CN104817664A - Amphiphilic segmented copolymers, nanoparticles containing amphiphilic segmented copolymers, preparation method of amphiphilic segmented copolymers and use of nanoparticles - Google Patents
Amphiphilic segmented copolymers, nanoparticles containing amphiphilic segmented copolymers, preparation method of amphiphilic segmented copolymers and use of nanoparticles Download PDFInfo
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- CN104817664A CN104817664A CN201510155268.7A CN201510155268A CN104817664A CN 104817664 A CN104817664 A CN 104817664A CN 201510155268 A CN201510155268 A CN 201510155268A CN 104817664 A CN104817664 A CN 104817664A
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- segmented copolymers
- amphipathic nature
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Abstract
The invention discloses amphiphilic segmented copolymers, nanoparticles containing the amphiphilic segmented copolymers, a preparation method of the amphiphilic segmented copolymers and a use of the nanoparticles. The amphiphilic segmented copolymers are shown in the structural formulas (I) to (IV). In the structural formulas (I) to (IV), R1 represents H or methyl, R2, R3 and R5 represent H, alkyl or aryl, R4 represents hydroxyl or methyl, x is an integer of 3-20, Spacer is a spacer arm, the spacer arm is optional, and when the spacer arm exists, the spacer arm is one of an ester group, an alkyl group, a triazole group and a thioether group. The nanoparticle obtained by chelating of the amphiphilic segmented copolymer and rare earth ions has good biocompatibility, has excellent fluorescence imaging performances and MRI imaging performances and has a wide application value in the field of medicine and biology and especially in breast cancer diagnosis and treatment.
Description
Technical field
The invention belongs to synthesis of polymer material and biomedicine field, be specifically related to the application of a kind of fluorescence-MRI double function probe and carrier and biomedicine field thereof.
Background technology
Closely during the last ten years, molecular image (molecular imaging) technical development is rapid, its can not only organize level, cell levels even molecular level carry out living imaging to show its biological behaviour, qualitative and quantitative study (Nature can also be carried out to specific molecular, 2008,452 (7187): 580-590).In current different kinds of molecules image technology, MRI is one of image technology of maturation the most at present, and it is a kind of non-invasive medical diagnosis technology, can provide high-resolution organizational information and three-dimensional structure imaging on sub-millimeter level.At present for the T of clinical magnetic resonance imaging diagnosis
1contrast medium is mainly the title complex of gadolinium, but its residence time in blood circulation is short, can be excreted rapidly, and this just needs relatively high injection volume and frequency of injection just can reach the imaging effect of expection.By Gd
3+ion induces one high molecular system, forms macromolecular contrast agent, can fall low molecular speed of rotation, and can relatively long-time in the blood vessel in keep stable concentration, be conducive to angiography (WO.9700087,1998-01-03).
Fluorescent rare earth complex has high fluorescence intensity, long fluorescence lifetime, the luminosities such as the narrow and Stokes displacement of emission peak is large, are widely used in the field such as fluorescent probe, biomarker (Luminescence 2012,132 (8): 2005-2011).In view of fluorescence imaging method, there is high sensitivity and the advantage of real-time multi-target imaging can be carried out, the deficiency of MRI technology can be made up preferably, therefore MRI is combined with optical image technology, high-resolution structure organization information can be provided, high-sensitive function assessment video picture can be realized again simultaneously, qualitative and the quantitative display of minimal disease and pathology target molecule is made to become possibility, finally reach the double goal (Chem.Comm. of function assessment video picture and structure organization video picture, 2008, (48): 6591-6593.).Compared with independent imaging, mixed mode has obvious superiority, adds the accuracy of diagnosis.
By rare earth compounding being used for the fluorescence imaging of the tumour cells such as mammary cancer, and bag medicine carrying thing is in its hydrophobic core, the biocompatibility utilizing it good, the controllability of drug release and this nanoparticle are to the targeting of the tumour cells such as mammary cancer, and combined with MRI technology will improve clinical diagnosis and treatment integration effect.In addition, this contrast medium also can be used for preoperative location, MRI guide under intervene operation or the field such as fluorescence surgical navigation.
