CN102440961B - Targeting polymer micelle containing acid-sensitive subsurface, and preparation method thereof - Google Patents
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Abstract
The invention relates to a targeting polymer micelle containing acid-sensitive subsurface, and a preparation method thereof. The polymer micelle is in a core-shell structure. Targeting ligand molecules are grafted on a hydrophilic segment terminal of the shell, and a hydrophobic anti-tumor medicine is loaded by a hydrophobic segment of the core. An acid-sensitive response subsurface exists between the core and the shell, wherein the acid-sensitive response subsurface can load negative electricity quantum dots. When a pH value is 5.0, the acid-sensitive layer is hydrophobic; when the pH value is 7.4, the acid-sensitive layer is hydrophobic. According to the invention, with the targeting molecules, the tumor targeting function of the micelle is realized; with the quantum dots loaded on the acid-sensitive subsurface, a visualizing function is realized; with low-pH-responsibility of the acid-sensitive subsurface, a controlled-release function can be realized, and the release of the medicine can be controlled. With the micelle system provided by the invention, the release behavior of the medicine is improved, and a premature-release problem of the micelle system in blood circulation ispossible to be solved. With an internalization behavior mediated by the targeting molecules, the treatment effect of the medicine is enhanced. With the loaded quantum dots, monitoring during the treatment process can be realized.
Description
Technical field
The present invention relates to chemistry, pharmacy and biomedical engineering field, is a kind of preparation and application aspect medicine control release and video picture with target polymer micelle of acid-sensitive subsurface stratum specifically.
Background technology
The mankind's health in the tumor serious threat, and mortality rate is high.Oncotherapy has three kinds of Main Means: surgical operation, radiotherapy and chemotherapy (chemotherapy), wherein first two is the locality treatment, usually takes chemotherapy when tumor diffusion occurs and shifts.And chemotherapeutics commonly used mostly is the hydrophobic type medicine clinically, the dissolubility in water is very low, has limited their application.During clinical use, adopt the micromolecule surfactants to hydrophobic type chemotherapeutics emulsifying solubilising, but this dosage form blood stability is very poor, the adjuvant toxic and side effects of interpolation is also very large, and release behavior is not easy to control more.Polymer micelle is a kind of novel pharmaceutical carrier, and it is that autonomous dress forms in water by amphipathic copolymer take the nucleocapsid structure that the hydrophobicity section is shell as the kernel hydrophilic section, and the drug loading of hydrophobic type is in the core of micelle.The dissolubility of polymer micelle energy load hydrophobic drug and significantly increasing medicament, improve the bioavailability of medicine.
Conventional micelle medicine carrying system exists two large problems, the one, medicine too early release in blood circulation, the 2nd, the tumor locus medicine assemble and release concentration inadequate.Medicine too early release in blood circulation makes effective dose reduce, and the whole body toxic and side effects also can increase simultaneously, and medicine can make curative effect of medication greatly reduce in tumor locus gathering or release concentration not.The release of medicine while therefore reducing body-internal-circulation, discharging medicine after control medicine-carried system arrival tumor cell is a kind of effective means that reduces the whole body toxic and side effects, improves the therapeutic effect of medicine again.
Avoid the too early release of medicine in blood, will improve the drug release behavior, this respect environmental response type micelle advantage is fairly obvious.Wherein acid-sensitive type micelle is very important a kind of.The physiological environment pH value that has been found that blood in medical research is about 7.4, and in cell, lysosomal pH value is 4.5 ~ 5.0.According to the difference of pH value under this two states, design acid-sensitive type micelle and can accomplish that micelle is stablized medicine in blood and do not discharge, and, after arriving Cytolysosome, medicine just starts to discharge the performance curative effect.So just can reduce medicine and discharge the toxic and side effects of bringing in blood, avoid again the loss of medicine in blood circulation.
