CN113134083A - 声敏剂及其应用 - Google Patents
声敏剂及其应用 Download PDFInfo
- Publication number
- CN113134083A CN113134083A CN202110254571.8A CN202110254571A CN113134083A CN 113134083 A CN113134083 A CN 113134083A CN 202110254571 A CN202110254571 A CN 202110254571A CN 113134083 A CN113134083 A CN 113134083A
- Authority
- CN
- China
- Prior art keywords
- black phosphorus
- modified
- microbubbles
- sound
- microbubble
- 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
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 69
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 27
- 239000002105 nanoparticle Substances 0.000 claims abstract description 21
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000002135 nanosheet Substances 0.000 claims abstract description 17
- 210000004881 tumor cell Anatomy 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 25
- 150000003904 phospholipids Chemical class 0.000 claims description 17
- 229960002685 biotin Drugs 0.000 claims description 15
- 239000011616 biotin Substances 0.000 claims description 15
- 239000002202 Polyethylene glycol Substances 0.000 claims description 12
- 229920001223 polyethylene glycol Polymers 0.000 claims description 12
- 229920002873 Polyethylenimine Chemical class 0.000 claims description 9
- 235000021355 Stearic acid Nutrition 0.000 claims description 9
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 9
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 9
- 239000003642 reactive oxygen metabolite Substances 0.000 claims description 9
- 239000008117 stearic acid Substances 0.000 claims description 9
- 230000003993 interaction Effects 0.000 claims description 8
- 239000004698 Polyethylene Substances 0.000 claims description 7
- -1 polyethylene Polymers 0.000 claims description 7
- 229920000573 polyethylene Polymers 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- 230000001225 therapeutic effect Effects 0.000 claims description 6
- 201000010099 disease Diseases 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 230000036571 hydration Effects 0.000 claims description 4
- 238000006703 hydration reaction Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 239000007858 starting material Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 2
- 239000010409 thin film Substances 0.000 claims description 2
- 239000002055 nanoplate Substances 0.000 claims 3
- 230000001678 irradiating effect Effects 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 15
- 238000006243 chemical reaction Methods 0.000 abstract description 12
- 239000000463 material Substances 0.000 abstract description 12
- 239000000758 substrate Substances 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 238000006555 catalytic reaction Methods 0.000 abstract description 3
