CN116327985A - 一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 - Google Patents
一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 Download PDFInfo
- Publication number
- CN116327985A CN116327985A CN202310147628.3A CN202310147628A CN116327985A CN 116327985 A CN116327985 A CN 116327985A CN 202310147628 A CN202310147628 A CN 202310147628A CN 116327985 A CN116327985 A CN 116327985A
- Authority
- CN
- China
- Prior art keywords
- mesoporous silica
- hollow
- drug
- hollow mesoporous
- vesicle
- 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
- 239000003814 drug Substances 0.000 title claims abstract description 78
- 229940079593 drug Drugs 0.000 title claims abstract description 66
- 238000003384 imaging method Methods 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 40
- 238000002604 ultrasonography Methods 0.000 title claims abstract description 31
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 265
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 130
- 239000002502 liposome Substances 0.000 claims abstract description 61
- 150000002632 lipids Chemical class 0.000 claims abstract description 15
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 12
- 238000012377 drug delivery Methods 0.000 claims abstract description 12
- 230000004087 circulation Effects 0.000 claims abstract description 6
- 238000001125 extrusion Methods 0.000 claims abstract description 5
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 52
- 239000002245 particle Substances 0.000 claims description 49
- 239000011148 porous material Substances 0.000 claims description 49
- 239000000725 suspension Substances 0.000 claims description 43
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 35
- 235000012000 cholesterol Nutrition 0.000 claims description 26
- 239000007789 gas Substances 0.000 claims description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 239000007853 buffer solution Substances 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 238000002156 mixing Methods 0.000 claims description 18
- 229910052757 nitrogen Inorganic materials 0.000 claims description 17
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 16
- 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 claims description 16
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 16
- 229910052710 silicon Inorganic materials 0.000 claims description 16
- 239000010703 silicon Substances 0.000 claims description 16
- 150000003863 ammonium salts Chemical class 0.000 claims description 15
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical group [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 14
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 239000003570 air Substances 0.000 claims description 11
- 239000012528 membrane Substances 0.000 claims description 11
- KSXTUUUQYQYKCR-LQDDAWAPSA-M 2,3-bis[[(z)-octadec-9-enoyl]oxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCC(=O)OCC(C[N+](C)(C)C)OC(=O)CCCCCCC\C=C/CCCCCCCC KSXTUUUQYQYKCR-LQDDAWAPSA-M 0.000 claims description 10
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 10
- 238000011068 loading method Methods 0.000 claims description 10
- 239000002244 precipitate Substances 0.000 claims description 10
- 239000000047 product Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- 229920001400 block copolymer Polymers 0.000 claims description 8
- 229960004679 doxorubicin Drugs 0.000 claims description 8
- 238000011049 filling Methods 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 7
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 7
- -1 DOPE Chemical compound 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000443 aerosol Substances 0.000 claims description 6
- 235000019270 ammonium chloride Nutrition 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 238000002601 radiography Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- BYWPQQOQVPVCTC-UHFFFAOYSA-N 2-[3-[2-[2-[2-[2-[bis[3-[2-(2-methyl-3-octylsulfanylpropanoyl)oxyethoxy]-3-oxopropyl]amino]ethylamino]ethyl-[3-[2-(2-methyl-3-octylsulfanylpropanoyl)oxyethoxy]-3-oxopropyl]amino]ethylamino]ethyl-[3-[2-(2-methyl-3-octylsulfanylpropanoyl)oxyethoxy]-3-oxopropyl]amino]propanoyloxy]ethyl 2-methyl-3-octylsulfanylpropanoate Chemical compound CCCCCCCCSCC(C)C(=O)OCCOC(=O)CCN(CCNCCN(CCC(=O)OCCOC(=O)C(C)CSCCCCCCCC)CCC(=O)OCCOC(=O)C(C)CSCCCCCCCC)CCNCCN(CCC(=O)OCCOC(=O)C(C)CSCCCCCCCC)CCC(=O)OCCOC(=O)C(C)CSCCCCCCCC BYWPQQOQVPVCTC-UHFFFAOYSA-N 0.000 claims description 5
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 5
