CN109395092A - 一种基于主客体相互作用的载体及其应用 - Google Patents
一种基于主客体相互作用的载体及其应用 Download PDFInfo
- Publication number
- CN109395092A CN109395092A CN201811264940.6A CN201811264940A CN109395092A CN 109395092 A CN109395092 A CN 109395092A CN 201811264940 A CN201811264940 A CN 201811264940A CN 109395092 A CN109395092 A CN 109395092A
- Authority
- CN
- China
- Prior art keywords
- carrier
- pcl
- hpg
- molecule
- pei600
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000003993 interaction Effects 0.000 title claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 31
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 20
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 19
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 19
- 229920000642 polymer Polymers 0.000 claims abstract description 15
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 230000008878 coupling Effects 0.000 claims abstract description 4
- 238000010168 coupling process Methods 0.000 claims abstract description 4
- 238000005859 coupling reaction Methods 0.000 claims abstract description 4
- 150000001875 compounds Chemical class 0.000 claims description 21
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims description 19
- 108020004414 DNA Proteins 0.000 claims description 14
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 239000013612 plasmid Substances 0.000 claims description 7
- 239000001116 FEMA 4028 Substances 0.000 claims description 6
- 229960004853 betadex Drugs 0.000 claims description 6
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 claims description 3
- 229930012538 Paclitaxel Natural products 0.000 claims description 3
- DMLAVOWQYNRWNQ-UHFFFAOYSA-N azobenzene Chemical compound C1=CC=CC=C1N=NC1=CC=CC=C1 DMLAVOWQYNRWNQ-UHFFFAOYSA-N 0.000 claims description 3
- 229960003668 docetaxel Drugs 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 3
- 229960001592 paclitaxel Drugs 0.000 claims description 3
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 claims description 3
- ORILYTVJVMAKLC-UHFFFAOYSA-N adamantane Chemical compound C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 claims 2
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 claims 1
- 238000001890 transfection Methods 0.000 abstract description 24
- 231100000135 cytotoxicity Toxicity 0.000 abstract description 12
- 230000003013 cytotoxicity Effects 0.000 abstract description 12
- 229940079593 drug Drugs 0.000 abstract description 10
- 210000002966 serum Anatomy 0.000 abstract description 8
- 230000004044 response Effects 0.000 abstract description 5
- 230000002209 hydrophobic effect Effects 0.000 abstract description 4
- 230000004048 modification Effects 0.000 abstract description 3
- 238000012986 modification Methods 0.000 abstract description 3
- 230000009466 transformation Effects 0.000 abstract description 2
- 229920002873 Polyethylenimine Polymers 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 17
- 229920001610 polycaprolactone Polymers 0.000 description 17
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin 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(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 16
- 108090000623 proteins and genes Proteins 0.000 description 15
- 230000006872 improvement Effects 0.000 description 14
- 230000015572 biosynthetic process Effects 0.000 description 11
- 238000003786 synthesis reaction Methods 0.000 description 11
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 206010028980 Neoplasm Diseases 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 239000000693 micelle Substances 0.000 description 6
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- 229920001577 copolymer Polymers 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000004632 polycaprolactone Substances 0.000 description 5