Solve medicine and not usually adopt the medicine transmission system of targeting modification at the aggregate concentration of tumor locus.Introduce tumor cell surface or tumor tissues blood vessel surface on the polymer micelle surface and cross the part of its receptor of expression as targeted molecular (as folic acid, polypeptide or antibody etc.), strengthen the endocytosis of tumor cell to medicine-carried system, thereby improve aggregate concentration and the burst size of medicine at tumor locus.
In addition, in oncotherapy, realize that Real-Time Monitoring will have to the treatment of tumor very great meaning.When surgical intervention, Real-Time Monitoring can, to accurate positioning tumor position, make excision more thorough.In chemotherapy, if realize Real-Time Monitoring, convenient chemotherapy effect is estimated.In at present clinical, Real-Time Monitoring is carried out in many employing magnetic resonance imagings or ultrasonoscopy.But magnetic resonance imaging or ultrasonoscopy are just estimated tissue, the appearance of Imaging-PAM can be studied people on cellular level to tumor, conveniently cell inner structure, cell-cell interaction, cellular signal transduction are carried out to more deep understanding.The R and D of Imaging-PAM and fluorescent probe have two large difficulties: the one, can't effectively overcome covering that cell signals to labelled molecule at the autofluorescence of visible region; The 2nd, photobleaching easily occurs in the fluorescence organic dye molecule, can't realize preferably the long-time fluorescent labeling of research molecule is observed.Quantum dot has unique optical property as novel fluorometric reagent: wide excitation wavelength range and emission wavelength is very narrow, fluorescence lifetime is long, do not allow to be subject to other signal disturbs size that can be by controlling it by anti-photobleaching, emission wavelength and form to adjust and its size characteristics also makes it be very suitable for doing fluorescent marker in the bioluminescence imaging.
Summary of the invention
The object of the invention is to overcome the too early release And Spread of Solute of medicine-carried system in blood circulation, a kind of target polymer micelle with acid-sensitive subsurface stratum is provided.
In order to overcome the deficiencies in the prior art, the present invention adopts following technical scheme:
Target polymer micelle with acid-sensitive subsurface stratum of the present invention is nucleocapsid structure, the hydrophilic section tip of shell is connected to the targeting ligand molecular, the hydrophobic section load hydrophobic anticancer drug of kernel, but exist the acid-sensitive response subsurface stratum of one deck load negative electricity quantum dot between kernel and shell, when pH 5.0, acid-sensitive layer is hydrophilic, and when pH 7.4, acid-sensitive layer is hydrophobic.
The triblock copolymer that the Polyethylene Glycol hydrophilic section that micelle polymer used is the folic acid functionalization, acid-sensitive interlude and hydrophobic section form.When pH 5.0 by the hydrophobic type drug loading in micelle core, now acid-sensitive section hydrophilic, carrying medicament not; PH is adjusted to at 7.4 o'clock again, acid-sensitive section hydrophobic formation layer protective layer, stop medicine to discharge when pH 7.4, plays the problem that prevents that medicine from discharging too early in blood circulation.After medicine-carried system enters tumor cell by folate-mediated endocytosis, under the sour environment of the lysosomal pH 4.5 ~ 5.0 of tumor cell, acid-sensitive layer is hydrophilic, and medicine starts to discharge from micelle core, the performance therapeutical effect.
Described acid-sensitive interlude can be poly-aminoacyl diisopropyl ethylenediamine or polyacrylamide diisopropyl ethylenediamine, and hydrophobic section can be the hydrophobic type molecules such as polyester or cholic acid.Adopt folic acid polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine-cholic acid triblock copolymer (FA-PEG-P (Asp-dip)-CA) in the embodiment of the present invention.
Described hydrophobic type medicine can be the hydrophobic type antitumor drug such as paclitaxel (PTX).
Target polymer micelle with acid-sensitive subsurface stratum of the present invention can provide real time monitoring function in chemotherapy, is the medicine-carried system that has image displaying function concurrently.The image displaying function of medicine-carried system utilizes acid-sensitive intermediate layer this feature of positively charged when pH 5.0 to realize.In pH 5.0 medicine carryings, add electronegative quantum dot, by electrostatic interaction, quantum dot is loaded to acid-sensitive layer, then pH is adjusted to 7.4, the acid-sensitive section hydrophobic skin at carrier micelle core forms layer protective layer, and the negative electricity quantum dot is wrapped in to protective layer.But acid-sensitive intermediate layer load negative electricity quantum in anti-release is too early lighted the function of Real-Time Monitoring fluorescence imaging.