- 239000002243 precursor Substances 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000001354 calcination Methods 0.000 claims description 4
- 239000012159 carrier gas Substances 0.000 claims description 4
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 239000004417 polycarbonate Substances 0.000 claims description 4
- 229920000515 polycarbonate Polymers 0.000 claims description 4
- 238000009417 prefabrication Methods 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 4
- CPUDPFPXCZDNGI-UHFFFAOYSA-N triethoxy(methyl)silane Chemical compound CCO[Si](C)(OCC)OCC CPUDPFPXCZDNGI-UHFFFAOYSA-N 0.000 claims description 4
- MWWSFMDVAYGXBV-RUELKSSGSA-N Doxorubicin hydrochloride Chemical compound Cl.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 MWWSFMDVAYGXBV-RUELKSSGSA-N 0.000 claims description 3
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 claims description 3
- 229930012538 Paclitaxel Natural products 0.000 claims description 3
- 108020004459 Small interfering RNA Proteins 0.000 claims description 3
- ABBQHOQBGMUPJH-UHFFFAOYSA-M Sodium salicylate Chemical compound [Na+].OC1=CC=CC=C1C([O-])=O ABBQHOQBGMUPJH-UHFFFAOYSA-M 0.000 claims description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 3
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 claims description 3
- 229960004562 carboplatin Drugs 0.000 claims description 3
- 190000008236 carboplatin Chemical compound 0.000 claims description 3
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 claims description 3
- 229960004316 cisplatin Drugs 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 229960002918 doxorubicin hydrochloride Drugs 0.000 claims description 3
- 229960002949 fluorouracil Drugs 0.000 claims description 3
- 229960001592 paclitaxel Drugs 0.000 claims description 3
- 239000000137 peptide hydrolase inhibitor Substances 0.000 claims description 3
- 230000001105 regulatory effect Effects 0.000 claims description 3
- 239000002924 silencing RNA Substances 0.000 claims description 3
- 229960004025 sodium salicylate Drugs 0.000 claims description 3
- 239000002904 solvent Substances 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
- LFQCEHFDDXELDD-UHFFFAOYSA-N tetramethyl orthosilicate Chemical compound CO[Si](OC)(OC)OC LFQCEHFDDXELDD-UHFFFAOYSA-N 0.000 claims description 3
- SZEMGTQCPRNXEG-UHFFFAOYSA-M trimethyl(octadecyl)azanium;bromide Chemical compound [Br-].CCCCCCCCCCCCCCCCCC[N+](C)(C)C SZEMGTQCPRNXEG-UHFFFAOYSA-M 0.000 claims description 3
- 102000003390 tumor necrosis factor Human genes 0.000 claims description 3
- 239000002451 tumor necrosis factor inhibitor Substances 0.000 claims description 3
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 2
- NLMKTBGFQGKQEV-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-hexadecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO NLMKTBGFQGKQEV-UHFFFAOYSA-N 0.000 claims description 2
- HXLAEGYMDGUSBD-UHFFFAOYSA-N 3-[diethoxy(methyl)silyl]propan-1-amine Chemical compound CCO[Si](C)(OCC)CCCN HXLAEGYMDGUSBD-UHFFFAOYSA-N 0.000 claims description 2
- ZYAASQNKCWTPKI-UHFFFAOYSA-N 3-[dimethoxy(methyl)silyl]propan-1-amine Chemical compound CO[Si](C)(OC)CCCN ZYAASQNKCWTPKI-UHFFFAOYSA-N 0.000 claims description 2
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 claims description 2
- 239000000032 diagnostic agent Substances 0.000 claims description 2
- 229940039227 diagnostic agent Drugs 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- BFXIKLCIZHOAAZ-UHFFFAOYSA-N methyltrimethoxysilane Chemical compound CO[Si](C)(OC)OC BFXIKLCIZHOAAZ-UHFFFAOYSA-N 0.000 claims description 2
- 150000003904 phospholipids Chemical class 0.000 claims description 2
- 239000011257 shell material Substances 0.000 claims description 2
- FZHAPNGMFPVSLP-UHFFFAOYSA-N silanamine Chemical compound [SiH3]N FZHAPNGMFPVSLP-UHFFFAOYSA-N 0.000 claims description 2
- IZRJPHXTEXTLHY-UHFFFAOYSA-N triethoxy(2-triethoxysilylethyl)silane Chemical compound CCO[Si](OCC)(OCC)CC[Si](OCC)(OCC)OCC IZRJPHXTEXTLHY-UHFFFAOYSA-N 0.000 claims description 2
- QQQSFSZALRVCSZ-UHFFFAOYSA-N triethoxysilane Chemical compound CCO[SiH](OCC)OCC QQQSFSZALRVCSZ-UHFFFAOYSA-N 0.000 claims description 2
- LFRDHGNFBLIJIY-UHFFFAOYSA-N trimethoxy(prop-2-enyl)silane Chemical compound CO[Si](OC)(OC)CC=C LFRDHGNFBLIJIY-UHFFFAOYSA-N 0.000 claims description 2
- 239000008186 active pharmaceutical agent Substances 0.000 claims 1
- UBAZGMLMVVQSCD-UHFFFAOYSA-N carbon dioxide;molecular oxygen Chemical compound O=O.O=C=O UBAZGMLMVVQSCD-UHFFFAOYSA-N 0.000 claims 1
