CN111729094A - Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof - Google Patents
Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof Download PDFInfo
- Publication number
- CN111729094A CN111729094A CN202010717482.8A CN202010717482A CN111729094A CN 111729094 A CN111729094 A CN 111729094A CN 202010717482 A CN202010717482 A CN 202010717482A CN 111729094 A CN111729094 A CN 111729094A
- Authority
- CN
- China
- Prior art keywords
- contrast agent
- phospholipid
- block copolymer
- amphiphilic block
- 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.)
- Granted
Links
- 239000002961 echo contrast media Substances 0.000 title claims abstract description 158
- 229920000469 amphiphilic block copolymer Polymers 0.000 title claims abstract description 101
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 230000008859 change Effects 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 27
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 25
- YHHSONZFOIEMCP-UHFFFAOYSA-O phosphocholine Chemical compound C[N+](C)(C)CCOP(O)(O)=O YHHSONZFOIEMCP-UHFFFAOYSA-O 0.000 claims abstract description 24
- 229950004354 phosphorylcholine Drugs 0.000 claims abstract description 24
- 238000003745 diagnosis Methods 0.000 claims abstract description 9
- 230000001225 therapeutic effect Effects 0.000 claims abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 42
- 229920001577 copolymer Polymers 0.000 claims description 38
- 229920001223 polyethylene glycol Polymers 0.000 claims description 36
- 239000000126 substance Substances 0.000 claims description 35
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 26
- TXEYQDLBPFQVAA-UHFFFAOYSA-N tetrafluoromethane Chemical compound FC(F)(F)F TXEYQDLBPFQVAA-UHFFFAOYSA-N 0.000 claims description 24
- 239000002202 Polyethylene glycol Substances 0.000 claims description 23
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 22
- 238000002604 ultrasonography Methods 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 17
- 239000007908 nanoemulsion Substances 0.000 claims description 17
- 229960004692 perflenapent Drugs 0.000 claims description 15
- 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 description 15
- 239000000725 suspension Substances 0.000 claims description 15
- 238000003384 imaging method Methods 0.000 claims description 14
- 230000004044 response Effects 0.000 claims description 14
- 238000001338 self-assembly Methods 0.000 claims description 13
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 13
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 11
- 238000001727 in vivo Methods 0.000 claims description 11
- 239000012046 mixed solvent Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 230000009471 action Effects 0.000 claims description 9
- 238000011282 treatment Methods 0.000 claims description 9
- ZSZRUEAFVQITHH-UHFFFAOYSA-N 2-(2-methylprop-2-enoyloxy)ethyl 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CC(=C)C(=O)OCCOP([O-])(=O)OCC[N+](C)(C)C ZSZRUEAFVQITHH-UHFFFAOYSA-N 0.000 claims description 8
- 238000000265 homogenisation Methods 0.000 claims description 8
- 238000000338 in vitro Methods 0.000 claims description 8
- 230000010355 oscillation Effects 0.000 claims description 7
- PAPBSGBWRJIAAV-UHFFFAOYSA-N ε-Caprolactone Chemical compound O=C1CCCCCO1 PAPBSGBWRJIAAV-UHFFFAOYSA-N 0.000 claims description 6
- 238000010560 atom transfer radical polymerization reaction Methods 0.000 claims description 5
- 239000006185 dispersion Substances 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- 239000003054 catalyst Substances 0.000 claims description 4
- 210000000987 immune system Anatomy 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 4
- 229920000936 Agarose Polymers 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- QYSGYZVSCZSLHT-UHFFFAOYSA-N octafluoropropane Chemical compound FC(F)(F)C(F)(F)C(F)(F)F QYSGYZVSCZSLHT-UHFFFAOYSA-N 0.000 claims description 3
- KAVGMUDTWQVPDF-UHFFFAOYSA-N perflubutane Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)F KAVGMUDTWQVPDF-UHFFFAOYSA-N 0.000 claims description 3
- 229950003332 perflubutane Drugs 0.000 claims description 3
- 229960004065 perflutren Drugs 0.000 claims description 3
- 230000000379 polymerizing effect Effects 0.000 claims description 3
- 230000027756 respiratory electron transport chain Effects 0.000 claims description 3
- 230000007704 transition Effects 0.000 claims description 3
- 238000012661 block copolymerization Methods 0.000 claims description 2
- 229960004624 perflexane Drugs 0.000 claims description 2
- LGUZHRODIJCVOC-UHFFFAOYSA-N perfluoroheptane Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)F LGUZHRODIJCVOC-UHFFFAOYSA-N 0.000 claims description 2
- ZJIJAJXFLBMLCK-UHFFFAOYSA-N perfluorohexane Chemical compound FC(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)F ZJIJAJXFLBMLCK-UHFFFAOYSA-N 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 239000004698 Polyethylene Substances 0.000 claims 1
- 229920013747 hydroxypolyethylene Polymers 0.000 claims 1
- 229920000573 polyethylene Polymers 0.000 claims 1
- 239000002245 particle Substances 0.000 abstract description 9
- 238000009826 distribution Methods 0.000 abstract description 6
- 230000007547 defect Effects 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 2
- 239000002872 contrast media Substances 0.000 description 32
- 229920000642 polymer Polymers 0.000 description 19
- SFZCNBIFKDRMGX-UHFFFAOYSA-N sulfur hexafluoride Chemical compound FS(F)(F)(F)(F)F SFZCNBIFKDRMGX-UHFFFAOYSA-N 0.000 description 17
- 229960000909 sulfur hexafluoride Drugs 0.000 description 17
- 230000000694 effects Effects 0.000 description 9
- 239000007789 gas Substances 0.000 description 5
- 210000004185 liver Anatomy 0.000 description 5
- 229920001427 mPEG Polymers 0.000 description 5
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 229920002946 poly[2-(methacryloxy)ethyl phosphorylcholine] polymer Polymers 0.000 description 3
- 238000006116 polymerization reaction Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- YOCIJWAHRAJQFT-UHFFFAOYSA-N 2-bromo-2-methylpropanoyl bromide Chemical compound CC(C)(Br)C(Br)=O YOCIJWAHRAJQFT-UHFFFAOYSA-N 0.000 description 2
- KCBWAFJCKVKYHO-UHFFFAOYSA-N 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-[[4-[1-propan-2-yl-4-(trifluoromethyl)imidazol-2-yl]phenyl]methyl]pyrazolo[3,4-d]pyrimidine Chemical compound C1(CC1)C1=NC=NC(=C1C1=NC=C2C(=N1)N(N=C2)CC1=CC=C(C=C1)C=1N(C=C(N=1)C(F)(F)F)C(C)C)OC KCBWAFJCKVKYHO-UHFFFAOYSA-N 0.000 description 2
- 101710141544 Allatotropin-related peptide Proteins 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 2
- 238000005422 blasting Methods 0.000 description 2
- 229920001400 block copolymer Polymers 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000003759 clinical diagnosis Methods 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- NBVXSUQYWXRMNV-UHFFFAOYSA-N fluoromethane Chemical compound FC NBVXSUQYWXRMNV-UHFFFAOYSA-N 0.000 description 2
- 229920001519 homopolymer Polymers 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N methylene chloride Substances ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 125000001095 phosphatidyl group Chemical group 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- SRLOHQKOADWDBV-NRONOFSHSA-M sodium;[(2r)-2,3-di(octadecanoyloxy)propyl] 2-(2-methoxyethoxycarbonylamino)ethyl phosphate Chemical compound [Na+].CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCCNC(=O)OCCOC)OC(=O)CCCCCCCCCCCCCCCCC SRLOHQKOADWDBV-NRONOFSHSA-M 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229940126585 therapeutic drug Drugs 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 229910021589 Copper(I) bromide Inorganic materials 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 229910018503 SF6 Inorganic materials 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000002583 angiography Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 229940039231 contrast media Drugs 0.000 description 1
