CN103333301B - Amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer and preparation method thereof - Google Patents
Amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of high molecular polymer materials for biomedicine, and discloses an amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer of which the number-average molecular weight is 30000-54000g/mol. The invention also discloses a preparation method of the amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer, which comprises the following steps: mixing epsilon-caprolactone, stannous octoate and low molecular initiator, reacting at 110-140 DEG C for 24-48 hours, performing reduced pressure distillation, precipitating, filtering, and drying to obtain polycaprolactone high molecular initiator; dissolving the polycaprolactone high molecular initiator, 2-diethylaminoethyl methacrylate, hexamethyltriethylenetetramine and copper bromide in toluene, stirring, then adding stannous octoate, and reacting at 60-90 DEG C for 5-12 hours; and adding methoxy polyoxyethylene methacrylate, continuously polymerizing for 5-12 hours, removing a catalyst, filtering, and performing posttreatment on the filtrate to obtain the amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer. The amphiphilic pH-responsive 4/6 heteroarm star-shaped copolymer is used for preparation of a micelle system for carrying water insoluble medicaments.
Description
Technical field
The invention belongs to biological medicine macromolecule polymer material technical field, particularly the amphipathic pH of one responds 4/6 Miktoarm star copolymer and preparation method thereof.
Background technology
Cancer is 21 century three one of disease greatly, and the health of the mankind in serious harm.The global cancer report display of the World Health Organization: the annual life seizing more than 700 ten thousand people in the whole world at present, along with world population aging increasingly, estimates that whole world number of cancer deaths also will rise fast, may by more than 1,310 ten thousand to the year two thousand thirty.Due to the heterogeneity, multidrug resistance etc. of cancer cell, the treatment of cancer medicine is still faced with huge challenge.As one of most important instrument of resistance cancer, chemotherapy plays very important effect for the treatment of cancer.Desirable chemotherapeutics should only play a role to the cancer cells of diseased region, and or can not produce toxic side effect to normal cell less.In order to reduce toxic side effect, improve the availability of medicine, reduce degraded and the loss of medicine, people are the various Drug Carrier Systems of Devoting Major Efforts To Developing in recent years.These Drug Carrier Systems can change medicine and enter the mode of human body and distribution in vivo, control the release rate of medicine and conduct drugs to target organs.In order to find suitable pharmaceutical carrier, people to various carrier system as polymer micelle, hydrogel, vesica, microballoon, liposome, microemulsion etc. have carried out large quantifier elimination.Wherein, polymer micelle becomes the focus of research at present with its peculiar excellent properties.
Biodegradable amphiphilic polymkeric substance forms the micella of core/shell structure in aqueous by self-assembly, it is more and more subject to the common concern of various countries investigator as anti-cancer medicament carrier.It has the following advantages: (1) polymericular weight is large, and medicine can be made when using as carrier at lesions position stay longer; The permeability cell of tumor locus is in hyperfunction state, and the time that cancer therapy drug exists in blood circulation is longer, and the chance entering tumor locus is more; (2) medicine reaches the object of slowly-releasing or controllable release in polymer micelle by diffusion or the degraded of polymkeric substance self; (3) some functional components with targeting or Drug controlled release can be attached to polymer beads sub-surface by the mode of chemical bonding; (4) biodegradable polymer materials, after can avoiding drug release, solid support material is built up and produces toxic side effect in human organ tissue.
Star-type polymer refers to the polymkeric substance of at least drawing three polymeric arms in a core.Generally being divided into two large classes: a class is the star-type polymer (AB) that every arm all has identical chemical structure, every arm may be homopolymer, multipolymer or triblock copolymer; Another kind of is the assorted arm star polymkeric substance that at least two arms chemical structure is not identical, common are A
2b
2, A
3b
3, A
2b, A
3b, ABC, A
5b, AB
2c
2, ABCD etc.The chemical structure of its uniqueness of star polymer makes it have some special physical and chemical performances, as little in hydrodynamic radius, limiting viscosity is low, degree of crystallinity is low, be easy to form stable micella etc., will be widely used in more field.Because have the topological framework similar to polymer micelle, can improve the thermodynamic stability of micella, star polymer is highly suitable for the Co ntrolled release of cancer therapy drug.
As the Typical Representative of branch-shape polymer, star polymer synthesizes mainly through core first (core-first) or arm first (arm-first) mode.Core first needs first to synthesize multi-functional initiator, and the consistent and arm number of the brachium of the star-type polymer of synthesis is determined, accurately can control the polymerization degree.Arm first needs first to synthesize monofunctional active's linear macromolecule, and the consistent but arm number of the brachium of the star-type polymer of synthesis is difficult to determine.
