CN109966514A - 一种相转变靶向纳米泡、制备方法及应用 - Google Patents
一种相转变靶向纳米泡、制备方法及应用 Download PDFInfo
- Publication number
- CN109966514A CN109966514A CN201910342099.6A CN201910342099A CN109966514A CN 109966514 A CN109966514 A CN 109966514A CN 201910342099 A CN201910342099 A CN 201910342099A CN 109966514 A CN109966514 A CN 109966514A
- Authority
- CN
- China
- Prior art keywords
- phase
- nanometer
- bubble
- perflenapent
- ultrasonic
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 230000007704 transition Effects 0.000 title claims abstract description 15
- 239000002101 nanobubble Substances 0.000 title claims abstract description 10
- 238000003384 imaging method Methods 0.000 claims abstract description 37
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 29
- 229960004692 perflenapent Drugs 0.000 claims abstract description 29
- NJCBUSHGCBERSK-UHFFFAOYSA-N perfluoropentane Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)F NJCBUSHGCBERSK-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229920000747 poly(lactic acid) Polymers 0.000 claims abstract description 28
- 239000004626 polylactic acid Substances 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 239000003337 fertilizer Substances 0.000 claims abstract description 14
- 239000000945 filler Substances 0.000 claims abstract description 9
- 239000002775 capsule Substances 0.000 claims abstract description 8
- 239000011229 interlayer Substances 0.000 claims abstract description 3
- 239000002245 particle Substances 0.000 claims description 20
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 17
- 239000002872 contrast media Substances 0.000 claims description 17
- 239000000839 emulsion Substances 0.000 claims description 17
- 239000005457 ice water Substances 0.000 claims description 8
- 239000002504 physiological saline solution Substances 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- 238000009210 therapy by ultrasound Methods 0.000 claims description 8
- 239000006185 dispersion Substances 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 230000001376 precipitating effect Effects 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 5
- 238000013019 agitation Methods 0.000 claims description 4
- 230000000118 anti-neoplastic effect Effects 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 4
- 239000003921 oil Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 229940079593 drug Drugs 0.000 claims description 3
- 229920000151 polyglycol Polymers 0.000 claims 1
- 239000010695 polyglycol Substances 0.000 claims 1
- 238000002604 ultrasonography Methods 0.000 abstract description 16
- 230000000694 effects Effects 0.000 abstract description 12
- 238000007920 subcutaneous administration Methods 0.000 abstract description 5
- 230000009471 action Effects 0.000 abstract description 3
- 229910021642 ultra pure water Inorganic materials 0.000 abstract description 3
- 239000012498 ultrapure water Substances 0.000 abstract description 3
- 230000008859 change Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 17
- 238000012360 testing method Methods 0.000 description 16
- 239000007789 gas Substances 0.000 description 14
- 230000008685 targeting Effects 0.000 description 10
- 229920001577 copolymer Polymers 0.000 description 8