- 229940088679 drug related substance Drugs 0.000 claims 1
- 230000008685 targeting Effects 0.000 abstract description 12
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 5
- 210000000056 organ Anatomy 0.000 abstract description 5
- 239000008280 blood Substances 0.000 abstract description 4
- 210000004369 blood Anatomy 0.000 abstract description 4
- 230000017531 blood circulation Effects 0.000 abstract description 2
- 238000003745 diagnosis Methods 0.000 abstract description 2
- 238000009792 diffusion process Methods 0.000 abstract description 2
- 238000004090 dissolution Methods 0.000 abstract description 2
- 238000004581 coalescence Methods 0.000 abstract 1
- 238000004880 explosion Methods 0.000 abstract 1
- 238000004806 packaging method and process Methods 0.000 abstract 1
- 238000002560 therapeutic procedure Methods 0.000 abstract 1
- 208000006411 Hereditary Sensory and Motor Neuropathy Diseases 0.000 description 56
- 208000021995 hereditary motor and sensory neuropathy Diseases 0.000 description 56
- 239000002953 phosphate buffered saline Substances 0.000 description 26
- 229920001223 polyethylene glycol Polymers 0.000 description 20
- 230000002209 hydrophobic effect Effects 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 18
- 238000001179 sorption measurement Methods 0.000 description 16
- 239000012071 phase Substances 0.000 description 12
- 238000001988 small-angle X-ray diffraction Methods 0.000 description 12
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 8
- 239000002872 contrast media Substances 0.000 description 7
- 238000003917 TEM image Methods 0.000 description 6
- 239000000872 buffer Substances 0.000 description 6
- 230000002147 killing effect Effects 0.000 description 6
- 239000002202 Polyethylene glycol Substances 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 239000010410 layer Substances 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 239000001569 carbon dioxide Substances 0.000 description 4
- 229910002092 carbon dioxide Inorganic materials 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 238000003795 desorption Methods 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 239000002961 echo contrast media Substances 0.000 description 4
- 239000002101 nanobubble Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000004094 surface-active agent Substances 0.000 description 4
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 210000000805 cytoplasm Anatomy 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 201000005202 lung cancer Diseases 0.000 description 3
- 208000020816 lung neoplasm Diseases 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012285 ultrasound imaging Methods 0.000 description 3
- 238000012800 visualization Methods 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000011261 inert gas Substances 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 230000009871 nonspecific binding Effects 0.000 description 2
- 238000012758 nuclear staining Methods 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000001338 self-assembly Methods 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 229910018503 SF6 Inorganic materials 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 230000008497 endothelial barrier function Effects 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 238000000024 high-resolution transmission electron micrograph Methods 0.000 description 1
- 238000002173 high-resolution transmission electron microscopy Methods 0.000 description 1
- 230000005661 hydrophobic surface Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000013335 mesoporous material Substances 0.000 description 1
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 1
- 238000009828 non-uniform distribution Methods 0.000 description 1
- QYSGYZVSCZSLHT-UHFFFAOYSA-N octafluoropropane Chemical compound FC(F)(F)C(F)(F)C(F)(F)F QYSGYZVSCZSLHT-UHFFFAOYSA-N 0.000 description 1
- 229960004065 perflutren Drugs 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000001464 small-angle X-ray scattering data Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- SFZCNBIFKDRMGX-UHFFFAOYSA-N sulfur hexafluoride Chemical compound FS(F)(F)(F)(F)F SFZCNBIFKDRMGX-UHFFFAOYSA-N 0.000 description 1
- 229960000909 sulfur hexafluoride Drugs 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 229940046728 tumor necrosis factor alpha inhibitor Drugs 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
Images
Classifications