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000007423 decrease Effects 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 238000012637 gene transfection Methods 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 229920006317 cationic polymer Polymers 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- 239000002053 C09CA06 - Candesartan Substances 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 102000004144 Green Fluorescent Proteins Human genes 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 108091027967 Small hairpin RNA Proteins 0.000 description 2
- 108700005077 Viral Genes Proteins 0.000 description 2
- 150000001335 aliphatic alkanes Chemical class 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 125000003785 benzimidazolyl group Chemical group N1=C(NC2=C1C=CC=C2)* 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- SGZAIDDFHDDFJU-UHFFFAOYSA-N candesartan Chemical compound CCOC1=NC2=CC=CC(C(O)=O)=C2N1CC(C=C1)=CC=C1C1=CC=CC=C1C1=NN=N[N]1 SGZAIDDFHDDFJU-UHFFFAOYSA-N 0.000 description 2
- 229960000932 candesartan Drugs 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000012876 carrier material Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000001427 coherent effect Effects 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 239000005090 green fluorescent protein Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 2
- WEVYAHXRMPXWCK-UHFFFAOYSA-N methyl cyanide Natural products CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 238000011580 nude mouse model Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 229920000729 poly(L-lysine) polymer Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004043 responsiveness Effects 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000000954 titration curve Methods 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- NVUJWPQINQUNNM-UHFFFAOYSA-N 1h-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1.C1=CC=C2NC=NC2=C1 NVUJWPQINQUNNM-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 1
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 102000053028 CD36 Antigens Human genes 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- -1 DOX compound Chemical class 0.000 description 1
- 241000555268 Dendroides Species 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 1
- 108091005487 SCARB1 Proteins 0.000 description 1
- 102100031463 Serine/threonine-protein kinase PLK1 Human genes 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical class CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 229920003118 cationic copolymer Polymers 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000012650 click reaction Methods 0.000 description 1
- 229940000425 combination drug Drugs 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000007334 copolymerization reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 229960004679 doxorubicin Drugs 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001476 gene delivery Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000000111 isothermal titration calorimetry Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 108010056274 polo-like kinase 1 Proteins 0.000 description 1
- 229920000223 polyglycerol Polymers 0.000 description 1
- 150000004032 porphyrins Chemical group 0.000 description 1
- 229910001414 potassium ion Inorganic materials 0.000 description 1
- BDAWXSQJJCIFIK-UHFFFAOYSA-N potassium methoxide Chemical compound [K+].[O-]C BDAWXSQJJCIFIK-UHFFFAOYSA-N 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002109 single walled nanotube Substances 0.000 description 1
- 239000004055 small Interfering RNA Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 230000005747 tumor angiogenesis Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000010148 water-pollination Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
-
- 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
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种基于主客体相互作用的载体及其应用,载体包括主体分子和客体分子,主体分子为聚合物接枝改性的环糊精,客体分子可以容纳于环糊精的环内,客体分子共价偶联在PCL‑HPG的表面。本发明的载体,细胞毒性低,对疏水性药物分子和核酸序列均具有很好的载运能力,对血清具有很好的耐受能力和细胞相容性,同时可以很好地提高转染效率。通过客体分子的变换,可以获得不同pH响应的载体。