The form of the aggregate that the star block copolymer reported at present is formed through self-assembly has spherical, tubulose, vesica shape, vermiform and composite-like etc., thus facilitate the potential application of star block copolymer at numerous areas, as medicament slow release, separation, photoelectric material and catalysis etc.In medicament slow release, (the Journal of Colloid and Interface Science such as Lang, 2011,364:92.) adopt straight chain polyoxyethylene glycol-b-polycaprolactone (PEG-b-PCL) and six arm star polycaprolactone-b-polymethyl acrylic acid lignocaine ethyl ester (S (PCL-b-PDEAEMA)) to prepare a kind of mixed micelle, effectively improve the stability of micella.Wrapped after carrying nifedipine, utilized PDEAEMA hydrophilic and hydrophobic with the change of pH value to control the rate of release of nifedipine.(the Biomacromolecules such as Cao, 2011,12 (7): 2697.) one has been prepared with polylactide (PLLA) for kernel, polyoxyethylene glycol (PEG) is hydrophilic outer shell, the star unimolecular micelle of surface coupling folic acid is used for the target administration of Zorubicin, this micella rate of release when pH7.4 is very slow, and rate of release is obviously accelerated when pH5.0.Patent application WO2003078489-A1 discloses a kind of preparation method preparing wetting ability two block, three blocks and star-type polymer; under protection of inert gas, be polymerized preparation by least one vinyl monomer, macromole degradability chain-transfer agent and initiator and obtain.Patent application 200810107054.2 discloses a kind of three arm star multipolymer tri-arm P (NIPAAm-co-DMAEMA) and self-assembled micelles thereof, utilizes the controlled release of outside or intrinsic in body environmental change realization to medicine.Patent application 200910024899.X discloses a kind of star type block acid sensitive nano micelle, it causes D by cage modle eight oligomeric silsesquioxane, the active ring-opening polymerization of L-rac-Lactide, again with N, N-dimethylaminoethyl-methacrylic ester (DMAEMA) carries out synthesized by ATRP reaction, by realizing the controlled release to medicine to temperature and pH sensitivity.
Summary of the invention
In order to overcome the shortcoming of above-mentioned prior art with not enough, primary and foremost purpose of the present invention is to provide a kind of amphipathic pH to respond 4/6 Miktoarm star copolymer.
This star copolymer structure is: with tetramethylolmethane or Dipentaerythritol for core, connect hydrophobic group successively respectively, and pH responds group and hydrophilic radical and forms amphipathic pH and respond 4/6 assorted arm radial copolymer.
Another object of the present invention is the preparation method providing a kind of above-mentioned star-type polymer.
The method is for first with acylating agent, two or three hydroxyls of tetramethylolmethane (or Dipentaerythritol) being carried out acidylate obtains assorted functional group initiator; then carry out the ROP polymerization of caprolactone, then the ARGET ATRP polymerization adopting continuous polymerization to carry out pH response monomer and hydrophilic macromonomer successively obtains the assorted arm star polymer of amphipathic pH response 4/6.
Still a further object of the present invention is to provide above-mentioned star-type polymer to load the application in poorly water soluble drugs micellar system in preparation.
Adopting dialysis method to prepare internal layer is hydrophobic grouping, and middle layer is the nano grade polymer micella that pH responds group, shell is hydrophilic radical, and wraps up cancer therapy drug.After this nano-medicament carrier enters cell, the sour environment in tumour cell can be utilized to impel its middle pH response layer to occur protonated, thus realize the quick controllable release of medicine in tumour cell.The assorted arm radial copolymer structure of this pH response can improve the pH response sensitivity of micella under the prerequisite maintaining high drug load, and more effective Drug controlled release, improves the therapeutic efficiency of medicine.
Object of the present invention is realized by following proposal:
A kind of amphipathic pH responds 4/6 Miktoarm star copolymer, has structure shown in formula (1) or formula (2):
R
1for:
R
2for:
x=17～53，y=10～41，z=12～29。
The number-average molecular weight that described amphipathic pH responds 4/6 Miktoarm star copolymer is 30000 ~ 54000g/mol.
Described amphipathic pH responds the preparation method of 4/6 Miktoarm star copolymer, comprises following concrete steps:
(1) prepare polycaprolactone macromole evocating agent: 6-caprolactone, stannous octoate and small molecules initiator are mixed, react 24 ~ 48h at 110 ~ 140 DEG C, underpressure distillation, precipitation, filtration, drying, obtain polycaprolactone macromole evocating agent;
(2) prepare amphipathic pH and respond 4/6 arm Miktoarm star copolymer: polycaprolactone macromole evocating agent step (1) prepared, diethylaminoethyl methacrylate, hexamethyl Triethylenetetramine (TETA), cupric bromide are dissolved in toluene, add stannous octoate after stirring, at 60 ~ 90 DEG C, react 5 ~ 12h; Add methacrylic acid mono methoxy polyethylene glycol ester successive polymerization 5 ~ 12h again, remove catalyzer, filter, filtrate aftertreatment, obtains amphipathic pH and responds 4/6 Miktoarm star copolymer.
In described step (1), the parts by weight formula of reactant is as follows:
Small molecules initiator 2.35 ~ 13.43 parts
6-caprolactone 86.51 ~ 97.52 parts
Stannous octoate 0.06 ~ 0.13 part;
In described step (2), the parts by weight formula of reactant is as follows:
The consumption of described toluene is that every 1g polycaprolactone macromole evocating agent uses 2 ~ 4mL toluene.