- 239000011258 core-shell material Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- LQZZUXJYWNFBMV-UHFFFAOYSA-N dodecan-1-ol Chemical compound CCCCCCCCCCCCO LQZZUXJYWNFBMV-UHFFFAOYSA-N 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000002592 echocardiography Methods 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 238000010874 in vitro model Methods 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 230000002794 monomerizing effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 238000002601 radiography Methods 0.000 description 1
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- -1 small-molecule phospholipid Chemical class 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 239000005720 sucrose Substances 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
- 230000001960 triggered effect Effects 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000001132 ultrasonic dispersion Methods 0.000 description 1
- 238000012285 ultrasound imaging Methods 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
- A61K49/221—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
- 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
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F230/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and containing phosphorus, selenium, tellurium or a metal
- C08F230/02—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and containing phosphorus, selenium, tellurium or a metal containing phosphorus
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F293/00—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule
- C08F293/005—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N29/00—Investigating or analysing materials by the use of ultrasonic, sonic or infrasonic waves; Visualisation of the interior of objects by transmitting ultrasonic or sonic waves through the object
- G01N29/02—Analysing fluids
- G01N29/032—Analysing fluids by measuring attenuation of acoustic waves
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N29/00—Investigating or analysing materials by the use of ultrasonic, sonic or infrasonic waves; Visualisation of the interior of objects by transmitting ultrasonic or sonic waves through the object
- G01N29/04—Analysing solids
- G01N29/06—Visualisation of the interior, e.g. acoustic microscopy
- G01N29/0654—Imaging
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F2438/00—Living radical polymerisation
- C08F2438/01—Atom Transfer Radical Polymerization [ATRP] or reverse ATRP
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Acoustics & Sound (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Pathology (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Radiology & Medical Imaging (AREA)
- Inorganic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention belongs to the field of ultrasonic image diagnosis, and particularly relates to a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and a preparation method thereof. The invention provides a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, which comprises a shell and an inner core, wherein the shell is an amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine, and the inner core is an ultrasonic responder. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent prepared by the method has uniform particle size distribution, has stability obviously superior to that of the phospholipid-based ultrasonic contrast agent, and is more suitable for ultrasonic diagnosis and therapeutic research; and the defects of pressure resistance and mechanical index change resistance of the conventional ultrasonic contrast agent can be overcome, and the application range of the ultrasonic contrast agent is expanded.
Description
Technical Field
The invention belongs to the field of ultrasonic image diagnosis, and particularly relates to a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and a preparation method thereof.
Background
The ultrasonic contrast agent can enhance the contrast effect of a super image, remarkably improve the ultrasonic diagnosis precision, be widely applied to the field of clinical diagnosis and have great application potential in ultrasonic-mediated therapy. However, most of the currently applied ultrasound contrast agents in clinical use are composed of small-molecule phospholipid or albumin encapsulated inert gas, which has the disadvantages of large polydispersity or short half-life (<10min), and so on, and limits further application in imaging and therapy.
To enhance the stability of ultrasound contrast agents, materials with higher stiffness than phospholipids have been developed to stabilize the gaseous core in ultrasound contrast agents, which are referred to as hard shell contrast agents. The hard shell contrast agent exhibits little volume expansion and remains intact under low sound pressure conditions. But above a certain pressure threshold, the shell of the hard shell ultrasound contrast agent will also rupture. The polymer has higher rigidity than phospholipid, and the polymer-based ultrasonic contrast agent prepared based on the polymer shell can greatly improve the acoustic behavior of the ultrasonic contrast agent. Furthermore, by adjusting the chemical composition and relative molecular weight of the polymer, the acoustic properties of polymer-based ultrasound contrast agents can also be controlled. The polymer shell ultrasound contrast agent not only has better acoustic stability, but also has greatly improved pressure resistance and mechanical index change resistance under ultrasound. In addition, the grafted or encapsulated therapeutic drug for drug delivery can also be used for ultrasound image-guided diagnosis and treatment integrated preparation, so that the versatility of the multi-modal ultrasound contrast agent is increased, and the application range of the ultrasound contrast agent is further expanded.
Disclosure of Invention
The invention aims to provide a novel polymer-based ultrasonic contrast agent and a preparation method thereof, wherein the obtained contrast agent is a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, so that the defects of insufficient pressure resistance and mechanical index change resistance of the conventional ultrasonic contrast agent are solved, and the application range of the ultrasonic contrast agent is expanded.
The technical scheme of the invention is as follows:
the first technical problem to be solved by the invention is to provide a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, which comprises an outer shell and an inner core, wherein the outer shell is an amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine (PCL-b-PMPC), and the inner core is an ultrasonic responder.
Further, the amphiphilic block copolymer is a copolymer obtained by block copolymerization of Caprolactone (CL) and 2-Methacryloyloxyethyl Phosphorylcholine (MPC). Further, the arrangement of the hydrophilic segment/hydrophobic segment (A/B) in the amphiphilic block copolymer is AB type.
Further, the shell in the ultrasound contrast agent further comprises a modifying substance M, wherein the modifying substance M is: the shell is provided with a substance containing PEG chain segments which can avoid being cleared by the immune system in vivo, so as to increase the circulation time of the ultrasonic contrast agent.