- 239000013589 supplement Substances 0.000 abstract description 3
- 125000004122 cyclic group Chemical group 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 25
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 14
- 239000001301 oxygen Substances 0.000 description 14
- 229910052760 oxygen Inorganic materials 0.000 description 14
- 230000009471 action Effects 0.000 description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 239000007789 gas Substances 0.000 description 9
- 239000002904 solvent Substances 0.000 description 8
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 7
- 230000008685 targeting Effects 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 125000002091 cationic group Chemical group 0.000 description 5
- 238000009214 sonodynamic therapy Methods 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- 238000002604 ultrasonography Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 238000000527 sonication Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 3
- 108010090804 Streptavidin Proteins 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- NRJAVPSFFCBXDT-HUESYALOSA-N 1,2-distearoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCCCCCC NRJAVPSFFCBXDT-HUESYALOSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 239000002122 magnetic nanoparticle Substances 0.000 description 2
- 239000002064 nanoplatelet Substances 0.000 description 2
- QYSGYZVSCZSLHT-UHFFFAOYSA-N octafluoropropane Chemical compound FC(F)(F)C(F)(F)C(F)(F)F QYSGYZVSCZSLHT-UHFFFAOYSA-N 0.000 description 2
- 230000010355 oscillation Effects 0.000 description 2
- 229960004065 perflutren Drugs 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- SFZCNBIFKDRMGX-UHFFFAOYSA-N sulfur hexafluoride Chemical compound FS(F)(F)(F)(F)F SFZCNBIFKDRMGX-UHFFFAOYSA-N 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- VQVUBYASAICPFU-UHFFFAOYSA-N (6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(OC(C)=O)C=C1OC1=C2C=C(Cl)C(OC(=O)C)=C1 VQVUBYASAICPFU-UHFFFAOYSA-N 0.000 description 1
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 1
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 1
- ZKSVYBRJSMBDMV-UHFFFAOYSA-N 1,3-diphenyl-2-benzofuran Chemical compound C1=CC=CC=C1C1=C2C=CC=CC2=C(C=2C=CC=CC=2)O1 ZKSVYBRJSMBDMV-UHFFFAOYSA-N 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 229910018503 SF6 Inorganic materials 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 238000005054 agglomeration Methods 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000009830 antibody antigen interaction Effects 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- BQRGNLJZBFXNCZ-UHFFFAOYSA-N calcein am Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=C(OC(C)=O)C=C1OC1=C2C=C(CN(CC(=O)OCOC(C)=O)CC(=O)OCOC(=O)C)C(OC(C)=O)=C1 