility and
It is widely studied as pharmaceutical carrier.Small peptide is due to its adjustable function, biodegradability and good biocompatibility
And it is widely deployed.Peptide amphiphile is received more and more attention based on its self assembly characteristic and as the component of MOLECULE DESIGN,
It has a wide range of applications in Three-dimensional cell culture, hemostasis and drug delivery etc..So far, it has been proposed that several
System based on MSN, using small peptide as the nano-valve for control release, for example, existing at present have enzyme response (such as
Elastoser, thermolysin) small peptide-MSN systems and with redox response (be typically glutathione)
Small peptide-MSN systematic accounts, if these small peptides-MSN systems are used in biologic applications, it is necessary to certain trigger agent
Triggering could realize the release for containing object, but this triggering based on enzyme or redox reaction may be only applicable for some lifes
Some local locations of object either organism, it is difficult to accomplish effectively controllable release.Martelli constructs to crimp spiral shell
The mesoporous silica nano-particle that peptide motif is thermo-responsive valve is revolved, can release using conventional heating at a temperature of 80 DEG C
The dyestuff loaded in the nano particle, if being applied in organism, 80 DEG C of temperature can cause the destruction and death of normal cell,
The running temperature of the system be still considered as be biologic applications a relatively harsh condition.The above-mentioned state of the art of base, if energy
Develop that release temperature is more mild, the better drug of release process control passes based on mesoporous silica nano-particle
Delivery system, for promoting mesopore silicon dioxide nano material that will generate positive meaning in the practical application of pharmaceutical carrier field.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of nucleocapsids of controllable temperature-sensitive peptide nanometer valve modification
Mesoporous silica nano-particle and preparation method thereof and the nucleocapsid mesoporous silicon oxide of the controllable temperature-sensitive peptide nanometer valve modification
Application of the nano particle as pharmaceutical carrier to reduce the drug release temperature of existing small peptide-MSN systems, and effectively promotes medicine
Object discharges process control, solves the problems, such as that existing small peptide-MSN systems are difficult to apply in organism.
The nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification provided by the invention, by hud typed Jie
Hole nano SiO 2 particle and the amphiphilic that hud typed mesoporous silica nano-particle surface is grafted on by long chain amino
Property small peptide composition, the hud typed mesoporous silica nano-particle by superparamagnetic nano particle core and package superparamagnetism
The mesoporous silicon oxide shell composition of nano particle core, hud typed mesoporous silicon dioxide nano is grafted on by long chain amino
The amphipathic small peptide on grain surface, which can respond the variation self assembly of temperature or unlock self assembly, to be made on mesoporous silicon oxide shell
Duct be in closure or opening state, the amino acid sequence of the amphipathic small peptide is Boc-Phe Phe Gly Gly-
The structural formula of the amphipathic small peptides of COOH isLong chain amino is Or
In the nucleocapsid mesoporous silica nano-particle of above-mentioned controllable temperature-sensitive peptide nanometer valve modification, amphipathic small peptide is grafted on
Shown in the structural formula of the group formed on long chain amino such as formula (I)~(III), in the structural formula shown in formula (I)~(III), with Si
Connected O is connected to form Si-O keys with the Si on hud typed mesoporous silica nano-particle.
In the nucleocapsid mesoporous silica nano-particle of above-mentioned controllable temperature-sensitive peptide nanometer valve modification, hud typed meso-porous titanium dioxide
The mass ratio of nano silicon particles, long chain amino and amphipathic small peptide is preferably 95:(5~7):(4~10).
In the technical solution of the nucleocapsid mesoporous silica nano-particle of above-mentioned controllable temperature-sensitive peptide nanometer valve modification, superparamagnetic
Property nano particle core be preferably to adulterate the superparamag-netic iron oxide of manganese and cobalt.It is highly preferred that the magnetism of doping manganese and cobalt
In ferric oxide nanometer particle, manganese, iron, the molar ratio of cobalt are 1:4:1.
In the technical solution of the nucleocapsid mesoporous silica nano-particle of above-mentioned controllable temperature-sensitive peptide nanometer valve modification, superparamagnetic
Property nano particle core grain size for 8.5~13.5nm, the grain size of hud typed mesoporous silica nano-particle for 32.5~
65.5nm。
The present invention also provides a kind of nucleocapsid mesoporous silica nano-particles of above-mentioned controllable temperature-sensitive peptide nanometer valve modification
Preparation method, step is as follows:
(1) superparamag-netic iron oxide for adulterating manganese and cobalt is scattered in CHCl3Middle formation dispersion liquid A, by dispersion liquid A
It is sufficiently mixed with cetyl trimethylammonium bromide solution, then heating evaporation removal CHCl3, water is added to adjust to cetyl three
The concentration of methyl bromide ammonium is 1.8~2.2mg/mL, and then adjusting pH value to 11.0~11.5 obtains mixed liquid B, 68~72
DEG C stirring 30~60min, be added dropwise tetraethyl orthosilicate, add ethyl acetate, 68~72 DEG C react 3~4h, by reaction product
It is washed with water and ethyl alcohol, obtains hud typed mesoporous silica nano-particle;
In mixed liquid B, manganese and the superparamag-netic iron oxide of cobalt and the quality of cetyl trimethylammonium bromide are adulterated
Than for 1:(18~22), mixed liquid B, tetraethyl orthosilicate, the volume ratio of ethyl acetate are 100:1:(5~15).
(2) under nitrogen protection, hud typed mesoporous silica nano-particle is scattered in ethyl alcohol, then adds in (3-
Aminopropyl) trimethoxy silane, N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines or diethylenetriamine base propyl trimethoxy
Base silane is heated to reflux being stirred to react 10~14h under nitrogen protection, reaction product ethyl alcohol and water washing is subsequently placed in
Ammonium nitrate concn is that removal cetyl trimethylammonium bromide is heated to reflux in 4%~8% ammonium nitrate ethanol solution, uses ethyl alcohol
The hud typed mesoporous silica nano-particle of surface amino groups modification is obtained after being washed;(3- aminopropyls) trimethoxy silicon
Alkane, N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines or diethylenetriamine base propyl trimethoxy silicane with it is hud typed mesoporous
The mass ratio of nano SiO 2 particle is 5%~8%;
(3) it is amphipathic small peptide, 1- (the 3- dimethylaminos third of Boc-Phe Phe Gly Gly-COOH by amino acid sequence
Base) -3- ethyl-carbodiimide hydrochlorides and n-hydroxysuccinimide sulfonate sodium be according to (6~10):10:5 mass ratio is molten
Solution makes the concentration of n-hydroxysuccinimide sulfonate sodium mixed liquor C be obtained, by table for 9~12mg/mL in MES buffer solutions
The amino modified hud typed mesoporous silica nano-particle in face, which is scattered in, obtains concentration as 8~15mg/mL's in PBS buffer solution
Dispersion liquid D, by mixed liquor C and dispersion liquid D according to (0.8~1.2):1 volume ratio mixing, is reacting at room temperature 10~18h, will be anti-
Product is answered to be washed with water to get controllable temperature-sensitive peptide nanometer valve modification nucleocapsid mesoporous silica nano-particle.
In the above method, the preparation method for adulterating the superparamag-netic iron oxide of manganese and cobalt is as follows:
(1) by Fe (acac)3、Mn(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:(9~10):(5~
7):The molar ratio of (5~7), which is dissolved in benzylic ether, makes Fe (acac)3Concentration for 0.08~0.12mmol/mL, protected in nitrogen
190~210 DEG C are heated under shield and stirring condition, then keeps the temperature 2~2.5h in 190~210 DEG C under nitrogen protection, is reheated
Gained reaction solution is cooled to room temperature by 0.5~2h of back flow reaction, is added in ethyl alcohol and is precipitated to get the magnetic iron oxide of doping manganese
Nano particle;
(2) by Fe (acac)3、Co(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:(9~10):2:2
Molar ratio be dissolved in benzylic ether and make Fe (acac)3Concentration for 0.08~0.12mmol/mL, obtain mixed liquor E, will adulterate
The superparamag-netic iron oxide of manganese, which is scattered in n-hexane, makes MnFe2O4The concentration of nano particle is formed for 15~20mg/mL divides
Dispersion liquid F, by mixed liquor E and dispersion liquid F according to 10:The volume ratio mixing of (5~6), heating evaporation fall n-hexane, are protected in nitrogen
With 190~210 DEG C are heated under stirring, then keep the temperature 1~1.5h in 190~210 DEG C under nitrogen protection, it is anti-to reheat reflux
0.5~2h is answered, gained reaction solution is cooled to room temperature, precipitation in ethyl alcohol is added in and is received to get the magnetic iron oxide of doping manganese and cobalt
Rice grain.
In the above method, the pH value of MES buffer solutions is 6.0, concentration is 50~100mmol/L, and the pH value of PBS buffer solution is
7.4th, concentration is 50~100mmol/L.