- 238000001727 in vivo Methods 0.000 description 7
- 229960000304 folic acid Drugs 0.000 description 6
- 239000011724 folic acid Substances 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- OVBPIULPVIDEAO-LBPRGKRZSA-N Folic acid Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 5
- 241000699660 Mus musculus Species 0.000 description 5
- 239000002246 antineoplastic agent Substances 0.000 description 5
- 229940041181 antineoplastic drug Drugs 0.000 description 5
- 210000004204 blood vessel Anatomy 0.000 description 5
- 235000013339 cereals Nutrition 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 238000003745 diagnosis Methods 0.000 description 5
- 238000009826 distribution Methods 0.000 description 5
- 238000011580 nude mouse model Methods 0.000 description 5
- 238000000935 solvent evaporation Methods 0.000 description 5
- 102000009027 Albumins Human genes 0.000 description 4
- 108010088751 Albumins Proteins 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 230000017531 blood circulation Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000003902 lesion Effects 0.000 description 4
- -1 phosphatide Substances 0.000 description 4
- 238000012285 ultrasound imaging Methods 0.000 description 4
- OBWDYOYJIGYDSU-YDALLXLXSA-N CC(O)C(O)=O.N1=C2C(=O)NC(N)=NC2=NC=C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 Chemical compound CC(O)C(O)=O.N1=C2C(=O)NC(N)=NC2=NC=C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OBWDYOYJIGYDSU-YDALLXLXSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000002616 MRI contrast agent Substances 0.000 description 3
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 3
- 241000209094 Oryza Species 0.000 description 3
- 235000007164 Oryza sativa Nutrition 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 235000019152 folic acid Nutrition 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 3
- 235000009566 rice Nutrition 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 238000010008 shearing Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 230000036760 body temperature Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 210000003022 colostrum Anatomy 0.000 description 2
- 235000021277 colostrum Nutrition 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000013399 early diagnosis Methods 0.000 description 2
- 239000002961 echo contrast media Substances 0.000 description 2
- 210000003038 endothelium Anatomy 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000002349 favourable effect Effects 0.000 description 2
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 239000011261 inert gas Substances 0.000 description 2
- QWTDNUCVQCZILF-UHFFFAOYSA-N isopentane Chemical compound CCC(C)C QWTDNUCVQCZILF-UHFFFAOYSA-N 0.000 description 2
- 229960000485 methotrexate Drugs 0.000 description 2
- 239000004005 microsphere Substances 0.000 description 2
- 239000007908 nanoemulsion Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 229910052697 platinum Inorganic materials 0.000 description 2
- 229920001610 polycaprolactone Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000013558 reference substance Substances 0.000 description 2
- 230000003014 reinforcing effect Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000002626 targeted therapy Methods 0.000 description 2