Summary of the invention
The invention discloses a kind of amphipathic nature block polymer, comprise the nanoparticle of this multipolymer, its preparation method and application, this nanoparticle good biocompatibility, there is superior fluorescence imaging performance and MRI imaging performance simultaneously.
A kind of amphipathic nature block polymer, structure is as the one in formula I ~ formula IV:
In formula I ~ (IV), R
1for H or methyl; R
2, R
3, R
5for H or alkyl, aryl; R
4for hydroxyl or methyl; The integer of x=3 ~ 20; Spacer is optional spacerarm (namely spacerarm can exist, or does not exist, and when not existing, the group at two ends is directly connected), comprises the one in ester group, alkyl, triazole group, sulfide group;
N is the integer of 20 ~ 450, and m is the integer of 10 ~ 450.
Present invention also offers a kind of polymer nano-particle, obtain after carrying out chelatropic reaction by described amphipathic nature block polymer and rare earth ion.
This polymer nano-particle is coreshell type structure, is made up of the rare earth ion of the amphipathic nature block polymer containing diketone function fragment and doping, and kernel is the double function probe with fluorescence and MRI imaging performance, and shell has biocompatible hydrophilic segment.This polymer nano-particle make use of the higher relaxation rate of rare earth compounding and better image contrast, sub-millimeter level provides high-resolution organizational information and three-dimensional structure imaging, and rely on the slightly pointed absorption of rare earth ion tool and emission peak, long lifetime, the superior fluorescence property such as high-quantum efficiency realizes optical imagery.
As preferably, described rare earth ion comprises a kind of ion in gadolinium (Gd) ion and following rare earth element:
Lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu), terbium (Tb), dysprosium (Dy), holmium (Ho), erbium (Er), thulium (Tm), ytterbium (Yb), lutetium (Lu).
As preferably, described amphipathic nature block polymer (in dione unit) is 10 ~ 1:1 with the mol ratio of rare earth ion, is preferably 3 ~ 1:1;
The mol ratio of gadolinium (Gd) ion and another kind of rare earth ion is 100 ~ 0.01:1.
As preferably, the diameter of described polymer nano-particle is between 10 ~ 500nm.
Present invention also offers a kind of preparation method of described amphipathic nature block polymer, comprise the following steps:
(1) under the effect of micromolecule chain transfer agent, containing (methyl) acrylate-type monomer polymerization reaction take place of diketone function fragment, (methyl) acrylate based polymer containing diketone function fragment is obtained;
(2) under initiator exists, (methyl) acrylate based polymer containing diketone function fragment step (1) obtained and hydrophilic second comonomer carry out polyreaction or carry out linked reaction with hydrophilic homopolymer, obtain described amphipathic nature block polymer.
This preparation method uses RAFT synthesizing amphipathic segmented copolymer, and synthesis condition is simple, and step is easy, and synthesized polymer architecture is clearly controlled.
In step (1); as preferably, the described acrylate-type monomer containing dione unit comprises vinylformic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (CAS:157174-83-9), methacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (CAS:157174-85-1).
In step (1), as preferably, described chain-transfer agent is selected from the one in the compound shown in following formula:
As preferably, the polyreaction of step (1) is carried out under the existence of initiator;
The mol ratio of described (methyl) acrylate-type monomer containing diketone function fragment, initiator and micromolecule chain transfer agent is 50 ~ 500:0.3:1.Wherein, initiator is Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile).
As preferably, the polyreaction of step (1) is carried out in reaction solvent, and described (methyl) acrylate-type monomer volumetric molar concentration containing diketone function fragment is 0.1 ~ 10mol/L; Reaction solvent is the one in DMF, N,N-dimethylacetamide, toluene, methyl-phenoxide, dioxane, methyl-sulphoxide, acetonitrile, chloroform, N-Methyl pyrrolidone water and tetrahydrofuran (THF).
As preferably, in step (1), the temperature of polyreaction is 60 ~ 90 DEG C, and the time of polyreaction is 3 ~ 48h.