Described negative electricity quantum dot can be carboxylated CdSe/ZnS or other electronegative quantum dot.
Another object of the present invention is to provide the above-mentioned preparation method with target polymer micelle of acid-sensitive subsurface stratum, concrete steps are as follows: the polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine of 10 weight portion folic acid end-blockings-cholic acid triblock copolymer and 1 weight portion hydrophobic anticancer drug are dissolved in oxolane, slowly add to acidic buffer under ultrasonication in, oxolane is vapored away naturally, the hydrophobic drug aggregation is removed in filtration, with centrifugal ultrafiltration pipe (MWCO=100 kD) centrifugal ultrafiltration 3 times, add negative electricity quantum dot CdSe/ZnS (100 μ L, 8 μ moL/L), mix latter static 30 minutes, regulate pH to 7.4, the centrifugal quantum dot of removing not load of 6000 r/min.The gained micelle shape is even, and it is 53.4 ± 0.8 nm that mean diameter is measured in light scattering, and it is 40 nm left and right that transmission electron microscope is measured particle diameter.
Compared with prior art, the present invention has following beneficial effect:
Under the condition of pH 5.0, self assembly forms the triblock copolymer of the folic acid functionalization that the target polymer micelle with acid-sensitive subsurface stratum that we design is is acid-sensitive section by interlude.Utilize micelle core load hydrophobic drug; Exist the intermediate layer of one section acid-sensitive sense between micelle core and shell; under the condition of pH 5.0; acid-sensitive section positively charged can pass through electrostatic interaction load elecrtonegativity quantum dot; under the environment of pH 7.4; the acid-sensitive section hydrophobic skin at micelle core forms layer protective layer; the negative electricity quantum dot is wrapped in to protective layer, and this layer can play the effect that prevents that medicine from discharging under the condition of pH 7.4 simultaneously.After mediating by folate-targeted when micelle and entering tumor cell, the acid-sensitive section protonated hydrophilic that becomes under the condition of the low pH 5.0 of Cytolysosome, now medicine can discharge the performance curative effect from micelle core.This system is the collection Multifunction micelle medicine carrying system that initiatively targeting, video picture and anti-medicine discharge too early.Target polymer micelle with acid-sensitive subsurface stratum of the present invention can not only solve the too early release And Spread of Solute of medicine-carried system in blood, has improved aggregate concentration and the rate of release of medicine at tumor locus, can also realize Real-Time Monitoring simultaneously.Oncotherapy and novel pharmaceutical formulation aspect are had to very large researching value and application prospect.
The accompanying drawing explanation
Fig. 1 is the dynamic light scattering scattergram of micelle particle diameter in embodiment 3.Result shows PEG-P (Asp-dip)
17the micelle particle diameter that-CA forms is 53.4 ± 0.8 nm.
Fig. 2 is micelle transmission electron microscope shape appearance figure in embodiment 3.Quantum dot during pH 7.4 is evenly distributed on the skin of micelle kernel, and the micelle particle diameter is (40 nm) evenly, has illustrated that result shows that quantum dot, by the loading in the micelle sample of success, has also proved that we have acid-sensitive subsurface stratum by the micelle of design really simultaneously.
Fig. 3 is micelle vitro drug release figure in embodiment 4, in embodiment, the acid-sensitive interlude ground number of repeat unit of polymer used is 17, the paclitaxel loaded rear sample of PEG-P (Asp-dip)-CA burst size in the buffer of pH 7.4 is very low, and all shows quick release behavior in the buffer of pH 5.0.The drug release in vitro result shows that designed micelle volume has the function that prevents that medicine from discharging too early in pH 7.4.