- HAWSQZCWOQZXHI-FQEVSTJZSA-N 10-Hydroxycamptothecin Chemical compound C1=C(O)C=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 HAWSQZCWOQZXHI-FQEVSTJZSA-N 0.000 description 1
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 1
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102000008857 Ferritin Human genes 0.000 description 1
- 108050000784 Ferritin Proteins 0.000 description 1
- 238000008416 Ferritin Methods 0.000 description 1
- 108010063045 Lactoferrin Proteins 0.000 description 1
- 102100032241 Lactotransferrin Human genes 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- 241000549556 Nanos Species 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- PRPAGESBURMWTI-UHFFFAOYSA-N [C].[F] Chemical class [C].[F] PRPAGESBURMWTI-UHFFFAOYSA-N 0.000 description 1
- XGCDHPDIERKJPT-UHFFFAOYSA-N [F].[S] Chemical class [F].[S] XGCDHPDIERKJPT-UHFFFAOYSA-N 0.000 description 1
- MXZROTBGJUUXID-UHFFFAOYSA-I [Gd+3].[O-]C(=O)CN(CC([O-])=O)CCN(CC(=O)[O-])CCN(CC([O-])=O)C(C([O-])=O)COCC1=CC=CC=C1 Chemical compound [Gd+3].[O-]C(=O)CN(CC([O-])=O)CCN(CC(=O)[O-])CCN(CC([O-])=O)C(C([O-])=O)COCC1=CC=CC=C1 MXZROTBGJUUXID-UHFFFAOYSA-I 0.000 description 1
- XJJWWOUJWDTXJC-UHFFFAOYSA-N [Mn].N1C(C=C2N=C(C=C3NC(=C4)C=C3)C=C2)=CC=C1C=C1C=CC4=N1 Chemical compound [Mn].N1C(C=C2N=C(C=C3NC(=C4)C=C3)C=C2)=CC=C1C=C1C=CC4=N1 XJJWWOUJWDTXJC-UHFFFAOYSA-N 0.000 description 1
- 229940009456 adriamycin Drugs 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229920002988 biodegradable polymer Polymers 0.000 description 1
- 239000004621 biodegradable polymer Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 190000008236 carboplatin Chemical compound 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000012412 chemical coupling Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000002059 diagnostic imaging Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- AFABGHUZZDYHJO-UHFFFAOYSA-N dimethyl butane Natural products CCCC(C)C AFABGHUZZDYHJO-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000007720 emulsion polymerization reaction Methods 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 229940031098 ethanolamine Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- NBVXSUQYWXRMNV-UHFFFAOYSA-N fluoromethane Chemical compound FC NBVXSUQYWXRMNV-UHFFFAOYSA-N 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 125000003929 folic acid group Chemical group 0.000 description 1
- 150000002251 gadolinium compounds Chemical class 0.000 description 1
- GFSTXYOTEVLASN-UHFFFAOYSA-K gadoteric acid Chemical compound [Gd+3].OC(=O)CN1CCN(CC([O-])=O)CCN(CC([O-])=O)CCN(CC([O-])=O)CC1 GFSTXYOTEVLASN-UHFFFAOYSA-K 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 description 1
- 229940078795 lactoferrin Drugs 0.000 description 1
- 235000021242 lactoferrin Nutrition 0.000 description 1
- 108010071397 lactoferrin receptors Proteins 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 150000002697 manganese compounds Chemical class 0.000 description 1
- 230000010358 mechanical oscillation Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229920000867 polyelectrolyte Polymers 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009774 resonance method Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 229960001603 tamoxifen Drugs 0.000 description 1
