CN103333301A - 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 a kind of amphipathic pH response 4/6 assorted arm star multipolymer and preparation method thereof.
Background technology
Cancer is 21 centurys three one of big diseases, and human beings'health in serious harm.The global cancer report of the World Health Organization shows: the annual life that seizes more than 700 ten thousand people in the whole world at present, along with world population aging day by day, estimate whole world cancer mortality number also with fast rise, and may surpass 1,310 ten thousand to the year two thousand thirty.Because the heterogeneity of cancer cell, multidrug resistance etc., the treatment of cancer medicine still is faced with great challenge.As resisting one of most important instrument of cancer, chemotherapy plays important effect for treatment for cancer.Desirable chemotherapeutics should play a role by a cancer cells to diseased region, and can or not lack normal cell is produced toxic side effect.In order to reduce toxic side effect, improve the availability of medicine, reduce degraded and the loss of medicine, people develop various pharmaceutical carrier systems in recent years energetically.These pharmaceutical 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 seek suitable pharmaceutical carrier, people have carried out a large amount of research to various carrier systems such as polymer micelle, hydrogel, vesica, microballoon, liposome, microemulsion etc.Wherein, polymer micelle becomes the focus of present research with its peculiar excellent properties.
The biodegradable amphiphilic polymkeric substance forms the micella of core/shell structure by self-assembly in the aqueous solution, it more and more is subjected to various countries investigator's common concern as anti-cancer medicament carrier.It has the following advantages: (1) polymericular weight is big, can make medicine stop the long period at lesions position when using as carrier; The permeability cell of tumor locus is in hyperfunction state, and the time that cancer therapy drug exists in blood circulation is more long, and the chance that enters tumor locus is just more many; (2) medicine can reach the purpose of slowly-releasing or controllable release by the degraded of diffusion or polymkeric substance self in polymer micelle; (3) can be attached to the polymer beads sub-surface to some mode of functional component by chemical bonding with targeting or control drug release; (4) biodegradable polymeric, can avoid drug release after solid support material in the human organ tissue, build up and produce toxic side effect.
Star-type polymer refers to draw at least in the nuclear polymkeric substance of three polymeric arms.Generally be divided into two big classes: a class is the star-type polymer (AB) that every arm all has identical chemical structure, may be homopolymer, multipolymer or triblock copolymer on every arm; Another kind of is at least two assorted arm star polymkeric substance that the arm chemical structure is inequality, and common have an A
2B
2, A
3B
3, A
2B, A
3B, ABC, A
5B, AB
2C
2, ABCD etc.Its unique chemical structure of star polymer makes it have some special physicochemical performances, and is little as 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, the control that star polymer is highly suitable for cancer therapy drug discharges.
As typical case's representative of branch-shape polymer, star polymer is mainly synthetic by nuclear method (arm-first) mode after examining postbrachium method (core-first) or first arm earlier.Nuclear postbrachium method needs earlier synthetic multi-functional initiator earlier, and the brachium unanimity of synthetic star-type polymer and arm number are determined, can accurately be controlled the polymerization degree.The nuclear method needs the active linear macromolecule of earlier synthetic simple function behind elder generation's arm, and the brachium unanimity of synthetic star-type polymer but arm number are difficult to determine.
The star block copolymer of having reported at present has spherical, tubulose, vesica shape, vermiform and composite-like etc. through the form of the formed aggregate of self-assembly, thereby promoted the potential application of star block copolymer at numerous areas, as medicament slow release, separation, photoelectric material and catalysis etc.Aspect medicament slow release, (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 polycaprolactones-b-polymethyl acrylic acid lignocaine ethyl ester (S (PCL-b-PDEAEMA)) to prepare a kind of mixed micelle, effectively improve the stability of micella.Behind its bag year nifedipine, utilize the PDEAEMA hydrophilic and hydrophobic to control the rate of release of nifedipine with the variation of pH value.(Biomacromolecules such as Cao, 2011,12 (7): 2697.) having prepared a kind of is kernel with polylactide (PLLA), 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 has announced a kind of preparation method who prepares wetting ability two blocks, three blocks and star-type polymer; under protection of inert gas, got by at least a vinyl monomer, macromole degradability chain-transfer agent and initiator polymerization preparation.A kind of three arm star multipolymer tri-arm P (NIPAAm-co-DMAEMA) and self-assembled micelles thereof have been announced in patent application 200810107054.2, utilize intrinsic environmental change in outside or the body to realize controlled release to medicine.Patent application 200910024899.X has announced a kind of star type block acid sensitive nano micelle, it is to cause D by cage modle eight poly-silicious sesquioxanes, the active ring-opening polymerization of L-rac-Lactide, again with N, N-dimethylaminoethyl-methacrylic ester (DMAEMA) carries out that ATRP reaction synthesize, by to temperature and the responsive realization of the pH controlled release to medicine.