Precipitation described in step (1) all refer to 0 DEG C of volume fraction that after rotary evaporation solution adds 10 times of volumes be 50% methanol aqueous solution precipitate.
After removal catalyzer described in step (2) refers to that reaction product is dissolved in tetrahydrofuran (THF), cross neutral alumina chromatography column, tetrahydrofuran (THF) wash-out, removing catalyzer.
Filtrate aftertreatment described in step (2) refers to filtrate to be revolved evaporation, adds precipitation in the normal hexane of 10 times of volumes, filtration, drying.
When preparing amphipathic pH and responding 4 Miktoarm star copolymer, described small molecules initiator refers to 2-isobutyl bromide pentaerythritol diester, prepared by following methods: be the tetramethylolmethane of 1:2:2 by mol ratio, 2-bromine isobutyl acylbromide, triethylamine be dissolved in tetrahydrofuran (THF) (THF), 4 ~ 6h is reacted in ice-water bath, then at room temperature 16 ~ 30h is reacted, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide pentaerythritol diester.
The consumption of described tetrahydrofuran (THF) is that every 1g tetramethylolmethane uses 20 ~ 25mL THF.
When preparing amphipathic pH and responding 6 Miktoarm star copolymer, described small molecules initiator refers to 2-isobutyl bromide Dipentaerythritol three ester, prepared by following methods: be the Dipentaerythritol of 1:3:3 by mol ratio, 2-bromine isobutyl acylbromide, triethylamine be dissolved in THF, 4 ~ 6h is reacted in ice-water bath, then at room temperature 16 ~ 30h is reacted, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide Dipentaerythritol three ester.
The consumption of described THF is that every 1g Dipentaerythritol uses 10 ~ 15mL THF.
Above-mentioned amphipathic pH responds the application of 4/6 Miktoarm star copolymer in preparation loading poorly water soluble drugs micellar system.
Described loading poorly water soluble drugs micellar system is prepared by following methods: amphipathic pH is responded 4/6 Miktoarm star copolymer and poorly water soluble drugs is dissolved in organic solvent, stirred at ambient temperature 4 ~ 6h, deionized water dialysis 24 ~ 48h, lyophilize, obtains loading poorly water soluble drugs micellar system.
Described poorly water soluble drugs refers to that solubleness is less than or equal to the medicine of 1g in 1L water.
Described organic solvent refers at least one in dimethyl formamide and dimethyl sulfoxide (DMSO).
Described loading poorly water soluble drugs micellar system can control the medicine slow releasing when healthy tissues (pH7.4) loaded, quick controllable release under tumour cell solutions of weak acidity (pH5 ~ 6).
Mechanism of the present invention is:
Amphipathic pH disclosed by the invention responds that 4/6 Miktoarm star copolymer is excellent performance, the medicine carrying material with pH susceptibility.Polycaprolactone (PCL) as the hydrophobic inner core of micella, for solubilising insoluble cancer therapy drug.Be polymerized the polymethyl acrylic acid lignocaine ethyl ester structure (PDEAEMA) obtained and respond middle layer for pH, be hydrophobic state under physiological pH (7.4), is contracted in PCL surface common as kernel, can effectively prevents burst drug release phenomenon; The amino generation protonation positively charged of side chain under tumor tissues solutions of weak acidity, easily with electronegative cytolemma generation adsorption, and then by endocytosis in cell; After entering lysosomal strong acidic environment, protonated further, micella occurs swelling, by drug release in tenuigenin and nucleus.PPEGMA is as hydrophilic outer layer, be conducive to improving micellar shell density, strengthen the wetting ability of micellar surface, anti-albumen and platelet adhesion ability, improve the stability of micella, thus extend micella cycling time in vivo, improve the controlled release properties of micelle medicine carrying system.Content by group each in telomerized polymer molecular material carrys out the rate of release of regulating medicine, meets the release request of different pharmaceutical.
The present invention has following advantage and beneficial effect relative to prior art:
(1) preparation method of the present invention is simple to operate, and reaction conditions is gentle, and be easy to the brachium that amphipathic pH responds 4/6 Miktoarm star copolymer, molecular weight is adjustable in wider scope.
(2) the amphipathic pH that the present invention prepares responds 4/6 Miktoarm star copolymer and is applied to preparation loading poorly water soluble drugs micellar system, can meet the release request of different pharmaceutical.
Accompanying drawing explanation
Fig. 1 is (PCL) in embodiment 1
2(PDEAEMA-b-PPEGMA)
2gPC elution curve.
Fig. 2 is (PCL) in embodiment 1
2(PDEAEMA-b-PPEGMA)
2's
1h NMR composes, and solvent is d-CDCl3.
Fig. 3 is (PCL) in embodiment 4
3(PDEAEMA-b-PPEGMA)
3gPC elution curve.
Fig. 4 is (PCL) in embodiment 4
3(PDEAEMA-b-PPEGMA)
3's
1h NMR composes, and solvent is d-CDCl3.
Fig. 5 is (PCL) in embodiment 8
3(PDEAEMA-b-PPEGMA)
3micelle-forming concentration test curve.