Further, the modifying substance M is selected from: dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 5000(DPPE-mPEG 5000), dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 2000(DPPE-mPEG2000), distearoylphosphatidylethanolamine-polyethylene glycol 2000(DSPE-MPEG2000), distearoylphosphatidylethanolamine-azidopolyethylene glycol 5000(DSPE-PEG5000-N3) Distearoyl phosphatidyl ethanolamine-azido polyethylene glycol 2000(DSPE-PEG 2000-N)3) Distearoyl phosphatidyl ethanolamine-polyethylene glycol-sulfhydryl cross-linked substance (DSPE-PEG5000-SH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-sulfhydryl cross-linked substance (DSPE-PEG2000-SH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-amino cross-linked substance (DSPE-PEG 5000-NH)2) Distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-amino cross-linked complex (DSPE-PEG 2000-NH)2) At least one of distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-carboxyl cross-linked complex (DSPE-PEG5000-COOH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-carboxyl cross-linked complex (DSPE-PEG2000-COOH) or distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-hydroxyl cross-linked complex (DSPE-PEG 5000-OH).
Preferably, the core of the ultrasound contrast agent is a liquid perfluorocarbon or a gaseous perfluorocarbon that is phase-changeable.
Further, the inner core of the ultrasound contrast agent comprises at least one of perfluoropropane, perfluorobutane, perfluoropentane, perfluorohexane, or perfluoroheptane.
Further, when the core of the ultrasonic contrast agent is liquid perfluorocarbon capable of changing phase, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is prepared into a nano-emulsion by polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic response substance in a self-assembly mode, and the obtained nano-emulsion is subjected to phase change to form the phospholipid-like amphiphilic block copolymer coated gaseous perfluorocarbon ultrasonic contrast agent.
Further, when the inner core of the ultrasonic contrast agent is gaseous perfluorocarbon, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is prepared from polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic response substance in a self-assembly mode.
Further, the self-assembly mode is one of the following modes: high shear homogenization, high pressure homogenization, high speed oscillation or ultrasonic sound vibration.
Further, the average particle size distribution range of the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is 5 +/-0.13 mu m.
Further, after the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is diluted by physiological saline, the shape is spherical, the dispersion is good, and the surface is smooth and bright when the ultrasonic contrast agent is observed under a laser confocal microscope.
The second technical problem to be solved by the present invention is to provide a preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent, wherein the preparation method comprises: the ultrasonic contrast agent of the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine coated ultrasonic responder is prepared by self-assembling the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and the ultrasonic responder.
Further, the self-assembly is performed in one of the following ways: high shear homogenization, high pressure homogenization, high speed oscillation or ultrasonic sound vibration.
Further, the ultrasonic responder is liquid perfluorocarbon or gaseous perfluorocarbon which can change phase; preferably a liquid perfluorocarbon. The liquid phase-changeable perfluorocarbon is selected, has better stability than gas, has longer storage time, and can be selected to be subjected to the phase change step before use; the direct use of gaseous preparations has a low success rate and is not conducive to storage.
Further, when the ultrasound response substance in the ultrasound contrast agent is gaseous perfluorocarbon, the preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent comprises the following steps: the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is prepared by directly performing ultrasonic vibration or high-speed oscillation on polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic responder.
Further, when the ultrasound response substance in the ultrasound contrast agent is liquid perfluorocarbon capable of phase transition, the preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent comprises the following steps: the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and the ultrasonic response substance are prepared into the nano-emulsion in a self-assembly mode, and the nano-emulsion is subjected to temperature-induced phase change or sound-induced phase change to form the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent. When the shell layer uses phospholipid-like amphiphilic block copolymer, high-temperature hydrophobic liquid perfluorocarbon which can change phase is wrapped in the shell layer, nano-emulsion, also called nano liquid particles, is prepared in a self-assembly mode by utilizing a similar compatibility principle and hydrophilic-hydrophobic interaction, and then the ultrasonic contrast agent of phospholipid-like amphiphilic copolymer wrapped with gaseous perfluorocarbon which can be subjected to ultrasonic contrast is formed through phase change.
Further, when the ultrasound response substance in the ultrasound contrast agent is liquid perfluorocarbon capable of phase change, the preparation method comprises the following steps:
(1) preparing a phospholipid-like amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine (PCL-b-PMPC);
(2) preparing a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent: dissolving the phospholipid-like amphiphilic block copolymer PCL-b-PMPC obtained in the step (1) and the modified substance M in a mixed solvent, mixing with an ultrasonic corresponding substance, preparing a nano-emulsion in a self-assembly mode, separating and purifying, and performing temperature-induced phase change or sound-induced phase change to prepare the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent; wherein the modifying substance M is: providing the shell with a material comprising a PEG segment that avoids clearance by the immune system in vivo; the mixed solvent is a mixed solvent of tetrahydrofuran and methanol, or: a mixed solvent of chloroform and methanol.
Further, in the step (2), the method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent after the nanoemulsion is separated and purified and undergoes temperature-induced phase change or sound-induced phase change comprises one of the following modes:
the first method is as follows: preparing the phospholipid-like amphiphilic block copolymer-based nano-emulsion suspension by using a water bath at 60-80 ℃ (preferably 70 ℃) for 5-15 min (preferably 10min) to obtain a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent;
the second method comprises the following steps: subjecting the prepared phospholipid-like amphiphilic block copolymer-based nanoemulsion suspension to ultrasonic action by an ultrasonic therapeutic apparatus to obtain the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, wherein the ultrasonic power is 1-3W/cm2(preferably 3W/cm)2) The duty ratio is 20-80% (preferably 50%), and the action time is 2-5 min (preferably 3 min).
Preferably, in the step (2), the mixed solvent is a mixture of solvents with a volume ratio of 2:1 of tetrahydrofuran and methanol.
Further, in the step (2) of the above method, the modifying substance M is selected from: dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 5000(DPPE-mPEG 5000), dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 2000(DPPE-mPEG2000), distearoylphosphatidylethanolamine-polyethylene glycol 2000(DSPE-MPEG2000), distearoylphosphatidylethanolamine-azidopolyethylene glycol 5000(DSPE-PEG 5000-N)3) Distearoyl phosphatidyl ethanolamine-azido polyethylene glycol 2000(DSPE-PEG 2000-N)3) Distearoyl phosphatidyl ethanolamine-polyethylene glycol-sulfhydryl cross-linked substance (DSPE-PEG5000-SH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-sulfhydryl cross-linked substance (DSPE-PEG2000-SH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-amino cross-linked substance (DSPE-PEG 5000-NH)2) Distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-amino cross-linked complex (DSPE-PEG 2000-NH)2) At least one of distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-carboxyl cross-linked complex (DSPE-PEG5000-COOH), distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-carboxyl cross-linked complex (DSPE-PEG2000-COOH) or distearoyl phosphatidyl ethanolamine-polyethylene glycol 5000-hydroxyl cross-linked complex (DSPE-PEG 5000-OH).