Summary of the invention
For the shortcoming and deficiency that overcomes above-mentioned prior art, primary and foremost purpose of the present invention is to provide a kind of amphipathic pH response 4/6 assorted arm star multipolymer.
This star copolymer structure is: be nuclear with tetramethylolmethane or Dipentaerythritol, connect hydrophobic group respectively successively, pH response group and hydrophilic radical form amphipathic pH response 4/6 assorted arm radial copolymer.
Another purpose of the present invention is to provide a kind of preparation method of above-mentioned star-type polymer.
This method obtains assorted functional group initiator for earlier with acylating agent two or three hydroxyls of tetramethylolmethane (or Dipentaerythritol) being carried out acidylate; carry out the ROP polymerization of caprolactone then, the ARGET ATRP polymerization of adopting continuous polymerization to carry out pH response monomer and hydrophilic macromonomer successively again makes amphipathic pH response 4/6 assorted arm star polymer.
Still a further object of the present invention is to provide the application of above-mentioned star-type polymer in preparation loading poorly water soluble drugs micellar system.
Adopting dialysis method to prepare internal layer is hydrophobic grouping, and the middle layer is that pH response group, shell are the nano grade polymer micella of hydrophilic radical, and the parcel cancer therapy drug.After this nano-medicament carrier enters cell, can utilize the sour environment in the tumour cell to impel its middle pH response layer generation protonated, thereby realize the fast controllable release of medicine in tumour cell.The assorted arm radial copolymer structure of this pH response can kept the pH response sensitivity that improves micella under the prerequisite of high drug load, more effective control drug release, the therapeutic efficiency of raising medicine.
Purpose of the present invention realizes by following proposal:
A kind of amphipathic pH response 4/6 assorted arm star multipolymer has structure shown in formula (1) or the formula (2):
R
1For:
R
2For:
x=17～53，y=10～41，z=12～29。
The number-average molecular weight of described amphipathic pH response 4/6 assorted arm star multipolymer is 30000～54000g/mol.
The preparation method of described amphipathic pH response 4/6 assorted arm star multipolymer comprises following concrete steps:
(1) preparation polycaprolactone macromole evocating agent: 6-caprolactone, stannous octoate and small molecules initiator are mixed, and 110～140 ℃ are reacted 24～48h down, and underpressure distillation, precipitation, filtration, drying obtain the polycaprolactone macromole evocating agent;
(2) the amphipathic pH of preparation responds the assorted arm star multipolymer of 4/6 arm: polycaprolactone macromole evocating agent, diethylaminoethyl methacrylate, hexamethyl Triethylenetetramine (TETA), the cupric bromide of step (1) preparation are dissolved in the toluene, stir the back and add stannous octoate, 60～90 ℃ are reacted 5～12h down; Add methacrylic acid mono methoxy polyethylene glycol ester successive polymerization 5～12h again, remove catalyzer, filter, the filtrate aftertreatment obtains amphipathic pH response 4/6 assorted arm star multipolymer.
The parts by weight of reactant prescription is as follows in the described step (1):
2.35～13.43 parts of small molecules initiators
86.51～97.52 parts of 6-caprolactones
0.06～0.13 part of stannous octoate;
The parts by weight of reactant prescription is as follows in the described step (2):
The consumption of described toluene is that every 1g polycaprolactone macromole evocating agent uses 2～4mL toluene.
The described precipitation of step (1) refers to that all 0 ℃ of volume fraction that behind rotary evaporation solution adds 10 times of volumes is that 50% methanol aqueous solution precipitates.
After the described removal catalyzer of step (2) refers to reaction product is dissolved in tetrahydrofuran (THF), cross the neutral alumina chromatography column, the tetrahydrofuran (THF) wash-out is removed catalyzer.
The described filtrate aftertreatment of step (2) refers to filtrate revolved precipitation, filtration, drying in the normal hexane that steams evaporation, adds 10 times of volumes.
When the arm star multipolymer is mixed in the amphipathic pH response 4 of preparation, described small molecules initiator refers to 2-isobutyl bromide pentaerythritol diester, prepared by following method: be that tetramethylolmethane, 2-bromine isobutyl acylbromide, the triethylamine of 1:2:2 is dissolved in the tetrahydrofuran (THF) (THF) with mol ratio, in ice-water bath, react 4～6h, at room temperature react 16～30h then, 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 the arm star multipolymer is mixed in the amphipathic pH response 6 of preparation, described small molecules initiator refers to 2-isobutyl bromide Dipentaerythritol three esters, prepared by following method: be that Dipentaerythritol, 2-bromine isobutyl acylbromide, the triethylamine of 1:3:3 is dissolved among the THF with mol ratio, in ice-water bath, react 4～6h, at room temperature react 16～30h then, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide Dipentaerythritol three esters.