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or 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
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
-
- 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
- A61K47/554—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 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
-
- 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
- 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
-
- 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
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- 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)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Acoustics & Sound (AREA)
- Radiology & Medical Imaging (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Dispersion Chemistry (AREA)
- Botany (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明公开了一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用。该载药递送系统包括中空介孔二氧化硅球和包覆在硅球表面的靶向性脂质体,其空心囊泡内装载有气体和药物。中空介孔二氧化硅球作为刚性载体封装气体用于超声造影成像;药物用于靶向递送治疗;靶向性脂质体能够促使囊泡向靶器官递送。其双层结构保证了囊泡结构的穿透性和稳定性,避免气泡在血液循环中被挤压爆破而导致气体扩散、溶解和聚结,延长超声囊泡在血液中的循环时间,保护药物分子在运输途中不被泄露。该载药递送系统作为实体瘤组织的靶向超声成像、诊断和药物递送试剂,在超声成像指导下对病灶部位进行定点成像和药物递送,使药物在特定窗口按需释放。
Description
技术领域
本发明属于靶向超声造影成像和抗癌药物靶向递送给药技术领域,具体涉及一种具有靶向超声造影成像功能的载药递送系统及其制备方法与应用。
背景技术
超声靶向微泡破坏(UTMD)技术是一种经可视化超声成像指导下确定载药微泡到达靶器官后,再给与微泡一定强度的超声辐照,致使微泡破裂并使得细胞膜产生小孔,最终将药物递送进细胞内的方法。传统的超声造影剂微泡直径多为1~10μm,尺寸较大不能穿过血管内皮屏障;早期研究中使用氧气、氮气等气体作为微泡的气核,但是这些气体在血液循环中受到压力易从薄层脂质膜中逸出并溶解于血液,使得微泡在血液中循环时间较短;目前商用微泡的气核多为高密度低溶解性的化学惰性气体,如常规临床使用的超声造影剂使用八氟丙烷或者六氟化硫作为囊泡气体,具有一定的生物毒性且生产成本较高。目前三种常用造影剂如 微泡尺寸分布从1μm到11μm不等,分布较广不均一,且没有靶向功能,缺乏对某种器官或组织的靶向性。其中,/>微泡选用人血清白蛋白作为封装气体的外壳,大大增加了药物的成本。基于目前微泡破坏技术存在的一系列问题,我们提出了利用空心二氧化硅球作为“微/纳泡”支撑体,在二氧化硅球表面包覆一层靶向性脂质体,形成靶向超声“微/纳泡”作为空心囊泡载药递送系统,达到靶向破坏癌细胞的目的。
发明内容
本发明的目的在于,针对现有技术的上述不足,提供了一种具有靶向超声造影成像功能的载药递送系统及其制备方法与应用。
为实现上述目的,本发明采用如下的技术方案:
本发明的第一目的是提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统,所述载药递送系统包括中空介孔二氧化硅球和包覆在硅球表面的靶向性脂质体,所述中空介孔二氧化硅球为空心球,包括球壳和空腔,球壳上有大量介孔,空腔内装载有气体和药物,所述脂质体包覆在所述球壳上。
进一步的,所述中空介孔二氧化硅球粒径为50~5000nm,球壳厚度为5~30nm,孔径为2~12nm。
进一步的,所述中空介孔二氧化硅球装载的气体包括空气、氮气、氧气、二氧化碳中的任一种。
进一步的,所述中空介孔二氧化硅球还装载有药物,所述药物为盐酸阿霉素、紫杉醇、喜树碱、顺铂、卡铂、多柔比星、5-氟尿嘧啶、siRNA、肿瘤坏死因子和蛋白酶抑制剂中的任一种或多种。
进一步的,所述靶向性脂质体为多种磷脂分子、胆固醇及PEG分子的组合。
进一步的,所述靶向性脂质体组成选自5A2-SC8/DOPE/胆固醇/DMG-PEG、5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP和5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA中的任一种。
本发明的第二目的是提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法,包括以下具体步骤:
步骤S1、中空介孔二氧化硅球的制备
步骤S11、前驱体溶液制备
水相,按摩尔比,称取介孔模板剂、铵盐和水,混合均匀,用稀盐酸调节pH不大于2;所述介孔模板剂、铵盐和水的摩尔比范围为(0.2~0.8):(0.5~4):100;
醇相,按摩尔比,称取硅源和乙醇,混合均匀;所述硅源和乙醇的摩尔比范围为(1~10):100;
步骤S12,将步骤S1分别制得的水相和醇相混合搅拌均匀后静置5~29h,然后分批加入雾化器,被雾化成微纳米尺寸的气溶胶小液滴进入高温管式炉内,设定温度为200~500℃,在高温炉出口端滤膜上维持80~85℃收集产物;将产物在500~600℃高温空气中煅烧6~8h,得到中空介孔二氧化硅球;
步骤S2、装载药物
将步骤S1得到的中空介孔二氧化硅球溶解在PBS缓冲液中,加入PBS缓冲液溶解的药物分子,混合震荡后离心收集沉淀,并用PBS缓冲液离心洗涤,得到装载药物的中空介孔二氧化硅球;
步骤S3、脂质体包覆
步骤S31、脂质体悬液的预制
将商用5A2-SC8、DOPE、胆固醇、PEG、DOTAP或者18PA按预设摩尔比混合,溶解在氯仿中并蒸干溶剂,将干燥好的脂质膜用PBS缓冲液再水化得到脂质体悬液,使用配备有不同孔径的聚碳酸酯膜的脂质挤出器对脂质体悬液进行挤压循环,即得直径在特定值分布的脂质体悬液;所述预设摩尔浓度比为5A2-SC8/DOPE/胆固醇/DMG-PEG的摩尔浓度比为5:5:10:1、5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP的摩尔浓度比为5:5:10:1:21和5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA的摩尔浓度比为5:5:10:1:9.1中的任一种;
步骤S32、载药中空介孔二氧化硅囊泡的制备
将载药中空介孔二氧化硅球与脂质体悬液经过多次移液混合,离心除去上清液,再用PBS缓冲液离心洗涤沉淀,最后将沉淀分散在PBS缓冲液中冷藏保存,得载药中空介孔二氧化硅囊泡悬浮液;
步骤S4、装载气体
将步骤S3得到的载药中空介孔二氧化硅囊泡悬浮液冷冻干燥抽真空后装入无菌容器中,并将气体充入至无菌容器中,密封,冷藏保存,即可得到基于中空介孔二氧化硅囊泡的载气载药递送系统。
进一步的,所述介孔模板剂选自十六烷基三甲基溴化铵(CTAB)、十八烷基三甲基溴化铵、水杨酸钠、嵌段共聚物P123、嵌段共聚物F127、嵌段共聚物F-108、Brij-56、Brij-58、Brij-93和Brij-98中的任一种。
进一步的,所述铵盐为氯化铵或硫酸铵,所述硅源选自硅酸四乙酯、硅酸四甲酯、甲基三乙氧基硅烷、甲基三甲氧基硅烷、1,2-二(三乙氧基甲硅烷基)乙烷、氨基硅烷、3-氨丙基三乙氧基硅烷、3-氨丙基甲基二乙氧基硅烷、3-氨丙基甲基二甲氧基硅烷、Y-巯丙基三甲氧基硅烷、Y-巯丙基三乙氧基硅烷、烯丙基三乙氧基硅烷、烯丙基三甲氧基硅烷、三乙氧基乙烯基硅烷等中的任一种。
本发明的第三个目的是提供一种治疗实体瘤的药物或诊断试剂,采用上述的空心囊泡载药递送系统制备得到。
与现有技术比较,本发明提供的技术方案带来的有益效果是:
(1)本发明提供的一种具有靶向超声造影成像功能的空心囊泡载药递送系统,利用中空介孔二氧化硅球作为载体封装药物分子和气体。用粒径和孔径可控的单分散介孔氧化硅球作为支撑体,包覆一层靶向性脂质体,使得微泡具有足够的刚性(机械强度)和稳定性,被封装在硅球内的囊泡内气体不容易逸出并溶解于血液。囊泡内还可以灌装药物、磁性材料、荧光等材料,赋予其多功能性,使之成为一种新型的“靶向超声微/纳泡”,用于超声造影指导下的靶向药物递送至肿瘤组织,可实现超声治疗和药物治疗一体化;
(2)针对目前市面上超声造影剂气核为惰性气体的成本高毒性大,以及微泡尺寸大、分布不均一的问题,本发明采用具有一定刚性的中空介孔二氧化硅球包载空气、氧气、氮气、二氧化碳,成本低且安全性更高。二氧化硅球可以做到微米、亚微米及纳米尺寸,微泡的穿透性更强,亦可以满足全尺寸的“微/纳泡”的制备及应用。而且中空介孔二氧化硅球较大的孔容可以装载各种药物试剂,满足多种疾病治疗的需求;
(3)本发明提供的一种中空介孔二氧化硅球,采用气溶胶辅助自组装法合成,可获得粒径、厚度、孔径和孔容可控的中空介孔二氧化硅球,粒径为50~5000nm,孔径为2~12nm,壳层厚度为5~30nm;
(4)中空介孔二氧化硅颗粒可大规模生产,作为载体成本较低,同时具有良好的生物相容性、较大的孔容、开放的孔结构、均匀可调的孔径以及较好的化学和机械稳定性,可作为药物、基因、生物传感器、细胞内标记物和生物活性分子的有效载体;
(5)不同靶向性脂质体的修饰,如权利要求书里列出的三种脂质体组成分别靶向至肝脏、肺、脾,可以满足囊泡对多种实体瘤组织的靶向超声微泡破坏治疗;
(6)本发明提供表面亲疏水可调的二氧化硅球的制备方法,满足囊泡制备的多样性需求;亦可通过二氧化硅球表面改性,包覆双层脂质体或者化学修饰单层脂质体,满足超声造影在不同场景下的可控调节;