The present invention also provides a kind of nucleocapsid mesoporous silica nano-particles of above-mentioned controllable temperature-sensitive peptide nanometer valve modification
Application as pharmaceutical carrier.The present invention is grafted amphiphilic on hud typed mesoporous silica nano-particle surface by long chain amino
Property small peptide, carrying medicament and discharge drug schematic diagram it is as shown in Figure 1.In application, the core that controllable temperature-sensitive peptide nanometer valve is modified
Shell mesoporous silica nano-particle is dispersed in drug solution, and 2~4h is placed at 50~60 DEG C so that drug enters institute
In the pore passage structure of mesoporous silicon oxide shell for stating nano particle, then lead in the 10~15h that is stored at room temperature no more than 40 DEG C
A nanometer valve can be self-assembly of by crossing long chain amino and being grafted on the amphipathic small peptide on hud typed mesoporous silica nano-particle surface
The duct closure of goalkeeper's mesoporous silicon oxide shell, thus by drug loading in the nano particle.
Experiments have shown that under conditions of oscillating magnetic field or temperature is not applied no more than 50 DEG C, controllable temperature-sensitive peptide is supported on
Drug in the nucleocapsid mesoporous silica nano-particle of nanometer valve modification hardly leaks, and without cytotoxicity, and receives
Also without apparent cytotoxicity, hud typed mesoporous silica nano-particle surface is grafted on by long chain amino in itself for rice grain
Amphipathic small peptide can also be oscillated magnetic field triggering to unlock self assembly, so as to fulfill drug by the nanometer valve being self-assembly of
Release, and the oscillating magnetic field applied will not generate any toxicity to exposed cell.Based on this, controlled thermal provided by the invention
It, can be according to practical application after the nucleocapsid mesoporous silica nano-particle of quick peptide nanometer valve modification is as pharmaceutical carrier carrying medicament
Needs, apply oscillating magnetic field in target site and realize the controlled release of drug.
Compared with prior art, present invention produces technique effects beneficial below:
1. the present invention provides a kind of nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification, by core
Shell mould mesoporous silica nano-particle and hud typed mesoporous silica nano-particle surface is grafted on by long chain amino
Amphipathic small peptide composition, hud typed mesoporous silica nano-particle by superparamagnetic nano particle core and package superparamagnetic
Property nano particle core mesoporous silicon oxide shell composition, since the length of long chain amino is appropriate, be grafted by long chain amino
Amphipathic small peptide on hud typed mesoporous silica nano-particle surface can respond the variation self assembly of temperature or unlock from group
Dress makes the duct on mesoporous silicon oxide shell be in closure or opening state, and is received in the nano particle with superparamagnetism
Rice grain, therefore, the amphipathic small peptide that hud typed mesoporous silica nano-particle surface is grafted on by long chain amino pass through
The nanometer valve being self-assembly of can be oscillated magnetic field triggering and realize the release for containing object, and not only release conditions are mild, can meet
The condition of organism practical application, and can realize the controlled release for containing object, thus the nano particle is in pharmaceutical carrier and medicine
Substance delivery system field has important actual application value.
2. for the existing small peptide-MSN systems with enzyme response and with redox response, this
The variation that the nucleocapsid mesoporous silica nano-particle for the controllable temperature-sensitive peptide nanometer valve modification that invention provides can respond temperature is realized
The controlled release of object is contained, and the variation of this temperature is realized by applying controllable oscillatory magnetic field, thus applying
Shi Buhui is limited be subject to certain enzyme or oxidation-reduction quality environment, and universality is more preferable, compared with it is existing in conventional heating to 80 DEG C
For the small peptide-MSN systems for containing object could be discharged, the release conditions that object is contained in nano particle of the invention substantially more heat
With.The nano particle has no apparent cytotoxicity in itself, under conditions of oscillating magnetic field is not applied, containing in nano particle
Object hardly leaks, and will not generate cytotoxicity, and the oscillating magnetic field applied will not be any to the generation of exposed cell
Toxicity.This characteristic combination infiltration-Noninvasive stimulation can make the nano particle become the ideal candidate that following object delivers.
Description of the drawings
It is amphipathic short that Fig. 1 is that the present invention is grafted on hud typed mesoporous silica nano-particle surface by long chain amino
Peptide, carrying medicament and the schematic diagram for discharging drug, wherein, figure A, B, C, D represent hud typed mesoporous silicon dioxide nano respectively
Grain is grafted amphipathic small peptide, carrying medicament and the schematic diagram for discharging drug by long chain amino, and figure E, F are respectively the office for scheming A, B
Portion's enlarged drawing, figure G, H are the partial enlarged view before the amphipathic small peptide self assembly in figure C and after self assembly, and figure I is the office of figure D
Portion's enlarged drawing, in figure,The long chain amino for being grafted amphipathic small peptide is represented,Represent super-paramagnetism nano
Grain core,Represent drug.
Fig. 2 is MnFe2O4The TEM figures of nano particle.
Fig. 3 is MnFe2O4The grain size distribution of nano particle.
Fig. 4 is the TEM figures of SPNC.
Fig. 5 is the grain size distribution of SPNC.
Fig. 6 is hysteresis loops of the SPNC in 300K.
Fig. 7 is the high angle X-ray diffractogram of SPNC.
Fig. 8 is the TEM figures of SPNC@MSN.
Fig. 9 is the grain size distribution of SPNC@MSN.
Figure 10 is amphipathic small peptide (curve (a)), SPNC@MSN (curve (b)) and SPNC@MSN-PEP (curve (c))
Infrared spectrogram.
Figure 11 be amphipathic small peptide 25 DEG C (filled squares), 40 DEG C (hollow squares), 50 DEG C (black triangles),
The interior circular dichroism for returning to 25 DEG C (hollow circle) at a temperature of 60 DEG C (hollow triangles), 80 DEG C (solid circles) and when 8 is small
Spectrogram.
Figure 12 is that amphipathic small peptide is schemed in aqueous solution with the TEM after the concentration self assembly of 0.2mg/mL.
Figure 13 is SPNC@MSN, SPNC@MSN-NH2With Zetas of the SPNC@MSN-PEP in the deionized water of pH=7.0
Potentiometric analysis result.
Figure 14 is SPNC@MSN (curve (a)), SPNC@MSN-NH2(curve (b)) and SPNC@MSN-PEP (curve (c))
Dynamic light scattering measurement result.
Figure 15 is SPNC@MSN (curve (a)), SPNC@MSN-NH2(curve (b)) and SPNC@MSN-PEP (curve (c))
Thermal gravimetric analysis curve.
Figure 16 is the nitrogen adsorption and desorption isotherm of SPNC MSN (square) and SPNC MSN-PEP (filled circles).
Figure 17 is the pore size distribution curve calculated with NLDFT methods, and wherein solid line represents SPNC@MSN, and dotted line represents SPNC@
MSN-PEP。
Figure 18 is the curve of the fluorescein sodium discharged by conventional heating at 50 DEG C from MSN-PEP, and in figure, arrow represents
1h lines of demarcation.
The fluorescein sodium release conditions figure that Figure 19 is SPNC@MSN-PEP in superparamagnetism heating, block diagram generation therein
Table superparamagnetism heats.
Figure 20 is the SPNC@MSN-PEP (black column) and DNR@SPNC@MSN-PEP (grey column) of various concentration to PANC-
The cytotoxicity assay result of 1 cell.
Figure 21 is that SPNC@MSN-PEP and DNR@SPNC@MSN-PEP live to cell under with and without magnetic field condition
Property analysis, wherein black column and grey column difference representative sample " not exposing " and " being exposed to " magnetic field, " control " represent do not add in
Nano particle.
Specific embodiment
The nucleocapsid mesoporous silicon oxide of controllable temperature-sensitive peptide nanometer valve modification provided by the invention is received by the following examples
Rice grain and preparation method and application is described further.It is necessary to note that following embodiment be served only for making the present invention into
One step explanation, it is impossible to be interpreted as limiting the scope of the invention, one of ordinary skill in the art are right according to foregoing invention content
The present invention makes some nonessential modifications and adaptations and is embodied, and still falls within the scope of invention protection.
The information of the chemical reagent used in detailed description below is as follows:
Praseodynium iron (Fe (acac)3, 97%), manganese acetylacetonate (II) (Mn (acac)2, 21%-23%Mn), second
Acyl acetone cobalt (II) (Co (acac)2, 97%), oleic acid (90%), oleyl amine (70%), 1,2- dodecanediol (90%), benzyl oxide
(98%), tetraethyl orthosilicate (TEOS, 99%), cetyl trimethylammonium bromide (CTAB, 98%), ammonium nitrate, absolute ethyl alcohol
(EtOH, Aldrich), chloroform (CHCl3), diethylenetriamine base propyl trimethoxy silicane (TSPDT, 99%), (3- ammonia
Propyl) trimethoxy silane ((CH3O)3Si(CH2)3NH2, 99%), N- [3- (trimethoxy silicon substrate) propyl] ethylenediamine
((CH3O)3Si(CH2)3NHCH2CH2NH2, 99%), n-hydroxysuccinimide sulfonate sodium (Sulfo-NHS, 98%), 2-
One water object of (N- morpholines) ethanesulfonic acid (MES Buffer, 99%), ethyl acetate (EA, 99%), fluorescein sodium (90%), hydrochloric acid assistant
It is soft than star (DNR,>95%) it is purchased from Sigma-Aldrich.
1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC.HCl) come from CreoSalus Inc.