Fig. 6 is (PCL) in embodiment 9
3(PDEAEMA-b-PPEGMA)
3scanning electron microscope (SEM) figure of carrier micelle.
Fig. 7 is (PCL) in embodiment 10
2(PDEAEMA-b-PPEGMA)
2(embodiment 2 product) carries the In-vitro release curves of Zorubicin micella.
Fig. 8 is (PCL) in enforcement 10
3(PDEAEMA-b-PPEGMA)
3(embodiment 4 product) carries the In-vitro release curves of Zorubicin micella.
Fig. 9 is (PCL) in enforcement 11
3(PDEAEMA-b-PPEGMA)
3the vitro cytotoxicity of blank micella.
Figure 10 is (PCL) in enforcement 11
2(PDEAEMA-b-PPEGMA)
2(PCL)
3-(PDEAEMA-b-PPEGMA)
3carry the vitro cytotoxicity of Zorubicin micella.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1: amphipathic pH responds the preparation of 4 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take 6.08g(0.05mol in 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 10.12g(0.1mol) triethylamine (TEA, AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 4h, then at room temperature 25 DEG C stirring reaction 30h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 56%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, building-up reactions formula is shown in formula (3), obtains 2-isobutyl bromide pentaerythritol diester.
(2) polycaprolactone macromole evocating agent ((PLC)
2-Br
2) preparation: 2-isobutyl bromide pentaerythritol diester 0.43g step (1) prepared adds in flask; soft rubber ball seals; vacuumize-Tong argon gas 3 times, under argon shield, add 6g 6-caprolactone (ε-CL, Sigma-Aldrich) and 0.01g stannous octoate (Sn (Oct)
2, traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 24h is reacted in 130 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve, add the cold methanol aqueous solution that 500mL volume fraction is 50% with 50mLTHF, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder, and building-up reactions formula is shown in formula (4).Productive rate is 83%, M
n=5007, PDI=1.49.
(3) amphipathic pH responds 4 Miktoarm star copolymers ((PCL)
2(PDEAEMA-b-PPEGMA)
2) preparation: get (PLC) that 4.8g step (2) prepares
2-Br
2, cupric bromide (CuBr
2shanghai is newly precious) in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL toluene, monomer methacrylic acid lignocaine ethyl ester (DEAEMA, TCI-EP), part hexamethyl Triethylenetetramine (TETA) (HMTETA, Sigma-Aldrich) injects reaction flask, stirs 10min and catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.26g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 70 DEG C of oil baths after stirring 5min and react 7h; Reinject methacrylic acid mono methoxy polyethylene glycol ester (PEGMA, M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 7h, be cooled to room temperature, add 50mLTHF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder, and building-up reactions formula is shown in formula (5), utilizes GPC to measure its molecular weight, and carries out nmr analysis, see Fig. 1 and Fig. 2.Productive rate is 93%, M
n=28200, PDI=1.28.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 2: amphipathic pH responds the preparation of 4 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take 6.08g(0.05mol in 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 10.12g(0.1mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 6h, then at room temperature 25 DEG C stirring reaction 16h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 56%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide pentaerythritol diester.
(2) (PCL)
2-Br
2preparation: 2-isobutyl bromide pentaerythritol diester 0.6g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 9g ε-CL(Sigma-Aldrich under argon shield) and 0.01g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 24h is reacted in 140 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 96%, M
n=8633, PDI=1.53.
(3) amphipathic pH responds 4 Miktoarm star copolymers ((PCL)
2(PDEAEMA-b-PPEGMA)
2) preparation: get (PLC) that 7.2g step (2) prepares
2-Br
2, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.32g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 60 DEG C of oil baths after stirring 5min and react 5h; Reinject PEGMA(M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 98%, M
n=28524, PDI=1.43.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 3: amphipathic pH responds the preparation of 4 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take 6.08g(0.05mol in 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 10.12g(0.1mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h, then at room temperature 25 DEG C stirring reaction 24h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 56%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide pentaerythritol diester.
(2) (PCL)
2-Br
2preparation: 2-isobutyl bromide pentaerythritol diester 0.29g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 6g ε-CL(Sigma-Aldrich under argon shield) and 0.01g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 48h is reacted in 120 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 96%, M
n=5741, PDI=1.38.
(3) amphipathic pH responds 4 Miktoarm star copolymers ((PCL)
2(PDEAEMA-b-PPEGMA)
2) preparation: get (PLC) that 4.8g step (2) prepares
2-Br
2, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.16g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 90 DEG C of oil baths after stirring 5min and react 10h; Reinject PEGMA(M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 8h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 76%, M
n=40470, PDI=1.37.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 4: amphipathic pH responds the preparation of 6 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three ester: take 12.72g(0.05mol in 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 4h, then at room temperature 25 DEG C stirring reaction 30h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 43%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide Dipentaerythritol three ester.
(2) (PCL)
3-Br
3preparation: 2-isobutyl bromide Dipentaerythritol three ester 0.93g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 6g ε-CL(Sigma-Aldrich under argon shield) and 0.01g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 24h is reacted in 130 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 100%, M
n=6090, PDI=1.28.