Through the reaction of step (1), the line polymer that structure is clear and definite can be obtained, the polymerization degree is 50 ~ 500 (being calculated by nuclear magnetic resonance result), and molecular weight distribution is 1.01 ~ 1.45 (gel permeation chromatographies), is preferably 1.1 ~ 1.3; It should be noted that when preparing polymkeric substance by different initiator, molecular weight and molecular weight distribution different.
As preferably, in step (2), the mol ratio containing (methyl) acrylate based polymer of diketone function fragment, initiator, second comonomer is 1:0.3:20 ~ 500; When with linked reaction synthetic segmented copolymer, the mol ratio containing (methyl) acrylate based polymer of diketone function fragment, hydrophilic polymer containing reactive terminal, coupling catalyst is 1:1.2:0.01 ~ 0.2.
As preferably, in step (2), the temperature of polyreaction is 60 ~ 90 DEG C, and the time of polyreaction is 2 ~ 24h; When with linked reaction synthetic segmented copolymer, the temperature of reaction is rt ~ 60 DEG C, and the time of reaction is 2 ~ 12h.
Through the reaction of step (2), the amphipathic nature block polymer that structure is clear and definite can be obtained, the polymerization degree is 70 ~ 1000 (being calculated by nuclear magnetic resonance result), the molecular weight distribution connecting hydrophilic segment is 1.1 ~ 1.45 (gel permeation chromatographies), is preferably 1.1 ~ 1.3.It should be noted that total polymerization degree is different with molecular weight distribution rangeability by different second comonomer or homopolymerization hydrophilic polymer.
In step (2), described initiator is Diisopropyl azodicarboxylate or 2,2'-Azobis(2,4-dimethylvaleronitrile); Second comonomer comprises at least one in the compound shown in following formula:
Wherein, the integer of y=1 ~ 10, R
6independently selected from H or methyl; R
7independently selected from hydroxyl or methyl; R
8independently selected from H, aryl or alkyl.
In step (2), when with linked reaction synthetic segmented copolymer, the hydrophilic polymer containing reactive terminal comprises at least one in the compound shown in following formula.
Wherein, the integer of n=20 ~ 450;
In step (2), coupling catalyst is the one in cuprous bromide, Diisopropyl azodicarboxylate, 2,2'-Azobis(2,4-dimethylvaleronitrile), triethylamine, 1,3-dicyclohexyl carbodiimide, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide, DMAP.
In step (2), reaction solvent is the one in DMF, N,N-dimethylacetamide, toluene, methyl-phenoxide, dioxane, methyl-sulphoxide, acetonitrile, chloroform, N-Methyl pyrrolidone water and tetrahydrofuran (THF).
Polymkeric substance in the present invention carries out purifying by the method for dissolution precipitation, and precipitation agent is the one in methyl alcohol, water, ether and normal hexane.
Present invention also offers a kind of preparation method of described polymer nano-particle, comprising: in the basic conditions, described amphipathic nature block polymer and rare-earth salts carry out chelatropic reaction, obtain described polymer nano-particle.
By the easy introducing of chelatropic reaction one step, there is gadolinium (Gd) ion of high relaxation rate in the basic conditions and have the rare earth ion of fluorescence property, process simple and effective, single stage method to achieve the two detection of fluorescence-MRI.
Described rare-earth salts is a kind of inorganic salt (one preferably in hydrochloride, nitrate, vitriol) in the inorganic salt (being preferably hydrochloride, nitrate, one in vitriol) of gadolinium and lanthanum (La), cerium (Ce), praseodymium (Pr), neodymium (Nd), samarium (Sm), europium (Eu), terbium (Tb), dysprosium (Dy), holmium (Ho), erbium (Er), thulium (Tm), ytterbium (Yb), lutetium (Lu).
As preferably, used alkali is the one in sodium carbonate/potassium, sodium hydroxide/potassium, sodium ethylate/potassium, sodium tert-butoxide/potassium, triethylamine, pyridine, 1,8-diazabicylo 11 carbon-7-alkene (DBU), DMAP.
In step (3), the mol ratio of block polymer (in dione unit), rare earth ion, alkali is 10 ~ 1:1:1.5; Gadolinium (Gd) is 100 ~ 0.01:1 with the content ratio of another kind of rare earth ion.