Further, the method for preparing the phospholipid-like amphiphilic block copolymer PCL-b-PMPC in the step (1) comprises the following steps: polymerizing Caprolactone (CL) and 2-Methacryloyloxyethyl Phosphorylcholine (MPC) by an atom transfer radical polymerization (ARGET ATRP) method of an electron transfer regeneration catalyst to obtain a phospholipid-like amphiphilic block copolymer PCL-b-PMPC.
Furthermore, the method for preparing the phospholipid-like amphiphilic block copolymer PCL-b-PMPC in the step (1) comprises the following steps:
1) preparing a macroinitiator PCL-Br:
taking-CL as a monomer and stannous octoate as a catalyst, carrying out ring-opening polymerization to obtain a homopolymer PCL, and then reacting the PCL with bromoisobutyryl bromide to obtain PCL-Br;
2) preparation of phospholipid-like amphiphilic block copolymer PCL-b-PMPC:
using PCL-Br obtained in 1) as a macroinitiator and MPC as a monomer, and polymerizing by an atom transfer radical polymerization (ARGET ATRP) method of an electron transfer regeneration catalyst to obtain a phospholipid-like amphiphilic block copolymer PCL-b-PMPC.
The third technical problem to be solved by the invention is to provide the application of the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, which can be applied to the field of ultrasonic image diagnosis and treatment and is used for in-vitro agarose model contrast imaging; or the medicine is used for in-vivo ultrasonic contrast imaging and treatment after being loaded with the medicine.
Compared with the prior art, the invention has the following beneficial technical effects:
(1) the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent prepared by the method has uniform particle size distribution, has stability obviously superior to that of a phospholipid-based ultrasonic contrast agent (a commercial Sonowei SonoVue contrast agent), and is more suitable for ultrasonic diagnosis and therapeutic research.
(2) The phase-change phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent prepared by the invention has a remarkable image enhancement effect in a high mechanical index mode of ultrasonic contrast, has a longer duration time than that of a phospholipid shell contrast agent, improves the short duration time of the phospholipid-based ultrasonic contrast agent in the high mechanical index contrast, and overcomes the defects that the shells of ultrasonic contrast agents prepared by partial high polymer materials are hard and difficult to develop.
(3) The phase-change type ultrasonic contrast agent prepared by coating liquid fluorocarbon with phospholipid-like amphiphilic block copolymer PCL-b-PMPC (namely PCL-b-PMPC is a shell and liquid fluorocarbon is a core) is explored to be feasible as the ultrasonic contrast agent; in vitro and in vivo experiments show that the PCL-b-PMPC-based ultrasonic contrast agent has good echo characteristics under various ultrasonic parameter conditions. More importantly, the imaging time of the PCL-b-PMPC-based ultrasound contrast agent is longer than that of the phospholipid-based ultrasound contrast agent under the same ultrasound parameters and concentrations, and the PCL-b-PMPC-based ultrasound contrast agent has great potential as a novel contrast agent in ultrasound imaging.
(4) The amphiphilic block copolymer in the phase-change amphiphilic phospholipid block copolymer-based ultrasonic contrast agent has the advantages of controllable structure, safety, no toxicity, obviously lower cost than synthetic phospholipid, and simple and convenient preparation process. In addition, the preparation method can be widely applied in the field of clinical ultrasonic imaging, can also be used as a carrier of various therapeutic drugs, and has a good clinical application prospect.
Description of the drawings:
FIG. 1 is a synthesis scheme of the phospholipid-like amphiphilic block copolymer PCL-b-PMPC and a schematic diagram of preparation of an ultrasonic contrast agent.
FIG. 2 shows the polymer PCL obtained in example 143、PCL43-Br and PCL43-b-PMPC25Nuclear magnetic spectrum of (1).
FIG. 3 shows the polymer PCL obtained in example 143、PCL43-Br and PCL43-b-PMPC25An infrared spectrum of (1).
FIG. 4 shows the polymer PCL obtained in example 143And PCL43-b-PMPC25DSC (a) temperature rise curve and (b) temperature fall curve of (a).
FIG. 5 shows the phospholipid-like amphiphilic block copolymer PCL obtained in example 143-b-PMPC25Ultrasound contrast agent based (a) CLSM pictures, (b) size distribution maps, and (c) size and zeta data.
FIG. 6 shows the phospholipid-like amphiphilic block copolymer PCL obtained in example 143-b-PMPC25The contrast agent is based on a contrast map of different ultrasound parameters in vitro and corresponding gray values-frequency (a, d), gray values-mechanical index MI (b, e) and gray values-concentration (c, f).
FIG. 7 shows the phospholipid-like amphiphilic block copolymer PCL obtained in example 143-b-PMPC25Variation of the in vitro contrast effect of the base ultrasound contrast agent over time ultrasound contrast map (a) and corresponding grey values (b).
FIG. 8a shows the phospholipid-like amphiphilic block copolymer PCL obtained in example 143-b-PMPC25The results of the in vivo contrast effect of animals based on ultrasound contrast agent, PBS and SonoVue contrast agent as a function of time, wherein the circles represent the liver imaging area of rabbits; FIG. 8b shows the phospholipid-like amphiphilic block copolymer PCL obtained in example 143-b-PMPC25Radical ultrasoundGrey values corresponding to contrast agent and SonoVue contrast agent.
Detailed Description
The first technical problem to be solved by the invention is to provide a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, which comprises an outer shell and an inner core, wherein the outer shell is an amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine (PCL-b-PMPC), and the inner core is an ultrasonic responder.
The second technical problem to be solved by the present invention is to provide a preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent, wherein the preparation method comprises: the ultrasonic contrast agent of the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine coated ultrasonic responder is prepared by self-assembling the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and the ultrasonic responder.
The third technical problem to be solved by the invention is to provide the application of the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, which can be applied to the field of ultrasonic image diagnosis and treatment and is used for in-vitro agarose model contrast imaging; or the medicine is used for in-vivo ultrasonic contrast imaging and treatment after being loaded with the medicine.
The following examples are given to further illustrate the embodiments of the present invention and are not intended to limit the scope of the present invention.
Example 1
1. The preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent comprises the following specific steps:
1) preparation of amphiphilic Block copolymer Polymer PCL43-b-PMPC25:
① PCL is first prepared by mixing lauryl alcohol (0.82g,0.004mol), -CL (10.13g,0.089mol) and Sn (Otc)2(4.05g,0.01mol) was added to the reaction vessel. Removing O therefrom2And residual water. The reaction was carried out at 120 ℃ for 24 hours under vacuum. The crude product was dissolved in CH2Cl2And purified by cold methanol precipitation to obtain PCL.