(3) amphipathic pH responds 6 Miktoarm star copolymers ((PCL)
3(PDEAEMA-b-PPEGMA)
3) preparation: get (PLC) that 4.8g step (2) prepares
3-Br
3, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.26g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 80 DEG C of oil baths after stirring 5min and react 8h; Reinject PEGMA(M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 6h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder, utilizes GPC to measure its molecular weight, and carries out nmr analysis, see Fig. 3 and Fig. 4.Productive rate is 67%, M
n=23100, PDI=1.31.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 5: amphipathic pH responds the preparation of 6 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three ester: take 12.72g(0.05mol in 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 6h, then at room temperature 25 DEG C stirring reaction 16h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 43%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide Dipentaerythritol three ester.
(2) (PCL)
3-Br
3preparation: 2-isobutyl bromide Dipentaerythritol three ester 0.93g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 12g ε-CL(Sigma-Aldrich under argon shield) and 0.01g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 48h is reacted in 110 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 92%, M
n=11031, PDI=1.36.
(3) amphipathic pH responds 6 Miktoarm star copolymers ((PCL)
3(PDEAEMA-b-PPEGMA)
3) preparation: get (PLC) that 6.36g step (2) prepares
3-Br
3, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.16g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 60 DEG C of oil baths after stirring 5min and react 12h; Reinject PEGMA(M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 5h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 81%, M
n=39270, PDI=1.43.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 6: amphipathic pH responds the preparation of 6 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three ester: take 12.72g(0.05mol in 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h, then at room temperature 25 DEG C stirring reaction 24h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 43%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide Dipentaerythritol three ester.
(2) (PCL)
3-Br
3preparation: 2-isobutyl bromide Dipentaerythritol three ester 0.07g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 12g ε-CL(Sigma-Aldrich under argon shield) and 0.01g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 36h is reacted in 130 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 88%, M
n=10500, PDI=1.45.
(3) amphipathic pH responds 6 Miktoarm star copolymers ((PCL)
3(PDEAEMA-b-PPEGMA)
3) preparation: get (PLC) that 6.36g step (2) prepares
3-Br
3, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.32g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 90 DEG C of oil baths after stirring 5min and react 5h; Reinject PEGMA(M
n=475, Sigma-Aldrich) carry out successive polymerization, after reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 45%, M
n=33410, PDI=1.45.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 7: amphipathic pH responds the preparation of 6 Miktoarm star copolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three ester: take 12.72g(0.05mol in 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar), add the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.After sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 DEG C of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h, then at room temperature 25 DEG C stirring reaction 24h.Reaction solution is proceeded in separating funnel, add 300mL ether, and successively with sodium hydrogen carbonate solution and the extraction of 200mL water of 200mL water, 200mL0.5M, filter after organic phase anhydrous magnesium sulfate drying, solution revolves steaming and obtains white solid (productive rate is 43%), at 40 DEG C, 35mb vacuum-drying 24h after Diethyl ether recrystallization twice, obtain 2-isobutyl bromide Dipentaerythritol three ester.
(2) (PCL)
3-Br
3preparation: 2-isobutyl bromide Dipentaerythritol three ester 0.29g step (1) prepared adds in flask, and soft rubber ball seals, and vacuumizes-Tong argon gas 3 times, adds 12g ε-CL(Sigma-Aldrich under argon shield) and 0.02g Sn (Oct)
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, be placed under argon shield after 48h is reacted in 120 DEG C of oil baths and use liquid nitrogen termination reaction.After underpressure distillation, dissolve with 50mL THF, add the cold methanol aqueous solution that 500mL volume fraction is 50%, filter after precipitation, product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 86%, M
n=10376, PDI=1.59.
(3) amphipathic pH responds 6 Miktoarm star copolymers ((PCL)
3(PDEAEMA-b-PPEGMA)
3) preparation: get (PLC) that 6.36g step (2) prepares
3-Br
3, CuBr
2(Shanghai is newly precious) is in eggplant-shape bottle, sealing, vacuumize-Tong argon gas 3 times, successively by 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA is formed.By Sn (Oct)
2(0.32g) be dissolved in 2mL toluene, inject reaction flask.Proceed in 80 DEG C of oil baths after stirring 5min and react 7h; Reinject PEGMA(M
n=475, Sigma-Aldrich)) carry out successive polymerization, after reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross neutral alumina pillar, remove catalyzer, obtain joining in 500mL normal hexane after reaction solution is concentrated into about 50mL precipitating, filter.Product, at 40 DEG C, 35mb vacuum-drying 24h, obtains white powder.Productive rate is 55%, M
n=37295, PDI=1.59.
Wherein, the parts by weight formula of each reactant is as follows:
Embodiment 8: what amphipathic pH responded 6 Miktoarm star copolymers closes on micellar concentration
The amphipathic pH utilizing fluorescence probe method testing example 4 to prepare responds 6 Miktoarm star copolymers (PCL)
3-(PDEAEMA-b-PPEGMA)
3close on micellar concentration.