(7)本发明制备得到的基于脂质体包覆中空介孔二氧化硅球的载药递送系统,能够在超声成像指导下对病灶部位进行定点成像和药物递送,使药物在特定窗口按需释放,可以达到最大化药物利用率。
附图说明
图1为具有靶向超声造影成像功能的空心囊泡载药递送系统示意图。空心囊泡的组成为:核心内部的气体和药物,中间层的中空介孔二氧化硅球以及最外层的靶向性脂质体。其中,脂质体中间嵌有胆固醇用以稳定双分子层结构,聚乙二醇用以保护囊泡并减小囊泡在体内循环中的非特异性结合;
图2为以CTAB和NH4Cl为模版制备的HMSNs的TEM图像;
图3为以CTAB和(NH4)2SO4为模版制备的HMSNs的TEM图像;
图4为以CTAB和(NH4)2SO4为模版制备的HMSNs性能分析图,其中a图为HMSNs的N2吸附脱附等温线;b图为HMSNs的小角X射线散射图;c图和d图为HMSNs的HRTEM图;
图5为平均粒径分别约为0.1μm(a)和1.0μm(b)的脂质体包覆中空介孔二氧化硅球的超声造影模式的成像图;
图6为脂质体包覆的亲水中空介孔二氧化硅球(a)和疏水中空介孔二氧化硅球(b)的超声造影模式的成像图;
图7为脂质体包覆疏水HMSNs在第3秒(a)和第40秒(b)时的超声造影模式和超声2D模式成像图;
图8为不同浓度脂质体包覆疏水HMSNs的实时静态超声2D模式成像图,其中a图HMSNs浓度为0.10mg/mL;b图HMSNs浓度为0.20mg/mL;c图HMSNs浓度为0.39mg/mL;d图HMSNs浓度为0.78mg/mL;e图HMSNs浓度为1.56mg/mL;f图HMSNs浓度为3.125mg/mL;g图HMSNs浓度为6.25mg/mL;h图HMSNs浓度为12.5mg/mL;i图HMSNs浓度为25mg/mL;j图HMSNs浓度为50mg/mL;
图9为载药中空介孔二氧化硅球的TEM图;
图10为中空介孔二氧化硅囊泡作为载体递送小分子进入癌细胞的能力研究结果图,其中a图为细胞核染色;b图表示荧光分子在细胞质中的分布;c图为A549细胞明场成像图;d为三者叠加图;
图11为装载阿霉素的中空介孔二氧化硅囊泡对肺癌细胞的杀伤作用研究结果图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面结合具体实施例和附图,对本发明的具体实施方式作进一步详细描述。实施例中未注明具体技术或条件的,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
本发明实施例中涉及的试剂和生物材料购买来源说明如下:硅酸四乙酯(C8H20O4Si),纯度≥98%,甲基三乙氧基硅烷,纯度≥98%及其他硅源购买自上海麦克林生化科技有限公司;硫酸铵((NH4)2SO4)、氯化铵(NH4Cl)、正己烷、环己烷、乙醇(均为分析纯)、盐酸(37%)购买自国药集团化学试剂有限公司;聚乙二醇(HO(CH2CH2O)nH)(Mn=400)购买自上海麦克林生化科技有限公司;聚乙二醇(HO(CH2CH2O)nH)(Mn=2000)购买自上海阿拉丁生化科技股份有限公司;十六烷基三甲基溴化铵(CTAB)、十八烷基三甲基溴化铵、水杨酸钠,纯度≥99%,购买自上海阿拉丁生化科技股份有限公司;嵌段共聚物表面活性剂和Brij系列表面活性剂、胆固醇、DMG-PEG2000、DOPE购买自Sigma-Aldrich;5A2-SC8购买自瀚香生物科技;磷酸盐缓冲液(PBS buffer)购买自生工生物工程(上海)股份有限公司,A549细胞购于中国科学院细胞库,目录号SCSP-503。
本发明实施例中涉及的仪器设备说明如下:高温管式炉德意生(080ST)、微型脂质体挤出器Avanti Polar Lipids Inc.、PhilipsEPIQ7彩色超声诊断系统、高压气流式雾化器TSI 9302A。
如图1所示,本发明制备的具有靶向超声造影成像功能的空心囊泡载药递送系统示意图,其包括中空介孔二氧化硅球和包覆在硅球表面的靶向性脂质体,中空介孔二氧化硅球为空心球,包括球壳和空腔,球壳上有大量介孔,空腔内装载有气体和药物,脂质体包覆在球壳上,具体制备过程如下:
步骤S1,中空介孔二氧化硅球的制备
步骤S11,前驱体溶液制备
水相,按摩尔比,称取介孔模板剂、铵盐和水,混合均匀,用稀盐酸调节pH不大于2;所述介孔模板剂、铵盐和水的摩尔比范围为(0.2~0.8):(0.5~4):100;
醇相,按摩尔比,称取硅源和乙醇,混合均匀;所述硅源和乙醇的摩尔比范围为(1~10):100;
步骤S12,将步骤S11分别制得的水相和醇相混合搅拌均匀后静置5~29h,然后分批加入雾化器,被雾化成微纳米尺寸的气溶胶小液滴进入高温管式炉内,设定温度为200~500℃,在高温炉出口端滤膜上维持80~85℃收集产物;将产物在500~600℃高温空气中煅烧6~8h,得到中空介孔二氧化硅颗粒。
前驱体溶液中不同原料起着不同的作用,十六烷基三甲基溴化铵(CTAB)是表面活性剂(模板剂),是HMSNs有序介孔结构的来源,氯化铵、硫酸铵等铵盐是颗粒空腔结构的来源,加酸使体系pH≤2目的抑制硅源的缩合反应,保证在静置过程中充分熟化,本发明的制备过程可控,制得的HMSNs粒径50~5000nm,孔径2~12nm,壳层厚度为5~30nm。
步骤S2、装载药物
将步骤S1得到的中空介孔二氧化硅用PBS缓冲液溶解,加入PBS缓冲液溶解的药物分子,在震荡器中震荡一段时间,再离心除去剩余的药物分子,并用PBS缓冲液离心洗涤沉淀2次,最终用PBS缓冲液重悬沉淀物,得到装载药物的中空介孔二氧化硅悬浮液;
在一些实施中,中空介孔二氧化硅颗粒内装载药物,药物可以为盐酸阿霉素、紫杉醇、喜树碱、顺铂、卡铂、多柔比星、5-氟尿嘧啶、siRNA、肿瘤坏死因子和蛋白酶抑制剂中的任一种或多种。
步骤S3、脂质体包覆
步骤S31、脂质体悬液的预制
将商用5A2-SC8、DOPE、胆固醇、PEG、DOTAP或者18PA按预设摩尔浓度比的配比溶解在氯仿中并蒸干溶剂,将干燥好的脂质膜用PBS缓冲液再水化得到脂质体悬液,使用配备有不同孔径的聚碳酸酯膜的脂质挤出器对脂质体悬液进行挤压循环,即得直径在特定值分布的脂质体悬液;
靶向性脂质体组成选自5A2-SC8/DOPE/胆固醇/DMG-PEG(摩尔浓度比为5:5:10:1)、5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP(摩尔浓度比为5:5:10:1:21)和5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA(摩尔浓度比为5:5:10:1:9.1)中的任一种;
步骤S32、载药中空介孔二氧化硅囊泡的制备
将载药中空介孔二氧化硅球与脂质体悬液经过多次移液混合,离心除去上清液,再用PBS缓冲液离心洗涤沉淀,最后将沉淀分散在PBS缓冲液中冷藏保存,得载药中空介孔二氧化硅囊泡悬浮液;
在实施中,不同的脂质体组成促使囊泡靶向至不同的器官组织;
在实施中,脂质体组成中的聚乙二醇可以保护中空介孔二氧化硅颗粒,作为屏蔽材料减少颗粒在生物体内的非特异性结合,并延长体内循环时间。
步骤S4,装载气体
将步骤S3得到的载药中空介孔二氧化硅囊泡悬浮液冷冻干燥抽真空后装在一个无菌小瓶中,将气体充入无菌小瓶中,密封,冷藏保存,即可得到基于中空介孔二氧化硅囊泡的载气载药递送系统,中空介孔二氧化硅囊泡装载的是空气、氮气、氧气、二氧化碳中的任一种。
在一些实施方式中,还可以将脂质体包覆中空介孔二氧化硅球的载药递送系统用于制备靶向超声造影剂,用于超声成像。
在一些实施方式中,还可以将脂质体包覆中空介孔二氧化硅颗粒的载药递送系统用于制备治疗实体瘤的药物或者治疗实体瘤的诊断试剂。
实施例1
本实施例提供一种中空介孔二氧化硅球的制备
步骤S1,前驱体溶液制备
称取4g十六烷基三甲基溴化铵(CTAB)、5g氯化铵和56g水,混合均匀,滴加稀盐酸调节pH=2,得水相;
称取10g硅酸四乙酯(TEOS)和30g乙醇,混合均匀,得醇相;
步骤S2,将步骤S1分别制得的水相和醇相混合搅拌均匀后静置8h,然后分批加入雾化器(1~3bar,10000~20000rpm/min),被雾化成微纳米尺寸的气溶胶小液滴进入高温管式炉内,设定温度为400℃,在高温炉出口端滤膜上维持80~85℃收集产物;产物在550℃高温空气中煅烧6h,得到中空介孔二氧化硅球。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为905m2/g,孔容为0.393cm3/g,孔径为2.2nm,平均粒径为938nm。
实施例2
本实施例的中空介孔二氧化硅球的制备方法与实施例1基本相同,不同之处在于:选用的铵盐为硫酸铵。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为949m2/g,孔容为0.407cm3/g,孔径为2.2nm,平均粒径为956nm。接触角测量仪测试其接触角为40°。
实施例3
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:选用的模板剂为嵌段共聚物P123。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为992m2/g,孔容为0.503cm3/g,孔径为6.4nm,平均粒径为793nm。
实施例4
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:选用的模板剂为B56。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为857m2/g,孔容为0.445cm3/g,孔径为4.5nm,平均粒径为956nm。
实施例5
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:选用的模板剂为CTAB和扩孔剂TMB。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为977m2/g,孔容为0.895cm3/g,孔径为11.8nm,平均粒径为513nm。
实施例6
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:步骤S2中,静置时间为29h。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为824m2/g,孔容为0.425cm3/g,孔径为2.2nm,平均粒径为967nm。