BQRGNLJZBFXNCZ-UHFFFAOYSA-N 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000004299 exfoliation Methods 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 125000002795 guanidino group Chemical group C(N)(=N)N* 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N hydrogen peroxide Substances OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
- 230000007954 hypoxia Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- YOBAEOGBNPPUQV-UHFFFAOYSA-N iron;trihydrate Chemical compound O.O.O.[Fe].[Fe] YOBAEOGBNPPUQV-UHFFFAOYSA-N 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000002715 modification method Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 238000002428 photodynamic therapy Methods 0.000 description 1
- 229920000333 poly(propyleneimine) Polymers 0.000 description 1
- 229920000867 polyelectrolyte Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 229960000909 sulfur hexafluoride Drugs 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000012876 topography Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000002341 toxic gas Substances 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Images
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
- A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
- A61K41/0033—Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0038—Radiosensitizing, i.e. administration of pharmaceutical agents that enhance the effect of radiotherapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/52—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/543—Lipids, e.g. triglycerides; Polyamines, e.g. spermine or spermidine
- A61K47/544—Phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/555—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound pre-targeting systems involving an organic compound, other than a peptide, protein or antibody, for targeting specific cells
- A61K47/557—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound pre-targeting systems involving an organic compound, other than a peptide, protein or antibody, for targeting specific cells the modifying agent being biotin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/221—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by the targeting agent or modifying agent linked to the acoustically-active agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/22—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations
- A61K49/222—Echographic preparations; Ultrasound imaging preparations ; Optoacoustic imaging preparations characterised by a special physical form, e.g. emulsions, liposomes
- A61K49/223—Microbubbles, hollow microspheres, free gas bubbles, gas microspheres