Description
技术领域
本发明涉及一种基于主客体相互作用的载体,同时还包括其应用。
背景技术
近年来,共递送化疗药物和基因用于癌症治疗成为了国内外的研究热点。药物和基因的联用可相互促进、相辅相成,减少化疗药物剂量,最终达到降低毒副作用、提高治疗效果的目的。该技术成功实施的关键在于一类既安全又高效的载体材料。目前,已经有多种载体材料被用于药物和基因的共递送,如无机纳米粒子、脂质体、胶束等。
Wang等[1]将药物坎地沙坦(CD)偶联在聚乙烯亚胺(PEI)上并修饰到单壁碳纳米管(SWNTs),然后再与血管内皮生长因子靶向的siRNA(siVEGF)发生静电复合,形成药物和基因的共递送体系(SWNT-PEI-CD/siVEGF),用于协同和靶向***血管生成。研究结果显示,SWNT-PEI-CD/siVEGF共递送体系具有肿瘤靶向特异性,并通过联用药物和基因可有效抑制肿瘤血管生成,治疗效果优于单独给药或基因。Wang和Chen[2]开发了一种脂蛋白衍生物纳米载体(rHDL),用于负载治疗基因(p53)和疏水性化疗药物(PTX),研究表明,经过优化的PTX-DODAB/p53-rHDL纳米粒子不仅可通过SR-BI介导实现优异的肿瘤靶向性,还对荷瘤裸鼠具有显著的抗肿瘤效果。Zhou课题组[3]在近期的一项研究中通过一锅法成功合成了pH敏感和氧化还原敏感的双亲性共聚物,该共聚物能形成胶束并对DOX和PLK1特异性shRNA进行有效负载,其敏感性可在肿瘤细胞微环境下被触发从而迅速释放包载的药物和基因;并以建立有U87胶质瘤的裸鼠为动物模型,研究了该体系对实体瘤的治疗效果,发现共递送治疗体系的治疗效率要高于单给DOX或单给shRNA治疗模式。为了将疏水药物DOX和功能基因pMMP-9有效地递送到肿瘤细胞,Ma[4]等设计合成了一类以卟啉以核、精氨酸修饰的聚(L-赖氨酸)为臂的星型阳离子聚合物(PP-PLLD-Arg)。研究表明,与单载药物或基因相比,PP-PLLD-Arg/DOC/pMMP-9复合物不仅可诱导更显著的细胞凋亡,而且显著降低了HNE-1细胞的侵袭能力。Liu[5]通过点击反应成功合成出由β-CD和树枝状聚(L-赖氨酸)构成的结构明确星型阳离子共聚物(CD-PLLD)。
科研工作者们在设计制备高效非病毒载体方面做了诸多努力,从阳离子脂质体到线性阳离子聚合物,然后到超支化阳离子聚合物,但如何平衡转染效率与细胞毒性之间的矛盾仍是一大挑战。以支化聚乙烯亚胺(PEI)为例,分子量为25000的PEI(PEI25k)是众所周知的具有稳定且高基因转染效率的基因载体,通常用作评价其他非病毒基因载体转染效率的黄金标准。然而,较高的细胞毒性和血液中的不稳定性限制了其在临床上的应用。低分子量PEI600(分子量为600的PEI)表现出低的细胞毒性和良好的稳定性,但基因转染效率却很低。
李晓慧.超支化聚己内酯/聚缩水甘油醚共聚物的合成及在基因传递载体方面的应用[D].中国协和医科大学,2009.公开了一种新型的超支化聚己内酯/聚缩水甘油醚共聚物(poly(epsilon-caprolactone)-b-hyperbranchedpolyglycidol,PCL-b-HPG)。PCL-b-HPG具有两亲性且带有大量羟基末端基团,能改善聚己内酯的亲水性和可修饰性。将PCL-b-HPG制备成纳米胶束,考察了其细胞毒性和基因转染效果,末端偶联RGD多肽,以期为开发新型靶向药物传递***奠定基础。通过TEM、SEM和粒径分析仪观察,所得纳米胶束为球形,形状均匀,粒径为(170±10.0)nm。采用MTT法,检测了PCL-b-HPG纳米胶束的细胞毒性,并制备了载pEGFP-C1基因纳米胶束,所制备的纳米胶束的平均粒径约为225nm,pEGFP-C1质粒的含量约占2%,基因包埋效率在85%以上。凝胶阻滞电泳实验,显示以纳米胶束的形式装载基因,泳道中出现了明显的拖带,表明PCL-b-HPG纳米胶束能有效的包裹质粒DNA分子,体外EA.hy926细胞转染实验证实PCL-b-HPG载基因纳米胶束能够转染质粒DNA,转染效率约在15%。其转染效率还是相对较低。
因此,如何设计具有高基因转染效率和低细胞毒性的非病毒基因载体是药物和基因共递送策略中的关键问题。
参考文献:
[1]Ding X F,Su Y J,Wang C,Zhang F R,Chen K R,Wang Y,Li M,WangW.Synergistic Suppression of Tumor Angiogenesis by the Co-delivering ofVascular Endothelial Growth Factor Targeted siRNA and Candesartan Mediated byFunctionalized Carbon Nanovectors[J].AcsAppl Mater Inter,2017,9(28):23353-23369.
[2]Wang W,Chen K,Su Y,Zhang J,Li M,Zhou J.Lysosome-IndependentIntracellular Drug/Gene Codelivery by Lipoprotein-Derived Nanovector forSynergistic Apoptosis-Inducing Cancer-Targeted Therapy[J].Biomacromolecules,2018,19(2):438-448.
[3]Wang P,Yu N,Wang Y,Sun H,Yang Z,Zhou S.Co-delivery of PLK1-specific shRNA and doxorubicin via core-crosslinked pH-sensitive and redoxultra-sensitive micelles for glioma therapy[J].J Mater Chem B,2018,6(1):112-124.
[4]Ma D,Lin Q M,Zhang L M,Liang Y Y,Xue W.Astar-shaped porphyrin-arginine functionalized poly(L-lysine)copolymer for photo-enhanced drug andgene co-delivery[J].Biomaterials,2014,35(14):4357-4367.
[5]Liu T,Xue W,Ke B,Xie M Q,Ma D.Star-shaped cyclodextrin-poly(L-lysine)derivative co-delivering docetaxel and MMP-9siRNA plasmid in cancertherapy[J].Biomaterials,2014,35(12):3865-3872.
[6]Appel E A,del Barrio J,Loh X J,Scherman O A.Supramolecularpolymeric hydrogels[J].ChemSoc Rev,2012,41(18):6195-6214.
[7]Guo Y J,Guo S J,Ren J T,Zhai Y M,Dong S J,Wang E K.CyclodextrinFunctionalized Graphene Nanosheets with High Supramolecular RecognitionCapability:Synthesis and Host-Guest Inclusion for Enhanced ElectrochemicalPerformance[J].Acs Nano,2010,4(9):5512-5512.
[8]Prochowicz D,Kornowicz A,Lewinski J.Interactions of NativeCyclodextrins with Metal Ions and Inorganic Nanoparticles:Fertile Landscapefor Chemistry and Materials Science[J].Chem Rev,2017,117(22):13461-13501.
[9]Zhang Z,Lv Q,Gao X Y,Chen L,Cao Y,Yu S J,He C L,Chen X S.pH-Responsive Poly(ethylene glycol)/Poly(L-lactide)Supramolecular Micelles Basedon Host-Guest Interaction[J].AcsAppl Mater Inter,2015,7(16):8404-8411.
[10]Xue M,Zhong X,Shaposhnik Z,Qu Y Q,Tamanoi F,Duan X F,Zink J I.pH-Operated Mechanized Porous Silicon Nanoparticles[J].J Am ChemSoc,2011,133(23):8798-8801.