(Louisville,KY,USA).N-hexane (99.9%) and hydrochloric acid (37.5%), sodium hydroxide (99%), phosphate-buffered is molten
From generation is flown, you are scientific and technological for liquid (PBS, pH=7.4) purchase.All chemicals all use without further purification.All is organic
Reaction under the inert atmosphere of drying nitrogen, is carried out in dry glassware using standard Schlenk technique.
The amino acid sequence of amphipathic small peptide is Boc-Phe Phe Gly Gly-COOH, and structural formula isMolecular weight is 526.6, purity 92%, by GenScript USA Inc.
Synthesis, is stored in -20 DEG C, is used without being further purified.
The instrument and characterization conditional information used when being characterized in detailed description below is as follows:
Fourier transform infrared spectroscopy (FTIR) is operated by JASCO FT/IR-420 spectrometers.With 1cm-1Resolution ratio note
Record 4000cm-1To 400cm-1Wave number.By mixing KBr and nano particle under a certain pressure come tabletting sample preparation.
Transmission electron microscope (TEM) image is obtained on JEM1200-EX (JEOL) with the accelerating potential of 80kV.It will be dense
It spends and is ultrasonically treated 30s for the sample suspension of 1mg/mL, then drop on copper mesh.It is ultrasonically treated MnFe2O4And MnFe2O4@CoFe2O4
Nano-particle is disperseed in hexane with the concentration of 0.05mg/mL.The TecnaiT12 electron microscopes that operating voltage is 120kV are used
In MnFe2O4And MnFe2O4@CoFe2O4The detection of nano-particle.
Nitrogen adsorption and desorption isotherm are in the Autosorb-iQ (Quantachrome that temperature is 77K
Instruments carried out on).Data pass through N2NLDFT balance models calculate.
It records Zeta potential in deionized water with ZetaSizer Nano (Malvern Instr Ltd., Britain) and moves
State light scattering (DLS), sample concentration 0.1mg/mL.
Thermogravimetric analysis (TGA) is 200mL/min's in flow velocity by Perkin-Elmer Pyris Diamond TG/DTA
It is operated under air.Usually 10mg samples are put into crucible, sample is kept the temperature into 10min at 50 DEG C, with the heating speed of 5 DEG C/min
Degree is heated to 550 DEG C from 50 DEG C, then keeps the temperature 10min at 550 DEG C.
With Shimadzu ICPE-9000 instruments, surveyed using inductively coupled plasma emission spectrography (ICP-OES)
Determine MnFe2O4Middle Mn:Fe ratios and MnFe2O4@CoFe2O4The Mn of nano particle:Co:Fe ratios.In general, by 3mg powderies
MnFe2O4And MnFe2O4@CoFe2O4It is dissolved in respectively in 0.5mL hydrochloric acid solutions.Solution is further diluted to carry out quantitative survey
Amount.
Hysteresis loop is measured using superconducting quantum interference device (SQUID) (SQUID) magnetometer (Quantum Design MPMS XL).
Powder x-ray diffraction (XRD) collection of illustrative plates with Cu-Ka by radiatingPANalytical X'Pert
PRO diffractometers obtain.Operating current and voltage are respectively 40mA and 45kV.
Circular dichroism spectra (CD) is obtained by 400 type spectrometers of AVIV (AVIV Associates, Lakewood, NJ).By sample
Product keep 10min to reach balance under different test temperatures, then obtain spectrum from 185nm to 260nm with 1nm intervals.
Pass through Acton Spectra Pro 2300i CCD and CUBE 445-40C laser (Coherent Inc., Santa
Clara, CA, USA) collect fluorescence spectrum.The standard curve of dyestuff is obtained at a temperature of 25 DEG C and 50 DEG C.
Superparamagnetism heating is generated by magnetic thermal therapy system (MSI Automation, Inc).Copper wire loop diameter 5cm, five circle lines
Enclose high 5cm.Working frequency is 370kHz, induced power 5kW.
Embodiment 1
In the present embodiment, the nucleocapsid mesoporous silica nano-particle (SPNC of controllable temperature-sensitive peptide nanometer valve modification is prepared
MSN-PEP), step is as follows:
(1) magnetic iron oxide (MnFe of doping manganese is prepared2O4) nano particle
By 2mmol Fe (acac)3、1mmol Mn(acac)2, 10mmol 1,2- dodecanediols, 6mmol oleic acid and
6mmol oleyl amines are dissolved in 20mL benzylic ethers, and 200 DEG C are heated under nitrogen atmosphere and intense agitation, then at 200 DEG C
2h is kept the temperature, then in 298 DEG C of back flow reaction 1h, is obtained containing MnFe2O4The reaction solution is cooled to room by the reaction solution of nano particle
Temperature adds in 40mL ethyl alcohol and precipitates MnFe2O4Nano particle, by centrifuging MnFe2O4Nano particle further separates, and lays equal stress on
New be dispersed in 10mL n-hexanes preserves.
MnFe is detected by inductively coupled plasma emission spectrography (ICP-OES)2O4Manganese and iron rubs in nano particle
You are than being 1:2.MnFe prepared by the step2O4The TEM figures of nano particle are as shown in Fig. 2, Fig. 2 shows MnFe2O4Nano particle is in
Spherical shape, MnFe2O4The particle diameter distribution situation of nano particle is as shown in figure 3, MnFe2O4The particle diameter distribution of nano particle for 8.1 ±
0.9nm。
(2) magnetic iron oxide (MnFe of doping manganese and cobalt is prepared2O4@CoFe2O4) nano particle
MnFe2O4@CoFe2O4Nano particle is by MnFe2O4CoFe is grown on nano particle2O4Come what is prepared.
By 2mmol Fe (acac)3、1mmol Co(acac)2, 10mmol 1,2- dodecanediols, 2mmol oleic acid and
2mmol oleyl amines, which are dissolved in the benzylic ether of 20mL, obtains mixed liquor E, by 180mg MnFe2O4Nano particle be dispersed in 10mL just oneself
Dispersion liquid F is formed in alkane, mixed liquor E is mixed with dispersion liquid F, is heated to 90 DEG C and in temperature 30min to remove just
Hexane is heated to 200 DEG C under nitrogen atmosphere and intense agitation, and 1h is then kept the temperature at 200 DEG C, then anti-in 298 DEG C of reflux
1h is answered, is obtained containing MnFe2O4@CoFe2O4The reaction solution is cooled to room temperature, adds by the reaction solution of nano particle (abbreviation SPNC)
Enter in 40mL ethyl alcohol and precipitate SPNC, SPNC is further separated by centrifuging, and is dispersed in 10mL n-hexanes and preserves again.
By inductively coupled plasma emission spectrography (ICP-OES) detect SPNC in manganese, iron, cobalt molar ratio be 1:
4:1.The TEM figures of SPNC prepared by the step are as shown in figure 4, Fig. 4 shows that SPNC is spherical in shape, and the particle diameter distribution situation of SPNC is as schemed
Shown in 5, the particle diameter distribution of SPNC is 11.3 ± 0.8nm.
Fig. 6 show under conditions of 300K as measured by superconducting quantum interference device (SQUID) mgnetic observations with field
Relevant magnetization curve, it will be appreciated from fig. 6 that SPNC has superparamagnetism, saturation magnetization 117emu/g, higher than Fe3O4It receives
The saturation magnetization 80emu/g of rice grain.Therefore, SPNC can provide higher magnetic thermal induction effect drug to be controlled to release
It puts.In order to measure electromagnetic heating efficiency, SPNC is dispersed in toluene and applies alternating magnetic field.
Fig. 7 shows high angle X-ray diffraction (XRD) figure of SPNC, and the diffraction maximum in figure confirms that SPNC is typically sharp
Spinel structure.
(3) hud typed mesoporous silica nano-particle (SPNC@MSN) is prepared
N-hexane is fallen to the n-hexane heating evaporation for being dispersed with SPNC, obtained SPNC is scattered in CHCl3In obtain
SPNC concentration is the SPNC dispersion liquids of 10mg/mL, and 800 μ L dispersion liquid additions are contained 20mL H2The water of O and 160mg CTAB
In solution, 10min is ultrasonically treated, then mild heating evaporation falls CHCl3, then add in 60mL water and with the hydrogen-oxygen of 2mol/L
Change sodium solution and adjust pH value to 11.0~11.5, stir 30min at 70 DEG C, 800 μ L TEOS are added dropwise, are then quickly added into
4.8mL ethyl acetate reacts 3h at 70 DEG C, and solid phase reaction product with water and ethyl alcohol is washed, obtained by separation of solid and liquid successively
SPNC@MSN are dispersed in 30mL n-hexanes and preserve by SPNC@MSN.
The TEM figures of SPNC@MSN prepared by the step on SPNC@MSN as shown in figure 8, have radially as can be seen from Figure 8
Passage, the particle diameter distribution situation of SPNC@MSN is as shown in figure 9, the particle diameter distribution of SPNC is 37.3 ± 4.8nm.
(4) hud typed mesoporous silica nano-particle (the SPNC@MSN-NH that surface amino groups are modified are prepared2)
Under nitrogen protection, 140mgSPNC@MSN are scattered in 30mL ethyl alcohol, 55 μ L TSPDT are then added in, in nitrogen
It is heated to reflux being stirred to react 12h under gas shielded, reaction product is used into ethyl alcohol and water washing successively, in order to remove in reaction product
Reaction product after washing is placed in the ammonium nitrate ethanol solution that ammonium nitrate concn is 6%, heats back by Surfactant CTAB
1h is flowed, isolates gained nano particle, adds in the ammonium nitrate ethanol solution that ammonium nitrate concn is 6% after being washed with ethyl alcohol again
In, 1h is heated to reflux, gained nano particle is isolated and is washed with ethyl alcohol, obtain SPNC@MSN-NH2, by SPNC@MSN-
NH2It is scattered in 10mL water and preserves.