实施例7
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:步骤S2中,高温管式炉内,设定温度为500℃。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为976m2/g,孔容为0.297cm3/g,孔径为2.2nm,平均粒径为878nm。
实施例8
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:硅源为甲基三乙氧基硅烷(MTES)。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为978m2/g,孔容为0.504cm3/g,孔径为2.1nm,平均粒径为335nm。接触角测量仪测试其接触角为125°。
实施例9
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:硅源为硅酸四甲酯。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为1193m2/g,孔容为0.516cm3/g,孔径为2.3nm,平均粒径为55nm。
实施例10
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:雾化器选用LPG-5,气体压力1~3bar,转速10000~20000rpm/min。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为458m2/g,孔容为0.602cm3/g,孔径为2.3nm,平均粒径为5000nm。
实施例11
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:雾化器选用TSI 3076,气体压力1~3bar,转速10000~20000rpm/min。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为591m2/g,孔容为0.452cm3/g,孔径为2.2nm,平均粒径为1250nm。
实施例12
本实施例的中空介孔二氧化硅球的制备方法与实施例2基本相同,不同之处在于:雾化器选用鱼跃超声雾化器403E,气体压力1~3bar,转速10000~20000rpm/min。采用TEM、小角XRD散射、低温氮气物理吸附仪和激光粒度仪记录了中空介孔二氧化硅球的比表面积为593m2/g,孔容为0.598cm3/g,孔径为2.2nm,平均粒径为3360nm。
为了探究铵盐种类的影响,通过TEM观察比较了实施例1和实施例2制备的HMSNs空腔形貌的差异。
如图2所示,以CTAB和NH4Cl为模版制备的HMSNs的TEM图像,如图3所示,以CTAB和(NH4)2SO4为模版制备的HMSNs的TEM图像,对比图2和图3可以看出用氯化铵合成的空腔没有用硫酸铵合成的空腔结构明显且规整,本发明实施例2-12是以硫酸铵作为铵盐。
如图4a所示,以CTAB和(NH4)2SO4为模版制备的HMSNs的N2吸附脱附等温线,可以观察到气溶胶辅助自组装法制备的HMSNs的N2吸附脱附等温线呈现介孔材料典型的IV型回滞环,孔径集中在2.5nm左右,具有高达949m2/g的比表面积和0.407cm3/g的孔容。
如图4b所示,HMSNs的小角X射线散射图,如图4c和4d所示,HMSNs的HRTEM图,从图4b-图4d可以看到HMSNs具有高度有序的介孔结构。
比较实施例2-12,可知,表面活性剂是HMSNs介孔结构的模板剂,不同模板剂的长度及结构组成决定了孔径的大小;不同的铵盐会影响二氧化硅球的空腔分散度;静置时间对HMSNs的比表面积、孔容、孔径并没有明显影响,但是随着静置时间的延长,颗粒粒径会有略微增长;管式炉温度对HMSNs的影响体现在铵盐的分解速率上,温度升高铵盐的分解速率加快,最终间接导致HMSNs孔容和比表面积减小,对孔径和粒径的影响不大;不同厂家的雾化器对颗粒粒径有较大影响。硅源种类对颗粒粒径和亲疏水性有影响,不同硅源分子上携带的基团亲水性越高,制备的中空介孔二氧化硅球则越亲水,反之,硅源分子上携带的基团疏水性越高,制备的中空介孔二氧化硅球则越疏水。
实施例13
本实施例提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备,
步骤S1,中空介孔二氧化硅球的制备
同实施例2。
步骤S2,装载药物
将步骤S1得到的中空介孔二氧化硅球用PBS缓冲液溶解得到1mg/mL的中空介孔二氧化硅悬浮液,加入药物分子使得药物分子终浓度为10mg/mL,在震荡器中震荡混合8h,然后离心除去剩余的药物分子,并用PBS缓冲液离心洗涤2次,离心沉淀物冷冻干燥后称重,再加入一定量的PBS缓冲液使得终浓度为2mg/mL,得到装载药物的中空介孔二氧化硅悬浮液。
步骤S3,脂质体包覆
步骤S31,脂质体悬液的预制
商用5A2-SC8、DOPE、胆固醇、DMG-PEG2000、DOTAP或者18PA在使用前在-20℃下冷冻储藏。使用时以10-25mg/mL的浓度溶解在氯仿中,将试样等分到玻璃小瓶中,脂质的总量被控制在每瓶2.5mg,在室温下真空干燥12小时蒸发小瓶中的氯仿。将干燥好的脂质膜放在室温下,加入1mL 0.5x磷酸缓冲盐溶液(PBS缓冲液)进行再水化,偶尔摇动至少1小时,形成混浊的脂质悬浮液。用Ananti Polar Lipids迷你脂质挤出机挤压悬浮液。使用孔径分别为0.1μm或1.0μm的聚碳酸酯膜,进行至少10个来回的挤压循环。得到的透明脂质体悬浮液分别储存在新的小瓶中,储存在4℃冰箱中备用。
步骤S32,载药中空介孔二氧化硅囊泡的制备:
将平均粒径分别为0.1μm和1.0μm的载药中空介孔二氧化硅称重并转移到不同的玻璃小瓶中。加入PBS缓冲液使得载药中空介孔二氧化硅的最终悬浮液浓度为25mg/mL。将相同体积(通常为2mL)和相同尺寸的载药中空介孔二氧化硅球悬浮液(25mg/mL)与脂质体悬液(2.5mg/mL)通过多次移液混合。混合物在室温下放置20分钟。将混合物在4000rpm下离心1分钟,然后除去上清液,除去多余的脂类。载药中空介孔二氧化硅囊泡随后用0.25x的200μL PBS缓冲液洗涤,最后分散在4mL 0.25x PBS缓冲液中,并冷冻干燥后储存于4℃的冰箱中备用,即得载药中空介孔二氧化硅囊泡悬浮液。
可以通过目测观察到,平均粒径为0.1μm的脂质体水溶液呈淡蓝色,而平均粒径为1.0μm的脂质体水溶液呈乳白色,这是不同粒径的脂质体对光的吸收和散射产生的现象。平均粒径为0.1μm和1.0μm的HMSNs悬液也有类似的现象。
步骤S4,装载气体
将步骤S3得到的载药中空介孔二氧化硅囊泡悬浮液冷冻干燥抽真空后装在一个无菌小瓶中,将气体充入无菌小瓶中,密封,冷藏保存,即可得到基于中空介孔二氧化硅囊泡的载气载药递送系统,中空介孔二氧化硅囊泡装载的是空气、氮气、氧气、二氧化碳中的任一种。
实施例14
本实施例提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法与实施例13基本相同,不同之处在于:脂质体组成为5A2-SC8/DOPE/胆固醇/DMG-PEG(摩尔浓度比为=5:5:10:1)。
实施例15
本实施例提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法与实施例13基本相同,不同之处在于:脂质体组成为5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP(摩尔浓度比为=5:5:10:1:21)。
实施例16
本实施例提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法与实施例13基本相同,不同之处在于:脂质体组成为5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA(摩尔浓度比为=5:5:10:1:9.1)。
实施例17
本实施例提供一种具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法与实施例15基本相同,不同之处在于:装载药物为阿霉素,制得的中空介孔二氧化硅囊泡装载阿霉素的装载率为36.7%。
为了阐述基于脂质体包覆中空介孔二氧化硅球的中空介孔二氧化硅囊泡超声造影性能,本发明进行如下研究:
超声造影采用PhilipsEPIQ7彩色超声诊断系统,配备eL18-4线阵探头,可以同时获得高帧频、高穿透力以及组织均匀一致性的图像。微泡造影剂悬液为现配现用,将微泡造影剂颗粒与生理盐水在专用配药瓶内混合,充分震荡后形成混悬液。首先选择浅表器官成像模式,记录纳米微泡造影剂的二维灰阶超声图像。观察微泡的分布是否均匀一致,调节增益及深度补偿,使图像可以清晰、稳定地显示。随后选择“contrast”模式,进行超声造影模式成像,超声造影成像原理为低机械指数的实时谐波成像,机械指数可低至0.06。采集微泡造影剂的实时动态图像,帧频为37Hz,至少采集1分钟的动态图像。
(1)HMSNs粒径对超声造影性能的影响研究
将制备的平均粒径约为0.1μm(如图5a所示)和平均粒径约1.0μm(如图5b所示)的中空介孔二氧化硅囊泡的PBS悬浮液,通过注射器分别注射进生理盐水中,同时用彩色超声诊断系统记录实时超声造影模式的成像图如图5中虚线框区域所示。可以看出1.0μm比0.1μm的中空介孔二氧化硅囊泡显示出来更好的超声对比增强信号。这与超声微泡体积越大成像效果越好的的结果一致。从记录的动态超声造影信号可以看出,平均粒径为1.0μm的中空介孔二氧化硅囊泡显示出很强的瞬时超声对比增强信号。
(2)HMSNs孔径对超声造影性能的影响研究
对实施例2-12中制得的孔径分别为2.2nm和6.4nm、平均粒径约为1.0μm的HMSNs进行实时超声造影模式,成像结果如图6所示。从图6中可以看出,孔径为2.2nm的HMSNs比孔径为6.4nm的HMSNs的信号更强,说明减小有序孔结构和孔径,从而可以有效减缓空腔中的气体扩散。
(3)亲疏水性硅源对超声造影性能的影响研究
采用实施例2制备的亲水HMSNs和实施例7制备的疏水HMSNs分别记为TEOS-1和MTES-2,将TEOS-1和MTES-2分别通过注射器直接注射进生理盐水中,进行实时超声造影模式成像如图6所示。可以看出,疏水HMSNs(图6b所示)比亲水HMSNs(图6a)表现出更好的超声对比增强信号,这是因为疏水的表面使外部的水更难跟空腔中的空气接触,从而防止空腔中的空气快速溶解,对比增强信号强度和持续时间都有明显的改善。