发明内容
本发明的目的在于克服现有技术的不足,提供一种基于主客体相互作用的载体。
本发明的另一个目的在于提供上述载体的应用。
本发明所采取的技术方案是:
一种基于主客体相互作用的载体,包括主体分子和客体分子,主体分子为聚合物接枝改性的环糊精,客体分子可以容纳于环糊精的环内,客体分子共价偶联在PCL-HPG的表面。
作为上述载体的进一步改进,PCL具有4条支链臂,为PCL-HPG的核心。
作为上述载体的进一步改进,HPG的数均分子量为45000~60000。
作为上述载体的进一步改进,PCL的数均分子量为14000~16000。
作为上述载体的进一步改进,环糊精接枝的聚合物为PEI。
作为上述载体的进一步改进,PEI的数均分子量为500~700。
作为上述载体的进一步改进,环糊精为β-环糊精,客体分子选自苯并咪唑、金刚烷、偶氮苯中的至少一种。
载体在制备疏水药物和/或核酸序列载体中的应用,其中,载体如上所述。
作为上述应用的进一步改进,疏水药物选自DOX、多西紫杉醇DOC、紫杉醇PTX;核酸序列为长度不超过5000bp的质粒、线性DNA及其复合物、RNA及其复合物。
一种载有疏水药物和/或核酸序列载体的制备方法,载体的主体分子和客体分子如上所述,包括:
1)将载体的主体分子和客体分子混合,反应得到载体;
2)将载体与疏水药物和/或核酸序列混合,去除未负载的疏水药物和/或核酸序列,得到载有疏水药物和/或核酸序列的载体。
本发明的有益效果是:
本发明的载体,细胞毒性低,对疏水性药物分子和核酸序列均具有很好的载运能力,对血清具有很好的耐受能力和细胞相容性,同时可以很好地提高转染效率。通过客体分子的变换,可以获得不同pH响应的载体。
附图说明
图1是载体的合成路线;
图2是客体分子的相关检测数据;
图3是PEI600、β-CD和β-CD-PEI600的1H NMR谱图;
图4是星型聚合物PCL-HPG-PEI600的结构示意图(A)和1H NMR谱图(B);
图5是不同浓度PCL-HPG-BM和β-CD-PEI600作用的谱图;
图6是β-CD-PEI600与PCL-HPG-BM相互作用的等温滴定曲线;
图7是不同转染条件下PEI25k/DNA和PCL-HPG-PEI600/DNA复合物的实验结果;
图8是PCL-HPG-PEI600在弱酸pH条件下的解组装行为结果;
图9是不同pH条件下PCL-HPG-PEI600的粒径。
具体实施方式
一种基于主客体相互作用的载体,包括主体分子和客体分子,主体分子为聚合物接枝改性的环糊精,客体分子可以容纳于环糊精的环内,客体分子共价偶联在PCL-HPG的表面。
作为上述载体的进一步改进,PCL具有4条支链臂,为PCL-HPG的核心。
作为上述载体的进一步改进,HPG的数均分子量为45000~60000。
作为上述载体的进一步改进,PCL的数均分子量为14000~16000。
作为上述载体的进一步改进,环糊精接枝的聚合物为PEI。
为避免PEI的细胞毒性过大,作为上述载体的进一步改进,PEI的数均分子量为500~700。
作为上述载体的进一步改进,环糊精为β-环糊精,客体分子选自苯并咪唑、金刚烷、偶氮苯中的至少一种。这种主客体分子可以很好地相互嵌合,形成更为稳定的主客体体系。利用不同客体分子在不同pH下的物化特性,可以得到不同pH响应的主客体体系。
载体在制备疏水药物和/或核酸序列载体中的应用,其中,载体如上所述。
作为上述应用的进一步改进,疏水药物包括但不限于DOX、多西紫杉醇DOC、紫杉醇PTX;核酸序列为长度不超过5000bp的质粒、线性DNA及其复合物、RNA及其复合物。
一种载有疏水药物和/或核酸序列载体的制备方法,载体的主体分子和客体分子如上所述,包括:
1)将载体的主体分子和客体分子混合,反应得到载体;
2)将载体与疏水药物和/或核酸序列混合,去除未负载的疏水药物和/或核酸序列,得到载有疏水药物和/或核酸序列的载体。
名词解释:
HPG:超支化聚缩水甘油醚Hyperbranched polyglycerol
BM:苯并咪唑Benzimidazole
β-CD:β-环糊精β-Cyclodextrin
PCL:聚己内酯Polycaprolactone
PEI:聚乙烯亚胺Polyethyleneimine
DOX:阿霉素Doxorubicin
下面结合实施例,进一步说明本发明的技术方案。
载体的合成
载体的合成路线如图1所示。图(A)PCL-HPG-BM的合成路线;(B)β-CD-PEI600的合成路线。(C)星型聚合物PCL-HPG-PEI600的结构示意图。
PCL-HPG的合成
首先称取3.0g(0.20mmol)干燥的PCL于50mL Schlenk烧瓶中,利用无水无氧双排管装置反复抽真空通氮气数次以保证反应体系中无氧气。然后加入0.8mL 25wt%甲醇钾溶液,边搅拌边逐渐升温至95℃。10min后,再次抽真空除去多余甲醇。随后,在氮气的保护下,用微量注射泵逐滴加入12g(162mmol)环氧丙醇至反应体系中。滴加完后,继续反应5h。反应结束后,用适量甲醇溶解产物,并加入酸化后的阳离子交换树脂以除去钾离子。旋蒸除去除溶剂得到粗产物后,用截留分子量为3000的透析袋透析后冻干得黄色透明粘稠液体。
PCL-HPG-BM的合成
将0.5g PCL-HPG和0.05g(0.3mmol)BM溶解于干燥的N,N-二甲基甲酰胺(DMF)中,随后加入0.02g(0.1mmol)DCC和0.005g(0.3mmol)DMAP至反应体系中并搅拌。室温反应48h后,将反应液用截留分子量为1000的透析袋透析后冻干得黄色透明粘稠液体。
图2是客体分子的相关检测数据,其中图(A)PCL-HPG的1H NMR谱图;(B)PCL-HPG-BM、PCL-HPG、4-arms-PCL和BM的1H NMR谱图。从图A可以看出化学位移1.3、1.6、2.3和3.99ppm分别为4-arms-PCL上不同位置的亚甲基质子吸收峰。而化学位移3.4ppm附近处的质子吸收峰归属于HPG的亚甲基和次甲基,4.6ppm处的吸收峰则归于HPG的羟基,这说明了PCL-HPG的成功合成。图显示的是原料BM和4-arms-PCL,产物PCL-HPG和PCL-HPG-BM的核磁氢谱图,从PCL-HPG-BM的核磁氢谱图中不仅可以找到4-arms-PCL和HPG中各个质子峰的归属,还可以在化学位移7.5-8.5ppm处看到BM的特征质子吸收峰。这说明客体分子PCL-HPG-BM的成功合成。