Present invention also offers a kind of described application of polymer nano-particle in biomedicine field, described polymer nano-particle is as fluorescence-MRI bimodal image reagent.
Invented nanoparticle is injected the mouse model of xenograft tumor, fluorescence and the imaging of mr bimodal can be realized, make single contrast medium injection can obtain more abundant Pathological Information; In contrast medium, be loaded into chemotherapeutics simultaneously, diagnosis and treatment integration can be realized; In addition this contrast medium is also applicable to the fields such as interventional navigation, preoperative location, fluorescence surgical navigational.Therefore this invention can become the multifunctional nano platform that video imaging, drug delivery, spike and surgical navigational, therapeutic evaluation are integrated.Comprehensive above feature, this nanoparticle not only can carry out the display of high resolving power, hypersensitivity to tumour, also being applicable to all respects through clinical cancer therapy such as the chemotherapy of tumors such as mammary cancer, tumor-localizing, surgical navigational, therapeutic evaluation, is that a class has the mr/fluorescent nano particles of target tracing to tumour.
As preferably, described fluorescence-MRI bimodal image reagent be used for viable cell with the fluorescence imaging of living animal and MRI radiography.
Cell in vitro pharmacodynamic experiment scheme of the present invention is as follows:
(1) cytotoxicity: adopt Thiazolyl blue method (MTT) to measure the cytotoxicity of polymkeric substance to breast cancer cell 48h, the absorbance of multi-functional microplate reader working sample at 570nm place.Each sample parallel tests 5 times, mapping of averaging (blank as a control group), calculates cell survival rate (per-cent);
(2) cellular uptake: laser confocal microscope is using 405nm as excitation wavelength, and 620nm observes at different time points 1h, 4h, 12h, 24h breast cancer cell its picked-up situation as accepting wavelength.
Compared with the existing technology, beneficial effect of the present invention is embodied in:
(1) in structure, novel fluorescence-MRI bimodal imaging nanoparticle provided by the invention, simple and stable structure, low cost of raw materials, can prepare on a large scale, fluorescence intensity is high, and fluorescence lifetime is long, and emission peak is narrow and Stokes displacement is large waits excellent fluorescence usefulness and good MRI resolving power.
(2) on bio-medical, novel fluorescence-MRI bimodal imaging nanoparticle provided by the invention, there is the advantages such as low cytotoxicity, cell absorbability, targeting, multi-modality imaging, can be used for the fields such as cell in vitro mark, living imaging, diagnosis and treatment integration, pathology location and navigation, curative effect evaluation.
Accompanying drawing explanation
Fig. 1 is the fluorescence picture of polymer nano-particle after breast cancer cell picked-up that embodiment 1 obtains;
Fig. 2 is the MRI imaging picture of the polymer nano-particle that embodiment 1 obtains.
Embodiment
Below in conjunction with specific embodiment, the present invention is further detailed.
Gained amphipathic nature block polymer molecular weight adopts nuclear-magnetism to follow the tracks of and calculates and SEC mensuration.Nuclear-magnetism Bruker ARX 400 (
1h:400MHz) instrument measures, with deuterated dimethyl sulfoxide (DMSO-d
6) or deuterochloroform (CDCl
3) as solvent, tetramethylsilane (TMS) is as interior mark.Number-average molecular weight is followed the tracks of transformation efficiency by nuclear-magnetism and is obtained.The relative molecular weight of polymkeric substance and molecular weight distribution adopt Waters gel chromatograph (Waters 1525HPLC configures Waters 2414RI detector), chromatographic column is Waters Styragel Columns HR4, HR3 and HR1, THF is moving phase, probe temperature is 40 DEG C, flow velocity is 1.0mL/min, and the relative molecular mass of polymkeric substance take polystyrene as standard calibration; And Waters 1515Isocratic efficient liquid-phase chromatographic pump, PLgel 5 μm of MIEXD-C chromatographic columns, containing the DMF of 0.05mol/L LiBr as moving phase, 60 DEG C, flow velocity is 1.0mL/min, and the relative molecular mass of polymkeric substance take polymethylmethacrylate as standard calibration.