- 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 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- KSBAEPSJVUENNK-UHFFFAOYSA-L tin(ii) 2-ethylhexanoate Chemical compound [Sn+2].CCCCC(CC)C([O-])=O.CCCCC(CC)C([O-])=O KSBAEPSJVUENNK-UHFFFAOYSA-L 0.000 description 1
- BENFPBJLMUIGGD-UHFFFAOYSA-I trisodium;2-[2-[carboxylatomethyl-[[3-hydroxy-2-methyl-5-(phosphonatooxymethyl)pyridin-4-yl]methyl]amino]ethyl-[[3-hydroxy-5-[[hydroxy(oxido)phosphoryl]oxymethyl]-2-methylpyridin-4-yl]methyl]amino]acetate;manganese(2+) Chemical compound [H+].[H+].[H+].[Na+].[Na+].[Na+].[Mn+2].CC1=NC=C(COP([O-])([O-])=O)C(CN(CCN(CC([O-])=O)CC=2C(=C(C)N=CC=2COP([O-])([O-])=O)[O-])CC([O-])=O)=C1[O-] BENFPBJLMUIGGD-UHFFFAOYSA-I 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000008215 water for injection Substances 0.000 description 1
- 230000010148 water-pollination Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/08—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by the carrier
- A61K49/10—Organic compounds
- A61K49/12—Macromolecular compounds
- A61K49/126—Linear polymers, e.g. dextran, inulin, PEG
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
- A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
- A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
- A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
- A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
- A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
- A61K49/1851—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule
- A61K49/1857—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an organic macromolecular compound, i.e. oligomeric, polymeric, dendrimeric organic molecule the organic macromolecular compound being obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. PLGA
-
- 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
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5146—Organic macromolecular compounds; Dendrimers obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
- A61K9/5153—Polyesters, e.g. poly(lactide-co-glycolide)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Medicinal Chemistry (AREA)
- Radiology & Medical Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Optics & Photonics (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Acoustics & Sound (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明提供一种相转变靶向纳米泡、制备方法及应用。纳米泡由囊心填充物、包膜材料和水组成,为夹层球状结构;所述囊心填充物为液态全氟戊烷;所述包膜材料为氨基封端聚乳酸;所述氨基封端聚乳酸含量为0.1~5.0wt%,液态全氟戊烷含量为1.0%~15.0wt%,余量为超纯水。所制备的纳米泡为乳白色,粒径318.4±5.1nm,体外3个月粒径变化较小,稳定性好,纳米泡在超声波的作用下,能增强超声成像的效果,并且能被超声波击破,在小鼠皮下瘤活体模型中,能显著地提高超声对肿瘤的成像效果。
Description
技术领域
本发明内容属于医药技术领域。涉及一种相转变纳米泡及其制备方法和性质,纳米泡采用乳化溶剂挥发法制备,可用于肿瘤病灶的超声成像、MRI成像诊断及肿瘤的靶向治疗,是一类诊疗一体化的新型多功能影像学纳米造影剂。
背景技术
微泡是一类能够显著增强医学超声检测信号的超声成像对比剂,微泡内气体和周围活体组织之间高的声阻抗差使微泡产生强反射,导致血液中的背向散射增强(在彩色和频谱多普勒模式下可高达27dB),从而达到增强超声图像效果的目的(JM Correas,etal.Ulreasound contrast agents:properties,principles of action,tolerance,andartifacts.Eur.Radiol.,2001,11:1316-1328.)。第一代微泡超声成像对比剂是含有自由气泡的生理盐水或乳液,以空气或氧气为主要成分,由于自由气体扩散很快而迅速失去声反射性,使其应用范围受到限制,仅能短暂地在右心系统超声显影。第二代微泡超声成像对比剂是以内部包裹空气的人血清白蛋白微泡和糖类微泡为代表,采用超声声振法制备,由于微泡外以固化的人血清白蛋白或糖类物质包膜,具有一定的稳定性,可实现左心系统超声显影,从而具有临床实用价值并实现商品化。1993年和1994年,Molecular Biosystems公司的包含空气声振白蛋白微泡产品首先在日本和美国上市,先灵葆雅公司包含空气的糖类微泡和产品也随后上市。但是,第二代微泡超声成像对比剂存在超声增强效果弱、血循环中持续时间短等缺点,2000年后逐渐退出市场。近十年来,随着表面活性剂、磷脂、聚电解质等新型微泡包膜材料和低扩散系数惰性气体(如氟碳类气体、氟硫类气体等)的应用,出现了第三代微泡超声成像对比剂。这类微泡较第二代微泡产品血液循环系统的稳定性有了一定提高,超声声波反射性能更强,可通过冠脉循环实现心肌超声显影。这类产品有Molecular Biosystems公司的先灵葆雅公司的Bracco公司的以及ImaRx Pharmaceutical公司的SonusPharmaceutical公司的等产品。