② PCL-Br was prepared next by mixing ① PCL-OH (7.00g,0.002mol) and triethylamine (0).96mL,0.007mmol) was dissolved in Tetrahydrofuran (THF); in N22-bromoisobutyryl bromide (0.85mL,0.0069mmol) was added dropwise to the above solution at-20 deg.C; and (3) carrying out reaction at room temperature for 48-72 hours, passing through a neutral alumina column to remove quaternary ammonium salt, and precipitating and evaporating a small amount of THF-containing crude product in cold methanol to obtain white powdery polymer PCL-Br.
③ Final preparation of PCL43-b-PMPC25PCL-Br (1.99g,0.596mmol) obtained in ②, MPC (6.15g,0.021mol) and Bpy (0.28g,1.79mmol) were dissolved in CH2Cl2Performing the following steps; general formula (N)2After 30 minutes, at N2CuBr (0.13g,0.894mmol) was added to the atmosphere; after 24 hours of reaction, purifying by a neutral alumina column; and further purified by dialysis with ultrapure water. The product PCL is obtained by freeze-drying43-b-PMPC25。
2) Preparing a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent: 3mg of PCL was weighed separately43-b-PMPC25Dissolving the block copolymer and 1mg of DPPE-mPEG 5000 in 567. mu.L of Tetrahydrofuran (THF) and 283. mu.L of methanol (MeOH) (2:1, v/v), dissolving by ultrasonic dispersion in a water bath, adding 2ml of PBS buffer (0.01M, pH 7.4, pure water or physiological saline is acceptable), and then adding 150. mu.L of perfluoropentane (PFP); ultrasonically emulsifying by using a probe under an ice-water bath environment, wherein ultrasonic parameters are as follows: the frequency is 24KHz, the power is 35W, the ultrasonic is switched on for 3s and then switched off for 6s, the diameter of a sound vibration probe is 3mm, the treatment time is 3min, a milky white suspension is formed, then the suspension is centrifuged for 5min under the centrifugal force of 3000g, the supernatant is discarded, and 6mL of PBS is added for heavy suspension. And (3) performing water bath action on the obtained copolymer-based nano-emulsion suspension for 10min at 70 ℃ to prepare the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent.
2. Characterization of phospholipid-like amphiphilic Block copolymers
FIG. 2 shows the synthesis of PCL43-b-PMPC25Nuclear magnetic spectra of various polymers in the copolymer process show that characteristic peaks (4.1 ppm;. 2.3 ppm;. 1.7 ppm;. 1.4ppm) of segments of PCL appear on the nuclear magnetic spectra, which indicates that PCL is successfully synthesized, and the polymerization degree of PCL is calculated by the area ratio of the peaks. By calculation, the inventionHomopolymers with the polymerization degrees of 43 are obtained; the appearance of a characteristic peak indicates that the PMPC chain segment is successfully grafted to the PCL chain segment; meanwhile, the degree of polymerization of the PMPC segment is determined by the area ratio of the peaks.
The structure of the polymer was further characterized by infrared, with the results shown in figure 3. 1728cm-1The peak at (A) is a stretching vibration peak of C ═ O, 2910cm-1Are continuous methylene absorption peaks; it can be seen that PCL-b-PMPC shows a characteristic absorption peak (1090 cm) related to the structure of the phosphatidylcholine in an infrared spectrum-1And 1230cm-1,-POCH2-;970cm-1,N+(CH3)3) Successful synthesis of PCL-b-PMPC copolymers was also demonstrated.
The thermal properties of the synthesized polymer were measured by DSC, and the results are shown in fig. 4. The PCL homopolymer has data such as highest crystallization temperature, melting temperature and crystallinity; compared with PCL, the PCL-b-PMPC copolymer is introduced with an amorphous PMPC chain segment, and when the PCL is crystallized, the regular arrangement of the chain segment is hindered, so that various data are reduced.
3. Characterization of microvesicles
The PCL-b-PMPC copolymer solution added with PBS and PFP becomes opaque milky white suspension after the sound vibration treatment; after the organic solvent is removed, the phase change of the PFP is triggered at the temperature, and the PFP is still a milky white suspension solution; the particle size change of the copolymer-based ultrasound contrast agent after phase transition is shown in fig. 5; as can be seen from FIG. 5, the particle size distribution of the PCL-b-PMPC copolymer and the ultrasound contrast agent is narrow, and the average particle size of the PCL-b-PMPC copolymer-based ultrasound contrast agent measured by a Malvern nanometer particle size analyzer is about 5 μm.
3. Stability test
The concentrations of the Sudong copolymer-based ultrasonic contrast agent and a reference phospholipid-based ultrasonic contrast agent (a commercial Sonowei SonoVue contrast agent)) measured by Coulter are diluted to ensure that the copolymer-based ultrasonic contrast agent and the phospholipid-based ultrasonic contrast agent have the same concentration, and the copolymer-based ultrasonic contrast agent and the phospholipid-based ultrasonic contrast agent are placed in a refrigerator at 4 ℃ and are kept still for 24 hours and then observed under a sampling microscope, so that the concentration of the phospholipid-based ultrasonic contrast agent is lower than that of the copolymer-based ultrasonic contrast agent, and the stability of the copolymer-based ultrasonic contrast agent is obviously higher than that of the phospholipid-based ultrasonic contrast agent.
4. In vitro radiography and blasting experiment
Adding 0.3mL of the copolymer-based ultrasonic contrast agent prepared in the step 2) into an agarose gel model, observing in a Cadence Contrast Agent Imaging (CCAI) mode (MI ═ 0.21) by using a Siemens color imaging system (SIEMENS Acuson Antares), wherein the echo in the model is obviously enhanced and is in a dense fine-dot-shaped echo, and observing the change of the echo intensity from the ultrasonic frequency of 4.0MHz to 10.0MHz, wherein the optimal developing frequency of the contrast agent is 5.71MHz (shown in figure 6); the contrast agent is broken by starting a blasting (MI is 0.67) mode, the echo in the model is instantaneously reduced, and the number of the fine-point echoes is sharply reduced.