Fig. 4 is the fluorescence release graphics of micelle when pH 5.0 of load FDA and QD in embodiment 5.Result shows when pH 5.0, extends in time, and FDA discharges gradually, and its fluorescence intensity strengthens gradually; QD is slowly cancellation of fluorescence gradually in sour environment, and its fluorescence intensity weakens gradually.
Fig. 5 is the fluorescence release graphics of micelle when pH 7.4 of load FDA and QD in embodiment 5.Result shows when pH 7.4, extends in time, and FDA and QD are stabilized in micelle core and do not discharge, and the fluorescence intensity of FDA and QD is substantially constant.
Fig. 6 is the toxotest figure of different micelle samples to the Bel-7402 cell in embodiment 6.Sample is respectively: FA-micelles, PTX/micelles, PTX/FA-micelles, PTX/FA-micelles/QD; Wherein FA-micelles is the hungry area bundle, and the experimental point series concentration containing paclitaxel in rear three's sample is (5 nM, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM), and incubation time is 36 h.Result shows the cytotoxicity minimum of hungry area bundle FA-micelles, the cytotoxicity of non-targeted group of PTX/micelles takes second place, more non-targeted group of the cytotoxicity of targeting group PTX/FA-micelles is high, simultaneously the sample P TX/FA-micelles/QD of load P TX and QD is substantially suitable with targeting group PTX/FA-micelles sample, illustrate that the toxicity of introducing sample after QD is less than increase.
The specific embodiment
Following examples are explained in detail the present invention.
The triblock copolymer FA-PEG-P (Asp-dip) with acid-sensitive interlude of modified with folic acid-CA's is synthetic
(1) one end is the synthetic of the pi-allyl other end Polyethylene Glycol (Allyl-PEG-OH) that is hydroxyl
By the tetrahydrofuran solution of potassium naphthalide (4 mL) with in dry reaction bulb, stir 15 minutes after propenyl (0.75 mL) is mixed homogeneously.Then the tetrahydrofuran solution (containing 1.5 g hexaoxacyclooctadecane-6-6 and 5 mL anhydrous tetrahydro furans) that adds anhydrous tetrahydro furan (20 mL) and hexaoxacyclooctadecane-6-6 under argon shield; stir after 15 minutes and mixture is placed in to cryosel and bathes cooling; and slowly pass into dry oxirane; maintain low temperature 24 hours so that polyreaction continues to carry out, continue reaction 72 hours under room temperature.
(2) one ends are that the pi-allyl other end is amino Polyethylene Glycol (Allyl-PEG-NH
2) synthetic
Take Allyl-PEG-OH(Mn=2000 Da, 20g) in dry there-necked flask (250 mL), 80 ℃ of dry 4 h of evacuation, then add anhydrous methylene chloride (100 mL), pyridine (50 mL) and paratoluensulfonyl chloride (TsCl, 5g), under room temperature, lucifuge is reacted 24 h.HC1(3 mol L-1 for reactant liquor) extracting and washing, organic layer NaHCO3(5g) washing, solution slowly is added drop-wise in a large amount of absolute ethers, vacuum drying obtains white powder PEG
2k-OTS.
Above-mentioned white powder (10 g) and strong aqua ammonia (500 mL, 25%) are placed in to the 1000mL container, and under room temperature, reaction is 5 days.CH with same volume
2c1
2after extraction by its with volume NaOH solution (1.0 mol L
-1) abundant mix and blend 2 h, organic layer is washed with water to neutrality, vacuum drying obtains Allyl-PEG
2k-NH
2.It is 80% that nuclear-magnetism calculates amino conversion ratio.
(3) benzyloxycarbonyl group aspartic acid anhydride (BLA-NCA) is synthetic
(250 mL) asparagine acid benzyl ester (25 g) of packing in dry there-necked flask, then add ethyl acetate (100 mL) stirring and dissolving of new steaming.After separately Triphosgene (13.2948 g) being dissolved by new ethyl acetate (40 mL) of steaming, be transferred in dry ground constant pressure funnel.After the ethyl acetate solution temperature of asparagine acid benzyl ester is risen to 40 ℃, stir and slowly drip the ethyl acetate solution of triphosgene, question response liquid turns transparent rear stirring ripening 1 hour, and logical argon bubbling is removed the hydrogen chloride that solution is produced.