(4)疏水中空介孔二氧化硅囊泡对超声造影性能的影响研究
将实施例7制备的疏水HMSNs称重并转移到玻璃小瓶中,加入含1%聚乙二醇(Mn=400)的去离子水,超声作用至少10分钟,使疏水HMSNs充分分散,最终水浓度为25mg/mL。将相同体积(通常为5mL)制备的疏水二氧化硅悬浮液(25mg/mL)与脂质体悬液(2.5mg/mL)过多次移液混合。混合物在室温下放置20分钟,偶尔搅拌。将混合物在4000rpm下离心1分钟,然后除去上清液,除去多余的脂类。疏水HMSNs囊泡随后用200μL的0.25x PBS缓冲液洗涤,最后分散在10mL 0.25x PBS缓冲液中,并冷冻干燥后储存于4℃的冰箱中备用。将上述方式冷冻干燥后的疏水HMSNs囊泡重新分在去离子水中,通过注射器直接注射进生理盐水中,进行实时超声造影和2D模式成像。
结果如图7所示,分别显示的是疏水HMSNs囊泡分散液注射进生理盐水第3秒(a)和第40秒(b)时的超声造影模式(a1,b1)和超声2D模式(a2,b2)成像。可以看出造影模式和2D模式对比增强信号区域的边界形状基本上是一致的,说明不仅可以用于病灶部位的显像,还可以根据显像区域的面积显示出病灶的大小,同时显像时间也能保证造影剂在外周注射后能够有足够的时间穿过肺毛细血管床,并在检查期间实现增强。
(5)HMSNs浓度对超声造影性能的影响研究
采用实施例7制备的疏水HMSNs囊泡,制备过程同“疏水中空介孔二氧化硅囊泡对超声造影性能的影响研究”。将疏水HMSNs囊泡按不同浓度分散在去离子水中,进行实时静态超声2D模式成像。
结构如图8所示,可以看出当浓度较低时(图8a-e)随着疏水HMSNs囊泡浓度的升高超声2D模式对比增强信号逐渐增强,其中1.56mg/mL浓度下的微泡造影剂显示出了清晰且均匀的对比增强信号,但当浓度过高时(图8f-j)疏水HMSNs囊泡在水相中逐渐开始团聚,甚至会吸收大部分超声波能量,导致大部分信号无法穿透,导致图像中这些区域呈现为黑色阴影。
(6)装载阿霉素的中空介孔二氧化硅囊泡对肺癌细胞的杀伤作用研究
以肺癌细胞(A549)为研究目标,将装载药物的中空介孔二氧化硅囊泡与细胞孵育,研究不同尺寸的二氧化硅囊泡携带的药物分子对癌细胞的杀伤作用。首先,利用中空介孔二氧化硅囊泡装载携带FITC(异硫氰酸荧光素)荧光分子,探究中空介孔二氧化硅囊泡作为载体递送小分子进入细胞的能力。图9所示为平均尺寸约200nm的二氧化硅囊泡装载荧光分子的TEM图。如图10所示,以平均尺寸约为500nm的中空介孔二氧化硅囊泡作为载体,可以大量地将荧光小分子递送至癌细胞中,荧光分子主要分布在细胞质中。其中图10a表示细胞核染色,图10b表示荧光分子在细胞质中的分布,图10c为A549细胞明场成像图,图10d为三者叠加图。
将荧光分子替换成阿霉素药物分子,装载在三种不同尺寸的中空介孔二氧化硅囊泡中:平均粒径分别为0.05μm(D10=26nm,D50=43nm,D90=51nm)、0.5μm(D10=364nm,D50=427nm,D90=502nm)和5μm(D10=3725nm,D50=4167nm,D90=5014nm),并保证装载的药量相同。将PBS缓冲液、HMSNs和载药HMSNs分别与A549细胞孵育6小时后,用CCK8试剂盒测试细胞存活率。如图11所示,HMSNs本身对细胞活性没有明显的影响,说明中空介孔二氧化硅材料本身不具有细胞毒性,而携带阿霉素的递送体系对细胞具有明显的杀伤作用,总体细胞存活率低于50%,说明中空介孔二氧化硅囊泡向癌细胞递送药物分子的可行性较高。而且,随着HMSNs尺寸增大,杀伤作用相对减少,说明细胞对体系的内吞速率相对减弱,但依然保持50%以上的杀伤作用。
在不冲突的情况下,本文中上述实施例及实施例中的特征可以相互结合。
以上所述仅为本发明的较佳实施例,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.一种具有靶向超声造影成像功能的空心囊泡载药递送系统,其特征在于,所述载药递送系统包括中空介孔二氧化硅球和包覆在硅球表面的靶向性脂质体,所述中空介孔二氧化硅球为空心球,包括球壳和空腔,球壳上有大量介孔,空腔内装载有气体和药物,所述脂质体包覆在所述球壳上。
2.如权利要求1所述的空心囊泡载药递送系统,其特征在于,所述中空介孔二氧化硅球粒径为50~5000nm,球壳厚度为5~30nm,介孔孔径为2~12nm。
3.如权利要求1所述的空心囊泡载药递送系统,其特征在于,所述中空介孔二氧化硅球装载的气体包括空气、氮气、氧气、二氧化碳中的任一种。
4.如权利要求3所述的空心囊泡载药递送系统,其特征在于,所述中空介孔二氧化硅球还装载有药物,所述药物为盐酸阿霉素、紫杉醇、喜树碱、顺铂、卡铂、多柔比星、5-氟尿嘧啶、siRNA、肿瘤坏死因子和蛋白酶抑制剂中的任一种或多种。
5.如权利要求4中任一项所述的空心囊泡载药递送系统,其特征在于,所述靶向性脂质体为多种磷脂分子、胆固醇及PEG分子的组合。
6.如权利要求5所述的空心囊泡载药递送系统,其特征在于,所述靶向性脂质体组成选自5A2-SC8/DOPE/胆固醇/DMG-PEG、5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP和5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA中的任一种。
7.一种如权利要求1-6中任一项所述的具有靶向超声造影成像功能的空心囊泡载药递送系统的制备方法,其特征在于,包括以下具体步骤:
步骤S1、中空介孔二氧化硅球的制备
步骤S11、前驱体溶液制备
水相,按摩尔比,称取介孔模板剂、铵盐和水,混合均匀,用稀盐酸调节pH不大于2;所述介孔模板剂、铵盐和水的摩尔比范围为(0.2~0.8):(0.5~4):100;
醇相,按摩尔比,称取硅源和乙醇,混合均匀;所述硅源和乙醇的摩尔比范围为(1~10):100;
步骤S12,将步骤S1分别制得的水相和醇相混合搅拌均匀后静置5~29h,然后分批加入雾化器,被雾化成微纳米尺寸的气溶胶小液滴进入高温管式炉内,设定温度为200~500℃,在高温炉出口端滤膜上维持80~85℃收集产物;将产物在500~600℃高温空气中煅烧6~8h,得到中空介孔二氧化硅球;
步骤S2、装载药物
将步骤S1得到的中空介孔二氧化硅球溶解在PBS缓冲液中,加入PBS缓冲液溶解的药物分子,混合震荡后离心收集沉淀,并用PBS缓冲液离心洗涤,得到装载药物的中空介孔二氧化硅球;
步骤S3、脂质体包覆
步骤S31、脂质体悬液的预制
将商用5A2-SC8、DOPE、胆固醇、PEG、DOTAP或者18PA按预设摩尔浓度比混合,溶解在氯仿中并蒸干溶剂,将干燥好的脂质膜用PBS缓冲液再水化得到脂质体悬液,使用配备有不同孔径的聚碳酸酯膜的脂质挤出器对脂质体悬液进行挤压循环,即得直径在特定值分布的脂质体悬液;所述预设摩尔浓度比为5A2-SC8/DOPE/胆固醇/DMG-PEG的摩尔浓度比为5:5:10:1、5A2-SC8/DOPE/胆固醇/DMG-PEG/DOTAP的摩尔浓度比为5:5:10:1:21和5A2-SC8/DOPE/胆固醇/DMG-PEG/18PA的摩尔浓度比为5:5:10:1:9.1中的任一种;
步骤S32、载药中空介孔二氧化硅囊泡的制备
将载药中空介孔二氧化硅球与脂质体悬液经过多次移液混合,离心除去上清液,再用PBS缓冲液离心洗涤沉淀,最后将沉淀分散在PBS缓冲液中冷藏保存,得载药中空介孔二氧化硅囊泡悬浮液;
步骤S4、装载气体
将步骤S3得到的载药中空介孔二氧化硅囊泡悬浮液冷冻干燥抽真空后装入无菌容器中,并将气体充入至无菌容器中,密封,冷藏保存,即可得到基于中空介孔二氧化硅囊泡的载气载药递送系统。
8.如权利要求7所述的制备方法,其特征在于,所述介孔模板剂选自十六烷基三甲基溴化铵、十八烷基三甲基溴化铵、水杨酸钠、嵌段共聚物P123、嵌段共聚物F127、嵌段共聚物F-108、Brij-56、Brij-58、Brij-93和Brij-98中的任一种。
9.如权利要求7所述的制备方法,其特征在于,所述铵盐为氯化铵或硫酸铵,所述硅源选自硅酸四乙酯、硅酸四甲酯、甲基三乙氧基硅烷、甲基三甲氧基硅烷、1,2-二(三乙氧基甲硅烷基)乙烷、氨基硅烷、3-氨丙基三乙氧基硅烷、3-氨丙基甲基二乙氧基硅烷、3-氨丙基甲基二甲氧基硅烷、Y-巯丙基三甲氧基硅烷、Y-巯丙基三乙氧基硅烷、烯丙基三乙氧基硅烷、烯丙基三甲氧基硅烷、三乙氧基乙烯基硅烷等中的任一种。
10.一种治疗实体瘤的药物或诊断试剂,其特征在于,采用如权利要求1-6中任一项所述的空心囊泡载药递送系统制备得到。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310147628.3A CN116327985A (zh) | 2023-02-21 | 2023-02-21 | 一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310147628.3A CN116327985A (zh) | 2023-02-21 | 2023-02-21 | 一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN116327985A true CN116327985A (zh) | 2023-06-27 |
Family
ID=86878190
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310147628.3A Pending CN116327985A (zh) | 2023-02-21 | 2023-02-21 | 一种具有靶向超声造影成像功能的空心囊泡载药递送系统及其制备方法与应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116327985A (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20150272885A1 (en) * | 2011-10-14 | 2015-10-01 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery including transdermal delivery of cargo and methods thereof |