Cell culture medium 1640 (Gibco BRL, USA), trypsin Trypin, Gibco BRL, USA), foetal calf serum (Hangzhou Sijiqing Biological Engineering Material Co., Ltd.), human breast cancer cell MCF-7 (Michigan Cancer Foundation-7, MCF-7, pharmaceutical preparation institute of Zhejiang University), Thiazolyl blue MTT (3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazoliumbromide, Sigma, USA), dimethyl sulfoxide (DMSO) (dimethyl sulfoxide, DMSO, the large biological company limited in sea, Wuxi), other solvent or chemical reagent are analytical pure.
CO2gas incubator (3110, Thermo Forma, USA), enzyme connection detector (Bio-Rad, Model 680, USA), laser confocal microscope (IX81-FV1000, Olympus, Japan), culture plate (Nalge Nunc International, Naperville, 1L, USA).
Embodiment 1
By 1.41g methacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (DKMA) (CAS:157174-85-1), 4.15mg chain-transfer agent dithiobenzoic acid-2-phenyl third-2-ester (CDB) (CAS:201611-77-0), 0.826mg AIBN, 10mL methyl-phenoxide adds in reaction flask, DKMA, the mol ratio of AIBN and CDB is 300:0.33:1, 12h is reacted at 70 DEG C, dissolution precipitation 3 removing small molecule monomers in 100mL ether, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (PDKMA), yield is 50%, it is 152 that nuclear-magnetism calculates the polymerization degree, SEC number-average molecular weight is 33.8kDa, molecular weight distribution is 1.28.
By 1.810g methacrylic acid oligomeric ethylene glycol ester (OEGMA), 0.564mg PDKMA, 1.519mg AIBN and 5mL methyl-phenoxide add in reaction flask, PDKMA, the mol ratio of AIBN and OEGMA is 1:0.33:130, 12h is reacted at 60 DEG C, after reaction terminates, 200mL ether sedimentation 3 removing monomers, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester-b-polymethyl acrylic acid oligomeric ethylene glycol ester (PDKMA-b-POEGMA), yield is 77.4%, the polymerization degree calculated by nuclear-magnetism is 310, SEC number-average molecular weight is 79.8kDa, molecular weight distribution is 1.33.Synthetic route is shown below:
Subsequently, under salt of wormwood action condition, by be synthesized amphipathic nature block polymer be dissolved in tetrahydrofuran (THF), and carry out chelatropic reaction with rare-earth salts, rear to after water dialysis 48h, obtain described polymer nano-particle.
Adopt the cytotoxic process of this material of Thiazolyl blue method (MTT) quantitative expedition as follows.First, according to 0.8-1 × 10
4the cell density of number/mL breast cancer cell liquid turns 96 porocyte plates, is placed in overnight incubation in 37 DEG C of 5% cell culture incubator.Secondly, prepare the polymers soln of a series of different concns, the volume that every hole adds copolymer solution is 200ul, parallel 5 multiple holes, and diluent media is 1640 high glucose mediums, the required concentration investigating polymkeric substance is followed successively by 0,50,100,200,300,400,500,600,800 units are ug/ml.Then, from incubator, take out 96 porocyte plates, suck nutrient solution, every hole 100ul PBS buffered soln rinses once, suck phosphate buffer soln again, the polymers soln of a series of different concns is joined in cell plate successively, be placed in 37 DEG C of 5% cell culture incubator and cultivate 48 hours.Then, from incubator, take out 96 porocyte plates, every hole adds 20ul MTT, then continues to put into incubator and hatch 4h; Take out cell plate after 4h, every hole adds 200 μ L dimethyl sulfoxide (DMSO), slightly shakes up 30min, treats that bluish voilet crystallisate first is praised (formazan) and dissolved completely, finally, and the absorbance of multi-functional microplate reader working sample at 570nm place.Each sample parallel tests 5 times, mapping of averaging (blank as a control group).Cell survival rate (per-cent) treats that the absorbance of test sample is compared Normocellular absorbance (namely the concentration of polymers soln is 0) and represented.
Relevant cytotoxic test result shows, synthesized novel nano particle does not have cytotoxicity in the concentration range of 0-800 μ g/mL, shows good cell compatibility.