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Radiology & Medical Imaging (AREA)
- Acoustics & Sound (AREA)
- Immunology (AREA)
- Inorganic Chemistry (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Biophysics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了声敏剂及其应用。本申请的第一方面,提供声敏剂,该声敏剂包括微泡,微泡的表面连接有黑磷纳米片和四氧化三铁纳米颗粒。根据本申请实施例的声敏剂,至少具有如下有益效果:本申请实施例提供一种具有更复杂结构的黑磷材料作为声敏剂使用,微泡表面修饰的四氧化三铁纳米颗粒不仅能够提高黑磷的靶向运载,减少载药微泡的使用剂量,而且可以与H2O2反应生成Fe2+与O2,循环催化芬顿反应,为黑磷的声动力治疗提供底物补充。对于肿瘤细胞而言,能够有效解决肿瘤内部缺氧状态的问题;同时提高空化作用,提高声动力治疗效果。
Description
技术领域
本申请涉及肿瘤治疗技术领域,尤其是涉及声敏剂及其应用。
背景技术
随着生物医药领域的快速发展以及与材料、化学等多学科的交叉融合,新兴的肿瘤治疗模式,肿瘤药物和载药系统快速发展,给肿瘤治疗带来了新的希望。近年来,基于活性氧(Reactive oxygen species/ROS,包括超氧阴离子O2 ·-、过氧化氢H2O2、单线态氧1O2以及羟基自由基·OH)的声动力疗法成为肿瘤治疗的新型疗法。它通过激活瘤体内的声敏剂,与周围的氧和水反应产生活性氧,诱导肿瘤细胞凋亡和坏死。声动力疗法因其无创性、毒副作用低、选择性好、适用性好、可控性好、可重复治疗等优点尤为引人注目。超声不仅可以穿透到更深的组织层(高达10cm),而且可以最大限度地减少对周围正常组织的损伤。此外,超声辐照产生的空化效应不仅能够增加肿瘤组织对声敏剂的摄取而且能够促进ROS的生成。由于这些优异的特征,声动力疗法在肿瘤治疗中表现出更大的应用潜力。
黑磷作为一种新兴的二维声敏剂,具有独特的性质。作为一种直接带隙材料,黑磷的带隙可根据厚度进行调节(0.3~2.0eV)。另外,黑磷具有更高的比表面积、高效的单线态-三线态交叉、受激三重态的寿命长(高达100μs),可以促进活性氧的产生。更重要的是,黑磷具有理想的生物降解性。这些独特优势使其不仅能够提高声动力治疗效果,而且能够增加声动力治疗的生物安全性,但是,黑磷应用于声动力治疗还存在两个关键问题亟待解决:第一,黑磷表面难以化学修饰,无法有效负载靶向配体,因此无法实现对肿瘤的靶向性治疗,这限制了其生物利用度和声动力治疗效率;第二,肿瘤微环境影响声动力治疗体系产生活性氧的能力。实体肿瘤内部常呈现缺氧状态(pO2<2.5mmHg),显著降低了声动力疗法的治疗效果。因此,有必要提供一种新的复合结构来提高黑磷作为声敏剂参与声动力治疗肿瘤的疗效。
发明内容
本申请旨在至少解决现有技术中存在的技术问题之一。为此,本申请提出一种在声动力治疗过程中具有更好疗效的声敏剂及其应用。
本申请的第一方面,提供声敏剂,该声敏剂包括微泡,微泡的表面连接有黑磷纳米片和四氧化三铁纳米颗粒。
根据本申请实施例的声敏剂,至少具有如下有益效果:
本申请实施例提供一种具有更复杂结构的黑磷材料作为声敏剂使用,微泡表面修饰的四氧化三铁纳米颗粒不仅能够提高黑磷的靶向运载,减少载药微泡的使用剂量,而且其中的Fe3+可以与H2O2反应生成Fe2+与O2,循环催化芬顿反应,为黑磷的声动力治疗提供底物补充,对于肿瘤细胞而言,能够有效解决肿瘤内部缺氧状态的问题;同时提高空化作用,提高声动力治疗效果。
其中,“微泡”是指内部具有气体形成的气核,并在气核外部形成壳层包裹的直径大致在微米尺度的气泡。形成的壳层的非限制性实例包括蛋白质、脂质体、表面活性剂、聚合物、聚电解质等本领域熟知的能够作为壳层使用的材料。“表面连接有”是指壳层在远离气核的外侧通过特定的作用力与黑磷纳米片或四氧化三铁纳米颗粒相连接。在一些情况下,连接所依靠的作用力的非限制性实例包括氢键、静电力、范德华力、生物分子的相互作用(例如链霉亲和素-生物素相互作用、酶-底物相互作用、抗体-抗原相互作用)等其中的至少一种。
在本申请的一些实施方式中,微泡的表面修饰有正电荷基团,黑磷纳米片通过静电力与正电荷基团相连接。黑磷纳米片表面具有一定量的负电荷,为了将黑磷纳米片与微泡连接,在微泡表面修饰上正电荷基团,使两者通过静电作用力相连接。其中,修饰的方法具体可以是在微泡制备完成后进行另外的修饰,或在制备过程中采用包含正电荷基团的原料参与微泡的形成。正电荷基团的非限制性实例包括氨基/胺基、胍基等其中至少一种。包含正电荷的原料包括聚烯化亚胺,如聚乙烯亚胺、聚丙烯亚胺等。
在本申请的一些实施方式中,四氧化三铁纳米颗粒通过链霉亲和素-生物素相互作用与微泡相连接。链霉亲和素-生物素相互作用是目前已知的结合力较强的相互作用力,为保证四氧化三铁纳米颗粒能够稳定地附着在微泡表面,提高活性氧水平,因而采用链霉亲和素-生物素相互作用将四氧化三铁纳米颗粒连接到微泡表面。
在本申请的一些实施方式中,微泡的制备原料包括磷脂、聚乙二醇修饰的磷脂和聚乙烯亚胺。其中,磷脂选自本领域熟知的能够形成微泡的磷脂类化合物或其混合物,磷脂类化合物的非限制性实例包括磷脂酰胆碱、磷脂酰甘油、磷脂酸、磷脂酰乙醇胺、磷脂酰丝氨酸等。
在本申请的一些实施方式中,微泡的制备原料还包括生物素化的聚乙二醇修饰的磷脂。
本申请的第二方面,提供声敏剂的制备方法,该制备方法包括以下步骤:
取磷脂、聚乙二醇修饰的磷脂、生物素化的聚乙二醇修饰的磷脂和硬脂酸修饰的聚乙烯亚胺,通过薄膜水化法制备得到微泡;
微泡与黑磷纳米片混合孵育,使黑磷纳米片连接到微泡的表面;
微泡与链霉亲和素修饰的四氧化三铁纳米颗粒混合孵育,使四氧化三铁纳米颗粒连接到微泡的表面。
其中,薄膜水化法是本领域制备微泡的一种常规方法,具体是将壳层原料溶于良溶剂中,除去溶剂后形成薄膜,薄膜重新在缓冲液中水化,及在形成气核的气体氛围中组装形成微泡结构。在其中一些实施方式中,良溶剂可以是易挥发的有机溶剂,如乙醚、氯仿等。除去溶剂的方法具体可以采用加热蒸发、通风干燥等本领域熟知的除去溶剂的方法中的至少一种。形成气核的气体的非限制性实例包括氮气、惰性气体或其他无毒且不参与化学反应的气体如六氟化硫(SF6)、全氟丙烷(C3H8)等。