β-CD-PEI600的合成
β-CD-PEI600的合成分为两步,第一步是单-6-对甲苯磺酰-β-环糊精(β-CD-OTs)的合成,称取31.2g(26.4mmol)β-CD于500mL单口圆底烧瓶中,加入250mL去离子水,搅拌至均匀分散;随后向混悬液中缓慢滴加10mL NaOH水溶液(8.2M)。滴加完待反应溶液澄清后,将反应体系冷却至0℃;然后向其中缓慢滴加20mL对甲苯磺酰氯乙腈溶液(1.3M),滴加结束后,在室温下继续反应2h。随后,用稀盐酸调节pH至6左右,离心收集沉淀得粗产物,用纯水重结晶两次后,真空干燥得白色粉末。第二步是β-CD-PEI600的合成,先将1.1g(1.9mmol)PEI600溶解于10mL干燥二甲基亚砜(DMSO)中,然后向体系中加入1.9g(1.5mmol)β-CD-OTs。将体系反应温度升至70℃后,在氮气的保护下反应3天。最后,将反应液用截留分子量为2000的透析袋透析后冻干得浅黄色透明液体。
图3是PEI600、β-CD和β-CD-PEI600的1H NMR谱图。如图所示,化学位移2.5-3.0ppm处的质子吸收峰归属于PEI,而化学位移5.0ppm和3.2-4.0ppm则分别归属于β-CD的1号氢和2-6号氢。1H NMR的结果表明β-CD-PEI600成功合成,同时通过积分β-CD和PEI的质子峰面积,计算得每个PEI接枝1.5个左右β-CD。
超分子共聚物PCL-HPG-PEI600的合成
先将0.1gβ-CD-PEI600和0.02g PCL-HPG-BM溶解5mL DMSO中,然后逐滴滴入到20mL PBS(pH 7.4)中。混合液在室温下搅拌过夜后,用截留分子量为3000的透析袋透析后冻干得超分子共聚物PCL-HPG-PEI600。
图4是星型聚合物PCL-HPG-PEI600的结构示意图(A)和1H NMR谱图(B)。如图所示,化学位移在1.3、1.6及2.3ppm峰为4-arms-PCL的特征质子峰;而化学位移在3.4-4.0ppm范围内的峰归属为HPG的特征峰;化学位移在2.5-3.0ppm范围内的峰归属于PEI600;化学位移在5.0ppm处的峰为典型的β-CD的特征峰;由于BM单元基本被β-CD组装,因此在7.5ppm处难以发现BM的特征峰。该核磁结果证实,组装体聚合物PCL-HPG-PEI600被成功制备。
产物的检测:
通过荧光光谱来研究β-CD与BM的主客体相互作用,进一步验证组装体PCL-HPG-PEI600的成功制备。如图5所示,图5中(A)PCL-HPG-BM在不同浓度的β-CD-PEI600溶液中的荧光发射光谱,PCL-HPG-BM的浓度为0.5mg/mL;(B)不同浓度的β-CD-PEI600溶液作用下PCL-HPG-BM在483nm处的荧光强度。PCL-HPG-BM中的BM分子的特征发射波长出现在483nm处,随着β-CD-PEI600的逐渐增加,体系荧光强度逐渐降低,说明越来越多的BM分子进入了β-CD的疏水空腔当中;而当β-CD-PEI600与PCL-HPG-BM的质量比达到10:1时,荧光强度几乎不再减少,荧光峰基本消失,这是由于β-CD分子与BM分子完全结合。通过荧光光谱的结果一方面验证了β-CD-PEI600和PCL-HPG-BM的成功组装,另一方面也确定了主体分子β-CD-PEI600与客体分子PCL-HPG-BM的组装最佳质量比为10:1。
此外,还通过等温滴定微量热法确定β-CD-PEI600和PCL-HPG-BM组装的实际质量比,滴定曲线如图6所示。通过消耗的β-CD-PEI600量计算确定了主体分子β-CD-PEI600与客体分子PCL-HPG-BM的组装质量比为7.9:1。
疏水药物和/或DNA复合物的装载
疏水药物以DOX为例,具体的包载步骤如下:
采用透析法制备负载DOX的PCL-HPG-PEI600复合物。称取5mg DOX·HCl和100mgPCL-HPG-PEI600溶解于5mL DMSO中,并加入5μL三乙胺中和DOX中的盐酸。在室温条件下避光搅拌过夜后,滴加至20mL去离子水中。所得溶液转移至截留分子量为500的透析袋中,避光透析48h以除去DMSO和未负载上的DOX,冻干得PCL-HPG-PEI600/DOX复合物。PCL-HPG-PEI600/DOX中DOX的含量采用紫外可见吸收光谱法(UV-VIS)进行测定,检测波长为483nm。
PCL-HPG-PEI//pMMP-9基因复合物的制备,将聚合物PCL-HPG-PEI600溶于超纯水中配成一定浓度的水溶液,经0.45μm的无菌滤头过滤后,将聚合物水溶液加入到pMMP-9溶液中,并充分混匀,室温下孵育20-30min。
转染实验中用到的DNA复合物也是参照上述步骤得到:将聚合物PCL-HPG-PEI600基因载体与DNA混合均匀后,于室温下孵育20-30min获得。
转染实验:
1)首先将MCF-7细胞按照每孔细胞数5×104的密度接种于24孔板中,待细胞融合率达70%时,吸弃培养基,更换成新鲜的含10%FBS的DMEM培养基;
2)置于37℃,5%CO2细胞培养箱中孵育2h后,加入不同质量比的PCL-HPG-PEI600/pMMP-9复合物(每孔pMMP-9的含量为2μg);
3)培养24h后采用倒置荧光显微镜定性观察绿色荧光蛋白的表达并拍照;
4)随后吸弃培养基,用PBS洗涤,并用胰酶消化,离心收集细胞;
5)最后,用适量PBS重悬细胞,使用FCM定量分析绿色荧光蛋白的表达百分比,并用Flow Jo7.6.1软件分析数据(每个样品做三组平行)。
PEI25k/pMMP-9(w/w=1.3)作为对照组。