The consumption of described THF is that every 1g Dipentaerythritol uses 10～15mL THF.
The application of above-mentioned amphipathic pH response 4/6 assorted arm star multipolymer in preparation loading poorly water soluble drugs micellar system.
Described loading poorly water soluble drugs micellar system is prepared by following method: amphipathic pH response 4/6 assorted arm star multipolymer and poorly water soluble drugs are dissolved in the organic solvent, stir 4～6h under the room temperature, deionized water dialysis 24～48h, lyophilize obtains loading the poorly water soluble drugs micellar system.
Described poorly water soluble drugs refers to that in 1L water solubleness is less than or equal to the medicine of 1g.
Described organic solvent refers at least a in dimethyl formamide and the dimethyl sulfoxide (DMSO).
The medicine that described loading poorly water soluble drugs micellar system can be controlled loading slowly discharges when healthy tissues (pH7.4), in tumour cell solutions of weak acidity (pH5～6) fast controllable release down.
Mechanism of the present invention is:
The medicine carrying material that it is excellent performance that amphipathic pH disclosed by the invention responds 4/6 assorted arm star multipolymer, have pH susceptibility.Polycaprolactone (PCL) is used for solubilising insoluble cancer therapy drug as the hydrophobic inner core of micella.The polymethyl acrylic acid lignocaine ethyl ester structure (PDEAEMA) that polymerization obtains is hydrophobic state for pH response middle layer under physiological pH (7.4), is contracted in the PCL surface jointly as kernel, can effectively prevent the prominent phenomenon of releasing of medicine; The amino generation protonation positively charged of side chain under the tumor tissues solutions of weak acidity, easily with electronegative cytolemma generation adsorption, and then by endocytosis in cell; After entering lysosomal strong acidic environment, further protonated, micella generation swelling, with drug release in tenuigenin and nucleus.PPEGMA is as hydrophilic outer layer, be conducive to improve micella shell density, strengthen wetting ability, anti-albumen and the thrombocyte adsorptive power of micellar surface, improve the stability of micella, thereby prolong micella cycling time in vivo, improve the controlled release properties of micelle medicine carrying system.Can come the rate of release of regulating medicine by the content of each group in the telomerized polymer molecular material, satisfy the release request of different pharmaceutical.
The present invention has following advantage and beneficial effect with respect to prior art:
(1) preparation method of the present invention is simple to operate, and the reaction conditions gentleness is easy to the brachium that amphipathic pH responds 4/6 assorted arm star multipolymer, and molecular weight is adjustable in wideer scope.
(2) the amphipathic pH response 4/6 assorted arm star multipolymer for preparing of the present invention is applied to preparation and loads the poorly water soluble drugs micellar system, can satisfy the release request of different pharmaceutical.
Description of drawings
Fig. 1 is (PCL) among the embodiment 1
2(PDEAEMA-b-PPEGMA)
2The GPC elution curve.
Fig. 2 is (PCL) among the embodiment 1
2(PDEAEMA-b-PPEGMA)
2 1H NMR spectrum, solvent is d-CDCl3.
Fig. 3 is (PCL) among the embodiment 4
3(PDEAEMA-b-PPEGMA)
3The GPC elution curve.
Fig. 4 is (PCL) among the embodiment 4
3(PDEAEMA-b-PPEGMA)
3 1H NMR spectrum, solvent is d-CDCl3.
Fig. 5 is (PCL) among the embodiment 8
3(PDEAEMA-b-PPEGMA)
3The micelle-forming concentration test curve.
Fig. 6 is (PCL) among the embodiment 9
3(PDEAEMA-b-PPEGMA)
3The scanning electron microscope of carrier micelle (SEM) figure.
Fig. 7 is (PCL) among the embodiment 10
2(PDEAEMA-b-PPEGMA)
2(embodiment 2 products) carry the release in vitro curve of Zorubicin micella.
Fig. 8 is for implementing in 10 (PCL)
3(PDEAEMA-b-PPEGMA)
3(embodiment 4 products) carry the release in vitro curve of Zorubicin micella.
Fig. 9 is for implementing in 11 (PCL)
3(PDEAEMA-b-PPEGMA)
3The vitro cytotoxicity of blank micella.
Figure 10 is for implementing in 11 (PCL)
2(PDEAEMA-b-PPEGMA)
2(PCL)
3-(PDEAEMA-b-PPEGMA)
3Carry the vitro cytotoxicity of Zorubicin micella.
Embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
Embodiment 1: the preparation of amphipathic pH response 4 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take by weighing 6.08g(0.05mol in the 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 10.12g(0.1mol) triethylamine (Jiangsu is China forever for TEA, AR).In 0 ℃ of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 4h is again at 25 ℃ of following stirring reaction 30h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 56%), twice back of ether recrystallization is at 40 ℃, 35mb vacuum-drying 24h, and the building-up reactions formula is seen formula (3), obtains 2-isobutyl bromide pentaerythritol diester.
(2) polycaprolactone macromole evocating agent ((PLC)
2-Br
2) preparation: the 2-isobutyl bromide pentaerythritol diester that 0.43g step (1) is prepared adds in the flask; the soft rubber ball sealing; vacuumize-Tong argon gas 3 times, add under the argon shield 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, use the liquid nitrogen termination reaction after under argon shield, placing 130 ℃ of oil baths reaction 24h.After the underpressure distillation, with the 50mLTHF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h, and the building-up reactions formula is seen formula (4).Productive rate is 83%, M
n=5007, PDI=1.49.
(3) amphipathic pH response 4 assorted arm star multipolymers ((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 with 18mL toluene, monomer diethylaminoethyl methacrylate (DEAEMA, TCI-EP), (HMTETA Sigma-Aldrich) injects reaction flask to part hexamethyl Triethylenetetramine (TETA), stirs 10min catalyst complexes Cu/HMTETA is formed.With Sn (Oct)
2(0.26g) be dissolved in the 2mL toluene, inject reaction flask.Change in 70 ℃ of oil baths behind the stirring 5min and react 7h; Methacrylic acid mono methoxy polyethylene glycol ester (PEGMA, M reinject
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 7h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h, and the building-up reactions formula is seen formula (5), utilizes GPC to measure its molecular weight, and carries out nmr analysis, sees Fig. 1 and Fig. 2.Productive rate is 93%, M
n=28200, PDI=1.28.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 2: the preparation of amphipathic pH response 4 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take by weighing 6.08g(0.05mol in the 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 10.12g(0.1mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 6h is again at 25 ℃ of following stirring reaction 16h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 56%), twice back of ether recrystallization obtains 2-isobutyl bromide pentaerythritol diester at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
2-Br
2Preparation: the 2-isobutyl bromide pentaerythritol diester that 0.6g step (1) is prepared adds in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, adds 9g ε-CL(Sigma-Aldrich) and 0.01g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 140 ℃ of oil baths reaction 24h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 96%, M
n=8633, PDI=1.53.
(3) amphipathic pH response 4 assorted arm star multipolymers ((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 with 18mL toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.32g) be dissolved in the 2mL toluene, inject reaction flask.Change in 60 ℃ of oil baths behind the stirring 5min and react 5h; PEGMA(M reinjects
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 98%, M
n=28524, PDI=1.43.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 3: the preparation of amphipathic pH response 4 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide pentaerythritol diester: take by weighing 6.08g(0.05mol in the 250mL there-necked flask) tetramethylolmethane (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 10.12g(0.1mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 22.98g(0.1mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h is again at 25 ℃ of following stirring reaction 24h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 56%), twice back of ether recrystallization obtains 2-isobutyl bromide pentaerythritol diester at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
2-Br
2Preparation: the 2-isobutyl bromide pentaerythritol diester that 0.29g step (1) is prepared adds in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, adds 6g ε-CL(Sigma-Aldrich) and 0.01g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 120 ℃ of oil baths reaction 48h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 96%, M
n=5741, PDI=1.38.
(3) amphipathic pH response 4 assorted arm star multipolymers ((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 with 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.16g) be dissolved in the 2mL toluene, inject reaction flask.Change in 90 ℃ of oil baths behind the stirring 5min and react 10h; PEGMA(M reinjects
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 8h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 76%, M
n=40470, PDI=1.37.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 4: the preparation of amphipathic pH response 6 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three esters: take by weighing 12.72g(0.05mol in the 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 4h is again at 25 ℃ of following stirring reaction 30h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 43%), twice back of ether recrystallization obtains 2-isobutyl bromide Dipentaerythritol three esters at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
3-Br
3Preparation: 2-isobutyl bromide Dipentaerythritol three esters that 0.93g step (1) is prepared add in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, add 6g ε-CL(Sigma-Aldrich) and 0.01g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 130 ℃ of oil baths reaction 24h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 100%, M
n=6090, PDI=1.28.