在本申请的一些实施方式中,磷脂为二硬脂酸磷脂酰胆碱,聚乙二醇修饰的磷脂为二硬脂酰基磷脂酰乙醇胺-聚乙二醇,生物素化的聚乙二醇修饰的磷脂为生物素化二硬脂酸磷脂酰乙醇胺-聚乙二醇。
在本申请的一些实施方式中,二硬脂酰磷脂酰乙醇胺-聚乙二醇-生物素中聚乙二醇(PEG)的分子量为2000~4000。PEG的分子量大于2000时能够保证微泡的长循环并且可以起到磁性靶向诊疗的效果,优选范围为2000~4000。
在本申请的一些实施方式中,硬脂酸修饰的聚乙烯亚胺(Stearic-PEI)占反应原料的摩尔百分比为30%~40%。Stearic-PEI含量增多可以增加微泡表面正电荷,从而结合更多的黑磷,但随着结合的越多,表面电荷下降就越多,发生聚集的可能性就越大,优选占比为30%~40%,更优选为36%。
在本申请的一些实施方式中,二硬脂酰磷脂酰乙醇胺-聚乙二醇-生物素占反应原料的摩尔百分比为5%~15%,更优选为9%。
在本申请的一些实施方式中,黑磷纳米片通过液相剥离法制得。液相剥离法的具体步骤包括:取黑磷材料加入到有机溶剂中,然后采用声波降解得到微米尺度的黑磷片材,再通过超声振荡处理使黑磷的尺度降低到纳米,在6000~8000rpm离心收集上清液,然后对上清液在高于14000rpm条件下离心取沉淀得到黑磷纳米片。
在本申请的一些实施方式中,链霉亲和素修饰的四氧化三铁纳米颗粒中链霉亲和素的摩尔数:二硬脂酰磷脂酰乙醇胺-聚乙二醇-生物素中生物素的摩尔数的比例范围为(2~10):1,优选地,比例范围为(4~10):1。当链霉亲和素与生物素的摩尔比例在4:1以上时能够有效地防止修饰后的微泡发生团聚现象。
本申请的第三方面,提供提高细胞的活性氧浓度的方法,该方法包括以下步骤:使上述的声敏剂与细胞接触,超声辐照;该法用于非疾病的诊断或治疗目的。
根据本申请实施例的方法,至少具有如下有益效果:
本申请实施例将上述的黑磷材料作为声敏剂使用,该声敏剂靶向到细胞后,在超声作用下,四氧化三铁纳米颗粒可以与H2O2反应生成Fe2+与O2,循环催化芬顿反应;同时,微泡的空化作用能够增加细胞膜的通透性,从而协同提高胞内的活性氧浓度。
其中,该方法的非疾病的诊断或治疗目的是指该方法不直接以有生命的人或动物体为对象,或者不直接用于消除病因或病灶。对于第一点,该方法的作用对象可以是离体细胞/组织/器官;并且当该离体细胞/组织/器官为包括肿瘤细胞的生物材料时,在作用后不会返回细胞/组织/器官所属的人或动物的主体以消除其病因病灶。对于第二点,该方法的作用对象可以是健康的人体或动物体,或者虽然是患有肿瘤等疾病的人体或动物体,但作用部位为非肿瘤部位,并且使用该方法的最终目的仅在于提高特定部位的活性氧浓度,或通过提高活性氧浓度实现非治疗目的,而不间接用于抑制肿瘤细胞的发生、发展或促进其凋亡等。例如,通过提高细胞的活性氧浓度来改变植物体或动物体的生长特性。
在本申请的一些实施方式中,细胞为肿瘤细胞。
在本申请的一些实施方式中,超声辐照的功率为1~3W/cm2。
本申请的附加方面和优点将在下面的描述中部分给出,部分将从下面的描述中变得明显,或通过本申请的实践了解到。
附图说明
图1为本申请实施例中微泡的形貌表征图。
图2为本申请实施例中微泡的性能检测结果。
图3为本申请实施例中微泡产生单线态氧的测试结果。
图4为本申请实施例中活性氧产率测试结果。
图5为本申请实施例中靶向声动力治疗效果测试的测试结果。
具体实施方式
以下将结合实施例对本申请的构思及产生的技术效果进行清楚、完整地描述,以充分地理解本申请的目的、特征和效果。显然,所描述的实施例只是本申请的一部分实施例,而不是全部实施例,基于本申请的实施例,本领域的技术人员在不付出创造性劳动的前提下所获得的其他实施例,均属于本申请保护的范围。
下面详细描述本申请的实施例,描述的实施例是示例性的,仅用于解释本申请,而不能理解为对本申请的限制。
在本申请的描述中,若干的含义是一个以上,多个的含义是两个以上,大于、小于、超过等理解为不包括本数,以上、以下、以内等理解为包括本数。如果有描述到第一、第二只是用于区分技术特征为目的,而不能理解为指示或暗示相对重要性或者隐含指明所指示的技术特征的数量或者隐含指明所指示的技术特征的先后关系。
本申请的描述中,参考术语“一个实施例”、“一些实施例”、“示意性实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本申请的至少一个实施例或示例中。在本说明书中,对上述术语的示意性表述不一定指的是相同的实施例或示例。而且,描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
下述实施例中使用的试剂信息如下:
二硬脂酰磷脂酰乙醇胺-聚乙二醇(DSPE-PEG2000),购于西安瑞禧生物科技有限公司。
二硬脂酰磷脂酰乙醇胺-聚乙二醇-生物素(DSPE-PEG2000-Biotin),购于西安瑞禧生物科技有限公司。
硬脂酸修饰的聚乙烯亚胺(Stearic-PEI)为通过化学反应,将硬脂酸接枝在PEI分子上,接枝步骤为:
取0.35gN,N′-羰基二咪唑(CDI)和0.6g硬脂酸分别溶解于10ml无水氯仿中,0.7g支链PEI600溶解于20ml无水氯仿中,分别得N,N′-羰基二咪唑溶液、硬脂酸溶液、支链PEI600溶液备用;
将上述制得的N,N′-羰基二咪唑溶液进行磁力搅拌,在不断磁力搅拌下,将硬脂酸溶液逐滴加入N,N′-羰基二咪唑溶液中,得混合物溶液一,将混合物溶液一在氩气保护下反应2小时后,再将其逐滴加入支链PEI600溶液中,得混合物溶液二备用;
将混合物溶液二在氩气保护下于室温环境中进一步搅拌24小时,产物在冷乙醚中沉淀纯化,然后放入大型离心机中清洗除去未反应的溶剂10min,收集下沉物得到纯化的Stearic-PEI600;然后将清洗收集后的Stearic-PEI600溶液放入真空干燥机中干燥数小时,完毕后置于-20℃储物箱中保存,制备微泡时将其融于无水氯仿中使用。
实施例1
本实施例提供了一种微泡,具体制备步骤如下所示:
(1)黑磷纳米片的制备:本实施例采用液相剥离法,在N-甲基吡咯烷酮(NMP)溶液中经过声波降解和超声震荡制备得到尺寸在纳米级别的黑磷纳米片,7000rpm离心20分钟,并将收集的上清液进一步以15000rpm离心5分钟。将获得的沉淀物分散在NMP溶液中,真空保存待用。