Cross the unreacted asparagine acid benzyl ester of elimination, in new steaming normal hexane, carry out reprecipitation, fully shake is put into refrigerator freezing at least 12 hours when having crystal to separate out.Freezing rear quick sucking filtration, with the normal hexane washing for several times, vacuum drying obtains the BLA-NCA crystal.
(4) PEG-PASP copolymer (PEG-PBLA) is synthetic
By PEG
2k-NH
2(1.0 g) adds in reaction bulb (50 mL), and 70 ℃ of vacuum drying 4 h, use DMF(10 mL) dissolve, add the DMF(2 mL containing 1.9 g BLA-NCA) solution.React 24 h after mix homogeneously under 35 ℃.By solution precipitation, in cold diethyl ether, filtration is drained, and obtains PEG
2k-PBLA
3.5k.
(5) PEG-PASP-cholic acid (PEG-PBLA-CA) is synthetic
Take cholic acid (0.05 g), EDC (0.03 g) and NHS(0.02 g) in reaction bulb (50 mL), add DMF/HCCl
3after (V:V=1:1,4 mL) dissolve, stir 30min, add PEG2k-PBLA3.5k(0.55 g), room temperature reaction 24h.Solution precipitation, in dehydrated alcohol, is filtered, washing, vacuum drying obtains PEG-PBLA-CA.
(6) polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine-cholic acid (PEG-P (Asp-dip)-CA) is synthetic
Take PEG-PBLA-CA(0.3 g) add in reaction bulb (50 mL), use DMF(1.0 mL) dissolve, add dip(4 mL), react 48h under 35 ℃ of stirrings.Solution precipitation, in absolute ether, is filtered, and vacuum drying obtains PEG-P (Asp-dip)-CA.
(7) the polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine-cholic acid (FA-PEG-P (Asp-dip)-CA) of folic acid functionalization is synthetic
Take PEG-P (Asp-dip)-CA(0.3 g, 1eq) be placed in two mouthfuls of flasks, 70 ℃ of vacuum drying 1 h, add DMF(5 mL) dissolve, then add mercaptoethylmaine hydrochlorate (80 mg, 15eq.) and azodiisobutyronitrile (6 mg, 0.8eq), at 70 ℃ of reaction 24 h, be deposited in cold absolute ether, obtain NH
2-PEG-P (Asp-dip)-CA.
Folic acid (0.044 g) adds in two-mouth bottle (50 mL), after vacuum drying 4 h, add dry DMSO(15 mL), then add successively N-hydroxy-succinamide 0.023g (2eq), N, N'-dicyclohexylcarbodiimide 0.041g (2eq), lucifuge is reacted 12 h, the centrifugal DCU that removes generation, then the folic acid of activation joins and is dissolved with NH
2-PEG-P (Asp-dip)-CA(0.3 g) in 5 ml DMF solution, then with triethylamine, pH value of reaction system is transferred to 8, lucifuge reaction 24h, with the bag filter of the MWCO=1k Da 48h that dialyses in distilled water, lyophilization obtains yellow powder shape product FA-PEG-P (Asp-dip)-CA.
Preparation with target polymer micelle of acid-sensitive subsurface stratum
(1) load QD micelle sample preparation: by FA-PEG-P (Asp-dip)-CA(2.0 mg) and PEG-P (Asp-dip)-CA(8.0 mg) mixture be dissolved in oxolane (THF, 1.0 mL), be added drop-wise to PBS(10 mL in ultrasonic situation, pH=5.0) in, stirring is spent the night, THF is vapored away naturally, add negative electricity quantum dot CdSe/ZnS (100 μ L after the membrane filtration that is 0.22 μ m with aperture, 8 μ moL/L), mix latter static 30 minutes, regulate pH to 7.4, the centrifugal quantum dot of removing not load of 6000r/min.