Fluorescence property detects: use the cellular uptake situation of this polymer nano-particle of laser confocal microscope qualitative observation as follows.First, according to 5 × 10
4the cell density of number/mL breast cancer cell liquid turns 24 porocyte plates, is placed in overnight incubation in 37 DEG C of 5% cell culture incubator (nutrient solution is RPMI-1640, and the concentration of contained polymer nano-particle is 100ug/ml).Secondly, add to every hole according to different time points 1h, 4h, 12h, 24h respectively, result shows that this multipolymer just has picked-up from 1h, and increases along with the prolongation picked-up of time point thereupon.Fig. 1 is shown in by the fluorescence picture obtained, and wherein, first row is fluorescence picture (excitation wavelength is 405nm, receives wavelength 620nm), and secondary series is light field-cell picture, and the 3rd row are superposition pictures.Namely this result shows that nanoparticle that embodiment 1 obtains is easily by cellular uptake, also shows to have good fluorescent effect simultaneously.
MRI Performance Detection: the nanoparticle obtain Gd-DTPA, embodiment 1 and water carry out MRI imaging respectively, and what obtain the results are shown in Figure in 2, figure, and the left side is the imaging results of Gd-DTPA, centre is the imaging results of nanoparticle, and the right is the imaging results of water.Result shows, the imaging function of nanoparticle will far above Gd-DTPA.
Chemotherapeutic agent can be wrapped up as taxol, Zorubicin, vincristine(VCR) etc. in this copolymer micelle hydrophobic core inside, and connect hydrophobic inner core and hydrophilic radical by disulfide linkage, be convenient to multipolymer and enter the object that disulfide linkage in body was reduced, reached drug controllable release, this external surface is carried polypeptide and can be realized this kind of material and study the active targeting of a certain specific tumors clone.
Embodiment 2
By 1.2g vinylformic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (DKA), 5.55mg chain-transfer agent dithiobenzoic acid-2-phenyl third-2-ester (CDB), 0.744mg AIBN, 5mL methyl-phenoxide adds in reaction flask, DKA, the mol ratio of AIBN and CDB is 200:0.33:1, 24h is reacted at 70 DEG C, dissolution precipitation 3 removing small molecule monomers in 200mL ether, vacuum-drying obtains polyacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (PDKA), yield is 20%, it is 51 that nuclear-magnetism calculates the polymerization degree, SEC number-average molecular weight is 15.8kDa, molecular weight distribution is 1.30.
By 0.8g methacrylic acid oligomeric ethylene glycol ester (OEGMA), 0.24g PDKA, 0.866mgAIBN and 5mL methyl-phenoxide adds in reaction flask, PDKA, the mol ratio of AIBN and OEGMA is 1:0.33:100, 18h is reacted at 60 DEG C, after reaction terminates, 200mL ether sedimentation 3 removing monomers, vacuum-drying obtains polyacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester-b-polymethyl acrylic acid oligomeric ethylene glycol ester (PDKA-b-POEGMA), yield is 54.3%, the polymerization degree calculated by nuclear-magnetism is 43, SEC number-average molecular weight is 34.8kDa, molecular weight distribution is 1.36.
Embodiment 3
By 1.45g methacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (DKMA), 4.15mg chain-transfer agent dithiobenzoic acid-2-phenyl third-2-ester (CDB), 0.826mg AIBN, 10mL methyl-phenoxide adds in reaction flask, DKMA, the mol ratio of AIBN and CDB is 310:0.33:1, 12h is reacted at 70 DEG C, dissolution precipitation 3 removing small molecule monomers in 300mL ether, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (PDKMA), yield is 53%, it is 168 that nuclear-magnetism calculates the polymerization degree, SEC number-average molecular weight is 34.8kDa, molecular weight distribution is 1.28.
By 0.52g NIPA (NIPAM), 0.373g PDKMA, 1.519mgAIBN and 5mL methyl-phenoxide adds in reaction flask, PDKMA, the mol ratio of AIBN and NIPAM is 1:0.33:728, 8h is reacted at 60 DEG C, after reaction terminates, 200mL ether sedimentation 3 removing monomers, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester-b-poly N-isopropyl acrylamide (PDKMA-b-NIPAM), yield is 50.4%, the polymerization degree calculated by nuclear-magnetism is 300, SEC number-average molecular weight is 43.1kDa, molecular weight distribution is 1.30.