(3) amphipathic pH response 6 assorted arm star multipolymers ((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 with 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.26g) be dissolved in the 2mL toluene, inject reaction flask.Change in 80 ℃ of oil baths behind the stirring 5min and react 8h; PEGMA(M reinjects
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 6h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h, utilizes GPC to measure its molecular weight, and carries out nmr analysis, sees Fig. 3 and Fig. 4.Productive rate is 67%, M
n=23100, PDI=1.31.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 5: the preparation of amphipathic pH response 6 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three esters: take by weighing 12.72g(0.05mol in the 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 6h is again at 25 ℃ of following stirring reaction 16h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 43%), twice back of ether recrystallization obtains 2-isobutyl bromide Dipentaerythritol three esters at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
3-Br
3Preparation: 2-isobutyl bromide Dipentaerythritol three esters that 0.93g step (1) is prepared add in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, add 12g ε-CL(Sigma-Aldrich) and 0.01g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 110 ℃ of oil baths reaction 48h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 92%, M
n=11031, PDI=1.36.
(3) amphipathic pH response 6 assorted arm star multipolymers ((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 with 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.16g) be dissolved in the 2mL toluene, inject reaction flask.Change in 60 ℃ of oil baths behind the stirring 5min and react 12h; PEGMA(M reinjects
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 5h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 81%, M
n=39270, PDI=1.43.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 6: the preparation of amphipathic pH response 6 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three esters: take by weighing 12.72g(0.05mol in the 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h is again at 25 ℃ of following stirring reaction 24h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 43%), twice back of ether recrystallization obtains 2-isobutyl bromide Dipentaerythritol three esters at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
3-Br
3Preparation: 2-isobutyl bromide Dipentaerythritol three esters that 0.07g step (1) is prepared add in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, add 12g ε-CL(Sigma-Aldrich) and 0.01g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 130 ℃ of oil baths reaction 36h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 88%, M
n=10500, PDI=1.45.
(3) amphipathic pH response 6 assorted arm star multipolymers ((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 with 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.32g) be dissolved in the 2mL toluene, inject reaction flask.Change in 90 ℃ of oil baths behind the stirring 5min and react 5h; PEGMA(M reinjects
n=475, Sigma-Aldrich) carry out successive polymerization, behind the reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 45%, M
n=33410, PDI=1.45.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 7: the preparation of amphipathic pH response 6 assorted arm star multipolymers
(1) preparation of small molecules initiator 2-isobutyl bromide Dipentaerythritol three esters: take by weighing 12.72g(0.05mol in the 250mL there-necked flask) Dipentaerythritol (Alfa-Aesar) adds the anhydrous THF of 150mL, logical argon gas 10min deoxygenation.Behind the sealed flask, inject 15.17g(0.15mol) TEA(AR, Jiangsu is China forever).In 0 ℃ of ice-water bath, dropwise inject 34.48g(0.15mol) 2-bromine isobutyl acylbromide (Alfa-Aesar), stirring reaction 5h is again at 25 ℃ of following stirring reaction 24h of room temperature.Reaction solution is changed in the separating funnel, add the 300mL ether, and use the sodium hydrogen carbonate solution of 200mL water, 200mL0.5M and 200mL water to extract successively, organic phase anhydrous magnesium sulfate drying after-filtration, solution revolves to steam and obtains white solid (productive rate is 43%), twice back of ether recrystallization obtains 2-isobutyl bromide Dipentaerythritol three esters at 40 ℃, 35mb vacuum-drying 24h.
(2) (PCL)
3-Br
3Preparation: 2-isobutyl bromide Dipentaerythritol three esters that 0.29g step (1) is prepared add in the flask, and the soft rubber ball sealing vacuumizes-Tong argon gas 3 times, add 12g ε-CL(Sigma-Aldrich) and 0.02g Sn (Oct) under the argon shield
2(traditional Chinese medicines group), with liquid nitrogen carry out three times freezing-bleed-ramp cycle after, use the liquid nitrogen termination reaction after under argon shield, placing 120 ℃ of oil baths reaction 48h.After the underpressure distillation, with 50mL THF dissolving, adding 500mL volume fraction is 50% the cold methanol aqueous solution, and post precipitation filters, and product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 86%, M
n=10376, PDI=1.59.
(3) amphipathic pH response 6 assorted arm star multipolymers ((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 with 18mL dry toluene, monomer DEAEMA(TCI-EP), part HMTETA(Sigma-Aldrich) inject reaction flask, stir 10min catalyst complexes Cu/HMTETA formed.With Sn (Oct)
2(0.32g) be dissolved in the 2mL toluene, inject reaction flask.Change in 80 ℃ of oil baths behind the stirring 5min and react 7h; PEGMA(M reinjects
n=475, Sigma-Aldrich)) carry out successive polymerization, behind the reaction 12h, be cooled to room temperature, add 50mL THF and stirring and dissolving, cross the neutral alumina pillar, remove catalyzer, obtain reaction solution and be concentrated into to join in the 500mL normal hexane behind about 50mL and precipitate filtration.Product obtains white powder at 40 ℃, 35mb vacuum-drying 24h.Productive rate is 55%, M
n=37295, PDI=1.59.