(2) load QD and the sample preparation of paclitaxel micelle: by FA-PEG-P (Asp-dip)-CA(2.0 mg) and PEG-P (Asp-dip)-CA(8.0 mg) mixture and PTX(1.0 mg) be dissolved in oxolane (THF, 1.0 mL), be added drop-wise to PBS(10 mL in ultrasonic situation, pH=5.0) in, stirring is spent the night, THF is vapored away naturally, the membrane filtration that is 0.45 μ m with aperture is removed the PTX aggregation, with centrifugal ultrafiltration pipe (MWCO=100 kD) centrifugal ultrafiltration 3 times, add negative electricity quantum dot CdSe/ZnS (100 μ L, 8 μ moL/L), mix latter static 30 minutes, regulate pH to 7.4.The centrifugal quantum dot of removing not load of 6000r/min.
(3) load QD and fluorescein(e) diacetate (FDA) micelle sample preparation: by FA-PEG-P (Asp-dip)-CA(2.0 mg) and PEG-P (Asp-dip)-CA(8.0 mg) mixture and FDA(1.0 mg) be dissolved in oxolane (THF, 1.0 mL), be added drop-wise to PBS(10 mL in ultrasonic situation, pH=5.0) in, stirring is spent the night, THF is vapored away naturally, the membrane filtration that is 0.45 μ m with aperture is removed the FDA aggregation, with centrifugal ultrafiltration pipe (MWCO=100 kD) centrifugal ultrafiltration 3 times, add negative electricity quantum dot CdSe/ZnS (100 μ L, 8 μ moL/L), mix latter static 30 minutes, regulate pH to 7.4.The centrifugal quantum dot of removing not load of 6000r/min.
embodiment 3particle diameter and pattern testing experiment with target polymer micelle of acid-sensitive subsurface stratum:
The size of load QD micelle adopts the dynamic light scattering system to be measured, and test result is shown in Fig. 1; Its form is observed definite by transmission electron microscope, test result is shown in Fig. 2.
Tablets in vitro test with target polymer micelle (take paclitaxel loaded micelle as example) of acid-sensitive subsurface stratum
Do the extracorporeal releasing experiment of paclitaxel by the method for dialysis.The paclitaxel loaded PEG-P (Asp-dip) that newly does-CA micelle is divided into to two parts, and it is 5.0 that portion maintains its pH value, and another part is adjusted to 7.4.Again the sample of each pH value is divided into to three parts (parallel laboratory tests) and installs in the bag filter that molecular cut off is 14000 Da, bag filter is put into to the PBS buffer of the identical pH value of 100 mL.Be placed in 37
℃shaking table in, under the rotating speed of 75 r/min, the point in time sampling of setting, add the fresh buffer of equal volume afterwards.Detect the concentration of sample with HPLC, calculate the cumulative release amount of different time points.The Ultimate that wherein HPLC chromatographic column model used is welch materials company
?aQ-C18,5 μ m, 4.6 * 250mm, mobile phase is acetonitrile and water (V:V=50:50), flow velocity is 1.0 mL min
-1, column temperature is 40 ℃, the injected sample amount is 20 μ L.With the time m-cumulative release amount be figure, obtain the curve that external PTX discharges from micelle, test result is as Fig. 3.
External fluorescence release test with target polymer micelle (take load FDA and QD micelle be example) of acid-sensitive subsurface stratum
The load FDA that to get pH be 7.4 and the 1.0 mg mL of QD
-1micellar solution 1 mL, regulating pH is 5.0, is settled to 2 mL.With fluorescent spectrophotometer assay fluorescence intensity temporal evolution figure, the excitation wavelength of fluorescent scanning is 430 nm, excites slit to be set to 10.0 nm, and the emission slit is set to 10.0 nm, and scanning speed is 500 nm min
-1.Test result is as Fig. 4.