Embodiment 4
By 1.41g methacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (DKMA), 4.15mg chain-transfer agent dithiobenzoic acid-2-phenyl third-2-ester (CDB), 0.826mg AIBN, 10mL methyl-phenoxide adds in reaction flask, DKMA, the mol ratio of AIBN and CDB is 300:0.33:1, 12h is reacted at 70 DEG C, dissolution precipitation 3 removing small molecule monomers in 100mL ether, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (PDKMA), yield is 50%, it is 152 that nuclear-magnetism calculates the polymerization degree, SEC number-average molecular weight is 33.8kDa, molecular weight distribution is 1.28.
By 0.92g NVP (NVP), 0.564mg PDKMA, 1.519mg AIBN and 5mL methyl-phenoxide add in reaction flask, PDKMA, the mol ratio of AIBN and OEGMA is 1:0.33:680, 24h is reacted at 70 DEG C, after reaction terminates, 200mL ether sedimentation 3 removing monomers, vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester-b-poly N-vinyl pyrrolidone (PDKMA-b-PNVP), yield is 12.4%, the polymerization degree calculated by nuclear-magnetism is 89, SEC number-average molecular weight is 37.5kDa, molecular weight distribution is 1.32.
Embodiment 5
By 1.39g methacrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (DKMA), 4.24mg chain-transfer agent dithiobenzoic acid-2-propynyloxy acyl group third-2-ester (CAS:1027346-89-9), 0.826mg AIBN, 10mL methyl-phenoxide adds in reaction flask, DKMA, the mol ratio of AIBN and dithiobenzoic acid-2-propynyloxy acyl group third-2-ester is 295:0.33:1, 12h is reacted at 70 DEG C, dissolution precipitation 3 removing small molecule monomers in 100mL ether, vacuum-drying obtains equal polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester (PDKMA) that lightpink is alkynyl containing end group, yield is 50%, it is 120 that nuclear-magnetism calculates the polymerization degree, SEC number-average molecular weight is 26.7kDa, molecular weight distribution is 1.29.
By 2.0g polyethyleneglycol nitrine (PEG-N
3m
n=2000g/moL), 0.564gPDKMA, 1.519mg AIBN and 5mL methyl-phenoxide adds in reaction flask, PDKMA and PEG-N
3mol ratio be 1:1.2; cuprous iodide reaction 12h is added under rt; after reaction terminates; 200mL ether sedimentation 3 times; vacuum-drying obtains polymethyl acrylic acid-4-(3-oxo-3-PHENYLPROPIONYL) phenyl ester-b-polyoxyethylene glycol (PDKMA-b-PEG); yield is 80.4%, SEC number-average molecular weight is 28.5kDa, and molecular weight distribution is 1.33.
Claims (9)
1. an amphipathic nature block polymer, is characterized in that, structure is as shown in the one in formula I ~ formula IV:
In formula I ~ (IV), R
1for H or methyl; R
2, R
3, R
5for H or alkyl, aryl; R
4for hydroxyl or methyl; The integer of x=3 ~ 20; Spacer is optional spacerarm, comprises the one in ester group, alkyl, triazole group, sulfide group;
N is the integer of 20 ~ 450, and m is the integer of 10 ~ 450.
2. a polymer nano-particle, is characterized in that, obtains after carrying out chelatropic reaction by amphipathic nature block polymer according to claim 1 and rare earth ion.
3. polymer nano-particle according to claim 2, is characterized in that, described rare earth ion comprises a kind of ion in gadolinium ion and following rare earth element:
Lanthanum, cerium, praseodymium, neodymium, samarium, europium, terbium, dysprosium, holmium, erbium, thulium, ytterbium or lutetium.
4. polymer nano-particle according to claim 2, is characterized in that, described amphipathic nature block polymer (in dione unit) is 10 ~ 1:1 with the mol ratio of rare earth ion;
The mol ratio of gadolinium ion and another kind of rare earth ion is 100 ~ 0.01:1.