(2)微泡的制备:将二硬脂酰基磷脂酰胆碱(DSPC)、二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-PEG2000)、二硬脂酰磷脂酰乙醇胺-聚乙二醇-生物素(DSPE-PEG2000-Biotin,)和硬脂酸修饰低分子量的聚乙烯亚胺(Stearic-PEI600)按摩尔比49:9:9:36溶解于氯仿和甲醇混合溶液中,并用磁搅拌器搅拌均匀后,在60℃水浴锅中真空旋转蒸发2小时。待有机溶剂挥发,试管壁上形成均匀的薄膜,再放入真空烘箱中干燥2小时,使有机溶剂挥发彻底。取出试管后向其中加入一定量的脱气Tris缓冲液水化,60℃加热15分钟,并用水浴超声得到一定浓度的磷脂溶液。将其分装到西林瓶中,置换空气为全氟丙烷(C3F8),再用机械振荡器震荡30秒,获得微泡(生物素化阳离子微泡)。
(3)微泡的修饰
取制备得到的微泡,先与黑磷纳米片孵育20分钟,用去离子水清洗2次,得到黑磷微泡;
按亲和素和生物素摩尔比2:1的比例加入链酶亲和素修饰的四氧化三铁纳米颗粒,孵育15分钟后,用去离子水离心清洗2次,获得最终产品声敏剂(磁性黑磷微泡)。将声敏剂重悬于PBS溶液中,置于4℃冰箱中备用。
图1是本实施例制备得到的生物素化阳离子微泡、黑磷微泡和声敏剂的形貌表征结果,图中比例尺为5微米,其中(A)和(A′)分别表示生物素化阳离子微泡的照片和光学图片,(B)和(B′)分别表示黑磷微泡的照片和光学图片,(C)和(C′)分别表示终产品声敏剂的照片和光学图片。从图中可以看出,和黑磷连接后形成的黑磷微泡的颜色由白色变为黑色,而声敏剂的粒径较小并且能够在磁铁作用下移动。
使用马尔文Zetasizer Nano纳米粒度电位仪分别对生物素化阳离子微泡、黑磷微泡和声敏剂的表面电位和尺寸进行测量,结果如图2的a和b所示。从图2的a中可以看出,制备出的生物素化阳离子微泡表面带有正电荷,负载黑磷后的黑磷微泡的表面电荷由正电荷变为负值,表明黑磷成功连接在微泡表面,而在负载磁性纳米颗粒以后制得的声敏剂的表面电荷为负值但绝对值下降。而从b中可以看出,负载黑磷和磁性纳米颗粒后,微泡尺寸增加。上述结果表明,黑磷纳米片和四氧化三铁纳米颗粒已经成功负载到微泡上。
实施例2
稳定性测试
分别取实施例1中的黑磷微泡和声敏剂在PBS缓冲液中储存20、40和60分钟后测试其浓度,结果如图2的c所示,黑磷微泡的下清液中的吸光度测试结果如图2的d所示(由上到下分别为0min、20min、40min和60min的曲线)。从图中可以看出,黑磷微泡在1小时内浓度无明显变化,而声敏剂的浓度随时间减少,该结果表明,四氧化三铁纳米颗粒的负载会使微泡的稳定性下降,但1小时后浓度仍然在80%以上。黑磷微泡的下清液中1小时内没有检测到明显的黑磷掉落,表明黑磷稳定的负载在微泡的表面。
另外,对声敏剂的磁靶向性测试结果表明,在磁场作用下声敏剂会聚集到磁场作用的位置,这说明声敏剂具有磁靶向性。另外,声敏剂聚集部分的超声造影图像强度相比于未聚集部分的超声造影图像强度增加了3倍。
实施例3
单线态氧产率测试
分别取0.5mL的PBS溶液、黑磷纳米片溶液、黑磷微泡溶液和声敏剂溶剂,加入0.5mL的3-二苯基异苯并呋喃(DPBF),施加超声辐照(1MHz,1W/cm2,40%占空比)3分钟,离心去除材料后检测410nm处的吸光值。
结果如图3所示,吸光值越小表示产生的单线态氧越多,从图中可以看出,声敏剂溶剂相比于其它的黑磷材料所产生的单线态氧要明显更高。
实施例4
活性氧产率测试
以MCF-7细胞(人体乳腺癌细胞)为模型细胞,将其接种于35mm培养皿中进行常规培养24小时。随后分别加入对照组黑磷和实验组实施例1中的声敏剂,在磁场作用5分钟后,施加超声辐照(1MHz,1W/cm2,40%占空比)3分钟,然后用2′,7′-二氯荧光素二乙酸酯(DCFH-DA)替换细胞培养液再孵育30分钟,用PBS清洗3次。采用共聚焦激光扫描显微镜观察MCF-7细胞中的活性氧浓度和分布情况。
结果如图4所示,第一排为黑磷作用下超声后细胞的分布和活性氧情况,第二排为磁性黑磷微泡作用下超声后的细胞的分布和活性氧情况,图中比例尺为200μm。在磁场作用5分钟后,磁性黑磷微泡被靶向到MCF-7细胞上。在超声作用前,声敏剂几乎不产生活性氧,但是在超声作用后,声敏剂爆破,活性氧产率显著增强。而对照组所用的单独的黑磷在相同情况下活性氧产率极低,即使孵育3小时后再施加相同的超声作用,其活性氧产率仍然远低于实施例1中的声敏剂。
实施例5
靶向声动力治疗效果测试
以MCF-7细胞(人体乳腺癌细胞)为模型细胞,将其接种于35mm培养皿中进行常规培养24小时,与磁性黑磷微泡在磁铁作用(≈1.2T)下孵育5分钟,采用倒置荧光显微镜观察比较其细胞摄取情况。然后施加超声辐照(1MHz,1W/cm2,40%占空比)3分钟,处理结束后,细胞放入培养箱中继续培养6小时后弃细胞培养液。然后加入钙黄绿素(calcein-AM)和碘化丙啶(PI)溶液染色活细胞(绿色)和死细胞(红色)15分钟。用倒置荧光显微镜定性地观察和拍照。接着用ImageJ软件计数活死细胞数分析凋亡情况。
结果如图5所示,A为无超声作用时,B为超声作用后的情况,图中比例尺为400μm。从图中可以看到,采用磁性黑磷微泡磁场作用5分钟、超声作用3分钟后,MCF-7细胞的存活率下降为不到40%。而相同条件下,黑磷纳米片或黑磷微泡作用后MCF-7细胞的存活率下降明显要小得多。
综合上述结果可以看到,本申请实施例所提供的声敏剂在表面修饰的四氧化三铁纳米颗粒不仅能够提高黑磷的靶向运载,减少载药微泡的使用剂量,而且可以与H2O2反应生成Fe2+与O2,循环催化芬顿反应,为黑磷的声动力治疗提供底物补充,对于肿瘤细胞而言,能够有效解决肿瘤内部缺氧状态的问题;同时提高空化作用,提高声动力治疗效果。而利用其靶向作用和空化作用能够缩短给药时间,实现精准靶向肿瘤诊疗。
上面结合实施例对本申请作了详细说明,但是本申请不限于上述实施例,在所属技术领域普通技术人员所具备的知识范围内,还可以在不脱离本申请宗旨的前提下作出各种变化。此外,在不冲突的情况下,本申请的实施例及实施例中的特征可以相互组合。
Claims (9)
1.声敏剂,其特征在于,包括微泡,所述微泡的表面连接有黑磷纳米片和四氧化三铁纳米颗粒。
2.根据权利要求1所述的声敏剂,其特征在于,所述微泡的表面修饰有正电荷基团,所述黑磷纳米片通过静电力与所述正电荷基团相连接。
3.根据权利要求1所述的声敏剂,其特征在于,所述四氧化三铁纳米颗粒通过链霉亲和素-生物素相互作用与所述微泡相连接。