(5) the nucleocapsid mesoporous silica nano-particle (SPNC MSN-PEP) of controllable temperature-sensitive peptide nanometer valve modification is prepared
By 30mg amino acid sequences be the amphipathic small peptide of Boc-Phe Phe Gly Gly-COOH, 50mg EDC.HCl and
25mg Sulfo-NHS add in 2.5mL MES buffer solutions (pH=6.0,100mM)) in, 10min is stirred at room temperature and obtains mixed liquor
C, by 30mg SPNC@MSN-NH2It is scattered in 2.5mLPBS buffer solutions (pH=7.4,100mM) and obtains dispersion liquid D, will mix
Liquid C and dispersion liquid D mixing, in room temperature reaction 12h, reaction process, SPNC@MSN-NH2The nitrogen of amino on surface is as parent
Core reagent, the carbon attacked on the carboxyl of amphipathic small peptide form C-N keys, reaction product are washed with water 3 times to get SPNC@
In MSN-PEP, SPNC@MSN-PEP, amphipathic small peptide is grafted on structural formula such as formula (I) institute of the group formed on long chain amino
Show, in the structural formula shown in formula (I), the Si on O and SPNC@MSN being connected with Si is connected to form Si-O keys.
Figure 10 is amphipathic small peptide (curve (a)), SPNC@MSN (curve (b)) and SPNC@MSN-PEP (curve (c))
Infrared spectrogram, in figure, amide I bands of a spectrum (v=1676cm-1) C=O stretching vibrations are represented, this shows that amphipathic small peptide forms β foldings
Stack structure, amide II (v=1546cm-1) it is as caused by the N-H bending vibrations in plane and C-N stretching vibrations.
With the amphipathic small peptide that circular dichroism spectra (CD) analysis of amino acid sequence is Boc-Phe Phe Gly Gly-COOH
Secondary structure, Figure 11 are amphipathic small peptides in 25 DEG C (filled squares), 40 DEG C (hollow squares), 50 DEG C of (triangles
Shape), 60 DEG C (hollow triangles), at a temperature of 80 DEG C (solid circles) and the interior circle for returning to 25 DEG C (hollow circle) when 8 is small
Two chromatograms.As shown in Figure 11, the spectrum at 25 DEG C shows negative peak ([190]=- 6,630 ° of every cm2.dmol-1),
It is entirely different with two phenylalanines there are one small peak near 220nm.By with 50 reference proteins in CD Pro programs into
Row compares, and calculates the Secondary Structure Content of amphipathic small peptide, calculation shows that, which has up to 35.5% β pieces
Layer structure content is the basic second grade structure that nanofiber is formed.In order to obtain the thermal stability of amphipathic small peptide, two are measured
The circular dichroism spectra of parent's property small peptide at different temperatures.Statistics indicate that from room temperature (25 DEG C) be heated to 40 DEG C when, spectral intensity does not have
Notable difference.When temperature is increased to 50 DEG C, detects that apparent decline occurs in the negative peak near 190nm, show amphipathic short
Certain variation has occurred in the secondary structure of peptide.The spectrum of amphipathic small peptide at 60 DEG C is almost identical with 50 DEG C of spectrum.When
At 80 DEG C, spectral intensity reduces temperature stabilization, and signal-to-noise ratio becomes quite big.Then temperature is restored to 25 DEG C, after 8h again
Sample is measured, compared with the original spectrum at 25 DEG C, amphipathic small peptide recovers in 8h.The result shows that amphipathic small peptide
Peptide has thermal sensitivity, has quick self assembly ability, and assembles temperature at 50 DEG C or so.
Figure 12 is that amphipathic small peptide is schemed in aqueous solution with the TEM after the concentration self assembly of 0.2mg/mL, it can be seen that
Amphipathic small peptide forms fiber-like nanostructures after self assembly in aqueous solution, this is pass of the peptide as the nano-valve of MSN
Keyness matter.
SPNC@MSN, SPNC@MSN-NH are recorded in deionized water with ZetaSizer Nano2With SPNC@MSN-PEP's
Zeta potential and dynamic light scattering (DLS), sample concentration 0.1mg/mL.SPNC@MSN, SPNC@MSN-NH2With SPNC@MSN-
Zeta potential analysis results of the PEP in the deionized water of pH=7.0 is shown in Figure 13.Figure 14 be SPNC@MSN (curve (a)),
SPNC@MSN-NH2The dynamic light scattering measurement of (curve (b)) and SPNC@MSN-PEP (curve (c)) as a result, as shown in Figure 14,
SPNC@MSN have the larger hydrodynamic size of about 120nm, SPNC@MSN-NH2Hydrodynamic diameter be about
114nm is not changed significantly compared with SPNC MSN, after Amphiphilic peptide is modified, the hydrodynamics of SPNC MSN-PEP
Diameter is about 68nm, illustrates that SPNC@MSN-PEP have preferable dispersiveness in water.
Figure 15 is SPNC@MSN (curve (a)) manufactured in the present embodiment, SPNC@MSN-NH2(curve (b)) and SPNC@
The thermal gravimetric analysis curve of MSN-PEP (curve (c)).
With reference to infrared spectrogram, thermal gravimetric analysis curve and Zeta potential analysis result can verify that amphipathic small peptide successfully connects
Branch is on the surface of nucleocapsid mesoporous silica nano-particle.In infrared spectrum, 3200~3550 and 1000~1100cm-1Near
Absorption band represent O-H stretching vibrations and Si-O-Si asymmetric stretching vibrations respectively, with SPNC@MSN, SPNC@MSN-PEP in v
=1653cm-1The bands of a spectrum at place are caused by C=O stretching vibrations (amide I), and the appearance of the key shows that amphipathic small peptide has succeeded
It is grafted on mesoporous silicon oxide.The infrared spectrum of SPNC@MSN-PEP is in v=2925cm-1(alkane C-H vibrations), v=
2854cm-1(alkane C-H vibrations), v=3060cm-1The new bands of a spectrum that (aromatic ring) occurs also demonstrate this point.Due to N-H groups
Bending vibration and C-N groups stretching vibration, amide II (v=1539cm-1) it is conformation sensitization.It is red with amphipathic small peptide
External spectrum is compared, and some absworption peaks of SPNC@MSN-PEP disappear, such as v=1411cm-1(C=O stretches in COO-) and
1742cm-1(- COOH stretching vibrations), due to not having COO- or COOH in SPNC@MSN-PEP, this also illustrates amphipathic short
Peptide successfully connects branch and has arrived on mesoporous silicon oxide.Thermogravimetric curve shows mass loss in air from the 5% of sample SPNC@MSN
Increase to SPNC@MSN-NH212% and 16% with SPNC@MSN-PEP, show in each functionalisation step successfully
Grafting, with sample SPNC@MSN-NH2It compares, it is weightless corresponding to the two of grafting with after amphipathic small peptide condensation reaction increased 4%
Parent's property small peptide.In the deionized water of pH=7.0, sample SPNC@MSN, SPNC@MSN-NH2With SPNC@MSN-PEP go from
Zeta potential in sub- water (pH=7.0) is respectively -33mV ,+42mV and+39mV, SPNC@MSN and SPNC@MSN-NH2Between
Significant difference be because most of electronegative silanol be grafted to it is positively charged on the surface of SPNC@MSN
Amino shields.
The results are shown in Table 1 for nitrogen adsorption-desorption, and as shown in Table 1, the surface area of sample SPNC@MSN-PEP is 482m2/
G, aperture are 3.5nm (being calculated by DFT method), compared with sample SPNC@MSN, are slightly reduced, but for as pharmaceutical carrier
For, the surface area of SPNC@MSN-PEP is enough.Figure 16 is that SPNC@MSN (square) and SPNC@MSN-PEP are (solid
Circle) nitrogen adsorption and desorption isotherm.Figure 17 is the pore size distribution curve calculated with NLDFT methods, and wherein solid line represents SPNC@
MSN, dotted line represent SPNC@MSN-PEP.
1 nitrogen adsorption of table-desorption data
Comparative example 1
In this comparative example, short peptide modified mesoporous silica nano-particle (MSN-PEP) is prepared, step is as follows:
(1) mesoporous silica nano-particle (MSN) is prepared
The round bottom that 250mg CTAB and 875 μ L concentration are added to 120mL water for the sodium hydroxide solution of 2mol/L is burnt
In bottle, 80 DEG C are heated to, the TEOS of 1200 μ L is added dropwise under stiring, react 2h, separation of solid and liquid, by solid phase reaction product at 80 DEG C
It is washed successively with water and ethyl alcohol, obtains MSN, MSN is dispersed in 30mL ethyl alcohol and is preserved.