Wherein, the parts by weight of each reactant prescription is as follows:
Embodiment 8: amphipathic pH responds the micellar concentration that closes on of 6 assorted arm star multipolymers
The amphipathic pH response 6 assorted arm star multipolymers (PCL) that utilize fluorescent probe method test implementation example 4 to prepare
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 by weighing 10mg (PCL)
3(PDEAEMA-b-PPEGMA)
3Be dissolved in the 5mL acetone, dropwise join in the 100mL deionized water, obtain the solution of 0.1mg/mL behind the volatilization acetone, (concentration range is 0.0001～0.1mg/mL) to be diluted to a series of concentration.Get 20 10mL volumetric flasks, add the pyrene solution of 0.1mL step (1) configuration respectively, add the copolymer solution constant volume of above-mentioned different concns then respectively, shake up, obtain sample solution.The concentration of pyrene is 12 * 10 in the sample solution
-7M.
(3) fluorescence spectrum test: as emission wavelength, specimen solution is at the excitation spectrum of 300～350nm with 373nm, and getting wavelength is the intensity rate (I of 339nm and 336nm
339/ I
336) to concentration logarithm logC mapping, see Fig. 5, curve break is the 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: the preparation of amphipathic pH response 6 assorted arm star copolymer carrier micelles
Adopt dialysis method to prepare carrier micelle: to take by weighing 20mg Zorubicin (DOX, Beijing Hua Fenglianbo) and be dissolved in the 20mL dimethyl formamide (DMF), add the TEA20 μ L of 2 times of molar weights, stir 12h.Take by weighing the amphipathic pH response 6 assorted arm star multipolymers (PCL) that 40mg embodiment 4 prepares
3(PDEAEMA-b-PPEGMA)
3Be dissolved in the 20mL dimethyl sulfoxide (DMSO) (DMSO), two kinds of solution are mixed, stir 4h, deionized water dialysis 24h, the every 4h of preceding 12h changes water one time, and 6h changes water one time subsequently.Freeze-drying after the 0.45m filtering head filters namely obtains DOX carrier micelle powder.Adopt SEM to observe its pattern for spherical, particle size range is 100～180nm, sees Fig. 6.
Embodiment 10: the release in vitro of carrier micelle
According to embodiment 9 preparation methods, the amphipathic pH response 4 assorted arm star multipolymers (PCL) that utilize embodiment 2 to prepare
2(PDEAEMA-b-PPEGMA)
2Prepare DOX carrier micelle I.Embodiment 9 prepares DOX carrier micelle II.
Extracorporeal releasing test: take by weighing 5mg DOX carrier micelle I and DOX carrier micelle II respectively and be scattered in (acetate buffer and phosphate buffered saline buffer) in the 5mL damping fluid, the pH of buffer value is respectively 5.0,6.5 and 7.4.Above-mentioned being scattered in placed dialysis tubing, change in the damping fluid of the identical pH value of 40mL, place medicament dissolution instrument, at 37 ℃, carry out release in vitro under the 110rpm rotating speed.Timing sampling 3mL carries out ultra-violet analysis, and adds the 3mL fresh buffer simultaneously.Release DOX concentration in the fragrant liquid with the determined by ultraviolet spectrophotometry different time, draw the release in vitro curve, see Fig. 7 and Fig. 8.
As seen from Figure 7, under the pH7.4 of healthy tissues environment, the rate of release of DOX is very slow, and the cumulative release amount of 24h has only about 22%, and the cumulative release amount of 96h is 35%.Be near the condition tumor tissues when pH is reduced to 6.5() time, the rate of release of DOX is accelerated, and the cumulative release amount of 24h and 96h is compared with pH7.4 have been increased respectively about 5% and 10%.And under the pH5.0 of tumour cell endosome environment, the rate of release of DOX is obviously accelerated, and 24h cumulative release amount is increased to 50%, 48h and reaches 62%, 96h and discharged 91%.
In like manner, as seen from Figure 8, under the pH7.4 of healthy tissues environment, the rate of release of DOX is very slow, and the cumulative release amount of 24h has only about 25%, and the cumulative release amount of 96h is 38%.Be near the condition tumor tissues when pH is reduced to 6.5() time, the rate of release of DOX is accelerated, and the cumulative release amount of 24h and 96h is compared all with pH7.4 and has been increased about 10%.And under the pH5.0 of tumour cell endosome environment, the rate of release of DOX is obviously accelerated, and 24h cumulative release amount is increased to 65%, 48h and reaches 80%, 96h and discharged 96%.