上述商品化的微泡超声对比剂一般采用冷冻干燥等方法将包膜材料制成粉末,再向包装瓶内灌充惰性气体,使气体渗透到包膜材料粉末的空心结构中。使用时向瓶中注入注射用水,进行机械振荡产生微泡。这种制备方法难以准确控制微泡粒径大小,粒径分布非常宽,导致声衰减明显,微泡血液循环时间短,体内有效超声增强时间有待进一步延长。而且,上述微泡超声成像对比剂均为微米尺度(1~10μm),属于血池显影,仅对血管丰富的器官有较好的显影增强效果,无法穿越血管内皮进入组织间隙,增强血管外病灶组织超声显像效果。
近年来,纳米尺度含气微泡—纳米泡作为新型超声成像对比剂的研究引起人们的重视。纳米泡具有纳米尺度的粒径,可以穿越血管内皮进入组织间隙,使血管外靶组织(如肿瘤组织)显像成为可能。中国发明专利CN101683272A公开了一种超声敏感载药纳米泡,但其实质上是采用透析法结合超声共振法制备的载氟碳化合物的聚合物胶束,并不是真正意义上的包膜纳米泡,且没有动物在体试验或细胞试验数据证明其作用效果。中国发明专利CN101954096B公开了一种能够增强超声、CT和MRI成像的含壳核结构纳米粒的脂质乳剂,但其实质是载液体氟碳化合物和纳米磁球的脂质乳剂,不是真正意义上的包膜纳米泡,也不具有靶向性。该脂质乳剂采用传统的薄膜水化-分散乳化-超声制备工艺,难以准确控制粒径,粒径分布宽,制备工艺重现性差。中国发明专利CN100563719C公开了一种包载常规磁共振磁造影剂和氟碳气体的高分子微球,采用超声波声振空化法制备,其粒径为1~5μm,未给出粒径分布数据。其靶向修饰只是简单的物理混合过程,同样也没有动物在体试验或细胞试验数据证明其作用效果。
目前,已公开的超声微泡多采用白蛋白、磷脂、淀粉、纤维素等天然材料制备,或采用聚丙交酯(PLA)、乙交酯-丙交酯共聚物(PLGA)、聚己内酯(PCL)等可生物降解高分子材料制备。前者制备的微泡存在体内抗压性差、声衰减明显、稳定性差等缺点,后者存在亲水性差、体内降解时间长、分子中缺乏活性位点难以靶向修饰等问题。已公开的超声微泡制备方法主要有声振空化法、机械匀化法、薄膜水化法、冷冻干燥法、喷雾干燥法以及乳液聚合法等(戴志飞等,仿生膜材料与技术,科学出版社,2010年)。但这些方法在实际应用中存在许多问题,如:难以得到纳米尺度的微泡,制得的微泡粒径大且粒径均一性差,制备工艺重现性差,制备过程中剧烈的机械作用和/或高温可能导致多肽、蛋白质、抗体等靶向因子失去生物活性等。
同时,随着医学影像学的发展,对影像对比剂提出了更高的要求,能够同时应用于超声、MRI等不同影像学技术,尤其用于肿瘤等重大疾病影像学早期诊断和靶向治疗的诊疗一体化的新型多功能影像学纳米对比剂,将在临床上有良好的应用前景。同时,有必要发展新的纳米对比剂制备技术。
发明内容
为了解决上述技术问题,本发明的目的在于提供一种相转变纳米泡、制备方法及应用。
本发明所制备的纳米泡粒径均一可控,体内外稳定性好,能增强超声成像,可用于影像学诊断的纳米对比剂和造影剂。本发明还提供了该纳米泡的制备方法,其制备条件温和,制备工艺重现性好。
本发明提供的方案如下:
一种相转变靶向纳米泡,由囊心填充物、包膜材料和水组成,为夹层球状结构;
所述囊心填充物为液态全氟戊烷;
所述包膜材料为氨基封端聚乳酸;
所述氨基封端聚乳酸含量为0.1wt%~5.0wt%,液态全氟戊烷含量为1.0%~15.0wt%,余量为超纯水。优选的,所述氨基封端聚乳酸含量为0.1wt%~2wt%,液态全氟戊烷含量为1.0wt%~10.0wt%,余量为水相。
上述纳米泡粒径范围为30~1000nm。优选的,纳米泡粒径范围为200~400nm,多分散指数PDI≤0.35。纳米泡泡心填充物质为全氟戊烷(C5F12),全氟戊烷常温下为液体,沸点29.5℃,在体内发生液-气相转变成为气体。
上述聚乳酸的数均分子量为5000~30000。聚乳酸可通过化学偶联肿瘤特异性靶向因子实现肿瘤靶向,肿瘤特异性靶向因子为叶酸、乳铁蛋白、乳铁蛋白受体单链抗体、转铁蛋白、转铁蛋白受体单链抗体、甲胎蛋白(AFP)受体单抗、RGD肽或各种癌细胞的单克隆抗体等。
本发明的另一目的在于提供上述相转变靶向纳米泡的制备方法,其特征在于,包括以下步骤:
(1)将聚乳酸作为包膜材料溶于二氯甲烷中,成为油1即O1相;液态全氟戊烷作为油2即O2相;在冰水浴中以高剪切混合两相,使O2相均匀分散在O1相中,形成O2/O1初乳;
(2)将O2/O1初乳在冰水浴的磁力搅拌下逐滴加入水相中,得到稳定预复乳O2/O1/W,其中W相是指水相;
(3)将形成的预复乳进行超声处理,直至得到粒径均一的O2/O1/W复乳;
(4)将O2/O1/W复乳在冰水浴的条件下磁力搅拌过夜,固化形成纳米泡;得到的纳米泡溶液进行离心,弃去上清液,沉淀重新分散于生理盐水中,得到载液态全氟戊烷的纳米泡水分散体。
上述步骤(1)中,高剪切的剪切速率5000~20000rpm。
上述步骤(3)中所述超声处理采用间歇式工作的方式,超声处理1~10s,间隔1~10s,重复1~20次,超声功率设定为30~300W。优选的,超声处理2~8s,间隔2~8s,重复5~15次,超声的功率设定50~300W。通过控制超声条件来控制粒径大小。
本发明还提供了上述相转变靶向纳米泡在肿瘤诊断的影像造影剂和抗肿瘤靶向药物中的应用。
纳米泡可负载的MRI对比剂作为影像造影剂,所述MRI对比剂包括超顺磁性纳米Fe3O4、超顺磁性纳米Fe2O3、钆化合物(如Gd-DTPA、Gd-DOTA或Gd-BOPTA等)或锰化合物(Mn-DPDP、卟啉锰等)等。MRI对比剂可占纳米泡水分散体总质量的0.05~3.0wt%,优化用量为0.1~1.5wt%。
纳米泡亦可负载的抗肿瘤药物作为抗肿瘤靶向药物,抗肿瘤药物包括紫杉醇、多西他赛、羟基喜树碱、阿霉素、丝裂霉素、他莫昔芬、5-氟尿嘧啶、甲氨蝶呤、阿糖胞苷、环磷酰胺、或铂类药物(顺铂、卡铂或奥沙利铂)等临床常用的抗肿瘤药物。
除此以外,纳米泡水分散体系中还可添加注射剂常用的添加剂以增加纳米泡的性能,如防腐剂叠氮钠、硫柳汞、苯酚等,添加剂可占纳米泡水分散体系总质量的0.5~2.0wt%。
本发明的有益效果:
(1)包膜材料选用氨基封端聚乳酸,具有生物相容性好,易于与靶向因子偶联等优点,通过选择适宜的分子量,可以调控其力学性能、降解时间等,从而制备力学性能适宜(韧性好、抗压性适中)、稳定性好、和降解时间适宜的纳米泡对比剂;
(2)囊心填充物选用液态全氟戊烷,其在体温下发生液-气相转变,可形成含气纳米泡,并在超声作用下聚集合并为微泡,增强肿瘤病灶的超声成像效果;
(3)与常规超声微泡对比剂比较,纳米泡可以穿越肿瘤血管内皮,并在靶向因子作用下直达特异性靶向肿瘤病灶;
(4)所制备的纳米泡可以实现功能性负载;可以负载常规MRI对比剂,提高微小肿瘤病灶MRI成像的准确性和灵敏性,改善肿瘤影像学早期诊断效果;亦可以负载抗肿瘤药物,用于肿瘤的靶向治疗;