(2) mesoporous silica nano-particle (MSN-NH that surface amino groups are modified is prepared2)
Under nitrogen protection, 140mg MSN are scattered in 30mL ethyl alcohol, then add in 55 μ L TSPDT, protected in nitrogen
The lower reflux of shield is stirred to react 12h, reaction product is used ethyl alcohol and water washing successively, in order to remove the surface-active in reaction product
Reaction product after washing is added in the ammonium nitrate ethanol solution that ammonium nitrate concn is 6%, is heated to reflux 1h by agent CTAB, point
Gained nano particle is separated out, adds in the ammonium nitrate ethanol solution that ammonium nitrate concn is 6%, heats back again after being washed with ethyl alcohol
1h is flowed, isolate gained nano particle and is washed with ethyl alcohol, obtains MSN-NH2, by MSN-NH2It is scattered in 10mL water and preserves.
(3) short peptide modified mesoporous silica nano-particle (MSN-PEP) is prepared
By 30mg amino acid sequences be the amphipathic small peptide of Boc-Phe Phe Gly Gly-COOH, 50mg EDC.HCl and
25mg Sulfo-NHS add in 2.5mL MES buffer solutions (pH=6.0,100mM)) in, 10min is stirred at room temperature and obtains mixed liquor
G, by 30mg MSN-NH2Be scattered in 2.5mLPBS buffer solutions (pH=7.4,100mM) and obtain dispersion liquid H, by mixed liquor G and
Dispersion liquid H is mixed, in room temperature reaction 12h, reaction process, MSN-NH2The nitrogen of amino on surface is as nucleopilic reagent, attack
Carbon on the carboxyl of amphipathic small peptide forms C-N keys, and reaction product is washed with water 3 times to get MSN-PEP.
Embodiment 2
In the present embodiment, temperature-sensitive release behaviors of the MSN-PEP of the preparation of comparative example 1 under normal heating conditions is investigated, with
And embodiment 1 prepare SPNC@MSN-PEP and the temperature-sensitive release behavior under superparamagnetic heating condition.
(1) MSN-PEP of 15mg is added in into the aqueous solution of the fluorescein sodium of 1.5mL 1mmol/L, ultrasonic disperse is uniform,
In view of amphipathic small peptide self-assembly property at room temperature and CD data, 50 DEG C of temperature is set to open and contaminate for opening of the channel
Material loading, 2h is kept the temperature so that fluorescein sodium to be born by the test tube equipped with MSN-PET and fluorescein sodium aqueous solution in 50 DEG C of water-bath
It is loaded on MSN-PEP, test tube is then being stored at room temperature 12h to ensure the amphipathic small peptide self assembly again being grafted on MSN, is being obtained
To the MSN-PEP for loading drug.
By 3mg load drug MSN-PEP be dispersed in 250 μ L water, be then charged into both ends open and opening covered with
In the container of dialysis membrane (14kDa MWCO), before release, container immersion is persistently changed in the deionized water of water several days, and
It is detected by ultraviolet spectra, it is ensured that unsupported drug is washed off.Then the container is immersed into the cuvette equipped with deionized water
In, start release experiment, first discharge 1h in room temperature, then heat to 50 DEG C and carry out heating release, during which survey at regular intervals
Fluorescent emission intensity of the solution in the range of 490nm~540nm in a fixed cuvette, according to the standard of fluorescein sodium concentration
Song calculates the total concentration of the fluorescein sodium released in cuvette, as a result as shown in figure 18.
As shown in Figure 18, load the MSN-PEP of drug when being placed at room temperature for, the concentration of fluorescein sodium almost without increase,
Illustrate the system at room temperature almost without leakage.When outside, heating makes temperature from when being increased to 50 DEG C for 25 DEG C, due to fluorescein
The release of sodium, the significant increase of concentration of fluorescein sodium.20 it is small when after also measure load drug MSN-PEP room temperature release
Situation does not observe apparent leakage yet.Illustrate to block MSN by the self assembly energy range of the amphipathic small peptide of MSN surface graftings
Duct, the drug loaded at room temperature in duct hardly leaks, and amphipathic small peptide nanometer valve can be in relatively mild temperature
It triggers under the conditions of 50 DEG C of degree and discharges drug.
(2) the SPNC@MSN-PEP of 15mg, ultrasonic disperse are added in into the aqueous solution of the fluorescein sodium of 1.5mL 1mmol/L
Uniformly, the test tube equipped with SPNC@MSN-PEP and fluorescein sodium aqueous solution is kept the temperature to 2h in 50 DEG C of water-bath with by fluorescein sodium
It is supported on SPNC@MSN-PEP, it is then that test tube is amphipathic to ensure to be grafted on SPNC@MSN-PEP being stored at room temperature 12h
Small peptide self assembly again obtains loading the SPNC@MSN-PEP of drug.
The SPNC@MSN-PEP that 3mg is loaded to drug are dispersed in 250 μ L water, are then charged into both ends open and opening is covered
It is stamped in the container of dialysis membrane (14kDa MWCO), before release, container immersion is persistently changed several in the deionized water of water
My god, and detected by ultraviolet spectra, it is ensured that unsupported drug is washed off.Then the container is immersed into the ratio equipped with deionized water
In color ware, 1h is being stored at room temperature, during which every the solution in a 10min cuvette of measure in the range of 490nm~540nm
Fluorescent emission intensity, then by cuvette in superparamagnetism magnetic field, AMF is heated in 5 circle copper coil (power:5kW;Frequently
Rate:30 minutes (heating in 10 minutes and 1 minute cooling cycle are three times) is carried out under 370kHZ).It is kept away using the circulating cooling of ice bath
Exempt from continuous magnetic heating and generate amount of heat.After superparamagnetism heating, monitoring release at room temperature (measures 0 in 30 minutes respectively
Minute, the mean fluorecence emissive porwer after 15 minutes and 30 minutes 3 times cycle periods).4 such superparamagnetics have been carried out to add
Heat/monitoring Xun Huan.According to the bent amount for calculating the fluorescein sodium discharged under different temperatures of the standard of fluorescein sodium concentration, as a result as schemed
Shown in 49.It appears from figure 19 that the amphipathic small peptide for being grafted on SPNC@MSN surfaces can also by the nanometer valve being self-assembly of
It is oscillated magnetic field triggering.
Embodiment 3
In the present embodiment, Daunomycin (DNR) is loaded on SPNC@MSN-PEP, step is as follows:
The SPNC@MSN-PEP of 15mg are added in into the DNR aqueous solutions of 1.5mL 2mg/mL, ultrasonic disperse is uniform, will be equipped with
The test tube of SPNC@MSN-PEP and DNR aqueous solutions keeps the temperature 2h so that DNR is supported on SPNC@MSN-PEP in 50 DEG C of water-bath,
Then test tube is being stored at room temperature 12h to ensure the amphipathic small peptide self assembly again being grafted on SPNC@MSN-PEP, is being filled
The SPNC@MSN-PEP of DNR are carried, are expressed as DNR@SPNC@MSN-PEP.
Embodiment 4
In the present embodiment, using daunorubicin (DNR) as the model anticancer drug of cell research, SPNC@MSN-PEP are investigated
With the cytotoxicity of DNR@SPNC@MSN-PEP.
SPNC@MSN-PEP and DNR@SPNC@are tested on PANC-1 cells using WST-8 Cell Proliferation Assay Kits
The cytotoxicity of MSN-PEP.By PANC-1 cells at 37 DEG C, 5%CO2Culture is extremely merged.100 μ L are contained 1 × 104A cell
Cell suspension inoculation into each hole of 16 hole microscopic slides, then respectively into hole add various concentration (6.125,
12.5,25,50,100 μ g/mL) SPNC@MSN-PEP or DNR@SPNC@MSN-PEP.By cell and SPNC@MSN-PEP or
DNR@SPNC@MSN-PEP cultivate 48h at 37 DEG C, then add in the WST-8 mixtures of 10 μ L into each hole respectively.By cell
At 37 DEG C, CO24h is cultivated in incubator.After being gently mixed, sample is measured under 450nm wavelength using microplate reader, is counted
Cell survival rate is calculated, as a result as shown in figure 20.
In Figure 20, black column represents SPNC@MSN-PEP test groups, and grey column represents DNR@SPNC@MSN-PEP test groups,
As shown in Figure 20, each group test group is respectively provided with higher cell survival rate, shows amphipathic short peptide modified nano particle in reality
No cytotoxicity under concentration is tested, apparent toxicity is not observed in DNR@SPNC@MSN-PEP test groups, this shows amphipathic short
The nano-valve that self-assembling peptide is formed can hold the agent in the passage of nano particle and hardly leak.
Embodiment 5
In the present embodiment, can investigate DNR@SPNC@MSN-PEP discharge drug by magnetic triggered, and verify that magnetic adds
Heat is to the security of exposed cell.