5. New Zealand white rabbit liver angiography
The method comprises the steps of fixing a male New Zealand white rabbit with the body weight of 2.5-3 kg on an experiment table, placing an indwelling needle into an ear edge vein, after the lumbar and the back of the right side are unhaired, respectively injecting a commercial Sonowev SonoVue contrast agent (a contrast sample) and the PCL-b-PMPC copolymer contrast agent obtained by the invention by adopting an ear edge vein bolus injection method, so as to obtain an ultrasonic contrast image of the liver. In the control experiment, only PBS buffered saline was injected, with dark areas in the ultrasound image, and the results are shown in FIG. 8. In contrast to the PBS control group, significant contrast enhancement began to occur in the liver region following intravenous injection of SonoVue and PCL-b-PMPC copolymer contrast agents, respectively. Bright ultrasound images of the liver indicate that PCL-b-PMPC copolymer contrast agent successfully passes through the pulmonary capillaries during blood circulation, which is necessary for the safety properties of intravenous contrast agents. As can be seen from FIG. 8, the ultrasound signal intensity of the SonoVue contrast agent is substantially comparable to the contrast image intensity of the PCL-b-PMPC copolymer contrast agent at 20 s; however, at 40s, the ultrasound signal of the SonoVue microbubbles decreased rapidly, and at 80s, the contrast agent substantially disappeared, while the PCL-b-PMPC copolymer contrast agent still had a contrast signal that was observable after 80 s. This shows that the PCL-b-PMPC phospholipid block polymer nano ultrasound contrast agent has longer in vivo duration compared with a SouoVue contrast agent, and proves that the PCL-b-PMPC phospholipid block polymer nano ultrasound contrast agent has great potential as an ultrasound contrast agent with novel diagnostic and therapeutic effects.
Example 2
A method different from that of example 1 was used to prepare a PCL-b-PMPC contrast medium by a shear method.
The specific preparation process of the PCL-b-PMPC phospholipid block polymer nano ultrasonic contrast agent prepared by the shearing method comprises the following steps:
3mg of PCL-b-PMPC copolymer and 1mg of DPPE-mPEG 5000 were each dissolved in 567. mu.L of Tetrahydrofuran (THF) and 283. mu.L of methanol (MeOH) (2:1, v/v), and after sufficient dispersion in a water bath, 2mL of PBS buffer (0.01M, pH 7.4) was added, followed by 150. mu.L of perfluoropentane (PFP). Under the condition of ice-water bath, adding liquid perfluoropentane, adopting an electric internally tangent homogenate method, wherein the homogenate rotating speed is 12000-30000rpm, and the homogenate time is 1-3min, and obtaining milky block polymer mixed solution wrapping the perfluoropentane after homogenate.
The mixed solution is subjected to phase change by a therapeutic ultrasonic instrument to form the PCL-b-PMPC phospholipid amphiphilic block copolymer-based ultrasonic contrast agent, and the parameter of the ultrasonic instrument is 3W/cm2The duty ratio is 50%, and the action time is 3 min.
EXAMPLE 3 Freeze-drying preparation of PCL-b-PMPC contrast media
The preparation process of the PCL-b-PMPC phospholipid amphiphilic block copolymer-based ultrasonic contrast agent by the freeze-drying method comprises the following steps:
3mg of PCL-b-PMPC copolymer and 1mg of DPPE-mPEG 5000 were each dissolved in 567. mu.L of Tetrahydrofuran (THF) and 283. mu.L of methanol (MeOH) (2:1, v/v), and after sufficient dispersion in a water bath, 2mL of PBS buffer (0.01M, pH 7.4) was added, followed by 150. mu.L of perfluoropentane (PFP). Performing ultrasonic vibration under ice salt water bath condition, wherein the probe frequency is 24KHz, the power is 35W, the ultrasonic is turned on for 3s and turned off for 6s, the action is performed for 3min, a milky white suspension is formed, then the suspension is centrifuged for 5min under the action of 3000g centrifugal force, the supernatant is discarded, and 6mL of sucrose solution containing 10 wt% is added respectively for heavy suspension. Subpackaging penicillin bottles with the specification of 10mL according to the volume of 2mL per bottle, placing the bottles in a refrigerator with the temperature of-20 ℃ for pre-freezing overnight, and freeze-drying the bottles for 24 hours by a freeze dryer, wherein the temperature of a cold trap is set to be-80 ℃.
Filling the prepared freeze-dried powder with gas such as perfluoropropane, perfluorobutane and sulfur hexafluoride through a ventilation device, recombining with 2mL of physiological saline after ventilation to obtain milky copolymer suspension, and mechanically oscillating the recombined microbubble suspension for 1min at the oscillation frequency of 75Hz to obtain the PCL-b-PMPC phospholipid amphiphilic block copolymer-based ultrasound contrast agent.
According to the three examples, the PCL-b-PMPC phospholipid amphiphilic block copolymer-based ultrasonic contrast agent prepared by different preparation methods is spherical in shape, good in dispersion, smooth and transparent in surface, uniform in size (shown in figure 5), and 5 +/-0.13 mu m in average particle size distribution range.
The mechanical index value (MI) has a great influence on the ultrasound contrast image, which substantially determines the intensity of the ultrasound because with higher energy ultrasound drives the contrast agent may reflect the echo signal more strongly, whereas when the MI is too large it may cause the destruction of the contrast agent and the loss of the contrast effect. Compared with the phospholipid-based contrast agent, the PCL-b-PMPC phospholipid amphiphilic block copolymer-based ultrasonic contrast agent can tolerate the action of higher ultrasonic mechanical index and keep an integral morphological structure.
The change of the PCL-b-PMPC copolymer-based ultrasound contrast signal intensity with time after ultrasound irradiation is shown in fig. 7 by acquiring ultrasound images of the PCL-b-PMPC copolymer-based ultrasound contrast agent and the SonoVue contrast agent at different time points, and analyzing the change of the average gray level of the copolymer-based ultrasound contrast agent and the SonoVue contrast image. As can be seen from the attached figure 7, the brightness of the ultrasonic image generated by the SonoVue microbubble within 1-3min is rapidly darkened along with the time, the intensity of the ultrasonic signal is rapidly reduced, and the contrast effect is almost lost after 4 min; the brightness of the ultrasonic contrast agent based on the PCL-b-PMPC copolymer shows good stability, the brightness of the image slowly becomes dark between 1 and 3min, the intensity of an ultrasonic signal slowly decreases, the ultrasonic signal tends to be stable after 6min, and the ultrasonic contrast agent still has a certain contrast effect in vitro for 20 min. However, the ultrasound signal generated by SonoVue ultrasound contrast agents drops sharply, to about 20% of its initial intensity, in a period of about 3min,at 20min there was only 2.5% of the initial intensity. However, the ultrasonic signal intensity of the PCL-b-PMPC copolymer-based ultrasonic contrast agent can be maintained for about 20min, and is reduced to 23% of the initial intensity at most and only 50% of the initial intensity at least compared with the initial ultrasonic signal intensity. The stronger stability of the PCL-b-PMPC copolymer-based ultrasound contrast agent is attributed to that the PCL-b-PMPC copolymer can form a stable core-shell structure in an aqueous solution, which provides a strong barrier for keeping PFP gas from diffusing and dissolving in the inner core under ultrasonic radiation, while the lipid shell of the SonoVue ultrasound contrast agent has a thickness of only about 4nm, and gas in the micro-bubbles can diffuse out through the shell quickly under ultrasonic radiation, so that the contrast capability of the SonoVue ultrasound contrast agent is reduced rapidly. Furthermore, gaseous PFP in PCL-b-PMPC copolymer-based ultrasound contrast agents compared to SF in SonoVue ultrasound contrast agents6Gas, has lower water solubility, which also helps to enhance the stability of PCL-b-PMPC copolymer-based ultrasound contrast agents. This allows the PCL-b-PMPC copolymer-based ultrasound contrast agents to have longer cycle periods, providing a longer imaging time window for clinical diagnosis.