(1) configuration of pyrene solution: with acetone solution pyrene (Sigma-Aldrich), configuration concentration is 12 × 10
-5the pyrene solution of M.
(2) configuration of sample solution: take 10mg (PCL)
3(PDEAEMA-b-PPEGMA)
3be dissolved in 5mL acetone, dropwise join in 100mL deionized water, obtain the solution of 0.1mg/mL after volatilization acetone, be diluted to a series of concentration (concentration range is 0.0001 ~ 0.1mg/mL).Get 20 10mL volumetric flasks, add the pyrene solution that 0.1mL step (1) configures respectively, then add the copolymer solution constant volume of above-mentioned different concns respectively, shake up, obtain sample solution.In sample solution, the concentration of pyrene is 12 × 10
-7m.
(3) fluorescence spectrum test: using 373nm as emission wavelength, test sample solution, at the excitation spectrum of 300 ~ 350nm, gets the intensity rate (I that wavelength is 339nm and 336nm
339/ I
336) log concentration logC is mapped, see Fig. 5, curve break is micelle-forming concentration value.Record (PCL) that embodiment 4 prepares
3-(PDEAEMA-b-PPEGMA)
3the micellar concentration that closes on be 3.4mg/L.
Embodiment 9: amphipathic pH responds the preparation of 6 Miktoarm star copolymer carrier micelles
Adopt dialysis method to prepare carrier micelle: to take 20mg Zorubicin (DOX, Beijing Hua Fenglianbo) and be dissolved in 20mL dimethyl formamide (DMF), add the TEA20 μ L of 2 times of molar weights, stir 12h.Take the amphipathic pH that 40mg embodiment 4 prepares and respond 6 Miktoarm star copolymers (PCL)
3(PDEAEMA-b-PPEGMA)
3be dissolved in 20mL dimethyl sulfoxide (DMSO) (DMSO), by two kinds of solution mixing, stir 4h, the every 4h of deionized water dialysis 24h, front 12h changes a water, and 6h changes a water subsequently.Freeze-drying after 0.45m filtering head filters, namely obtains DOX carrier micelle powder.It is spherical for adopting SEM to observe its pattern, and particle size range is 100 ~ 180nm, sees Fig. 6.
Embodiment 10: the release in vitro of carrier micelle
According to embodiment 9 preparation method, the amphipathic pH utilizing embodiment 2 to prepare responds 4 Miktoarm star copolymers (PCL)
2(PDEAEMA-b-PPEGMA)
2prepare DOX carrier micelle I.Embodiment 9 prepares DOX carrier micelle II.
Extracorporeal releasing test: take 5mg DOX carrier micelle I respectively and DOX carrier micelle II is scattered in (acetate buffer and phosphate buffered saline buffer) in 5mL damping fluid, pH of cushioning fluid is respectively 5.0,6.5 and 7.4.Above-mentioned being scattered in is placed in dialysis tubing, proceeds in the damping fluid of 40mL same pH, be placed in medicament dissolution instrument, at 37 DEG C, under 110rpm rotating speed, carry out release in vitro.Timing sampling 3mL carries out ultra-violet analysis, and adds 3mL fresh buffer simultaneously.Release DOX concentration in fragrant liquid with determined by ultraviolet spectrophotometry different time, draw In-vitro release curves, see Fig. 7 and Fig. 8.
As seen from Figure 7, under the pH7.4 environment of healthy tissues, slowly, the cumulative release amount of 24h only has the cumulative release amount of about 22%, 96h to be 35% to the rate of release of DOX.When pH is reduced to condition near 6.5(and tumor tissues) time, the rate of release of DOX is accelerated, and the cumulative release amount of 24h and 96h adds 5% and about 10% respectively compared with pH7.4.And under the pH5.0 environment of tumour cell endosome, the rate of release of DOX is obviously accelerated, 24h cumulative release amount is increased to 50%, 48h and reaches 62%, 96h and release 91%.
In like manner, as seen from Figure 8, under the pH7.4 environment of healthy tissues, slowly, the cumulative release amount of 24h only has the cumulative release amount of about 25%, 96h to be 38% to the rate of release of DOX.When pH is reduced to condition near 6.5(and tumor tissues) time, the rate of release of DOX is accelerated, and the cumulative release amount of 24h and 96h both increases about 10% compared with pH7.4.And under the pH5.0 environment of tumour cell endosome, the rate of release of DOX is obviously accelerated, 24h cumulative release amount is increased to 65%, 48h and reaches 80%, 96h and release 96%.
Embodiment 11: amphipathic pH responds the cytotoxicity test of 6 Miktoarm star copolymer micellas
(1) blank micella preparation: according to the method for embodiment 9, do not add Zorubicin, prepare the blank micella of embodiment 4 multipolymer.
(2) toxotest: get 96 hole flat bottomed tissue culture plates, will add 200 μ L cell culture mediums (DMEM) respectively as blank group in surrounding orifice plate.Middle Zhong Mei hole, 60 holes is with 1x10
4cell concn inoculation HepG2 cell (buying in ATCC) of cells/well (200 μ L), wherein 96 orifice plates in contrast, are placed into 37 DEG C, saturated humidity, 5%CO by the 2nd row
248h is cultivated in incubator.