不同条件的转染实验结果如图7所示。图7中,图(A)不同转染条件下PEI25k/DNA和PCL-HPG-PEI600/DNA复合物的体外转染效率(1:无血清;2:10%FBS,孵育4h;3:10%FBS,孵育24h);(B)有血清条件下不同质量比PCL-HPG-PEI600/DNA复合物的体外转染效率和细胞毒性(10%FBS,孵育24h);(C)有血清条件下不同质量比PCL-HPG-PEI600/DNA复合物转染后的荧光照片(10%FBS,孵育24h)。从图A中可以看出,血清的存在对PEI25k对照组的转染效率影响非常大,在含10%FBS培养基中与细胞共孵育4h后,其细胞转染效率从无血清条件下的31.5%显著降低至8.0%,下降约74%。而PCL-HPG-PEI600/pMMP-9的细胞转染效率也受到血清的影响,但在相同转染条件下仍有21%的细胞被成功转染,这一结果远高于PEI25k对照组,说明PCL-HPG-PEI600作为基因载体的血清耐受能力要优于PEI25k。
此外,PCL-HPG-PEI600/pMMP-9(120:1)复合物在含10%FBS培养基中与细胞共孵育24h后,相较于无血清条件下转染效率,其转染效率不仅没有下降反而高达50.5%,而PEI25k对照组的转染效率仅为15.5%。在此实验结果的基础上,还进一步的研究了不同质量比下PCL-HPG-PEI600/pMMP-9复合物在含10%FBS培养基中与细胞共孵育24h后的转染情况,并对转染过程中的细胞毒性进行了评估。如图B所示,当质量比仅为60:1时,就有32.5%的细胞被成功转染,而且随着质量比的增加,转染效率随之增加。为避免PCL-HPG-PEI600/pMMP-9复合物本身抑制细胞增殖对结果的干扰,转染过程中细胞毒性实验选取pEGFP作为模型质粒,图B结果显示,各质量比PCL-HPG-PEI600/pEGFP复合物在实验条件下并未有明显的细胞毒性,反观PEI25k/pEGFP对照组的细胞存活率仅为37%。各组对应的荧光图片如图C所示。上述结果表明,PCL-HPG-PEI600/pMMP-9复合物具有较好的血清耐受能力和细胞相容性。
pH响应的实验数据
为验证组装体PCL-HPG-PEI600的pH响应性,通过荧光光谱进一步研究PCL-HPG-PEI600在弱酸pH条件下的解组装行为。结果如图8所示,当pH≥6.5时,组装体PCL-HPG-PEI600的荧光强度几乎没有变化,但是当pH为6时,荧光强度显著减弱,这可能是因为在弱酸条件下,BM的亲水性增加,主客体识别作用减弱,使得BM分子从β-CD的疏水内腔中脱离,导致荧光强度的下降。
另外PCL-HPG-PEI600组装体在不同pH条件下的粒径通过纳米激光粒度仪测量,测量结果如图9所示。从图中可以看出,PCL-HPG-PEI600在pH值范围7.4-6.8条件下,粒径变化不大,保持约100-110nm左右;而随着体系中pH值的继续降低,当pH值为6.5时,PCL-HPG-PEI600的粒径陡然增大至160nm以上,且随pH值的继续下降粒径变化不大。荧光光谱和DLS结果表明PCL-HPG-PEI600在较低pH下,有较好的pH响应性,而且pH值响应范围在6.0-6.5之间,符合肿瘤细胞的微环境,这有望在肿瘤细胞中实现组装体聚合物的降解从而提高药物和基因的递送效率。
Claims (10)
1.一种基于主客体相互作用的载体,包括主体分子和客体分子,主体分子为聚合物接枝改性的环糊精,客体分子可以容纳于环糊精的环内,其特征在于:客体分子共价偶联在PCL-HPG的表面。
2.根据权利要求1所述的载体,其特征在于:PCL具有4条支链臂,为PCL-HPG的核心。
3.根据权利要求1所述的载体,其特征在于:HPG的数均分子量为45000~60000。
4.根据权利要求1所述的载体,其特征在于:PCL的数均分子量为14000~16000。
5.根据权利要求1~4任一项所述的载体,其特征在于:环糊精接枝的聚合物为PEI。
6.根据权利要求5所述的载体,其特征在于:PEI的数均分子量为500~700。
7.根据权利要求1~4任一项所述的载体,其特征在于:环糊精为β-环糊精,客体分子选自苯并咪唑、金刚烷、偶氮苯中的至少一种。
8.载体在制备疏水药物和/或核酸序列载体中的应用,其特征在于:载体如权利要求1~7任一项所述。
9.根据权利要求8所述的应用,其特征在于:疏水药物选自DOX、多西紫杉醇DOC、紫杉醇PTX;核酸序列为长度不超过5000bp的质粒、线性DNA及其复合物、RNA及其复合物。
10.一种载有疏水药物和/或核酸序列载体的制备方法,载体的主体分子和客体分子如权利要求1~7任一项所述,包括:
1)将载体的主体分子和客体分子混合,反应得到载体;
2)将载体与疏水药物和/或核酸序列混合,去除未负载的疏水药物和/或核酸序列,得到载有疏水药物和/或核酸序列的载体。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811264940.6A CN109395092A (zh) | 2018-10-29 | 2018-10-29 | 一种基于主客体相互作用的载体及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201811264940.6A CN109395092A (zh) | 2018-10-29 | 2018-10-29 | 一种基于主客体相互作用的载体及其应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN109395092A true CN109395092A (zh) | 2019-03-01 |
Family
ID=65469533
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811264940.6A Pending CN109395092A (zh) | 2018-10-29 | 2018-10-29 | 一种基于主客体相互作用的载体及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109395092A (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110438162A (zh) * | 2019-08-22 | 2019-11-12 | 广州晶欣生物科技有限公司 | 一种改良型非脂质体转染试剂盒及其应用方法 |