The 0.5 mg mL of the load FDA that is 7.4 with same parameter detecting pH and QD
-1the fluorescence intensity temporal evolution figure of micellar solution, test result is as Fig. 5.
embodiment 6in vitro toxicity test with polymer micelle of acid-sensitive subsurface stratum:
Detect the impact of micelle sample on the Bel-7402 ability of cell proliferation by tetrazolium salts colorimetry (MTT).All experimental result triplicates.By the Bel-7402 cell with 1 * 10
3the every hole of cell is inoculated in 96 orifice plates, then by the Bel-7402 cell at 100 μ L RPMI 1640(containing 10%FBS) in culture medium 37 ℃ cultivate 12h, then change the culture medium that does not contain FA and serum into, and add different sample systems to cultivate again 36 h, wherein sample sets has: 1) hungry area bundle (FA-PEG-P (Asp-dip)-CA) FA-micelles, 2) PEG-P (Asp-dip)-CA micelle paclitaxel loaded (PTX-micelles), 3) micelle of folate-targeted paclitaxel loaded (PTX/FA-micelles), 4) paclitaxel loaded and quantum dot (PTX/FA-micelles/QD) of the micelle of folate-targeted while.Finally, culture medium changed into to 150 μ L RPMI 1640 and add 5 mg mL
-1mTT solution 50 μ L, 37 ℃ are continued to hatch 4 h.Stop cultivating, culture supernatant in hole is drawn and discarded, add 150 μ L DMSO in every hole, shake 5 min, with enzyme-linked immunosorbent assay instrument, monitor the absorbance of each hole at OD570 nm.Take the blank group as benchmark, calculate cell proliferation rate.Test result is as Fig. 6.
Claims (3)
1. the target polymer micelle with acid-sensitive subsurface stratum, described target polymer micellar structure is nucleocapsid structure, the hydrophilic section tip that it is characterized in that shell is connected to the targeting ligand molecular, the hydrophobic section load hydrophobic anticancer drug of kernel, exist the acid-sensitive response subsurface stratum of one deck load negative electricity quantum dot between kernel and shell, when pH 5.0, acid-sensitive layer is hydrophilic, when pH 7.4, acid-sensitive layer is hydrophobic, described micelle consists of the Polyethylene Glycol of targeted molecular functionalization-acid-sensitive section-hydrophobic section triblock copolymer, acid-sensitive section is poly-aminoacyl diisopropyl ethylenediamine, hydrophobic section is gallbladder acid, the described preparation method with target polymer micelle of acid-sensitive subsurface stratum comprises the steps: the polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine of the polyethylene glycol-Radix Asparagi acyl diisopropyl ethylenediamine of the folic acid end-blocking of 2mg-cholic acid triblock copolymer and the 8mg-mixture of cholic acid triblock copolymer and the paclitaxel of 1mg are dissolved in the oxolane of 1ml, in the PBS that the pH that is added drop-wise to 10ml in ultrasonic situation is 5.0, stirring is spent the night, oxolane is vapored away naturally, the membrane filtration that is 0.45 μ m with aperture is removed the paclitaxel aggregation, use the centrifugal ultrafiltration pipe, MWCO=100 kD, centrifugal ultrafiltration 3 times, add negative electricity quantum dot CdSe/ZnS, 100 μ L, 8 μ moL/L, mix latter static 30 minutes, regulate pH to 7.4, the centrifugal quantum dot of removing not load of 6000r/min, obtain.
2. target polymer micelle as claimed in claim 1, is characterized in that described targeting ligand molecular is folic acid.
3. target polymer micelle as claimed in claim 1, is characterized in that described hydrophobic anticancer drug is paclitaxel.
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| CN103059297B (en) * | 2012-12-28 | 2014-11-05 | 武汉大学 | Multifunctional degradable polyasparaginate modified polymer and preparation method thereof |
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| CN107224589B (en) * | 2017-05-08 | 2021-03-26 | 中山大学 | MR imaging polymer micelle for delivering antibody for tumor immunotherapy and preparation method thereof |
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| 新型纳米材料量子点在生物及药学领域的研究进展;薛冰等;《中国医药工业杂志》;20101231;第41卷(第12期);第937-942页 * |
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