Subsequently free Zorubicin, blank micella, DOX carrier micelle powder I and DOX carrier micelle powder II DMEM is diluted to different polymer concentrations (blank micella 1 ~ 400mg/L) or drug level (free Zorubicin or carrier micelle, 0.1 ~ 20mg/L).Remove in 96 orifice plates from the 2nd row to the 11st row institute porose cell culture medium after, the 2nd arrange in add fresh developing medium, in contrast.From the 3rd row to the 10th row, in all holes, add the sample solution of 200 μ L respectively, the sample of each concentration joins in 6 holes and carries out repetition.
After the cultivation of 48h, siphon away all containing the supernatant liquor in the hole of cell, add the PBS rinse cell of 200 μ L, then siphon away PBS.From the 2nd row to the 11st row, respectively to the developing medium of the MTT solution and 180 μ L that add 20 μ L in each hole, then 96 orifice plates are positioned in incubator and cultivate 4h.Siphon away unreduced MTT solution and developing medium subsequently.Each hole PBS of 200 μ L washes one time, and siphons away PBS.The DMSO adding 200 μ L in each hole dissolves MTT crystallization.Whole 96 orifice plates are placed in 37 DEG C of shaking tables the 10min that vibrates, and then utilize microplate reader to measure the absorbancy in each hole, 490nm place, and then calculate cell survival rate.
Fig. 9 is the toxicity data of blank micella (embodiment 4 product), as we know from the figure, and (PCL)
3(PDEAEMA-b-PPEGMA)
3substantially nontoxic to HepG2 cell, under the high density of 400mg/L, cell survival rate is still up to 90%.
Figure 10 is the cytotoxicity result of free DOX, DOX carrier micelle I and DOX carrier micelle II.As we know from the figure, after 48h cultivates, the carrier micelle of lower concentration (0.1mg/L) just can kill cell, has obvious enhancement; When high density (20mg/L), the effect that carrier micelle and free DOX kill cell is similar, has the cell more than 80% to be killed.DOX carrier micelle I prepared by embodiment 4 product has close cytotoxicity with free Zorubicin, illustrates that Zorubicin can effectively keep its antitumour activity through Bao Zaihou.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (9)
1. amphipathic pH responds 4/6 Miktoarm star copolymer, it is characterized in that having structure shown in formula (1) or formula (2):
R
1for:
R
2for:
x＝17～53，y＝10～41，z＝12～29。
2. amphipathic pH according to claim 1 responds 4/6 Miktoarm star copolymer, it is characterized in that: the number-average molecular weight that described amphipathic pH responds 4/6 Miktoarm star copolymer is 30000 ~ 54000g/mol.
3. according to claim 1 or 2, amphipathic pH responds a preparation method for 4/6 Miktoarm star copolymer, it is characterized in that comprising following concrete steps:
(1) prepare polycaprolactone macromole evocating agent: 6-caprolactone, stannous octoate and small molecules initiator are mixed, react 24 ~ 48h at 110 ~ 140 DEG C, underpressure distillation, precipitation, filtration, drying, obtain polycaprolactone macromole evocating agent;
(2) prepare amphipathic pH and respond 4/6 arm Miktoarm star copolymer: polycaprolactone macromole evocating agent step (1) prepared, diethylaminoethyl methacrylate, hexamethyl Triethylenetetramine (TETA), cupric bromide are dissolved in toluene, add stannous octoate after stirring, at 60 ~ 90 DEG C, react 5 ~ 12h; Add methacrylic acid mono methoxy polyethylene glycol ester successive polymerization 5 ~ 12h again, remove catalyzer, filter, filtrate aftertreatment, obtains amphipathic pH and responds 4/6 Miktoarm star copolymer;
When preparing amphipathic pH and responding 4 Miktoarm star copolymer, described small molecules initiator refers to 2-isobutyl bromide pentaerythritol diester, prepared by following methods: be the tetramethylolmethane of 1:2:2 by mol ratio, 2-bromine isobutyl acylbromide, triethylamine be dissolved in tetrahydrofuran (THF), 4 ~ 6h is reacted in ice-water bath, then at room temperature 16 ~ 30h is reacted, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide pentaerythritol diester;
When preparing amphipathic pH and responding 6 Miktoarm star copolymer, described small molecules initiator refers to 2-isobutyl bromide Dipentaerythritol three ester, prepared by following methods: be the Dipentaerythritol of 1:3:3 by mol ratio, 2-bromine isobutyl acylbromide, triethylamine be dissolved in tetrahydrofuran (THF), 4 ~ 6h is reacted in ice-water bath, then at room temperature 16 ~ 30h is reacted, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide Dipentaerythritol three ester.
4. amphipathic pH according to claim 3 responds the preparation method of 4/6 Miktoarm star copolymer, it is characterized in that: in step (1), the parts by weight formula of reactant is as follows:
Small molecules initiator 2.35 ~ 13.43 parts
6-caprolactone 86.51 ~ 97.52 parts
Stannous octoate 0.06 ~ 0.13 part;
In step (2), the parts by weight formula of reactant is as follows:
The consumption of described toluene is that every 1g polycaprolactone macromole evocating agent uses 2 ~ 4mL toluene.