CN112011040A (zh) * | 2020-07-20 | 2020-12-01 | 广州医科大学 | 一种多重纳米传递***及其制备方法 |
CN115970066A (zh) * | 2022-12-29 | 2023-04-18 | 成都爱睿康乐医疗器械有限公司 | 基于主客体相互作用的载药纳米凝胶生物润滑剂及其制备方法和应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050276841A1 (en) * | 2004-06-07 | 2005-12-15 | California Institute Of Technology | Biodegradable drug-polymer delivery system |
CN102399369A (zh) * | 2010-09-14 | 2012-04-04 | 同济大学 | 对氨基酸敏感的超分子聚合物胶束药物载体的制备方法 |
CN105802106A (zh) * | 2016-04-22 | 2016-07-27 | 同济大学 | 一种温度、uv和还原剂三重响应的超分子纳米聚集体的制备方法 |
CN105920614A (zh) * | 2016-06-22 | 2016-09-07 | 佛山市高明绿化纳新材料有限公司 | 一种超分子水凝胶药物与基因双重载体材料及其制备方法 |
CN108659649A (zh) * | 2018-05-03 | 2018-10-16 | 南通大学 | 一种自修复可降解c3n4/主客体薄膜的制备方法 |
-
2018
- 2018-10-29 CN CN201811264940.6A patent/CN109395092A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050276841A1 (en) * | 2004-06-07 | 2005-12-15 | California Institute Of Technology | Biodegradable drug-polymer delivery system |
CN102399369A (zh) * | 2010-09-14 | 2012-04-04 | 同济大学 | 对氨基酸敏感的超分子聚合物胶束药物载体的制备方法 |
CN105802106A (zh) * | 2016-04-22 | 2016-07-27 | 同济大学 | 一种温度、uv和还原剂三重响应的超分子纳米聚集体的制备方法 |
CN105920614A (zh) * | 2016-06-22 | 2016-09-07 | 佛山市高明绿化纳新材料有限公司 | 一种超分子水凝胶药物与基因双重载体材料及其制备方法 |
CN108659649A (zh) * | 2018-05-03 | 2018-10-16 | 南通大学 | 一种自修复可降解c3n4/主客体薄膜的制备方法 |
Non-Patent Citations (1)
Title |
---|
XIAOYAN ZHOU,ET AL: ""Construction of a High-Efficiency Drug and Gene Co-Delivery System for Cancer Therapy from a pH-Sensitive Supramolecular Inclusion between Oligoethylenimine-graft-β-cyclodextrin and Hyperbranched Polyglycerol Derivative"", 《ACS APPLIED MATERIALS & INTERFACES 》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110438162A (zh) * | 2019-08-22 | 2019-11-12 | 广州晶欣生物科技有限公司 | 一种改良型非脂质体转染试剂盒及其应用方法 |
CN110438162B (zh) * | 2019-08-22 | 2021-06-25 | 广州晶欣生物科技有限公司 | 一种改良型非脂质体转染试剂盒及其应用方法 |
CN112011040A (zh) * | 2020-07-20 | 2020-12-01 | 广州医科大学 | 一种多重纳米传递***及其制备方法 |
CN112011040B (zh) * | 2020-07-20 | 2021-08-17 | 广州医科大学 | 一种多重纳米传递***及其制备方法 |
CN115970066A (zh) * | 2022-12-29 | 2023-04-18 | 成都爱睿康乐医疗器械有限公司 | 基于主客体相互作用的载药纳米凝胶生物润滑剂及其制备方法和应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Dai et al. | Controlled synthesis and surface engineering of Janus chitosan‐gold nanoparticles for photoacoustic imaging‐guided synergistic gene/photothermal therapy | |
Wei et al. | Dendrimer‐stabilized gold nanostars as a multifunctional theranostic nanoplatform for CT imaging, photothermal therapy, and gene silencing of tumors | |