(5)采用乳化溶剂挥发方法可以实现对纳米泡粒径的精确调控,制得的纳米泡粒径高度均一,单分散性好,体内外稳定性好,血液循环时间长,可以实现肿瘤病灶的重复检查、动态监测及疗效评估;
(6)乳化溶剂挥发方法制备条件温和,能够避免在制备中破坏多肽、蛋白质、抗体等靶向因子的生物活性;
(7)通过控制超声的条件,乳化溶剂挥发方法制备工艺重现性好,不同批次纳米泡的粒径、PDI值波动极小,且制备工艺可以按比例放大,易于实现大规模制备。
附图说明
图1是纳米泡的粒径和粒径分布图;
图2是4℃时纳米泡TEM图;
图3纳米泡4℃条件下放置稳定性试验结果;
图4是37℃水浴条件下纳米泡在乳胶手套中的成像图,其中4A为生理盐水,4B为耦合剂,4C为聚乳酸纳米泡;
图5是纳米泡在乳胶手套中击破之前与击破之后的成像图;
图6是乳胶手套模具中不同温度下PLA纳米泡的体外超声图像;
图7是裸鼠皮下肿瘤的超声成像图;7A为注射聚乳酸纳米气泡之前,7B为注射聚乳酸纳米气泡之后。
具体实施方式
下面通过借助实施例更加详细地说明本发明,但以下实施例仅是说明性的,本发明的保护范围并不受这些实施例的限制。
本发明所制备的纳米泡以可生物降解的聚乳酸为包膜材料,以可在体内发生液-气相转变的全氟戊烷为囊心填充物质,采用乳化溶剂挥发方法制备。其中,按质量百分比计,聚乳酸比例为0.1~5.0wt%,液态全氟戊烷为1.0wt%~15.0wt%,余量为超纯水。优选的,聚乳酸比例为0.1wt%~2wt%,液态全氟戊烷为1.0wt%~10.0wt%。所述聚乳酸的分子量为5000~30000。纳米泡粒径范围为30~1000nm;优选的,纳米泡粒径范围为200~400nm,多分散指数(PDI)≤0.35。
纳米泡经注射进入体内后,液态氟碳化合物在体温下发生液-气相转变,形成含气纳米泡,通过靶向因子与肿瘤细胞的特异性结合,纳米泡富集在肿瘤病灶部位,从而改善肿瘤病灶超声成像效果。纳米泡可以负载常规MRI对比剂,提高微小肿瘤病灶MRI成像的准确性和灵敏性,改善肿瘤影像学早期诊断效果。纳米泡还可以负载抗肿瘤药物,用于肿瘤的靶向治疗,作为抗肿瘤药物靶向输送材料,是一类新型的诊疗一体化的多功能影像学纳米对比剂。
纳米泡的基本制备过程如下:
(1)将氨基封端聚乳酸作为包膜材料溶于二氯甲烷溶剂中,成为油相1(O1相),液态全氟戊烷作为油相2(O2相),在冰浴中以高剪切混合两相,使O2相均匀分散在O1相中,形成O2/O1初乳;
(2)将上述两相在冰浴的磁力搅拌下逐渐滴加入水相(W相)中,得到稳定预复乳即O2/O1/W;
(3)将形成的预复乳进行超声处理,直至得到粒径均一的O2/O1/W复乳;
(4)将O2/O1/W复乳在冰浴的条件下磁力搅拌,过夜,固化形成纳米泡;得到的纳米泡溶液进行离心,弃去上清液,沉淀重新分散于生理盐水中,得到载液态全氟戊烷的纳米泡水分散体。
上述步骤(1)中,高剪切的剪切速率5000~20000rpm。
上述步骤(3)中所述超声处理采用间歇式工作的方式,超声处理1~10s,间隔1~10s,重复1~20次,超声功率设定为30~300W。优选的,超声处理2~8s,间隔2~8s,重复5~15次,超声的功率设定50~300W。
实施例1
聚乳酸-叶酸耦联的纳米泡的制备:
(1)制备聚乳酸共聚物
称取重结晶丙交酯适量,加入到50mL单口圆底烧瓶中,加辛酸亚锡、N-叔丁氧羰基乙醇胺适量,抽真空,充氮气两次,在氮气保护的条件下120℃反应3h。反应后生成的固体用二氯甲烷溶解,沉淀于无水乙醇中,沉淀在常温下真空干燥24h。
(2)制备聚乳酸-叶酸耦联的共聚物
将0.3g聚乳酸共聚物溶于10mL CH2Cl2中,叶酸0.088g溶于10mL无水DMSO中。将两者混溶后再加入0.041g DCC及0.1mL三乙胺,室温下搅拌反应24h,过滤、减压浓缩除去大部分溶剂。乙醚沉淀,沉淀物用CH2Cl2溶解,过滤以除去未反应的叶酸,旋蒸得产物。
(3)采用乳化溶剂挥发聚乳酸法制备纳米泡
(i)称取聚乳酸-叶酸耦联共聚物0.1g溶于10g二氯甲烷中配制成1wt%的溶液置于冰水浴中,然后迅速向溶液中加入质量分数为1g全氟戊烷(PFP),在5000rpm条件下,混合溶液高剪切3min,形成O1/O2型初乳;
(ii)将形成的初乳2mL逐滴滴加到12mL 0.3wt%的聚乙烯醇(PVA)水溶液中,在搅拌的作用下形成O1/O2/W型预复乳;
(iii)将形成的预复乳进行超声处理形成纳米乳,超声的条件为工作时间为1s,间隔1s,重复1次,超声的功率设定为300W;
(iv)在冰水浴的条件下,纳米乳搅拌过夜,固化形成纳米泡;得到的纳米泡溶液进行离心,弃去上清液,沉淀重新分散于生理盐水中,放置于4℃冰箱中保存备用。
粒径及微观形貌分析:
采用激光粒度仪(Zetasizer/Nano ZS90,Malvern公司)4℃时测定纳米泡粒径,测得平均粒径为318.4±5.1nm,PDI为0.167±0.020。图1为激光粒度仪测得的纳米泡粒径和粒径分布图。
采用透射电子显微镜(Tecnai G220,荷兰FEI公司)4℃时表征其微观形貌。结果见图2,纳米泡的泡壁边界清晰。
由激光粒度仪测试结果和TEM表征结果可知,纳米泡呈规则的圆泡形结构,单分散性好,粒径均一。
实施例2
制备方法同实施例1,,区别在于:
步骤(3)的(i)中乳酸-叶酸耦联共聚物用量为2g,全氟戊烷(PFP)用量为15g,剪切速率为15000rpm;
步骤(3)的(iii)中超声工作时间2s,间隔2s,重复5次,超声功率为30w。
实施例3
制备方法同实施例1,,区别在于:
步骤(3)的(i)中乳酸-叶酸耦联共聚物用量为3g,全氟戊烷(PFP)用量为20g,剪切速率为20000rmp;
步骤(3)的(iii)中超声工作时间8s,间隔8s,重复15次,超声功率为300w。
实施例4
制备方法同实施例1,,区别在于:
步骤(3)的(i)中乳酸-叶酸耦联共聚物用量为5g,全氟戊烷(PFP)用量为30g,剪切速率为20000rmp;
步骤(3)的(iii)中超声工作时间10s,间隔10s,重复20次,超声功率为50w。
实施例5
一、纳米泡放置稳定性试验
将实施例1制备的纳米泡置于4℃冰箱中保存,每隔一定时间取样采用激光粒度仪测定其粒径,评价纳米泡4℃条件下放置稳定性,试验结果见图3。由试验可知,纳米泡在4℃条件下放置3个月其粒径变化非常小,表明纳米泡在4℃条件下具有良好的稳定性。
二、37℃条件下纳米泡的超声成像试验
将实施例1制备的纳米泡及对照品生理盐水和耦合剂装入乳胶手套,置于37℃恒温水浴中,分别取样采用超声探头(GE-LOGIQ7;探头型号L10-5)的频率是10MHz,机械指数(MI)是0.4,成像模式是普通的B超模式。
结果见图4,在超声波的辐照下,纳米泡在手套中呈现出一个个亮点(如图4C所示),这些均匀密集的亮点形成了超声高信号,能增强超声成像,而对照品生理盐水和耦合剂中,未见到亮点(如图4AB所示)。
三、纳米泡体外击破试验
将实施例1制备的纳米泡置于乳胶手套中,手套放在37℃恒温水浴中,进行纳米泡体外超声成像试验。
仪器型号:Philips-EPIQ7C超声仪,5C探头。造影模式,Frq:5.0MHz。
试验结果:37℃超声条件下,用超声波(较强的机械指数)瞬间将聚乳酸纳米泡进行击破,击破之前和之后分别进行一次成像。如图5所示,通过两次图像的对比来评价纳米泡能否被超声波击破。试验结果表明,在击破之后的整个视野内,纳米泡出现的亮点明显减少,而且,在击破之后成像的图片上,明显发现乳胶手套的底部,有很多较大的亮点出现,其产生的原因是击破之后的纳米泡会释放出全氟戊烷气体,这些气体分散在液体中,会聚集增大,产生较大的气泡,所以成像的图片上亮点增大。