4.根据权利要求1所述的声敏剂,其特征在于,所述微泡的制备原料包括磷脂、聚乙二醇修饰的磷脂和聚乙烯亚胺。
5.根据权利要求4所述的声敏剂,其特征在于,所述微泡的制备原料还包括生物素化的聚乙二醇修饰的磷脂。
6.权利要求1至5任一项所述的声敏剂的制备方法,其特征在于,包括以下步骤:
取磷脂、聚乙二醇修饰的磷脂、生物素化的聚乙二醇修饰的磷脂和硬脂酸修饰的聚乙烯亚胺,通过薄膜水化法制备得到微泡;
所述微泡与黑磷纳米片混合孵育,使所述黑磷纳米片连接到所述微泡的表面;
所述微泡与链霉亲和素修饰的四氧化三铁纳米颗粒混合孵育,使所述四氧化三铁纳米颗粒连接到所述微泡的表面。
7.提高细胞的活性氧浓度的方法,其特征在于,包括以下步骤:使权利要求1至5任一项所述的声敏剂与所述细胞接触,超声辐照;所述方法用于非疾病的诊断或治疗目的。
8.根据权利要求7所述的方法,其特征在于,所述细胞为肿瘤细胞。
9.根据权利要求7所述的方法,其特征在于,所述超声辐照的功率为1~3W/cm2。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110254571.8A CN113134083B (zh) | 2021-03-09 | 2021-03-09 | 声敏剂及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110254571.8A CN113134083B (zh) | 2021-03-09 | 2021-03-09 | 声敏剂及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113134083A true CN113134083A (zh) | 2021-07-20 |
CN113134083B CN113134083B (zh) | 2023-07-07 |
Family
ID=76810966
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110254571.8A Active CN113134083B (zh) | 2021-03-09 | 2021-03-09 | 声敏剂及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113134083B (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114246947A (zh) * | 2021-12-27 | 2022-03-29 | 中国科学院苏州纳米技术与纳米仿生研究所 | 不依赖肿瘤微环境中氧气的无机声敏剂、制备方法及应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100155310A1 (en) * | 2007-07-06 | 2010-06-24 | M. Technique Co., Ltd. | Method for producing nanoparticles by forced ultrathin film rotary processing |
CN109529064A (zh) * | 2019-01-04 | 2019-03-29 | 北京大学深圳医院 | 可以实现肿瘤局部定点放射治疗的黑磷微泡药物载体及其制备方法 |
CA3145514A1 (en) * | 2018-07-31 | 2020-02-06 | Benjamin PINEDA OLVERA | Vaccination with microvesicles derived from tumour cells for cancer treatment |
-
2021
- 2021-03-09 CN CN202110254571.8A patent/CN113134083B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100155310A1 (en) * | 2007-07-06 | 2010-06-24 | M. Technique Co., Ltd. | Method for producing nanoparticles by forced ultrathin film rotary processing |
CA3145514A1 (en) * | 2018-07-31 | 2020-02-06 | Benjamin PINEDA OLVERA | Vaccination with microvesicles derived from tumour cells for cancer treatment |
CN109529064A (zh) * | 2019-01-04 | 2019-03-29 | 北京大学深圳医院 | 可以实现肿瘤局部定点放射治疗的黑磷微泡药物载体及其制备方法 |
Non-Patent Citations (5)
Title |
---|
李婷等: "磁性超声微泡的制备及基本性质检测", 《中国医学影像技术》 * |
李婷等: "磁性超声微泡的制备及基本性质检测", 《中国医学影像技术》, vol. 30, no. 6, 20 June 2014 (2014-06-20), pages 809 - 812 * |
葛金龙: "《金属有机骨架材料制备及其应用》", 3 September 2019, 中国科学技术大学出版社, pages: 95 * |
赵岳涛等: "二维黑磷的制备及表面修饰技术研究进展", 《科学通报》 * |
赵岳涛等: "二维黑磷的制备及表面修饰技术研究进展", 《科学通报》, vol. 62, no. 20, 20 July 2017 (2017-07-20), pages 2252 - 2261 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114246947A (zh) * | 2021-12-27 | 2022-03-29 | 中国科学院苏州纳米技术与纳米仿生研究所 | 不依赖肿瘤微环境中氧气的无机声敏剂、制备方法及应用 |
Also Published As
Publication number | Publication date |
---|---|