Embodiment 11: the cytotoxicity test of amphipathic pH response 6 assorted arm star copolymer micelles
(1) blank micella preparation: according to the method for embodiment 9, do not add Zorubicin, prepare the blank micella of embodiment 4 multipolymers.
(2) toxotest: get the flat tissue culturing plate in 96 holes, add 200 μ L cell culture mediums (DMEM) all around in the orifice plate respectively as blank group.Every hole is with 1x10 in middle 60 holes
4The cell concn inoculation HepG2 cell (buying in ATCC) of cells/well (200 μ L), wherein the 2nd row are placed into 37 ℃ with 96 orifice plates, saturated humidity, 5%CO in contrast
2Cultivate 48h in the incubator.
Zorubicin, blank micella, DOX carrier micelle powder I and the DOX carrier micelle powder II of will dissociating subsequently is diluted to different polymer concentrations (blank micella 1～400mg/L) or drug level (free Zorubicin or carrier micelle, 0.1～20mg/L) with DMEM.Behind the cell culture medium porose from the 2nd row to the 11st row institute in removing 96 orifice plates, the fresh developing medium of adding in the 2nd row, in contrast.To the 10th row, add the sample solution of 200 μ L from the 3rd row in all holes respectively, the sample of each concentration joins in 6 holes and carries out repetition.
Through after the cultivation of 48h, siphon away the supernatant liquor in all holes of containing cell, add the PBS rinse cell of 200 μ L, siphon away PBS then.To the 11st row, in each hole, add the MTT solution of 20 μ L and the developing medium of 180 μ L from the 2nd row respectively, 96 orifice plates are positioned over cultivate 4h in the incubator then.Siphon away unreduced MTT solution and developing medium subsequently.Each hole is washed one time with the PBS of 200 μ L, and siphons away PBS.The DMSO dissolving MTT crystallization that adds 200 μ L in each hole.Whole 96 orifice plates are placed on the 10min that vibrates in 37 ℃ of shaking tables, utilize microplate reader to measure the absorbancy in each hole, 490nm place then, and then calculate cell survival rate.
Fig. 9 is the toxicity result of blank micella (embodiment 4 products), as we know from the figure, and (PCL)
3(PDEAEMA-b-PPEGMA)
3Nontoxic substantially to the HepG2 cell, cell survival rate is still up to 90% under the high density of 400mg/L.
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 cultivating through 48h, the carrier micelle of lower concentration (0.1mg/L) just can cell killing, and tangible enhancement is arranged; When high density (20mg/L), the effect of carrier micelle and free DOX cell killing is similar, has above 80% cell to be killed.DOX carrier micelle I and the free Zorubicin of the preparation of embodiment 4 products have close cytotoxicity, illustrate that Zorubicin can effectively keep its antitumour activity through Bao Zaihou.
Above-described embodiment is preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. amphipathic pH response 4/6 assorted arm star multipolymer, the structure shown in (1) or the formula (2) that it is characterized in that having formula:
R
1For:
R
2For:
x=17～53，y=10～41，z=12～29。
2. amphipathic pH according to claim 1 responds 4/6 assorted arm star multipolymer, it is characterized in that: the number-average molecular weight of described amphipathic pH response 4/6 assorted arm star multipolymer is 30000～54000g/mol.
3. preparation method according to claim 1 or 2 described amphipathic pH response 4/6 assorted arm star multipolymers is characterized in that comprising following concrete steps:
(1) preparation polycaprolactone macromole evocating agent: 6-caprolactone, stannous octoate and small molecules initiator are mixed, and 110～140 ℃ are reacted 24～48h down, and underpressure distillation, precipitation, filtration, drying obtain the polycaprolactone macromole evocating agent;
(2) the amphipathic pH of preparation responds the assorted arm star multipolymer of 4/6 arm: polycaprolactone macromole evocating agent, diethylaminoethyl methacrylate, hexamethyl Triethylenetetramine (TETA), the cupric bromide of step (1) preparation are dissolved in the toluene, stir the back and add stannous octoate, 60～90 ℃ are reacted 5～12h down; Add methacrylic acid mono methoxy polyethylene glycol ester successive polymerization 5～12h again, remove catalyzer, filter, the filtrate aftertreatment obtains amphipathic pH response 4/6 assorted arm star multipolymer.
4. amphipathic pH according to claim 3 responds the preparation method of 4/6 assorted arm star multipolymer, it is characterized in that: the parts by weight of reactant prescription is as follows in the step (1):
2.35～13.43 parts of small molecules initiators
86.51～97.52 parts of 6-caprolactones
0.06～0.13 part of stannous octoate;
The parts by weight of reactant prescription is as follows in the step (2):
The consumption of described toluene is that every 1g polycaprolactone macromole evocating agent uses 2～4mL toluene.
5. the preparation method of amphipathic pH response 4/6 assorted arm star multipolymer according to claim 3 is characterized in that: the described precipitation of step (1) refers to that all behind the rotary evaporation 0 ℃ of volume fraction of 10 times of volumes of solution adding is that 50% methanol aqueous solution precipitates; After the described removal catalyzer of step (2) refers to reaction product is dissolved in tetrahydrofuran (THF), cross the neutral alumina chromatography column, the tetrahydrofuran (THF) wash-out is removed catalyzer; The described filtrate aftertreatment of step (2) refers to filtrate revolved precipitation, filtration, drying in the normal hexane that steams evaporation, adds 10 times of volumes.
6. amphipathic pH according to claim 3 responds the preparation method of 4/6 assorted arm star multipolymer, it is characterized in that: when the arm star multipolymer is mixed in the amphipathic pH response 4 of preparation, described small molecules initiator refers to 2-isobutyl bromide pentaerythritol diester, prepared by following method: be that tetramethylolmethane, 2-bromine isobutyl acylbromide, the triethylamine of 1:2:2 is dissolved in the tetrahydrofuran (THF) with mol ratio, in ice-water bath, react 4～6h, at room temperature react 16～30h then, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide pentaerythritol diester;
When the arm star multipolymer is mixed in the amphipathic pH response 6 of preparation, described small molecules initiator refers to 2-isobutyl bromide Dipentaerythritol three esters, prepared by following method: be that Dipentaerythritol, 2-bromine isobutyl acylbromide, the triethylamine of 1:3:3 is dissolved in the tetrahydrofuran (THF) with mol ratio, in ice-water bath, react 4～6h, at room temperature react 16～30h then, through rotary evaporation, precipitation, filtration, drying, obtain 2-isobutyl bromide Dipentaerythritol three esters.
7. amphipathic pH according to claim 6 responds the preparation method of 4/6 assorted arm star multipolymer, it is characterized in that: when the arm star multipolymer was mixed in the amphipathic pH response 4 of preparation, the consumption of described tetrahydrofuran (THF) was that every 1g tetramethylolmethane uses 20～25mL tetrahydrofuran (THF); When the arm star multipolymer was mixed in the amphipathic pH response 6 of preparation, the consumption of described tetrahydrofuran (THF) was that every 1g Dipentaerythritol uses 10～15mL tetrahydrofuran (THF).
8. the application of amphipathic pH response 4/6 assorted arm star multipolymer according to claim 1 and 2 in preparation loading poorly water soluble drugs micellar system.
9. the application of amphipathic pH response 4/6 assorted arm star multipolymer according to claim 8 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 method: amphipathic pH response 4/6 assorted arm star multipolymer and poorly water soluble drugs are dissolved in the organic solvent, stir 4～6h under the room temperature, deionized water dialysis 24～48h, lyophilize obtains loading the poorly water soluble drugs micellar system.
10. amphipathic pH response 4/6 assorted arm star multipolymer according to claim 9 loads application in the poorly water soluble drugs micellar system in preparation, it is characterized in that: described poorly water soluble drugs refers to that in 1L water solubleness is less than or equal to the medicine of 1g; Described organic solvent refers at least a in dimethyl formamide and the dimethyl sulfoxide (DMSO).
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2014206044A1 (en) * | 2013-06-29 | 2014-12-31 | 华南理工大学 | Amphiphilic 4/6 miktoarm star-shaped ph-responsive copolymer and preparation method thereof |
| CN105440229A (en) * | 2015-12-17 | 2016-03-30 | 华南理工大学 | pH/temperature sensitive amphiphilic polymer, and preparation method and applications thereof |
| CN105461750A (en) * | 2015-12-22 | 2016-04-06 | 华东师范大学 | PH sensitive phospholipid molecule and preparation method as well as application thereof |
| CN108359088A (en) * | 2018-04-10 | 2018-08-03 | 长春工业大学 | A kind of preparation method of star-like polycaprolactone-resveratrol polymer |
| CN108610462A (en) * | 2018-04-23 | 2018-10-02 | 广东工业大学 | Star-like amphipathic nature polyalcohol of a kind of pH responses and preparation method thereof |
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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 |
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| CN108610462A (en) * | 2018-04-23 | 2018-10-02 | 广东工业大学 | Star-like amphipathic nature polyalcohol of a kind of pH responses and preparation method thereof |
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| WO2024093657A1 (en) * | 2022-11-02 | 2024-05-10 | 香港中文大学（深圳） | Three-arm star-shaped organic spinning molecular initiator, homopolymer, and block copolymer, preparation method therefor, and polymer film |
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| WO2014206044A1 (en) | 2014-12-31 |
| CN103333301B (en) | 2015-04-22 |
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