CN106267248A (zh) * | 2016-09-08 | 2017-01-04 | 重庆医科大学 | 一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法 |
CN115227671A (zh) * | 2022-07-13 | 2022-10-25 | 中山大学附属第一医院 | 一种药物递送系统及其制备方法和应用 |
US20230355812A1 (en) * | 2020-09-08 | 2023-11-09 | Oregon Health & Science University | Stabilized hydrophobic nanoparticles for ultrasound imaging |
-
2023
- 2023-02-21 CN CN202310147628.3A patent/CN116327985A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20150272885A1 (en) * | 2011-10-14 | 2015-10-01 | Stc.Unm | Porous nanoparticle-supported lipid bilayers (protocells) for targeted delivery including transdermal delivery of cargo and methods thereof |
CN106267248A (zh) * | 2016-09-08 | 2017-01-04 | 重庆医科大学 | 一种载叶酸修饰介孔二氧化硅纳米粒的脂质超声微泡及其制备方法 |
US20230355812A1 (en) * | 2020-09-08 | 2023-11-09 | Oregon Health & Science University | Stabilized hydrophobic nanoparticles for ultrasound imaging |
CN115227671A (zh) * | 2022-07-13 | 2022-10-25 | 中山大学附属第一医院 | 一种药物递送系统及其制备方法和应用 |
Non-Patent Citations (3)
Title |
---|
CHENLU HUANG等: "A Dual-Model Imaging Theragnostic System Based on Mesoporous Silica Nanoparticles for Enhanced Cancer Phototherapy", 《ADVANCED HEALTHCARE MATERIALS》, no. 19, 12 September 2019 (2019-09-12), pages 1900840 * |
QIANG CHENG等: "Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR–Cas gene editing", 《NATURE NANOTECHNOLOGY VOLUME》, vol. 15, 6 April 2020 (2020-04-06), pages 313 - 320, XP093095557, DOI: 10.1038/s41565-020-0669-6 * |
付欣等: "原花青素-介孔二氧化硅纳米颗粒复合体制备及释放研究", 《食品科学技术学报》, vol. 38, no. 1, 25 January 2020 (2020-01-25), pages 67 - 73 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Li et al. | Charge-reversal biodegradable MSNs for tumor synergetic chemo/photothermal and visualized therapy | |
Yu et al. | Ultrasmall mesoporous organosilica nanoparticles: Morphology modulations and redox-responsive biodegradability for tumor-specific drug delivery | |
Wang et al. | Mitoxantrone-preloaded water-responsive phospholipid-amorphous calcium carbonate hybrid nanoparticles for targeted and effective cancer therapy | |
Song et al. | A facile synthesis of uniform hollow MIL-125 titanium-based nanoplatform for endosomal esacpe and intracellular drug delivery | |
US20130023714A1 (en) | Medical and Imaging Nanoclusters | |
Kuo et al. | Magnetically triggered nanovehicles for controlled drug release as a colorectal cancer therapy | |
CN102421418A (zh) | 在药物递送中有用的中空金纳米球(HAuNSs)和装载HAuNSs的微球体 | |
CN107019801A (zh) | 一种磁热释放的热敏脂质体 | |
Lin et al. | Doxorubicin loaded silica nanoparticles with dual modification as a tumor-targeted drug delivery system for colon cancer therapy | |
Wang et al. | Photo-responsive magnetic mesoporous silica nanocomposites for magnetic targeted cancer therapy | |
Sen et al. | Simple one-pot fabrication of ultra-stable core-shell superparamagnetic nanoparticles for potential application in drug delivery | |
Guo et al. | Gold nanoparticle-guarded large-pore mesoporous silica nanocomposites for delivery and controlled release of cytochrome c | |
Zhang et al. | Chitosan and dextran stabilized GO-iron oxide nanosheets with high dispersibility for chemotherapy and photothermal ablation | |
Jie et al. | Superparamagnetic iron oxide nanoparticles/doxorubicin-loaded starch-octanoic micelles for targeted tumor therapy | |
Tiburcius et al. | Egg-yolk core–shell mesoporous silica nanoparticles for high doxorubicin loading and delivery to prostate cancer cells | |
CN101874781B (zh) | 疏水改性葡聚糖修饰的长循环脂质体及其制备方法 | |
CN105233282B (zh) | 一种多功能纳米药物组合物及其制备方法 | |
Zielińska et al. | Mesoporous silica nanoparticles as drug delivery systems against melanoma | |
CN107970454A (zh) | 一种氧化石墨烯-脂质纳米复合材料的制备方法及应用 | |
US20230052618A1 (en) | Nanobowl-supported drug-loaded liposome, preparation method therefor and application thereof | |
Liu et al. | Hierarchical theranostic nanomedicine: MRI contrast agents as a physical vehicle anchor for high drug loading and triggered on-demand delivery | |
Liu et al. | Dual-responsive and controlled-release paclitaxel-loaded mesoporous silicon nanoparticles with cell membrane coating for homologous targeted therapy of tongue squamous cell carcinoma | |
Thakkar et al. | Vincristine-sulphate–loaded liposome-templated calcium phosphate nanoshell as potential tumor-targeting delivery system | |
Ge et al. | Walnut kernel-like mesoporous silica nanoparticles as effective drug carrier for cancer therapy in vitro | |
CN108888773B (zh) | 自组装球形药物纳米制剂及其制备方法与用途 |
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 |