From the above, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent obtained by the invention is applied to in-vitro model ultrasonic contrast imaging, and the concentration of the microbubbles is set to be 5.0 × 108After the volume/mL and the mechanical index MI are 0.2, an ultrasonic contrast image is obtained in a frequency range of 4-10 MHz, the change of the average gray value of the contrast image of the copolymer ultrasonic contrast agent along with the frequency is analyzed, and due to the difference between the resonance frequency of the ultrasonic contrast agent and the ultrasonic driving frequency, the brightness of the ultrasonic contrast image of part of the PCL-b-PMPC copolymer-based ultrasonic contrast agent is increased along with the increase of the frequency from 4MHz to 5.71 MHz. When the frequency is increased by 6.67MHz, the brightness of all the ultrasonic contrast images is rapidly reduced; the brightness of the image remains substantially unchanged by 7.27 MHz. When the ultrasound frequency is chosen at 5.71MHz, the contrast image gray value of the ultrasound contrast agent is maximal. Copolymer-based ultrasound contrast agents have a longer duration than SonoVue contrast agents. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is applied to New ZealandThe ultrasonic contrast agent has a certain in-vivo contrast effect, and compared with a phospholipid-based contrast agent, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent has a prolonged contrast time under a high mechanical index.
Claims (10)
1. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is characterized by comprising a shell and an inner core, wherein the shell is amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine, and the inner core is an ultrasonic responder.
2. The phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent according to claim 1, wherein the amphiphilic block copolymer is a copolymer obtained by block copolymerization of caprolactone and 2-methacryloyloxyethyl phosphorylcholine;
further, the shell in the ultrasound contrast agent further comprises a modifying substance M, wherein the modifying substance M is: providing the shell with a material comprising a PEG segment that avoids clearance by the immune system in vivo;
still further, the modifying substance M is selected from: dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 5000, dipalmitoylphosphatidylethanolamine-methoxypolyethylene glycol 2000, distearoylphosphatidylethanolamine-polyethylene glycol 2000, distearoylphosphatidylethanolamine-azidopolyethylene glycol 5000, distearoylphosphatidylethanolamine-azidopolyethylene glycol 2000, distearoylphosphatidylethanolamine-polyethylene glycol-thiol cross-links, distearoylphosphatidylethanolamine-polyethylene glycol 2000-thiol cross-links, distearoylphosphatidylethanolamine-polyethylene glycol 5000-amino cross-links, distearoylphosphatidylethanolamine-polyethylene glycol 2000-amino cross-links, distearoylphosphatidylethanolamine-polyethylene glycol 5000-carboxyl cross-links, distearoylphosphatidylethanolamine-polyethylene glycol 2000-carboxyl cross-links, or distearoylphosphatidylethanolamine-polyethylene glycol 5000-hydroxypolyethylene glycol At least one of the group-crosslinked substances.
3. The phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent according to claim 1 or 2, wherein the core of the ultrasound contrast agent is a phase-changeable liquid perfluorocarbon or gaseous perfluorocarbon;
further, the inner core of the ultrasound contrast agent comprises at least one of perfluoropropane, perfluorobutane, perfluoropentane, perfluorohexane, or perfluoroheptane.
4. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent as claimed in claim 3, wherein when the core of the ultrasonic contrast agent is liquid perfluorocarbon capable of changing phase, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is prepared from polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic response substance in a self-assembly manner, and the obtained nanoemulsion forms the ultrasonic contrast agent with gaseous perfluorocarbon wrapped by the phospholipid-like amphiphilic block copolymer through phase change;
further, when the inner core of the ultrasonic contrast agent is gaseous perfluorocarbon, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is prepared from polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic response substance in a self-assembly mode;
further, the self-assembly mode is one of the following modes: high shear homogenization, high pressure homogenization, high speed oscillation or ultrasonic sound vibration.
5. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent according to any one of claims 1 to 4, wherein the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is diluted with physiological saline, and observed under a laser confocal microscope, the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is spherical in shape, good in dispersion and smooth and bright in surface.
6. The preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent as defined in any one of claims 1 to 5, wherein the preparation method comprises the following steps: self-assembling polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic responder to prepare an ultrasonic contrast agent of which the ultrasonic responder is coated by the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine;
further, the self-assembly is performed in one of the following ways: high shear homogenization, high pressure homogenization, high speed oscillation or ultrasonic sound vibration.
7. The method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent as claimed in claim 6, wherein the ultrasonic response substance is gaseous perfluorocarbon or liquid perfluorocarbon capable of phase change;
further, when the ultrasound response substance in the ultrasound contrast agent is gaseous perfluorocarbon, the preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent comprises the following steps: preparing a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent by directly performing ultrasonic vibration or high-speed oscillation on polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and an ultrasonic responder;
further, when the ultrasound response substance in the ultrasound contrast agent is liquid perfluorocarbon capable of phase transition, the preparation method of the phospholipid-like amphiphilic block copolymer-based ultrasound contrast agent comprises the following steps: the polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and the ultrasonic response substance are prepared into the nano-emulsion in a self-assembly mode, and the nano-emulsion is subjected to temperature-induced phase change or sound-induced phase change to form the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent.
8. The method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent according to claim 7, wherein when the ultrasonic response substance in the ultrasonic contrast agent is phase-changeable liquid perfluorocarbon, the method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent comprises the following steps:
(1) preparing a phospholipid-like amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine;
(2) preparing a phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent: dissolving the phospholipid-like amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine and the modified substance M obtained in the step (1) in a mixed solvent, mixing the dissolved phospholipid-like amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine with an ultrasonic response substance, preparing a nano emulsion in a self-assembly mode, separating and purifying, and preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent after temperature-induced phase change or sound-induced phase change; wherein the modifying substance M is: providing the shell with a material comprising a PEG segment that avoids clearance by the immune system in vivo; the mixed solvent is a mixed solvent of tetrahydrofuran and methanol, or: a mixed solvent of chloroform and methanol.