CN113134083B (zh) | 2023-07-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Liang et al. | Core-shell structured upconversion nanocrystal-dendrimer composite as a carrier for mitochondria targeting and catalase enhanced anti-cancer photodynamic therapy | |
Zhu et al. | Ru@ CeO2 yolk shell nanozymes: Oxygen supply in situ enhanced dual chemotherapy combined with photothermal therapy for orthotopic/subcutaneous colorectal cancer | |
Cai et al. | Organic molecules with propeller structures for efficient photoacoustic imaging and photothermal ablation of cancer cells | |
Cheng et al. | Polydopamine-coated manganese carbonate nanoparticles for amplified magnetic resonance imaging-guided photothermal therapy | |
Wang et al. | Lanthanide-doped upconversion nanoparticles electrostatically coupled with photosensitizers for near-infrared-triggered photodynamic therapy | |
Ding et al. | Conjugated polyelectrolyte–cisplatin complex nanoparticles for simultaneous in vivo imaging and drug tracking | |
Basuki et al. | Using fluorescence lifetime imaging microscopy to monitor theranostic nanoparticle uptake and intracellular doxorubicin release | |
Cui et al. | Amphiphilic chitosan modified upconversion nanoparticles for in vivo photodynamic therapy induced by near-infrared light | |
Erogbogbo et al. | In vivo targeted cancer imaging, sentinel lymph node mapping and multi-channel imaging with biocompatible silicon nanocrystals | |
Sohrabi et al. | Nanoscale Metal-Organic Frameworks: Recent developments in synthesis, modifications and bioimaging applications | |
US9616122B2 (en) | PH sensitive metal nanoparticle and preparation method | |
CN113456838B (zh) | 磁性黑磷微泡及其在制备超声诊断试剂和治疗乳腺癌药物中的应用 | |
Yang et al. | NIR-driven water splitting by layered bismuth oxyhalide sheets for effective photodynamic therapy | |
CN101390826B (zh) | 一种磁性肿瘤靶向聚合物纳米囊泡及其制备方法 | |
CN111671923B (zh) | 一种肽功能化载金属卟啉相变纳米粒及其制备方法和应用 | |
Cai et al. | A ph-activable chemo–photodynamic therapy based on cube-wrapped-cube α-naybf4: Tm@ caf2/nd@ zno nanoparticles mediated by 808 nm light | |
Wang et al. | Magnetic multi-walled carbon nanotubes for tumor theranostics | |
Wang et al. | HA targeted-biodegradable nanocomposites responsive to endogenous and exogenous stimulation for multimodal imaging and chemo-/photothermal therapy | |
CN114209829B (zh) | 负载荧光染料的光热脂质体及其制备方法和用途 | |
TWI619514B (zh) | 生物膜相變液滴、藥物載體及其用途 | |
Wang et al. | Cluster polyanions and surface-covered complexes: From synergistic self-assembly to bio-functionalization | |
CN113134083B (zh) | 声敏剂及其应用 | |
CN111632154A (zh) | 一种相转变纳米泡、其制备方法及用途 | |
CN106606778A (zh) | 含磷酸胆碱聚合物包覆的核壳式磁性复合粒子及其制备方法 | |
CN107970224B (zh) | 一种脂质修饰磁性氧化石墨烯复合材料的制备方法及应用 |
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 |