5. amphipathic pH according to claim 3 responds the preparation method of 4/6 Miktoarm star copolymer, it is characterized in that: the precipitation described in step (1) all refer to 0 DEG C of volume fraction that after rotary evaporation solution adds 10 times of volumes be 50% methanol aqueous solution precipitate; After removal catalyzer described in step (2) refers to that reaction product is dissolved in tetrahydrofuran (THF), cross neutral alumina chromatography column, tetrahydrofuran (THF) wash-out, removing catalyzer; Filtrate aftertreatment described in step (2) refers to filtrate to be revolved evaporation, adds precipitation in the normal hexane of 10 times of volumes, filtration, drying.
6. amphipathic pH according to claim 5 responds the preparation method of 4/6 Miktoarm star copolymer, it is characterized in that: when preparing amphipathic pH and responding 4 Miktoarm star copolymer, the consumption of described tetrahydrofuran (THF) is that every 1g tetramethylolmethane uses 20 ~ 25mL tetrahydrofuran (THF); When preparing amphipathic pH and responding 6 Miktoarm star copolymer, the consumption of described tetrahydrofuran (THF) is that every 1g Dipentaerythritol uses 10 ~ 15mL tetrahydrofuran (THF).
7. amphipathic pH according to claim 1 and 2 responds the application of 4/6 Miktoarm star copolymer in preparation loading poorly water soluble drugs micellar system.
8. amphipathic pH according to claim 7 responds the application of 4/6 Miktoarm star copolymer in preparation loading poorly water soluble drugs micellar system, it is characterized in that described loading poorly water soluble drugs micellar system is prepared by following methods: amphipathic pH is responded 4/6 Miktoarm star copolymer and poorly water soluble drugs is dissolved in organic solvent, stirred at ambient temperature 4 ~ 6h, deionized water dialysis 24 ~ 48h, lyophilize, obtains loading poorly water soluble drugs micellar system.
9. amphipathic pH according to claim 8 responds the application of 4/6 Miktoarm star copolymer in preparation loading poorly water soluble drugs micellar system, it is characterized in that: described poorly water soluble drugs refers to that solubleness is less than or equal to the medicine of 1g in 1L water; Described organic solvent refers at least one in dimethyl formamide and dimethyl sulfoxide (DMSO).
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| CN105440229B (en) * | 2015-12-17 | 2018-05-15 | 华南理工大学 | A kind of amphipathic copolymer of pH/ temperature Dual Sensitive and its preparation and application |
| CN105461750B (en) * | 2015-12-22 | 2017-05-24 | 华东师范大学 | PH sensitive phospholipid molecule and preparation method as well as application thereof |
| CN110227165A (en) * | 2018-03-06 | 2019-09-13 | 西南大学 | A kind of amphiphilic star-like camptothecin polymeric object prodrug and preparation method thereof |
| CN108359088B (en) * | 2018-04-10 | 2020-05-19 | 长春工业大学 | Preparation method of star-shaped polycaprolactone-resveratrol polymer |
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| CN109134870B (en) * | 2018-07-05 | 2021-05-25 | 广东工业大学 | pH response polymer carrier, micelle prepared from pH response polymer carrier, preparation method and application |
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| CN110772641A (en) * | 2019-11-29 | 2020-02-11 | 广州贝奥吉因生物科技股份有限公司 | Drug carrier and preparation method and application thereof |
| CN111635496B (en) * | 2020-05-13 | 2022-11-04 | 洛阳理工学院 | Temperature-sensitive cell culture plate and preparation method thereof |
| CN114258913B (en) * | 2021-12-13 | 2023-04-25 | 江苏耘农化工有限公司 | Temperature-sensitive controlled-release nano pesticide vesicle and preparation method and application thereof |
| CN114605600B (en) * | 2022-01-18 | 2023-01-06 | 华南理工大学 | Esterase-response amphiphilic linear polymer and preparation method and application thereof |
| CN115745747B (en) * | 2022-11-02 | 2024-01-19 | 香港中文大学（深圳） | Three-arm star-type organic spin molecular initiator, homopolymer, block copolymer, preparation method thereof and polymer film |
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| CN102702453A (en) * | 2012-06-11 | 2012-10-03 | 华南理工大学 | pH-responsive 6-arm star block copolymer and preparation method and application thereof |
| CN102702454A (en) * | 2012-06-11 | 2012-10-03 | 华南理工大学 | PH response four-arm star block copolymer and preparation method and application thereof |
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| CN102702454A (en) * | 2012-06-11 | 2012-10-03 | 华南理工大学 | PH response four-arm star block copolymer and preparation method and application thereof |
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| Synthesis, Characterization and pH-Responsive Self-Assembly Behavior of Amphiphilic Multiarm Star Triblock Copolymers Based on PCL,PDEAEMA, and PEG;You Qiang Yang et al.;《Macromolecular Research》;20131231;第21卷(第9期);1011-1020页 * |
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