9. The method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent according to claim 8, wherein in the step (2), the method for preparing the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent after the temperature-induced phase change or the sound-induced phase change of the separated and purified nano-emulsion is one of the following modes:
the first method is as follows: the prepared phospholipid-like amphiphilic block copolymer based nano-emulsion suspension is subjected to water bath at the temperature of 60-80 ℃ for 5-15 min to prepare the phospholipid-like amphiphilic block copolymer based ultrasonic contrast agent;
the second method comprises the following steps: subjecting the prepared phospholipid-like amphiphilic block copolymer-based nanoemulsion suspension to ultrasonic action by an ultrasonic therapeutic apparatus to obtain the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent, wherein the ultrasonic power is 1-3W/cm2The duty ratio is 20% -80%, and the action time is 2-5 min;
preferably, in the step (2), the mixed solvent is a mixture of solvents with a volume ratio of 2:1 of a mixed solvent of tetrahydrofuran and methanol;
further, the method for preparing the phospholipid-like amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine in the step (1) comprises the following steps: polymerizing caprolactone and 2-methacryloyloxyethyl phosphorylcholine by an atom transfer radical polymerization method of an electron transfer regenerated catalyst to obtain the prepared phospholipid amphiphilic block copolymer polycaprolactone-b-polymethacryloxyethyl phosphorylcholine.
10. The phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent can be used in the fields of ultrasonic image diagnosis and treatment or in-vitro agarose model contrast imaging; or the medicine is carried and then used for in vivo ultrasonic contrast imaging and treatment; wherein the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent is the phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent as defined in any one of claims 1 to 5 or the ultrasonic contrast agent prepared by the preparation method as defined in any one of claims 6 to 9.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010717482.8A CN111729094B (en) | 2020-07-23 | 2020-07-23 | Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010717482.8A CN111729094B (en) | 2020-07-23 | 2020-07-23 | Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN111729094A true CN111729094A (en) | 2020-10-02 |
CN111729094B CN111729094B (en) | 2021-07-02 |
Family
ID=72657437
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010717482.8A Active CN111729094B (en) | 2020-07-23 | 2020-07-23 | Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111729094B (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102391446A (en) * | 2011-09-05 | 2012-03-28 | 同济大学 | Method for preparing biocompatible polymer nano-vesicle in pure water |
WO2013176292A2 (en) * | 2012-05-23 | 2013-11-28 | Canon Kabushiki Kaisha | Polymer, contrast agent for nuclear magnetic resonance analysis or magnetic resonance imaging using the polymer, compound and method of nuclear magnetic resonance analysis and method of magnetic resonance imaging using the polymer |
CN107106692A (en) * | 2014-06-12 | 2017-08-29 | 埃文·C·昂格尔 | Phospholipid composite and micro-capsule and the emulsion formed using phospholipid composite and micro-capsule |
-
2020
- 2020-07-23 CN CN202010717482.8A patent/CN111729094B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102391446A (en) * | 2011-09-05 | 2012-03-28 | 同济大学 | Method for preparing biocompatible polymer nano-vesicle in pure water |
WO2013176292A2 (en) * | 2012-05-23 | 2013-11-28 | Canon Kabushiki Kaisha | Polymer, contrast agent for nuclear magnetic resonance analysis or magnetic resonance imaging using the polymer, compound and method of nuclear magnetic resonance analysis and method of magnetic resonance imaging using the polymer |
CN107106692A (en) * | 2014-06-12 | 2017-08-29 | 埃文·C·昂格尔 | Phospholipid composite and micro-capsule and the emulsion formed using phospholipid composite and micro-capsule |
Non-Patent Citations (3)
Title |
---|
K. OLOFSSON ET AL: "Acoustic formation of multicellular tumor spheroids enabling onchip functional and structural imaging", 《LAB ON A CHIP》 * |
赵晓莉 等: "超声造影剂外膜材料研究进展", 《中国医学影像技术》 * |
黄丹凤 等: "比较嵌段共聚物微泡与脂质微泡造影剂的体外稳定性及体内超声造影效果", 《中国介入影像与治疗学》 * |
Also Published As
Publication number | Publication date |
---|---|
CN111729094B (en) | 2021-07-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Kooiman et al. | Oil-filled polymer microcapsules for ultrasound-mediated delivery of lipophilic drugs | |
EP1784228B1 (en) | Gas-filled microvesicles composition for contrast imaging | |
AU703652B2 (en) | Gas-filled microspheres with fluorine-containing shells | |
Cui et al. | Preparation and evaluation of poly (L‐lactide‐co‐glycolide)(PLGA) microbubbles as a contrast agent for myocardial contrast echocardiography | |
US6110444A (en) | Gas-containing microcapsules useful as contrast agents for diagnostic imaging | |
JP5514798B2 (en) | Method for synthesizing hollow spheres | |
JP2000511510A (en) | Contrast agents or related improvements | |
BRPI0707150A2 (en) | Method and Equipment for Wireless Video Communication Error Resilience Algorithms | |
JPH09503202A (en) | Microbubbles encapsulated with polymeric surfactants and their use in ultrasound imaging | |
JP3843327B2 (en) | Improvements in or on contrast media | |
CN109364271B (en) | Ultrasound contrast agent, and preparation method and application thereof | |
Yuksel Durmaz et al. | Development of nanodroplets for histotripsy-mediated cell ablation | |
Houvenagel et al. | Comb-like fluorophilic-lipophilic-hydrophilic polymers for nanocapsules as ultrasound contrast agents | |
Picheth et al. | Echogenicity enhancement by end-fluorinated polylactide perfluorohexane nanocapsules: Towards ultrasound-activable nanosystems | |
Tsirkin et al. | Tailor-made single-core PLGA microbubbles as acoustic cavitation enhancers for therapeutic applications | |
CN111632154A (en) | Phase-transition nanobubble, preparation method and application thereof | |
CN111729094B (en) | Phospholipid-like amphiphilic block copolymer-based ultrasonic contrast agent and preparation method thereof | |
CN111671922B (en) | Amphiphilic comb-shaped block graft copolymer-based ultrasonic contrast agent and preparation method thereof | |
CN111760037B (en) | Phospholipid-like amphiphilic comb graft copolymer-based ultrasonic contrast agent and preparation method thereof | |
US20100221190A1 (en) | Method for producing a particle comprising a gas core and a shell and particles thus obtained | |
HUT74516A (en) | Gas-containing microparticles, agents containing them, their use in medical diagnosis by ultrasonic techniques and methods of producing said particles and agents | |
Kang et al. | Internal polymer scaffolding in lipid-coated microbubbles for control of inertial cavitation in ultrasound theranostics | |
WO2000012062A1 (en) | Local delivery of medications to the heart | |
Jeng | Synthesis of Multifunctional Microbubbles through Precursor Microfluidic Droplet Generation | |
Barmin et al. | Polymeric materials for ultrasound imaging and therapy |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |