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CN104231155A - Cholesterol modified amphiphilic pH response pennicuius copolymer as well as preparation and micelle of copolymer - Google Patents

Cholesterol modified amphiphilic pH response pennicuius copolymer as well as preparation and micelle of copolymer Download PDF

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CN104231155A
CN104231155A CN201410410038.6A CN201410410038A CN104231155A CN 104231155 A CN104231155 A CN 104231155A CN 201410410038 A CN201410410038 A CN 201410410038A CN 104231155 A CN104231155 A CN 104231155A
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amphipathic
cholesterol
copolymer
brush copolymer
response
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CN104231155B (en
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章莉娟
姚娜
张灿阳
林文静
赵斌
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South China University of Technology SCUT
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South China University of Technology SCUT
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Abstract

The invention belongs to the technical field of preparation of biomedical high molecular polymer materials and discloses a cholesterol modified amphiphilic pH response pennicuius copolymer and a preparation method and a micelle system prepared based thereof. The copolymer has a structure shown in the formula I in the specification, wherein x is 10-36, y is 15-40 and z is 8-30. The copolymer is obtained by virtue of irregular copolymerization of a hydrophilic block hydroxyethyl methylacrylate, hydrophobic cholesterol and pH response block methacrylic acid N, N-diethyl aminoethyl combined with hydrophilic poly(ethylene glycol) methyl ether methacrylate. The copolymer is dissolved in a solvent to obtain a nanoscale micelle system, wherein the inner layer is a cholesterol modified hydrophobic chain segment, the middle layer is a pH response chain segment and the shell is a hydrophilic chain segment, so that the function of high entrapment performance, stable existence of a neutral condition and quick release in a weak acidic condition is achieved. By adjusting the proportions of the blocks in the polymer, the release rates of medicines can be regulated to satisfy the release requirements on different drugs.

Description

Cholesterol is modified amphipathic pH and is responded brush copolymer and preparation and micella thereof
Technical field
The invention belongs to biomedical polymer polymer materials preparing technical field, particularly a kind of cholesterol modifies the micellar system that amphipathic pH responds brush copolymer and preparation method and prepares based on it.
Background technology
Cancer is a kind of disease of serious threat human health, and in current existing cancer treatment method, what effect was best surely belongs to chemical medicinal treatment.But chemotherapeutics is also a double-edged sword, it has irreplaceable effect in cancer therapy on the one hand, and it is while killing cancer cells on the other hand, also can produce very large toxic side effect to normal cell.Existing anticancer class chemicals major part exist poorly water-soluble, bioavailability low, in body the defect such as non-specific distribution.
In order to overcome above-mentioned defect, while making cancer therapy drug reach effective dose, reduce the toxic side effect of normal tissue, novel delivery system (Drug Delivery System, DDS) grows up gradually.Since 20th century, the sixties found that liposome can be used in drug conveying, nanotechnology creates tremendous influence to delivery system, and the organic/inorganic nano material as pharmaceutical carrier emerges like the mushrooms after rain.Nano-medicament carrier based on polymkeric substance is a study hotspot of delivery system.This type of pharmaceutical carrier can effective prolong drug cycling time in blood compared with simple drug conveying, increases the water-soluble of dewatering medicament, reduce bad immune response, thus reach the object reducing dosage and reduce toxic side effect.A kind of desirable nano-medicament carrier should possess following character simultaneously: simple preparation and method of purification, higher drug loading and encapsulation rate, excellent biological degradability, low toxicity or nontoxic, particle diameter suitably and circulation time in vivo longer etc.
Multipolymer is the polymkeric substance formed by two or more monomer polymerization.Have at least two kinds of structural units in the structure of multipolymer, connect between structural unit with chemical bond, these are different with the blend that two kinds of polymkeric substance machineries or physical mixed are formed.The multipolymer with difference in functionality all has a wide range of applications in fields such as chemical industry, biology, medicine, buildings.
PH responsive polymer micella, as a kind of newtype drug delivery vehicles having development prospect, has many excellent performance characteristics, if realize cancer therapy drug target administration, strengthen bioavailability, reduce the toxic side effect etc. of normal tissue.But this novel intelligent nano-medicament carrier is also faced with an a series of difficult problem, as solid support material technology of preparing aspect: how to obtain size tunable and the carrier micella of narrowly distributing; How to improve the pharmaceutical pack carrying capacity etc. of carrier; Carrier micelle controlled release properties aspect: how to ensure carrier micelle structural stability under difficult environmental conditions, how precisely the small pH change of response realizes the set point control release etc. of medicine.
Biomolecule cholesterol is due to the hydrophobicity sterol backbone of its height, and good biological degradability and biocompatibility, make it become one of optimal selection of many water-soluble polymers hydrophobically modifieds.The application aspect that the self-assembly of the amphipathic nature polyalcohol of cholesterol hydrophobically modified ties up to drug delivery also receives increasing concern in recent years.Prior art has carried out more research for an above-mentioned difficult problem, carries out combination copolymerization to obtain multi-functional multipolymer to multiple polymers monomer, but still is difficult to obtain desirable effect.
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 that providing a kind of cholesterol to modify amphipathic pH responds brush copolymer.The structure of this multipolymer is: hydrophilic block hydroxyethyl methylacrylate, hydrophobic cholesterol and pH respond block methacrylic acid N, the random copolymerization of N-lignocaine ethyl ester, and then combine with hydrophilic block polyethylene glycol methacrylate-styrene polymer and obtain amphipathic pH and respond brush copolymer.
Another object of the present invention is to provide a kind of above-mentioned cholesterol to modify the preparation method of amphipathic pH response brush copolymer.The method is by adopting transfer transport activating and regenerating Transfer Radical Polymerization (ARGET ATRP), with 2-isobutyl ethyl bromide (EBriB) for small molecules initiator, cause hydroxyethyl methylacrylate (HEMA) successively, pH responds monomer methacrylic acid N, N-lignocaine ethyl ester (DEA) and polyethylene glycol methacrylate-styrene polymer (PEGMA) carry out radical polymerization, gathered (hydroxyethyl methylacrylate-co-methacrylic acid N, N-lignocaine ethyl ester)-b-gathers polyethylene glycol methacrylate-styrene polymer (P (HEMA-co-DEA)-b-PPEGMA).Then, carry out alcoholysis reaction with cholesterol formyl chloride (Chol) and this polymkeric substance to prepare cholesterol and modifies and gather (hydroxyethyl methylacrylate-co-methacrylic acid N, N-lignocaine ethyl ester)-b-and gather polyethylene glycol methacrylate-styrene polymer (Chol-P (HEMA-co-DEA)-b-PPEGMA).
Still a further object of the present invention is that providing a kind of modifies based on above-mentioned cholesterol the micellar system that amphipathic pH responds brush copolymer.Modify amphipathic pH based on above-mentioned cholesterol and respond brush copolymer, to adopt dialysis method to prepare internal layer be hydrophobic segment, the middle layer that cholesterol is modified is the nano grade polymer micella that pH responds segment, shell is hydrophilic segment, realizes the loading to slightly water-soluble cancer therapy drug.Under tumor tissues solutions of weak acidity (pH 5 ~ 6), pH responds group generation protonation and shows as wetting ability, and micella dissociates, thus realizes quick, the controllable release that are loaded medicine.This cholesterol modifies amphipathic pH response brush copolymer structure under the prerequisite maintaining high drug load, can improve the pH response sensitivity of micella, more effective Drug controlled release, improves the therapeutic efficiency of medicine.
Another object of the present invention is that providing above-mentioned cholesterol to modify amphipathic pH responds the application of brush copolymer in loading poorly water soluble drugs.
Another object of the present invention is to provide above-mentioned micellar system loading the application in poorly water soluble drugs.
Object of the present invention is realized by following proposal:
A kind of cholesterol is modified amphipathic pH and is responded brush copolymer, has as shown in the formula structure shown in I:
Wherein, x=10 ~ 36, y=15 ~ 40, z=8 ~ 30.
It is Chol-P (HEMA-co-DEA)-b-PPEGMA that cholesterol of the present invention modifies amphipathic pH response brush copolymer.
Preferably, cholesterol of the present invention modifies the number-average molecular weight of amphipathic pH response brush copolymer is 11891 ~ 40460g/mol.
The invention provides a kind of described cholesterol and modify the preparation method that amphipathic pH responds brush copolymer, comprise following concrete steps:
(1) by Catalysts Cu Br 2, monomers hydroxy ethyl ester (HEMA) and methacrylic acid N, N-lignocaine ethyl ester (DEA), ligand 1,1,4,7,10,10-hexamethyl triethylene tetramine (HMTETA) is dissolved in solvent, then adds reductive agent Sn (Oct) 2initiator 2-isobutyl ethyl bromide (EBriB) is dropwise added after stirring, after reacting by heating, add monomer polyethylene glycol methacrylate-styrene polymer (PEGMA), continue isothermal reaction, obtain amphipathic pH and respond brush copolymer (P (HEMA-co-DEA)-b-PPEGMA);
(2) prepare cholesterol and modify amphipathic pH response brush copolymer: the amphipathic pH obtained by step (1) is responded brush copolymer and is dissolved in anhydrous methylene chloride (DCM), the triethylamine (TEA) and cholesteryl chloroformate that have been dissolved in anhydrous methylene chloride is added successively under ice bath, reaction 2 ~ 4h, room temperature continues reaction 24 ~ 48h, obtains cholesterol and modifies amphipathic pH response brush copolymer.
In described step (1), the molfraction formula of reactant is as follows:
In described step (2), the molfraction formula of reactant is as follows:
P (HEMA-co-DEA)-b-PPEGMA 1 part
Chol 9 ~ 15 parts
TEA 9 ~ 15 parts
Reacting by heating described in step (1) refers to react 5 ~ 7h at 60 ~ 90 DEG C.The time of described isothermal reaction is preferably 24 ~ 48h.
Preferably, after step (1) has been reacted, by reaction system cooling, purifying, drying, obtain the product after purifying.After described purifying refers to that reaction product is dissolved in tetrahydrofuran (THF), cross neutral alumina chromatography column, tetrahydrofuran (THF) wash-out, removing catalyzer, then by elutriant concentrated by rotary evaporation, the 0 DEG C of normal hexane adding 10 times of volumes precipitates, and filters and obtains product after purification.
Solvent in step (1) is preferably toluene.
After step (2) has been reacted, preferably by application system filtration, precipitation, drying, obtain the product after purifying.Described filtration is for removing insoluble triethylamine hydrochloride.Described precipitation refers to filtrate after filtration to revolve steaming, then the 0 DEG C of normal hexane adding 10 times of volumes precipitates.
Preferably, above-mentioned reaction is all carried out under protection of inert gas and anhydrous condition.
Its structural formula of described HEMA and DEA is as follows respectively:
The above-mentioned cholesterol of the present invention is modified amphipathic pH response brush copolymer and be can be applicable to load in poorly water soluble drugs.
Modify amphipathic pH based on above-mentioned cholesterol and respond the carrier micelle system of brush copolymer, respond brush copolymer and be dissolved in solvent system by cholesterol being modified amphipathic pH and obtain.
Above-mentioned micellar system can be applicable to load in poorly water soluble drugs.
Above-mentioned micellar system is applied to load in poorly water soluble drugs and comprises the following steps: poorly water soluble drugs is dissolved in organic solvent process of spending the night, cholesterol is modified amphipathic pH response brush copolymer is dissolved in same organic solvent simultaneously, micellar system is obtained after thing to be polymerized dissolves completely, this micellar system is mixed with poorly water soluble drugs solution, stirred at ambient temperature 4 ~ 6h, deionized water dialysis 24h, filtration, lyophilize, obtain 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 is preferably the wherein one in dimethyl sulfoxide (DMSO) and dimethyl formamide.
The medicine that described loading poorly water soluble drugs micellar system can control to load, at healthy tissues (pH 7.4) place's slow releasing, realizes quick, controllable release at tumour cell solutions of weak acidity (pH 5 ~ 6) place.
Above-mentioned based on cholesterol of the present invention modify amphipathic pH respond brush copolymer, to adopt dialysis method to prepare internal layer be hydrophobic segment, the middle layer that cholesterol is modified is the nano grade polymer micella that pH responds segment, shell is hydrophilic segment, realizes the loading to slightly water-soluble cancer therapy drug.Under tumor tissues solutions of weak acidity (pH 5 ~ 6), pH responds group generation protonation and shows as wetting ability, and micella dissociates, thus realizes quick, the controllable release that are loaded medicine.This cholesterol modifies amphipathic pH response brush copolymer structure under the prerequisite maintaining high drug load, can improve the pH response sensitivity of micella, more effective Drug controlled release, improves the therapeutic efficiency of medicine.
Mechanism of the present invention is:
The present invention utilizes hydrophilic block hydroxyethyl methylacrylate, hydrophobic cholesterol and pH respond block methacrylic acid N, the random copolymerization of N-lignocaine ethyl ester, and then to combine with hydrophilic block polyethylene glycol methacrylate-styrene polymer and obtain amphipathic pH and respond brush copolymer, be dissolved in solvent, obtaining internal layer is the hydrophobic segment that cholesterol is modified, middle layer is that pH responds segment, shell is the nano grade polymer micella of hydrophilic segment, the PPEGMA of micella outer layer hydrophilic has nontoxic, the advantage such as non-immunogenicity and no antigen, while increase micella stability, extend micella cycling time in blood, hydrophobic cholesterol kernel can strengthen and carries performance to the bag of insoluble medicine, the PDEA in middle layer shows as hydrophobicity when pH 7.4, and can jointly form the hydrophobic inner core of micella with cholesterol, this not only can prevent burst drug release, can strengthen the stability of micelle inner core simultaneously, under tumor tissues slightly acidic (pH 6.0) condition, in PDEA block side chain tertiary amine groups generation protonation and show as wetting ability, it is swelling that micella starts to occur to a certain degree, if carrier micelle enters in the endosome and lysosome with lower pH, PDEA will be completely protonated, and now micella degree of swelling becomes large, and micella starts to occur assembling behavior of even dissociating, thus by the drug release of package-contained in tumour cell.By the ratio of block each in telomerized polymer, can the rate of release of regulating medicine, meet the release request of different pharmaceutical.
The present invention, relative to prior art, has following advantage and beneficial effect:
(1) hydrophobic grouping and pH are responded group and be designed to random copolymerization form, both can maintain comparatively high drug load, and effectively can regulate again the pH responding range of its self-assembled micelle, and make micella not only can the change of response environment pH value rapidly, and effectively can alleviate prominent releasing, Drug controlled release.
(2) multipolymer after hydrophobic cholesterol is modified strengthens the bag loading capability of poorly water soluble drugs.
(3) cholesterol that the present invention prepares is modified amphipathic pH and is responded brush copolymer, is easy to the ratio of each block, is applied to preparation and loads poorly water soluble drugs micellar system, can meet the release request of different pharmaceutical.
Accompanying drawing explanation
Fig. 1 is the GPC elution curve of P (HEMA-co-DEA)-b-PPEGMA in embodiment 1.
Fig. 2 is embodiment 1 cholesterol modifying front and back polymkeric substance 1h NMR composes, and wherein, (A) is for P (HEMA-co-DEA)-b-PPEGMA's 1h NMR composes, and solvent is d-CDCl 3; (B) for Chol-P (HEMA-co-DEA)-b-PPEGMA's 1h NMR composes, and solvent is d-CDCl 3.
Fig. 3 is the GPC elution curve of Chol-P (HEMA-co-DEA)-b-PPEGMA in embodiment 1.
Fig. 4 is the micelle-forming concentration test curve of Chol-P (HEMA-co-DEA)-b-PPEGMA in embodiment 5.
Fig. 5 is DOX/DMSO typical curve in embodiment 6.
Fig. 6 is the potentiometric titration curve of Chol-P (HEMA-co-DEA)-b-PPEGMA self-assembled micelle in embodiment 7.
Fig. 7 is the relation of the particle diameter of Chol-P (HEMA-co-DEA)-b-PPEGMA self-assembled micelle in embodiment 7, zeta current potential and pH.
Fig. 8 is the In-vitro release curves carrying Zorubicin micella in embodiment 8.
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: cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA
(1) amphipathic pH responds the preparation of brush copolymer P (HEMA-co-DEA)-b-PPEGMA: in the dry eggplant-shape bottle of 50mL, load stirrer and cupric bromide (22.3mg, 1.0mmol), with anti-mouth soft rubber ball sealing, vacuumize-Tong argon gas three times.Successively with syringe by solvent toluene (20mL), monomer HEMA (1.30g, 10mmol), DEA (6.48g, 35mmol), part HMTETA (0.31mL, 1.2mmol) adds in bottle, stirs 10min and catalyst complexes is formed.Subsequently by reductive agent Sn (Oct) 2(0.40g, 1.0mmol) is dissolved in 2mL toluene and adds in reaction flask.Dropwise add initiator EBriB (0.15mL, 1.08mmol) fast with syringe after stirring 5min, in 60 DEG C of oil baths after stirring reaction 6h, add monomer PEGMA (4.75g, 10mmol), continue reaction 30h.Reaction solution is cooled to room temperature, add 50mL THF and stir make it dissolve, then use neutral alumina pillar Filtration of catalyst CuBr 2(making eluent with THF).Slowly join in the cold normal hexane of ten times amount (volume ratio) after the elutriant obtained is concentrated and precipitate.Precipitated product is at 35 DEG C, vacuum-drying 48h under 35mbar.Building-up reactions formula is shown in formula (1).Utilize GPC to measure its molecular weight, and carry out nmr analysis, see (A) in Fig. 1 and Fig. 2.Productive rate is 98%, M n=12481, M w/ M n=1.60.
(2) cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA: P (HEMA-co-DEA)-b-PPEGMA (5.68g step (1) prepared, 0.25mmol) be placed in 100mL eggplant shaped reaction device and seal with anti-mouth soft rubber ball, vacuumize-Tong argon gas three times, keeping in reactor is argon gas atmosphere.Add 20mL anhydrous methylene chloride, reactant is fully dissolved.Respectively triethylamine (2.1mL, 15mmol), Chol (3.86g, 10mmol) are dissolved in 5mL anhydrous methylene chloride, join successively in reactor with syringe under condition of ice bath, reaction 2h.Transfer room temperature condition to, continue reaction 30h.After having reacted, cross and filter insoluble triethylamine hydrochloride, precipitate three times with cold normal hexane, precipitation vacuum-drying (35mbar, 35 DEG C) 48h, obtain light yellow product, building-up reactions formula is shown in formula (2).Utilize GPC to measure its molecular weight, and carry out nmr analysis, see (B) in Fig. 2 and Fig. 3.M n=16341,?M w/M n=1.54。
Embodiment 2: cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA
(1) amphipathic pH responds the preparation of brush copolymer P (HEMA-co-DEA)-b-PPEGMA: in the dry eggplant-shape bottle of 50mL, load stirrer and cupric bromide (26.8mg, 0.8mmol), with anti-mouth soft rubber ball sealing, vacuumize-Tong argon gas three times.Successively with syringe by solvent toluene (20mL), monomer HEMA (1.30g, 10mmol), DEA (6.475g, 40mmol), part HMTETA (0.31mL, 1.2mmol) adds in bottle, stirs 10min and catalyst complexes is formed.Subsequently by reductive agent Sn (Oct) 2(0.486g, 1.2mmol) is dissolved in 2mL toluene and adds in reaction flask.Dropwise add initiator EBriB (0.17mL, 1.2mmol) fast with syringe after stirring 5min, in 70 DEG C of oil baths after stirring reaction 5h, add monomer PEGMA (14.25g, 30mmol), continue reaction 24h.Reaction solution is cooled to room temperature, add 50mL THF and stir make it dissolve, then use neutral alumina pillar Filtration of catalyst CuBr 2(making eluent with THF).Slowly join in the cold normal hexane of ten times amount (volume ratio) after the reaction solution obtained is concentrated and precipitate.Product is at 35 DEG C, vacuum-drying 48h under 35mbar.Productive rate is 89%, M n=20634, M w/ M n=1.51.
(2) cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA: P (HEMA-co-DEA)-b-PPEGMA (5.27g step (1) prepared, 0.25mmol) be placed in 100mL eggplant shaped reaction device and seal with anti-mouth soft rubber ball, vacuumize-Tong argon gas three times, keeping in reactor is argon gas atmosphere.Add 20mL anhydrous methylene chloride, reactant is fully dissolved.Respectively triethylamine (1.25mL, 9mmol), Chol (5.79g, 15mmol) are dissolved in 5mL anhydrous methylene chloride, join successively in reactor with syringe under condition of ice bath, reaction 2h.Transfer room temperature condition to, continue reaction 24h.After having reacted, cross and filter insoluble triethylamine hydrochloride, precipitate three times with cold normal hexane, vacuum-drying (35mbar, 35 DEG C) 48h, obtains light yellow product.M n=24494,M w/M n=1.47。
Embodiment 3: cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA
(1) amphipathic pH responds the preparation of brush copolymer P (HEMA-co-DEA)-b-PPEGMA: in the dry eggplant-shape bottle of 50mL, load stirrer and cupric bromide (26.8mg, 0.12mmol), with anti-mouth soft rubber ball sealing, vacuumize-Tong argon gas three times.Successively with syringe by solvent toluene (20mL), monomer HEMA (4.68g, 36mmol), DEA (2.775g, 15mmol), part HMTETA (0.31mL, 1.2mmol) adds in bottle, stirs 10min and catalyst complexes is formed.Subsequently by reductive agent Sn (Oct) 2(0.486g, 1.2mmol) is dissolved in 2mL toluene and adds in reaction flask.Initiator EBriB (0.17mL, 1.2mm is dropwise added fast with syringe after stirring 5min ol), in 90 DEG C of oil baths after stirring reaction 7h, add monomer PEGMA (14.25g, 30mmol), continue reaction 48h.Reaction solution is cooled to room temperature, add 50mL THF and stir make it dissolve, then use neutral alumina pillar Filtration of catalyst CuBr 2(making eluent with THF).Slowly join in the cold normal hexane of ten times amount (volume ratio) after the reaction solution obtained is concentrated and precipitate.Product is at 35 DEG C, vacuum-drying 48h under 35mbar.Productive rate is 96%, M n=21061, M w/ M n=1.48.
(2) cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA: P (HEMA-co-DEA)-b-PPEGMA (4.3g step (1) prepared, 0.25mmol) be placed in 100mL eggplant shaped reaction device and seal with anti-mouth soft rubber ball, vacuumize-Tong argon gas three times, keeping in reactor is argon gas atmosphere.Add 20mL anhydrous methylene chloride, reactant is fully dissolved.Respectively triethylamine (2.1mL, 15mmol), Chol (3.47g, 9mmol) are dissolved in 5mL anhydrous methylene chloride, join successively in reactor with syringe under condition of ice bath, reaction 4h.Transfer room temperature condition to, continue reaction 48h.After having reacted, cross and filter insoluble triethylamine hydrochloride, precipitate three times with cold normal hexane, vacuum-drying (35mbar, 35 DEG C) 48h, obtains light yellow product.M n=28781,M w/M n=1.46。
Embodiment 4: cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA
(1) amphipathic pH responds the preparation of brush copolymer P (HEMA-co-DEA)-b-PPEGMA: in the dry eggplant-shape bottle of 50mL, load stirrer and cupric bromide (17.84mg, 0.08mmol), with anti-mouth soft rubber ball sealing, vacuumize-Tong argon gas three times.Successively with syringe by solvent toluene (20mL), monomer HEMA (1.3g, 10mmol), DEA (2.775g, 15mmol), part HMTETA (0.2mL, 0.8mmol) adds in bottle, stirs 10min and catalyst complexes is formed.Subsequently by reductive agent Sn (Oct) 2(0.324g, 10.8mmol) is dissolved in 2mL toluene and adds in reaction flask.Dropwise add initiator EBriB (0.12mL, 0.8mmol) fast with syringe after stirring 5min, in 80 DEG C of oil baths after stirring reaction 6h, add monomer PEGMA (14.25g, 30mmol), continue reaction 36h.Reaction solution is cooled to room temperature, add 50mL THF and stir make it dissolve, then use neutral alumina pillar Filtration of catalyst CuBr 2(making eluent with THF).Slowly join in the cold normal hexane of ten times amount (volume ratio) after the reaction solution obtained is concentrated and precipitate.Product is at 35 DEG C, vacuum-drying 48h under 35mbar.Productive rate is 96%, M n=17187, M w/ M n=1.43.
(2) cholesterol modifies the preparation that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA: P (HEMA-co-DEA)-b-PPEGMA (4.3g step (1) prepared, 0.25mmol) be placed in 100mL eggplant shaped reaction device and seal with anti-mouth soft rubber ball, vacuumize-Tong argon gas three times, keeping in reactor is argon gas atmosphere.Add 20mL anhydrous methylene chloride, reactant is fully dissolved.Respectively triethylamine (1.25mL, 9mmol), Chol (3.47g, 9mmol) are dissolved in 5mL anhydrous methylene chloride, join successively in reactor with syringe under condition of ice bath, reaction 3h.Transfer room temperature condition to, continue reaction 36h.After having reacted, cross and filter insoluble triethylamine hydrochloride, precipitate three times with cold normal hexane, vacuum-drying (35mbar, 35 DEG C) 48h, obtains light yellow product.M n=21047,M w/M n=1.52。
Embodiment 5: cholesterol modifies the micelle-forming concentration CMC value that amphipathic pH responds brush copolymer
The cholesterol utilizing fluorescence probe method testing example 1 to prepare modifies the micelle-forming concentration that amphipathic pH responds brush copolymer Chol-P (HEMA-co-DEA)-b-PPEGMA.
(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 5mg Chol-P (HEMA-co-DEA)-b-PPEGMA and be dissolved in 10mL acetone, fast solution is joined in 50mL deionized water, stir 24h to vapor away acetone, obtain the polymer mother liquor that concentration is 0.1mg/mL, 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: take 373nm as emission wavelength, at the fluorescence excitation spectrum of 300 ~ 350nm scanning samples solution.Get the intensity rate (I that wavelength is 339nm and 336nm 339/ I 336) to the mapping of polymer concentration logarithm, as shown in Figure 5, the X-coordinate corresponding to curve catastrophe point is lg (CMC).The micelle-forming concentration recording Chol-P (the HEMA-co-DEA)-b-PPEGMA that embodiment 1 prepares is 6.92mg/L.
Embodiment 6: cholesterol modifies the preparation that amphipathic pH responds brush copolymer carrier micelle
Adopt dialysis method preparation to carry the polymer micelle of Zorubicin (DOX), concrete grammar is as follows: take 10mg DOXHCl and be dissolved in 20mL DMSO, add the TEA of 2 times of molar equivalents, and stirring spends the night obtains DOX alkali.Getting Chol-P (the HEMA-co-DEA)-b-PPEGMA that 40mg embodiment 1 prepares is dissolved in 20mL DMSO, after thing to be polymerized dissolves obtain micellar system completely, in the DOX solution prepared before micellar system is joined, after continuing to stir 6h, proceed to dialysis tubing to dialyse, every 2h changes a deionized water, and after 12h, every 6h changes once, and dialyse 24h altogether.After having dialysed, dialyzate 0.45 μm of filtering membrane is filtered, after filtered liquid lyophilize, obtain red powder solid and be DOX carrier micelle.
The preparation method of blank micella is identical therewith.
Ultraviolet visible spectrometry is adopted to detect drug loading LC and the encapsulation rate EE of DOX.Concrete steps are as follows: accurately take 1mg freeze-drying carrier micelle and be fully dissolved in 10mL DMSO, survey its uv-absorbing intensity at 480nm place, by DOX/DMSO typical curve, see Fig. 5, calculate DOX concentration, calculate drug loading and encapsulation rate respectively.The drug loading calculated is 10.8%, and encapsulation rate is 48.5%.
The method for drafting of DOX/DMSO typical curve is as follows: the DOX/DMSO standardized solution preparing a series of concentration.Take DMSO as blank, at 480nm wavelength place, optical density is measured to the DOX/DMSO solution of prepared series concentration.After deducting the absorbancy of blank DMSO solution, with the absorbance A recorded to concentration C mapping go forward side by side line linearity return, obtain the canonical plotting of DOX concentration and the absorbancy in DMSO solution as Fig. 5.
Dynamic light scattering method (DLS) is adopted to measure the particle diameter of blank micella and carrier micelle, distribution and zeta current potential.The particle diameter D of blank micella hbe 21.1mV for 155.6nm, PDI are 0.29, zeta current potential.The particle diameter D of carrier micelle hbe 19.7mV for 178.5nm, PDI are 0.31, zeta current potential.
Embodiment 7: cholesterol modifies the pH response self-assembly behavioral study that amphipathic pH responds brush copolymer micella
First the pK of PDEA block in potentiometric titration determination embodiment 1 polymkeric substance is adopted bvalue, concrete steps are as follows: Chol-P (HEMA-co-DEA)-b-PPEGMA prepared by 50mg embodiment 1 is fully dissolved in 20mL acetone, join fast under fast stirring in 50mL deionized water, stirred at ambient temperature 24h, to vapor away acetone, obtains the polymer micelle solution that ultimate density is 1mg/mL.Regulate the pH value of micellar solution with NaOH or HCl solution (0.1M), stir balance for some time and stablize to pH, read each pH value, as shown in Figure 6.Adopt DLS to test particle diameter and the zeta current potential of micella under different pH value, as shown in Figure 7 simultaneously.As shown in Figure 6, the pK of polymer micelle bvalue is 7.2.As shown in Figure 7, along with pH reduces, particle diameter and zeta current potential increase gradually, and are finally tending towards definite value.
Embodiment 8: the release in vitro of carrier micelle
The In-vitro release curves of DOX under different pH adopts medicament dissolution instrument test to obtain, and concrete steps are as follows: adopt method similarly to Example 5, prepare DOX carrier micelle, represent with PMs-1 with the polymkeric substance of embodiment 2.DOX carrier micelle prepared by embodiment 5 represents with PMs-2.Take 5mg DOX carrier micelle PMs-1 and PMs-2 to be respectively scattered in 5mL PBS damping fluid, pH of cushioning fluid is respectively 6.0 and 7.4.Above-mentioned solution 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 4mL carries out ultra-violet analysis, and adds 4mL fresh buffer simultaneously.By DOX concentration in determined by ultraviolet spectrophotometry different time release liquid, draw In-vitro release curves, as shown in Figure 8.
As shown in Figure 8, DOX carrier micelle PMs-1 with PMs-2 shows consistent release in vitro behavior.Under healthy tissues environment (pH 7.4), slowly, the cumulative release amount of 24h is no more than 35% to the rate of release of DOX, and rate of release subsequently tends to be steady substantially.And under subacidity (pH 6.0) condition of tumour cell, the rate of release of DOX is obviously accelerated, the cumulative release amount of 24h reaches more than 80%.And PMs-1 is relatively faster than the drug release rate of PMs-2.
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. cholesterol is modified amphipathic pH and is responded a brush copolymer, it is characterized in that having as shown in the formula structure shown in I:
Wherein, x=10 ~ 36, y=15 ~ 40, z=8 ~ 30.
2. cholesterol according to claim 1 is modified amphipathic pH and is responded brush copolymer, it is characterized in that its number-average molecular weight is 11891 ~ 40460g/mol.
3. the cholesterol described in any one of claim 1 ~ 2 modifies the preparation method that amphipathic pH responds brush copolymer, it is characterized in that comprising following concrete steps:
(1) prepare amphipathic pH and respond brush copolymer: by Catalysts Cu Br 2, monomers hydroxy ethyl ester and methacrylic acid N, N-lignocaine ethyl ester, ligand 1, Isosorbide-5-Nitrae, 7,10,10-hexamethyl triethylene tetramine is dissolved in solvent, then adds reductive agent Sn (Oct) 2, dropwise add initiator 2-isobutyl ethyl bromide after stirring, reacting by heating, add monomer polyethylene glycol methacrylate-styrene polymer, continue isothermal reaction, after having reacted, obtain amphipathic pH and respond brush copolymer;
(2) prepare cholesterol and modify amphipathic pH response brush copolymer: the amphipathic pH obtained by step (1) is responded brush copolymer and is dissolved in anhydrous methylene chloride, the triethylamine being dissolved in anhydrous methylene chloride and cholesteryl chloroformate is added successively under ice bath, reaction 2 ~ 4h, room temperature continues reaction 24 ~ 48h, obtains cholesterol and modifies amphipathic pH response brush copolymer.
4. cholesterol according to claim 3 modifies the preparation that amphipathic pH responds brush copolymer
Method, is characterized in that:
In step (1), the molfraction formula of each reactant is as follows:
In step (2), the molfraction formula of each reactant is as follows:
Amphipathic pH responds brush copolymer 1 part
Cholesteryl chloroformate 9 ~ 15 parts
Triethylamine 9 ~ 15 parts.
5. cholesterol according to claim 3 modifies the preparation method that amphipathic pH responds brush copolymer, it is characterized in that: the reacting by heating described in step (1) refers to react 5 ~ 7h at 60 ~ 90 DEG C; The time of described isothermal reaction is 24 ~ 48h.
6. a micellar system, is characterized in that modifying amphipathic pH response brush copolymer based on the cholesterol described in any one of claim 1 ~ 2 prepares.
7. micellar system according to claim 6 is loading the application in poorly water soluble drugs.
8. micellar system according to claim 7 is loading the application in poorly water soluble drugs, it is characterized in that comprising the following steps: poorly water soluble drugs is dissolved in organic solvent process of spending the night, cholesterol being modified amphipathic pH response brush copolymer is dissolved in same organic solvent, micellar system is obtained after thing to be polymerized dissolves completely, this micellar system is mixed with poorly water soluble drugs solution, stirred at ambient temperature 4 ~ 6h, deionized water dialysis 24h, filtration, lyophilize, obtain loading poorly water soluble drugs micellar system.
9. the cholesterol according to any one of claim 1 ~ 2 is modified amphipathic pH and is responded the application of brush copolymer in loading poorly water soluble drugs.
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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104856952A (en) * 2015-05-13 2015-08-26 江南大学 Preparation of poly(L-glutamic acid)-b-polyethylene glycol medicine carrying nano micelle
CN106565962A (en) * 2016-10-24 2017-04-19 东莞理工学院 Cholesterol grafted pH-response tri-block amphiphilic copolymer and preparation method and application thereof
CN106883360A (en) * 2017-02-14 2017-06-23 上海大学 Can functionalization stimulating responsive polymer and preparation method thereof
CN106957401A (en) * 2017-04-01 2017-07-18 苏州大学 The PEG methacrylate polymers and its synthetic method and methods for using them of a kind of cholesterol group grappling
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CN112851854A (en) * 2021-01-11 2021-05-28 上海市第十人民医院 Random copolymer based on natural cholesterol, preparation method and application thereof
CN113698556A (en) * 2021-08-18 2021-11-26 新乡医学院 Preparation method of redox-sensitive targeted drug-loaded polymer
CN113881002A (en) * 2021-11-17 2022-01-04 湘潭大学 Amphiphilic block copolymer and preparation thereof, and preparation and application of nano micelle
CN114507326A (en) * 2022-01-17 2022-05-17 华南理工大学 pH response crosslinking bond-based hydrophilic side chain-adjustable triblock polymer and preparation and application thereof
CN114605600A (en) * 2022-01-18 2022-06-10 华南理工大学 Esterase-response amphiphilic linear polymer and preparation method and application thereof
CN118370843A (en) * 2024-06-21 2024-07-23 吉林大学 Active oxygen responsive zwitterionic drug-loaded nano-particle, and preparation method and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102432783A (en) * 2011-09-13 2012-05-02 华南理工大学 PH response/hydrophobic group random copolymerization polymer and preparation method and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102432783A (en) * 2011-09-13 2012-05-02 华南理工大学 PH response/hydrophobic group random copolymerization polymer and preparation method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MASSOUMEH BAGHERI ET AL.: "pH-responsive stealth micelles composed of cholesterol-modified PLA as a nano-carrier for controlled drug release", 《PROGRESS IN BIOMATERIALS》 *

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