四、纳米泡在不同温度下的体外超声成像试验
将实施例1制备的纳米泡置于23℃水浴中,水浴缓慢升温,分别在23℃、25℃、27℃、29℃、31℃、33℃、35℃、37℃、39℃下进行超声成像,考察纳米泡不同温度条件下的变化趋势,试验结果见图6。由结果可知,随温度升高全氟戊烷发生气化,纳米泡逐渐膨胀,23℃-37℃成像的图片上亮点增大,39℃时亮点又减弱,说明37℃时候,纳米泡气化最完全,最适合超声成像。
五、纳米泡裸鼠在体超声成像试验
将实施例1制备的纳米泡注射至皮下瘤模型裸鼠,进行纳米泡在体超声成像试验。
仪器型号:GE LOGIQ 9型彩色多普勒超声仪。主要测试参数:线阵探头,频率9MHz,MI0.50,采用编码反向谐波成像技术(pulse inversion harmonic imaging,PIHI)。
试验结果:试验动物为皮下瘤模型裸鼠,由图7(7A为注射纳米泡之前,7B为注射纳米泡之后)可知,皮下瘤模型裸鼠尾静脉注射纳米泡造影剂后,肿瘤超声信号明显增强,显示了纳米泡良好的肿瘤超声增强成像作用。
以上所述,仅为本发明较佳的具体实施方式,但本发明保护的范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内所做的任何修改,等同替换和改进等,均应包含在发明的保护范围之内。
Claims (10)
1.一种相转变靶向纳米泡,其特征在于:
由包膜材料、囊心填充物和水相组成,为夹层球状结构;
所述包膜材料为氨基封端聚乳酸溶液,为油相O1相;
所述囊心填充物为液态全氟戊烷,为油相O2相;
由里向外依次为O2相、O1相和水相;
以O1、O2和水相的总质量为100%计,所述氨基封端聚乳酸含量为0.1wt%~5.0wt%,液态全氟戊烷含量为1.0wt%~15.0wt%,余量为水相。
2.根据权利要求1所述的纳米泡,其特征在于:所述氨基封端聚乳酸含量为0.1~2wt%,液态全氟戊烷含量为1.0wt%~10.0wt%,余量为水相。
3.根据权利要求1所述的纳米泡,其特征在于:所述纳米泡粒径范围为30~1000nm,多分散指数PDI≤0.35。
4.根据权利要求1所述的纳米泡,其特征在于:所述聚乳酸的数均分子量为5000~30000。
5.根据权利要求1所述的纳米泡,其特征在于:所述水相为聚乙二醇溶液,浓度为0.3wt%。
6.一种权利要求1~5任一项所述的相转变靶向纳米泡的制备方法,其特征在于,包括以下步骤:
(1)将聚乳酸作为包膜材料溶于二氯甲烷中,成为油1即O1相;液态全氟戊烷作为油2即O2相;在冰水浴中以高剪切混合两相,使O2相均匀分散在O1相中,形成O2/O1初乳;
(2)将O2/O1初乳在冰水浴的磁力搅拌下逐滴加入水相中,得到稳定预复乳O2/O1/W,其中W相是指水相;
(3)将形成的预复乳进行超声处理,直至得到粒径均一的O2/O1/W复乳;
(4)将O2/O1/W复乳在冰水浴的条件下磁力搅拌过夜,固化形成纳米泡;得到的纳米泡溶液进行离心,弃去上清液,沉淀重新分散于生理盐水中,得到载液态全氟戊烷的纳米泡水分散体。
7.根据权利要求6所述的制备方法,其特征在于:所述步骤(1)中,高剪切的剪切速率5000~20000rpm。
8.根据权利要求6所述的制备方法,其特征在于:所述步骤(3)中所述超声处理采用间歇式工作的方式,超声处理1~10s,间隔1~10s,重复1~20次,超声功率设定为30~300W。
9.根据权利要求8所述的制备方法,其特征在于:所述步骤(3)中,超声处理2~8s,间隔2~8s,重复5~15次,超声的功率设定50~300W。
10.权利要求1所述相转变靶向纳米泡在肿瘤诊断的影像造影剂和抗肿瘤靶向药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910342099.6A CN109966514A (zh) | 2019-04-26 | 2019-04-26 | 一种相转变靶向纳米泡、制备方法及应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910342099.6A CN109966514A (zh) | 2019-04-26 | 2019-04-26 | 一种相转变靶向纳米泡、制备方法及应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN109966514A true CN109966514A (zh) | 2019-07-05 |
Family
ID=67086447
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910342099.6A Pending CN109966514A (zh) | 2019-04-26 | 2019-04-26 | 一种相转变靶向纳米泡、制备方法及应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN109966514A (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111632154A (zh) * | 2020-06-01 | 2020-09-08 | 湖北科技学院 | 一种相转变纳米泡、其制备方法及用途 |
CN112979935A (zh) * | 2021-02-26 | 2021-06-18 | 湖北科技学院 | 线粒体靶向类高分子载体材料tpp-pla、荧光素纳米粒及制备方法和应用 |
CN115006555A (zh) * | 2022-05-07 | 2022-09-06 | 湖北科技学院 | 一种纳米级超声/磁共振双模态造影剂和其制备方法及应用 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102836446A (zh) * | 2012-05-21 | 2012-12-26 | 华中科技大学 | 体内相转变肿瘤靶向纳米泡及其制备方法和用途 |
CN106267241A (zh) * | 2015-06-26 | 2017-01-04 | 重庆医科大学 | 一种多功能多模态肿瘤特异性靶向相变型纳米微球光声造影剂及其应用 |
CN106581698A (zh) * | 2016-12-21 | 2017-04-26 | 中国人民解放军总医院 | 用于动脉粥样硬化不稳定斑块识别的超声荧光双模态纳米探针的制备方法 |
CN108114291A (zh) * | 2017-11-29 | 2018-06-05 | 重庆医科大学 | 靶向相变型多模态显像纳米造影剂及其制备方法 |
-
2019
- 2019-04-26 CN CN201910342099.6A patent/CN109966514A/zh active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102836446A (zh) * | 2012-05-21 | 2012-12-26 | 华中科技大学 | 体内相转变肿瘤靶向纳米泡及其制备方法和用途 |
CN106267241A (zh) * | 2015-06-26 | 2017-01-04 | 重庆医科大学 | 一种多功能多模态肿瘤特异性靶向相变型纳米微球光声造影剂及其应用 |
CN106581698A (zh) * | 2016-12-21 | 2017-04-26 | 中国人民解放军总医院 | 用于动脉粥样硬化不稳定斑块识别的超声荧光双模态纳米探针的制备方法 |