SPNC@MSN-PEP, DNR@SPNC@are investigated on PANC-1 cells using WST-8 Cell Proliferation Assay Kits
Influences of the MSN-PEP in magnetic field to cell.By PANC-1 cells at 37 DEG C, 5%CO2Culture extremely fusion P.1 × 10 will be contained4
100 μ L cell suspension inoculations of a cell are into each hole of 12 hole microscopic slides, then to being added in respectively into hole
SPNC@MSN-PEP or DNR@SPNC@MSN-PEP make the concentration of SPNC@MSN-PEP or DNR@SPNC@MSN-PEP reach 50 μ g/
ML, blank control group are added without SPNC@MSN-PEP or DNR@SPNC@MSN-PEP, cultivate 2h.Expose cells to oscillating magnetic field
Culture in (10 minutes of 6 Xun Huans are heated and 1 minute cooling cycle).In duplicate orifice plate, magnetic field is not exposed to
Cell is as a control group to compare cytotoxicity.Then 12 hole microscopic slides are cultivated at 37 DEG C 48 it is small when, respectively to
10 μ L WST-8 mixtures are added in each hole, at 37 DEG C, CO24h is cultivated in incubator, after being gently mixed, is existed using microplate reader
Each sample is carried out under 450nm wavelength can measurement.The test carries out three parts of parallel laboratory tests.The calculation cell survival being finally calculated
Rate result is as shown in figure 21.
In Figure 21, the situation in black column and grey column difference representative sample " not exposing " and " being exposed to " magnetic field, by Figure 21
It understands, exposure measures the significant difference between the effect for showing SPNC@MSN-PEP and DNR@SPNC@MSN-PEP:SPNC@MSN-
PEP is terminated with about 100% cell survival rate, and cell survival rate is reduced to about 33% by DNR@SPNC@MSN-PEP, data
The heat that showing the nanometer valve that amphipathic small peptide is self-assembly of can be generated by magnetic field triggers, and the drug of release successfully kills cancer
Cell, and magnetic field will not generate any toxicity to exposed cell.By blank control group and it is not exposed to the control group in magnetic field
Data understand, according to the test condition of the present embodiment in the case where not applying magnetic field, SPNC@MSN-PEP concentration be 50 μ
Do not have cytotoxicity during g/mL, DNR@SPNC@MSN-PEP wherein loaded when concentration is 50 μ g/mL almost without leakage,
It is consistent with the cytotoxicity testing result of embodiment 4.
Embodiment 6
In the present embodiment, the nucleocapsid mesoporous silica nano-particle (SPNC of controllable temperature-sensitive peptide nanometer valve modification is prepared
MSN-PEP), step is as follows:
(1) magnetic iron oxide (MnFe of doping manganese is prepared2O4) nano particle
By Fe (acac)3、Mn(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:9:5:5 molar ratio is molten
Solution makes Fe (acac) in benzylic ether3Concentration for 0.08mmol/mL, be heated to 210 DEG C under nitrogen protection and stirring condition,
Then 2h are kept the temperature in 210 DEG C under nitrogen protection, then in 298 DEG C of back flow reaction 0.5h, obtained containing MnFe2O4Nano particle
The reaction solution is cooled to room temperature by reaction solution, is added in ethyl alcohol and is precipitated MnFe2O4Nano particle, by centrifuging MnFe2O4It receives
Rice grain further separates, and is dispersed in n-hexane and preserves again.MnFe is detected by ICP-OES2O4In nano particle manganese with
The molar ratio of iron is 1:2.
(2) magnetic iron oxide (MnFe of doping manganese and cobalt is prepared2O4@CoFe2O4) nano particle
MnFe2O4@CoFe2O4Nano particle is by MnFe2O4CoFe is grown on nano particle2O4Come what is prepared.
By Fe (acac)3、Co(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:9:2:2 molar ratio is molten
Solution makes Fe (acac) in benzylic ether3Concentration for 0.08mmol/mL, obtain mixed liquor E, the magnetic iron oxide for adulterating manganese received
Rice grain, which is scattered in n-hexane, makes MnFe2O4The concentration of nano particle forms dispersion liquid F for 15mg/mL, by mixed liquor E with dividing
Dispersion liquid F is according to 10:6 volume ratio mixing, heating evaporation fall n-hexane, are heated to 210 DEG C in the case where nitrogen is protected and is stirred, then
1h are kept the temperature in 210 DEG C under nitrogen protection, then in 298 DEG C of back flow reaction 0.5h, are obtained containing MnFe2O4@CoFe2O4Nanometer
Grain (abbreviation SPNC) reaction solution, which is cooled to room temperature, add in ethyl alcohol in precipitate SPNC, by centrifugation by SPNC into
One step separates, and is dispersed in n-hexane and preserves again.By ICP-OES detect SPNC in manganese, iron, cobalt molar ratio be 1:4:
1。
(3) hud typed mesoporous silica nano-particle (SPNC@MSN) is prepared
N-hexane is fallen to the n-hexane heating evaporation for being dispersed with SPNC, obtained SPNC is scattered in CHCl3Middle formed is divided
Dispersion liquid A adds in dispersion liquid A in CTAB solution, is ultrasonically treated 15min, then mild heating evaporation removal CHCl3, add water
The concentration to CTAB is adjusted as 1.8mg/mL, pH value is then adjusted to 11.0~11.5 and obtains mixed liquid B, in mixed liquid B, SPNC
Mass ratio with CTAB is 1:18, mixed liquid B, TEOS, the volume ratio of ethyl acetate are 100:1:5,60min is stirred at 68 DEG C,
TEOS is added dropwise, adds ethyl acetate, reacts 4h at 68 DEG C, separation of solid and liquid washes solid phase reaction product with water and ethyl alcohol successively
It washs, obtains SPNC@MSN, SPNC@MSN are dispersed in n-hexane and are preserved.
N-hexane is fallen to the n-hexane heating evaporation for being dispersed with SPNC, obtained SPNC is scattered in CHCl3Middle formed is divided
Dispersion liquid A adds in dispersion liquid A in CTAB solution, is ultrasonically treated 20min, then mild heating evaporation removal CHCl3, add water
The concentration to CTAB is adjusted as 2.2mg/mL, pH value is then adjusted to 11.0~11.5 and obtains mixed liquid B, in mixed liquid B, SPNC
Mass ratio with CTAB is 1:22, mixed liquid B, TEOS, the volume ratio of ethyl acetate are 100:1:15,30min is stirred at 72 DEG C,
TEOS is added dropwise, adds ethyl acetate, reacts 3h at 72 DEG C, separation of solid and liquid washes solid phase reaction product with water and ethyl alcohol successively
It washs, obtains SPNC@MSN, SPNC@MSN are dispersed in n-hexane and are preserved.
(4) hud typed mesoporous silica nano-particle (the SPNC@MSN-NH that surface amino groups are modified are prepared2)
Under nitrogen protection, SPNC@MSN are scattered in ethyl alcohol, then add in N- [3- (trimethoxy silicon substrate) propyl]
The mass ratio of ethylenediamine, N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines and SPNC@MSN are 5%;It heats under nitrogen protection
Return stirring reacts 10h, and by reaction product ethyl alcohol and water washing, the reaction product after washing is placed in ammonium nitrate concn is
In 4% ammonium nitrate ethanol solution, 1h is heated to reflux, gained nano particle is isolated, adds in nitric acid after being washed with ethyl alcohol again
Ammonium concentration is in 4% ammonium nitrate ethanol solution, is heated to reflux 1h, isolates gained nano particle and is washed with ethyl alcohol,
Obtain SPNC@MSN-NH2, by SPNC@MSN-NH2It is dispersed in water preservation.
(5) the nucleocapsid mesoporous silica nano-particle (SPNC MSN-PEP) of controllable temperature-sensitive peptide nanometer valve modification is prepared
By amphipathic small peptide, EDC.HCl and the Sulfo-NHS that amino acid sequence is Boc-Phe Phe Gly Gly-COOH
According to 8:10:5 mass ratio is dissolved in MES buffer solutions (pH=6.0,100mM)) in make the concentration of Sulfo-NHS for 9mg/mL,
Mixed liquor C is obtained, by SPNC@MSN-NH2It is scattered in and point that concentration is 8mg/mL is obtained in PBS buffer solution (pH=7.4,50mM)
Dispersion liquid D, by mixed liquor C and dispersion liquid D according to 0.8:1 volume ratio mixing, in room temperature reaction 10h, reaction process, SPNC@
MSN-NH2For the nitrogen of amino on surface as nucleopilic reagent, the carbon attacked on the carboxyl of amphipathic small peptide forms C-N keys, will be anti-
Product is answered to be washed with water 3 times to get in SPNC@MSN-PEP, SPNC@MSN-PEP, amphipathic small peptide is grafted on long chain amino
Shown in the structural formula of the group of formation such as formula (II), in the structural formula shown in formula (II), on the O and SPNC@MSN that are connected with Si
Si is connected to form Si-O keys.
Embodiment 7
In the present embodiment, the nucleocapsid mesoporous silica nano-particle (SPNC of controllable temperature-sensitive peptide nanometer valve modification is prepared
MSN-PEP), step is as follows:
(1) magnetic iron oxide (MnFe of doping manganese is prepared2O4) nano particle
By Fe (acac)3、Mn(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:10:7:7 molar ratio
Being dissolved in benzylic ether makes Fe (acac)3Concentration for 0.12mmol/mL, be heated to 190 under nitrogen protection and stirring condition
DEG C, 2.5h then are kept the temperature in 190 DEG C under nitrogen protection, then in 298 DEG C of back flow reaction 2h, are obtained containing MnFe2O4Nano particle
Reaction solution, which is cooled to room temperature, add in ethyl alcohol in precipitate MnFe2O4Nano particle, by centrifuging MnFe2O4
Nano particle further separates, and is dispersed in n-hexane and preserves again.MnFe is detected by ICP-OES2O4Manganese in nano particle
Molar ratio with iron is 1:2.