Chen et al. | Redox-responsive supramolecular amphiphiles based on a pillar [5] arene for enhanced photodynamic therapy | |
Gu et al. | pH-triggered reversible “stealth” polycationic micelles | |
CN103435815B (zh) | 一种功能化聚酰胺-胺树状大分子及其纳米复合物用于基因转染的方法 | |
CN109395092A (zh) | 一种基于主客体相互作用的载体及其应用 | |
Huang et al. | Macrocycle-wrapped polyethylenimine for gene delivery with reduced cytotoxicity | |
Li et al. | Chitosan stabilized Prussian blue nanoparticles for photothermally enhanced gene delivery | |
CN107661504B (zh) | 一种树枝状大分子修饰的金纳米粒子及其制备方法和应用 | |
CN105175656B (zh) | 一种温度和氧化剂双重刺激响应性纳米聚集体制备方法和应用 | |
Xu et al. | Genetically multimodal therapy mediated by one polysaccharides-based supramolecular nanosystem | |
CN105199400A (zh) | 一种β-环糊精修饰的聚酰胺-树状大分子及其纳米金颗粒复合物的制备方法 | |
Zhang et al. | ROS-Responsive and active targeted drug delivery based on conjugated polymer nanoparticles for synergistic chemo-/photodynamic therapy | |
Chen et al. | Poly (N-isopropylacrylamide) derived nanogels demonstrated thermosensitive self-assembly and GSH-triggered drug release for efficient tumor Therapy | |
CN101864078B (zh) | 聚乙烯亚胺-壳聚糖-硬脂酸嫁接物及制备与应用 | |
Ding et al. | Tumor microenvironment responsive polypeptide-based supramolecular nanoprodrugs for combination therapy | |
Zhu et al. | Facile preparation of indocyanine green and tiny gold nanoclusters co-loaded nanocapsules for targeted synergistic sono-/photo-therapy | |
CN103110954A (zh) | 胆固醇修饰的生物可降解聚阳离子载体及制备方法和用途 | |
Zhao et al. | Leveraging a polycationic polymer to direct tunable loading of an anticancer agent and photosensitizer with opposite charges for chemo–photodynamic therapy | |
Zhang et al. | Enzyme-responsive polysaccharide supramolecular nanoassembly for enhanced DNA encapsulation and controlled release | |
Zhang et al. | General and facile syntheses of hybridized deformable hollow mesoporous organosilica nanocapsules for drug delivery | |
Shi et al. | Diselenide-containing nonionic gemini polymeric micelles as a smart redox-responsive carrier for potential programmable drug release | |
CN102250348B (zh) | 一种聚乙烯亚胺衍生物及其作为基因传递的载体 | |
Jiang et al. | A light and reduction dual sensitive supramolecular self-assembly gene delivery system based on poly (cyclodextrin) and disulfide-containing azobenzene-terminated branched polycations | |
Zhang et al. | GSH-triggered size increase of porphyrin-containing nanosystems for enhanced retention and photodynamic activity |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190301 |
|
RJ01 | Rejection of invention patent application after publication |