CN108114291A (zh) * | 2017-11-29 | 2018-06-05 | 重庆医科大学 | 靶向相变型多模态显像纳米造影剂及其制备方法 |
Non-Patent Citations (2)
Title |
---|
赵玉英 等: "新型多聚体超声造影剂的初步实验研究", 《中国超声医学杂志》 * |
黄宏杰 等: "高分子微泡超声对比剂制备条件的优化", 《中国组织工程研究》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111632154A (zh) * | 2020-06-01 | 2020-09-08 | 湖北科技学院 | 一种相转变纳米泡、其制备方法及用途 |
CN112979935A (zh) * | 2021-02-26 | 2021-06-18 | 湖北科技学院 | 线粒体靶向类高分子载体材料tpp-pla、荧光素纳米粒及制备方法和应用 |
CN112979935B (zh) * | 2021-02-26 | 2022-05-17 | 湖北科技学院 | 线粒体靶向类高分子载体材料tpp-pla、荧光素纳米粒及制备方法和应用 |
CN115006555A (zh) * | 2022-05-07 | 2022-09-06 | 湖北科技学院 | 一种纳米级超声/磁共振双模态造影剂和其制备方法及应用 |
CN115006555B (zh) * | 2022-05-07 | 2024-01-12 | 湖北科技学院 | 一种纳米级超声/磁共振双模态造影剂和其制备方法及应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8486444B2 (en) | Nanobubbles useful as an ultrasonic contrast agent for the lymphatic system | |
Chen et al. | Lipid/PLGA hybrid microbubbles as a versatile platform for noninvasive image-guided targeted drug delivery | |
Brismar et al. | Magnetite nanoparticles can be coupled to microbubbles to support multimodal imaging | |
CN100563718C (zh) | 用于反差成像的充气微囊组件 | |
Xu et al. | Uniform PEGylated PLGA microcapsules with embedded Fe3O4 nanoparticles for US/MR dual-modality imaging | |
US8940277B2 (en) | Intracellular microbubble for imaging an anatomical site | |
JP2001527547A (ja) | 超音波コントラスト剤として、および、血流への薬剤デリバリーのために有用な微小パーティクル | |
Teraphongphom et al. | Nanoparticle loaded polymeric microbubbles as contrast agents for multimodal imaging | |
CN109966514A (zh) | 一种相转变靶向纳米泡、制备方法及应用 | |
CN106267241A (zh) | 一种多功能多模态肿瘤特异性靶向相变型纳米微球光声造影剂及其应用 | |
Ohlerth et al. | Contrast ultrasound: general principles and veterinary clinical applications | |
CN103120799A (zh) | 用于反差成像的充气微泡组合物 | |
Cheng et al. | Ultrasound-triggered phase transition sensitive magnetic fluorescent nanodroplets as a multimodal imaging contrast agent in rat and mouse model | |
CN111632154A (zh) | 一种相转变纳米泡、其制备方法及用途 | |
Methachan et al. | Polymer-based materials in cancer treatment: from therapeutic carrier and ultrasound contrast agent to theranostic applications | |
Kim et al. | Radioprotective garment-inspired biodegradable polymetal nanoparticles for enhanced CT contrast production | |
Xu et al. | Ultrasound molecular imaging of breast cancer in MCF-7 orthotopic mice using gold nanoshelled poly (lactic-co-glycolic acid) nanocapsules: a novel dual-targeted ultrasound contrast agent | |
Barmin et al. | Optoacoustic/fluorescent/acoustic imaging probe based on air-filled bubbles functionalized with gold nanorods and fluorescein isothiocyanate | |
JP6292294B2 (ja) | ナノ粒子の製造方法 | |
Cruje et al. | Polymer assembly encapsulation of lanthanide nanoparticles as contrast agents for in vivo micro-CT | |
CN102836446B (zh) | 体内相转变肿瘤靶向纳米泡及其制备方法和用途 | |
Ding et al. | Lactoferrin-Conjugated Polylactic Acid Nanobubbles Encapsulated Perfluoropentane as a Contrast Agent for Ultrasound/Magnetic Resonance Dual-Modality Imaging | |
CN115006555B (zh) | 一种纳米级超声/磁共振双模态造影剂和其制备方法及应用 | |
Łopuszyńska et al. | Contrasting Properties of polymeric nanocarriers for mri-guided drug delivery | |
Mody et al. | Application of nanoparticles in diagnostic imaging via ultrasonography |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190705 |