(2) magnetic iron oxide (MnFe of doping manganese and cobalt is prepared2O4@CoFe2O4) nano particle
MnFe2O4@CoFe2O4Nano particle is by MnFe2O4CoFe is grown on nano particle2O4Come what is prepared.
By Fe (acac)3、Co(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:10:2:2 molar ratio
Being dissolved in benzylic ether makes Fe (acac)3Concentration for 0.12mmol/mL, obtain mixed liquor E, the magnetic iron oxide of manganese will be adulterated
Nano particle, which is scattered in n-hexane, makes MnFe2O4The concentration of nano particle for 20mg/mL formed dispersion liquid F, by mixed liquor E with
Dispersion liquid F is according to 10:5 volume ratio mixing, heating evaporation fall n-hexane, are heated to 190 DEG C in the case where nitrogen is protected and is stirred, so
1.5h are kept the temperature in 190 DEG C under nitrogen protection afterwards, then in 298 DEG C of back flow reaction 2h, are obtained containing MnFe2O4@CoFe2O4Nanometer
The reaction solution is cooled to room temperature by the reaction solution of particle (abbreviation SPNC), is added in ethyl alcohol and is precipitated SPNC, by centrifuging SPNC
Further separation, and be dispersed in n-hexane and preserve again.By ICP-OES detect SPNC in manganese, iron, cobalt molar ratio be 1:
4:1。
(3) hud typed mesoporous silica nano-particle (SPNC@MSN) is prepared
N-hexane is fallen to the n-hexane heating evaporation for being dispersed with SPNC, obtained SPNC is scattered in CHCl3Middle formed is divided
Dispersion liquid A adds in dispersion liquid A in CTAB solution, is ultrasonically treated 20min, then mild heating evaporation removal CHCl3, add water
The concentration to CTAB is adjusted as 2.2mg/mL, pH value is then adjusted to 11.0~11.5 and obtains mixed liquid B, in mixed liquid B, SPNC
Mass ratio with CTAB is 1:22, mixed liquid B, TEOS, the volume ratio of ethyl acetate are 100:1:15,30min is stirred at 72 DEG C,
TEOS is added dropwise, adds ethyl acetate, reacts 3h at 72 DEG C, separation of solid and liquid washes solid phase reaction product with water and ethyl alcohol successively
It washs, obtains SPNC@MSN, SPNC@MSN are dispersed in n-hexane and are preserved.
(4) hud typed mesoporous silica nano-particle (the SPNC@MSN-NH that surface amino groups are modified are prepared2)
Under nitrogen protection, SPNC@MSN are scattered in ethyl alcohol, then add in (3- aminopropyls) trimethoxy silane,
The mass ratio of (3- aminopropyls) trimethoxy silane and SPNC@MSN are 8%;It is heated to reflux being stirred to react under nitrogen protection
Reaction product after washing by reaction product ethyl alcohol and water washing, is placed in the ammonium nitrate second that ammonium nitrate concn is 8% by 14h
In alcoholic solution, 1h is heated to reflux, isolates gained nano particle, it is 8% to add in ammonium nitrate concn after being washed with ethyl alcohol again
In ammonium nitrate ethanol solution, 1h is heated to reflux, isolate gained nano particle and is washed with ethyl alcohol, obtains SPNC@MSN-
NH2, by SPNC@MSN-NH2It is dispersed in water preservation.
(5) the nucleocapsid mesoporous silica nano-particle (SPNC MSN-PEP) of controllable temperature-sensitive peptide nanometer valve modification is prepared
By amphipathic small peptide, EDC.HCl and the Sulfo-NHS that amino acid sequence is Boc-Phe Phe Gly Gly-COOH
According to 6:10:5 mass ratio is dissolved in MES buffer solutions (pH=6.0,100mM)) in make Sulfo-NHS concentration be 12mg/
ML obtains mixed liquor C, by SPNC@MSN-NH2It is scattered in PBS buffer solution (pH=7.4,100mM) and obtains concentration as 15mg/
The dispersion liquid D of mL, by mixed liquor C and dispersion liquid D according to 1.2:1 volume ratio mixing, in room temperature reaction 18h, reaction process,
SPNC@MSN-NH2For the nitrogen of amino on surface as nucleopilic reagent, the carbon attacked on the carboxyl of amphipathic small peptide forms C-N keys,
Reaction product is washed with water 3 times to get in SPNC@MSN-PEP, SPNC@MSN-PEP, amphipathic small peptide is grafted on long-chain ammonia
Shown in the structural formula of the group formed on base such as formula (III), in the structural formula shown in formula (III), the O and SPNC@MSN that are connected with Si
On Si be connected to form Si-O keys.
Claims (9)
1. the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification, which is characterized in that the nano particle by
Hud typed mesoporous silica nano-particle and hud typed mesoporous silica nano-particle table is grafted on by long chain amino
The amphipathic small peptide composition in face, the hud typed mesoporous silica nano-particle is by superparamagnetic nano particle core and package
The mesoporous silicon oxide shell composition of superparamagnetic nano particle core, hud typed meso-porous titanium dioxide is grafted on by long chain amino
The amphipathic small peptide on nano silicon particles surface, which can respond the variation self assembly of temperature or unlock self assembly, makes meso-porous titanium dioxide
Duct on silicon shell is in closure or opening state, and the amino acid sequence of the amphipathic small peptide is Boc-Phe Phe
Gly Gly-COOH, long chain amino areOr
2. the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification according to claim 1, feature
It is, in the nano particle, the mass ratio of hud typed mesoporous silica nano-particle, long chain amino and amphipathic small peptide is
95:(5~7):(4~10).
3. the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification according to claim 1 or claim 2, special
Sign is the superparamagnetic nano particle core for doping manganese and the superparamag-netic iron oxide of cobalt.
4. the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification according to claim 3, feature
It is, in the superparamag-netic iron oxide for adulterating manganese and cobalt, manganese, iron, the molar ratio of cobalt are 1:4:1.
5. the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification according to claim 4, feature
It is the grain size of superparamagnetic nano particle core for 8.5~13.5nm, the grain size of hud typed mesoporous silica nano-particle
For 32.5~65.5nm.
6. a kind of preparation method of the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification, it is characterised in that
Step is as follows:
(1) superparamag-netic iron oxide for adulterating manganese and cobalt is scattered in CHCl3Middle formation dispersion liquid A, by dispersion liquid A and ten
Six alkyl trimethyl ammonium bromide solution are sufficiently mixed, then heating evaporation removal CHCl3, water is added to adjust to cetyl trimethyl
The concentration of ammonium bromide is 1.8~2.2mg/mL, and then adjusting pH value to 11.0~11.5 obtains mixed liquid B, is stirred at 68~72 DEG C
30~60min is mixed, tetraethyl orthosilicate is added dropwise, adds ethyl acetate, 3~4h is reacted at 68~72 DEG C, by reaction product water
It is washed with ethyl alcohol, obtains hud typed mesoporous silica nano-particle;
In mixed liquid B, adulterate manganese and the superparamag-netic iron oxide of cobalt and the mass ratio of cetyl trimethylammonium bromide is
1:(18~22), mixed liquid B, tetraethyl orthosilicate, the volume ratio of ethyl acetate are 100:1:(5~15),
(2) under nitrogen protection, hud typed mesoporous silica nano-particle is scattered in ethyl alcohol, then adds in (3- ammonia third
Base) trimethoxy silane, N- [3- (trimethoxy silicon substrate) propyl] ethylenediamines or diethylenetriamine base propyl trimethoxy silicon
Alkane is heated to reflux being stirred to react 10~14h under nitrogen protection, by reaction product ethyl alcohol and water washing, is subsequently placed in nitric acid
Ammonium concentration is that removal cetyl trimethylammonium bromide is heated to reflux in 4%~8% ammonium nitrate ethanol solution, is carried out with ethyl alcohol
The hud typed mesoporous silica nano-particle of surface amino groups modification is obtained after washing;(3- aminopropyls) trimethoxy silane, N-
[3- (trimethoxy silicon substrate) propyl] ethylenediamine or diethylenetriamine base propyl trimethoxy silicane and hud typed mesoporous dioxy
The mass ratio of SiClx nano particle is 5%~8%;
(3) it is amphipathic small peptide, 1- (the 3- dimethylaminos third of Boc-Phe Phe Gly Gly-COOH by amino acid sequence
Base) -3- ethyl-carbodiimide hydrochlorides and n-hydroxysuccinimide sulfonate sodium be according to (6~10):10:5 mass ratio is molten
Solution makes the concentration of n-hydroxysuccinimide sulfonate sodium mixed liquor C be obtained, by table for 9~12mg/mL in MES buffer solutions
The amino modified hud typed mesoporous silica nano-particle in face, which is scattered in, obtains concentration as 8~15mg/mL's in PBS buffer solution
Dispersion liquid D, by mixed liquor C and dispersion liquid D according to (0.8~1.2):1 volume ratio mixing, is reacting at room temperature 10~18h, will be anti-
Product is answered to be washed with water to get controllable temperature-sensitive peptide nanometer valve modification nucleocapsid mesoporous silica nano-particle.