5. polymer nano-particle according to claim 2, is characterized in that, its diameter is between 10 ~ 500nm.
6. a preparation method for amphipathic nature block polymer as claimed in claim 1, is characterized in that, comprises the following steps:
(1) under the effect of micromolecule chain transfer agent, containing (methyl) acrylate-type monomer polymerization reaction take place of diketone function fragment, (methyl) acrylate based polymer containing diketone function fragment is obtained;
(2) under initiator exists, step (1) is obtained containing diketone function fragment (methyl) acrylate based polymer and hydrophilic second comonomer carries out polyreaction or hydrophilic homopolymer carries out linked reaction, obtain described amphipathic nature block polymer.
7. the preparation method of amphipathic nature block polymer according to claim 6, it is characterized in that, in step (1), the mol ratio of described (methyl) acrylate-type monomer containing diketone function fragment, initiator and micromolecule chain transfer agent is 50 ~ 500:0.3:1;
In step (1), polyreaction is carried out in reaction solvent, and described (methyl) acrylate-type monomer volumetric molar concentration containing diketone function fragment is 0.1 ~ 10mol/L; The temperature of polyreaction is 60 ~ 90 DEG C, and the time of polyreaction is 3 ~ 24h.
8. the application of the polymer nano-particle as described in any one of claim 2 ~ 5 in biomedicine field, is characterized in that, described polymer nano-particle is as fluorescence-MRI bimodal image reagent.
9. application according to claim 8, is characterized in that, described fluorescence-MRI bimodal image reagent is used for fluorescence imaging and the MRI radiography of viable cell and living animal.
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| CN113679854A (en) * | 2021-09-03 | 2021-11-23 | 苏州大学 | Magnetic resonance contrast agent and preparation and application thereof |
| WO2024034686A1 (en) * | 2022-08-10 | 2024-02-15 | 興和株式会社 | Polymer contrast agent |
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| CN112004848A (en) * | 2017-12-05 | 2020-11-27 | 国际商业机器公司 | Block copolymers and self-assembled nanoparticles formed therefrom |
| GB2583602B (en) * | 2017-12-05 | 2022-06-08 | Ibm | Block copolymers and self-assembling nanoparticles formed therefrom |
| US10376468B2 (en) | 2017-12-05 | 2019-08-13 | International Business Machines Corporation | Block copolymers and self-assembling nanoparticles formed therefrom |
| US10682313B2 (en) | 2017-12-05 | 2020-06-16 | International Business Machines Corporation | Block copolymers and self-assembling nanoparticles formed therefrom |
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| GB2583602A (en) * | 2017-12-05 | 2020-11-04 | Ibm | Block copolymers and self-assembling nanoparticles formed therefrom |
| WO2019111121A1 (en) * | 2017-12-05 | 2019-06-13 | International Business Machines Corporation | Block copolymers and self-assembling nanoparticles formed therefrom |
| CN112004848B (en) * | 2017-12-05 | 2023-07-21 | 国际商业机器公司 | Block copolymers and self-assembled nanoparticles formed therefrom |
| CN108893139B (en) * | 2018-08-01 | 2020-12-04 | 嘉兴笼列电子商务有限公司 | Method for desulfurizing fuel oil by using amphiphilic catalyst containing neodymium |
| CN108893139A (en) * | 2018-08-01 | 2018-11-27 | 李成霞 | A kind of method of use amphipathic catalyst fuel desulfuration containing neodymium |
| CN111603572A (en) * | 2020-06-02 | 2020-09-01 | 珠海市人民医院 | Nano contrast agent and preparation method and application thereof |
| CN111603572B (en) * | 2020-06-02 | 2022-03-29 | 珠海市人民医院 | Nano contrast medium and preparation method thereof |
| CN113679854B (en) * | 2021-09-03 | 2022-08-09 | 苏州大学 | Magnetic resonance contrast agent and preparation and application thereof |
| CN113679854A (en) * | 2021-09-03 | 2021-11-23 | 苏州大学 | Magnetic resonance contrast agent and preparation and application thereof |
| WO2024034686A1 (en) * | 2022-08-10 | 2024-02-15 | 興和株式会社 | Polymer contrast agent |
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