7. the preparation side of the nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification according to claim 6
Method, it is characterised in that the preparation method of the superparamag-netic iron oxide of doping manganese and cobalt is as follows:
(1) by Fe (acac)3、Mn(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:(9~10):(5~7):
The molar ratio of (5~7), which is dissolved in benzylic ether, makes Fe (acac)3Concentration for 0.08~0.12mmol/mL, in nitrogen protection and
190~210 DEG C are heated under stirring condition, then keeps the temperature 2~2.5h in 190~210 DEG C under nitrogen protection, reheats reflux
0.5~2h is reacted, gained reaction solution is cooled to room temperature, adds in ethyl alcohol and precipitates to get the magnetic ferric oxide nano of doping manganese
Particle;
(2) by Fe (acac)3、Co(acac)2, 1,2- dodecanediols, oleic acid and oleyl amine be according to 2:1:(9~10):2:2 rub
You make Fe (acac) than being dissolved in benzylic ether3Concentration for 0.08~0.12mmol/mL, obtain mixed liquor E, manganese will be adulterated
Superparamag-netic iron oxide, which is scattered in n-hexane, makes MnFe2O4The concentration of nano particle forms dispersion liquid for 15~20mg/mL
F, by mixed liquor E and dispersion liquid F according to 10:The volume ratio mixing of (5~6), heating evaporation fall n-hexane, protect and stir in nitrogen
It mixes down and is heated to 190~210 DEG C, then keep the temperature 1~1.5h in 190~210 DEG C under nitrogen protection, reheat back flow reaction
Gained reaction solution is cooled to room temperature by 0.5~2h, adds in ethyl alcohol the magnetic ferric oxide nano precipitated to get doping manganese and cobalt
Particle.
8. according to the system for the nucleocapsid mesoporous silica nano-particle that the controllable temperature-sensitive peptide nanometer valve of claim 6 or 7 is modified
Preparation Method, which is characterized in that the pH value of MES buffer solutions is 6.0, concentration is 50~100mmol/L, and the pH value of PBS buffer solution is
7.4th, concentration is 50~100mmol/L.
9. the nucleocapsid mesoporous silicon oxide of controllable temperature-sensitive peptide nanometer valve modification is received described in any claim in claim 1 to 5
Application of the rice grain as pharmaceutical carrier.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810136097.7A CN108042509B (en) | 2018-02-09 | 2018-02-09 | Controllable thermosensitive peptide nanometer valve modified core-shell mesoporous silica nanoparticle and preparation method and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810136097.7A CN108042509B (en) | 2018-02-09 | 2018-02-09 | Controllable thermosensitive peptide nanometer valve modified core-shell mesoporous silica nanoparticle and preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108042509A true CN108042509A (en) | 2018-05-18 |
CN108042509B CN108042509B (en) | 2020-12-01 |
Family
ID=62125882
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810136097.7A Expired - Fee Related CN108042509B (en) | 2018-02-09 | 2018-02-09 | Controllable thermosensitive peptide nanometer valve modified core-shell mesoporous silica nanoparticle and preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108042509B (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108743958A (en) * | 2018-05-31 | 2018-11-06 | 四川大学 | Drug molecule and the GSH response type nanometers of valve molecule synergy carry medicine particle and preparation method thereof |
CN114772606A (en) * | 2022-03-10 | 2022-07-22 | 四川大学 | Carbon-silicon dioxide core-shell composite nano material for electromagnetic wave absorption and preparation method thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103536935A (en) * | 2013-11-26 | 2014-01-29 | 上海师范大学 | Photosensitizer-modified core-shell structure magnetic nanocomposites and preparation method and application thereof |
CN103550793A (en) * | 2013-10-28 | 2014-02-05 | 苏州大学附属第一医院 | Magnetic nano material, preparation method and application of magnetic nano material in magnetic resonance contrast imaging |
CN106620729A (en) * | 2017-01-17 | 2017-05-10 | 上海大学 | Inorganic-inorganic nano hybrid material of bimodal mesoporous core-shell structure as well as preparation method and application of inorganic-inorganic nano hybrid material |
-
2018
- 2018-02-09 CN CN201810136097.7A patent/CN108042509B/en not_active Expired - Fee Related
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103550793A (en) * | 2013-10-28 | 2014-02-05 | 苏州大学附属第一医院 | Magnetic nano material, preparation method and application of magnetic nano material in magnetic resonance contrast imaging |
CN103536935A (en) * | 2013-11-26 | 2014-01-29 | 上海师范大学 | Photosensitizer-modified core-shell structure magnetic nanocomposites and preparation method and application thereof |
CN106620729A (en) * | 2017-01-17 | 2017-05-10 | 上海大学 | Inorganic-inorganic nano hybrid material of bimodal mesoporous core-shell structure as well as preparation method and application of inorganic-inorganic nano hybrid material |
Non-Patent Citations (1)
Title |
---|
G MARTELLI ET AL.: "Coiled-coil peptide motifs as thermoresponsive valves for mesoporous silica nanoparticles", 《ROYAL SOCIETY OF CHEMISTRY》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108743958A (en) * | 2018-05-31 | 2018-11-06 | 四川大学 | Drug molecule and the GSH response type nanometers of valve molecule synergy carry medicine particle and preparation method thereof |
CN108743958B (en) * | 2018-05-31 | 2021-09-28 | 四川大学 | GSH response type mesoporous silicon nano drug-loaded particle with combined action of drug molecules and valve molecules and preparation method thereof |
CN114772606A (en) * | 2022-03-10 | 2022-07-22 | 四川大学 | Carbon-silicon dioxide core-shell composite nano material for electromagnetic wave absorption and preparation method thereof |
CN114772606B (en) * | 2022-03-10 | 2023-09-19 | 四川大学 | Carbon-silicon dioxide core-shell composite nano material for electromagnetic wave absorption and preparation method thereof |
Also Published As
Publication number | Publication date |
---|---|
CN108042509B (en) | 2020-12-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Mahtab et al. | Fabrication of silica nanoparticles with both efficient fluorescence and strong magnetization and exploration of their biological applications | |
Wang et al. | A molecularly imprinted polymer-coated nanocomposite of magnetic nanoparticles for estrone recognition | |
Bohara et al. | Cancer cell extinction through a magnetic fluid hyperthermia treatment produced by superparamagnetic Co–Zn ferrite nanoparticles | |
CN101966344B (en) | Hollow core-shell nanometer mesoporous medicament carrying system with magnetism and luminescent performance, preparation method and application thereof | |
Arriortua et al. | RGD-Functionalized Fe3O4 nanoparticles for magnetic hyperthermia | |
Qin et al. | Highly water-dispersible TiO 2 nanoparticles for doxorubicin delivery: effect of loading mode on therapeutic efficacy | |
EP2184262B1 (en) | Process for production of surface-coated inorganic particles | |
Galli et al. | Superparamagnetic iron oxide nanoparticles functionalized by peptide nucleic acids | |
Benyettou et al. | Microwave assisted nanoparticle surface functionalization | |
Che et al. | CO 2-switchable drug release from magneto-polymeric nanohybrids | |
Šponarová et al. | The use of oligoperoxide-coated magnetic nanoparticles to label stem cells | |
Zhao et al. | Multifunctional superparamagnetic Fe3O4@ SiO2 core/shell nanoparticles: design and application for cell imaging | |
Yang et al. | A direct surface modification of iron oxide nanoparticles with various poly (amino acid) s for use as magnetic resonance probes | |
CN108815523A (en) | A kind of New Type of Mesoporous silicon ball is total to medicament-carried nano compound and preparation method thereof | |
CN108042509A (en) | Nucleocapsid mesoporous silica nano-particle of controllable temperature-sensitive peptide nanometer valve modification and preparation method and application | |
Shen et al. | Horseradish peroxidase-immobilized magnetic mesoporous silica nanoparticles as a potential candidate to eliminate intracellular reactive oxygen species | |
Luong et al. | Magnetothermal release of payload from iron oxide/silica drug delivery agents | |
Gao et al. | Dopamine coating as a general and facile route to biofunctionalization of superparamagnetic Fe 3 O 4 nanoparticles for magnetic separation of proteins | |
CN108743958A (en) | Drug molecule and the GSH response type nanometers of valve molecule synergy carry medicine particle and preparation method thereof | |
Niu et al. | Fabrication of uniform, biocompatible and multifunctional PCL-b-PAA copolymer-based hybrid micelles for magnetic resonance imaging | |
Liu et al. | Synthesis and applications of fluorescent-magnetic-bifunctional dansylated Fe 3 O 4@ SiO 2 nanoparticles | |
Khosroshahi et al. | Characterization and Cellular Fluorescence Microscopy of Superparamagnetic Nanoparticles Functionalized with Third Generation Nano-molecular Dendrimers: In-vitro Cytotoxicity and Uptake study. J Nanomater Mol Nanotechnol 5: 3 | |
CN108743960A (en) | A kind of mesoporous carbon nanoparticle and its application | |
Kim et al. | Synthesis of hybrid gold/iron oxide nanoparticles in block copolymer micelles for imaging, drug delivery, and magnetic hyperthermia | |
Cui et al. | Functional nanoscale metal–organic particles synthesized from a new vinylimidazole-based polymeric ligand and dysprosium ions |
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 | ||
CF01 | Termination of patent right due to non-payment of annual fee | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20201201 |