CN105367805A - H2O2-responded fluorescent single-molecule conjugated polymer and micelle, preparation method and application thereof - Google Patents
H2O2-responded fluorescent single-molecule conjugated polymer and micelle, preparation method and application thereof Download PDFInfo
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- CN105367805A CN105367805A CN201510818276.5A CN201510818276A CN105367805A CN 105367805 A CN105367805 A CN 105367805A CN 201510818276 A CN201510818276 A CN 201510818276A CN 105367805 A CN105367805 A CN 105367805A
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Abstract
The invention relates to the technical field of biological medicine, and especially relates to a hydrogen peroxide (H2O2)-responded fluorescent single-molecule conjugated polymer micelle used for discriminating tumor cells and normal cells, a preparation method and an application thereof. The fluorescent single-molecule conjugated polymer comprises a hydrophobic hyper branched conjugated polymer core and a hydrophobic polymer arm, and the hydrophobic hyper branched conjugated polymer core and the hydrophobic polymer arm are connected by using a H2O2-responded borate bond. According to the invention, a micelle solution formed by assembling the fluorescent single-molecule conjugated polymer can specially generate color change according to H2O2 response in the tumor cells, the fluorescent single-molecule conjugated polymer micelle has fluorescence imaging with different colors to tumor cells and H2O2 in the normal cells, and can reach a purpose of discrimination of the tumor cells and the normal cells.
Description
Technical field
The present invention relates to biomedicine technical field, particularly a kind of H
2o
2the fluorescence unit molecule conjugated polymers of response and micella, preparation method and application, described fluorescence unit molecule conjugated polymers micella can be used for differentiating tumour cell and normal cell.
Background technology
In recent years, relevant to cancer molecular imaging plays more and more important effect in the Diagnosis and Treat of cancer.H
2o
2second messenger as cell conductance participates in the adjustment of various physiological process.Any H caused because of environmental stress or transgenation
2o
2exception produce and accumulation, all can cause the generation of some diseases, comprise the diseases such as cancer, aging, cardiovascular disorder, A Cihaimo disease, Parkinson's disease.If can to H in cell
2o
2detect and imaging, will be of great significance medical diagnosis on disease, prevention and pathological study tool.
For realizing H
2o
2detect and imaging, lot of documents reports the small molecules H based on " fluorescence on-off " type
2o
2probe (
acc.Chem.Res.2011,44,793-804;
biosens.Bioelectron.2014,56,58-63;
j.Am.Chem.Soc.2011,133,10960-10965;
j.Am.Chem.Soc.2011,133,10629-10637 etc.), but, this small molecules " fluorescence on-off " type H
2o
2probe often causes data distortion because of serious photobleaching, high concentration dependent and the apparatus factor (illumination stability) of probe itself, the final sensitivity affecting probe.
Summary of the invention
In recent years, conjugated polymers because fluorescence quantum yield is high, the advantage such as the signal amplification of good light stability and " molecular wire ", it is made to become rapidly the study frontier in fluorescent optical sensor field, especially based on the conjugated polymers containing fluorescence " D-A " unit of FRET effect, under the effect of FRET, can construct become highly sensitive, " ratio fluorescent " probe that selectivity is strong.Therefore, in order to break through above-mentioned traditional small molecules " fluorescence on-off " type H
2o
2the limitation of probe, the present invention constructs a kind of " ratio fluorescent " type H based on conjugated polymers
2o
2probe, for H in viable cell
2o
2imaging and detection.
The invention provides a kind of H
2o
2the fluorescence unit molecule conjugated polymers of response and micella thereof, to solve traditional small molecules of the prior art " fluorescence on-off " type H
2o
2probe because of serious photobleaching, high concentration dependent and the apparatus factor (illumination stability) of probe itself and the data distortion caused, finally affects the technical matters such as probe sensitivity.
The present invention also provides above-mentioned H
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response.
The present invention also provides above-mentioned H
2o
2response fluorescence unit molecule conjugated polymers prepare intermediate and micella.
The present invention also provides above-mentioned H
2o
2the fluorescence unit molecule conjugated polymers micella of response is at H
2o
2h in detection, cell
2o
2the application of imaging, tumour cell and Normocellular discriminating aspect.
The object of the present invention is achieved through the following technical solutions:
A kind of H
2o
2the fluorescence unit molecule conjugated polymers of response, it comprises hydrophobic hyperbranched conjugated polymer core and hydrophilic polymeric arms, passes through H between described hydrophobic hyperbranched conjugated polymer core and hydrophilic polymeric arms
2o
2the boric acid ester bond of response connects.
Preferably, described hydrophobic hyperbranched conjugated polymer core primarily of FRET donor monomer (as
blue-fluorescence), FRET acceptor monomer (as
green fluorescence) and branched monomer prepare.
Preferably, described FRET donor monomer is the monomer of the bifunctional based on the brominated of fluorenes and/or boric acid base group, described FRET acceptor monomer is based on 2,1, the bifunctional monomer of the brominated and/or boric acid base group of 3-diazosulfide, described branched monomer is the monomer containing three or more bromines and/or boric acid base group functional group.
In some embodiments of the invention, described FRET donor monomer is selected from one or more of following monomer: 9,9-di-n-hexyl-2,7-hypoboric acid fluorenes, 9,9-di-n-hexyl-2,7-dibromo fluorenes, 9,9-di-n-octyl-2,7-hypoboric acid fluorenes,
n-(n-hexyl)-3,6-hypoboric acid carbazole,
n-(n-octyl)-3,6-hypoboric acid carbazole, 9,9-di-n-hexyl-2-bromo-7-boric acid fluorenes, 9,9-di-n-octyl-2-bromo-7-boric acid fluorenes,
n-(n-hexyl)-3-bromo-6-boric acid carbazole, or
n-(n-octyl)-3-bromo-6-boric acid carbazole etc., monomer like any associated class all should drop in the protection domain of the application.
In some embodiments of the invention, described FRET acceptor monomer is selected from one or both of following monomer: 4,7-bis-bromo-2; 1,3-diazosulfide or the bromo-7-boric acid-2,1 of 4-; 3-diazosulfide etc., monomer like any associated class all should drop in the protection domain of the application.
In some embodiments of the invention, described branched monomer is selected from one or both of following monomer: 1,3,5-tribromo-benzene or 1-boric acid-3,5-dibromobenzene etc., monomer like any associated class all should drop in the protection domain of the application.
Preferably, described hydrophilic polymeric arms comprises hydrophilic polyoxyethylene glycol.
Described H
2o
2the fluorescence unit molecule conjugated polymers of response self-assembly can form unimolecular micelle in water medium, and described unimolecular micelle is to be assembled into fluorescence unit molecule conjugated polymers micella further, this fluorescence unit molecule conjugated polymers micella can to H in water medium
2o
2respond specifically.
A kind of above-mentioned H
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, comprises the following steps:
(1) FRET donor monomer, FRET acceptor monomer and branched monomer are carried out Suzuki cross-coupling reaction under catalyst action, obtain hyperbranched conjugated polymer one; Wherein, ensure the total bromine in all raw materials and/or boric acid base group functional group equimolar ratio as far as possible;
Preferably, described in step (1), the molar feed ratio of FRET acceptor monomer is: acceptor monomer/(donor monomer+acceptor monomer)=5%; Relative to the donor monomer+acceptor monomer of 2mmol, the charging capacity of branched monomer is 0.1 ~ 0.5mmol, is preferably 0.1mmol.
(2) continue additionally to add catalyzer and the donor monomer containing two boric acid base groups as end-capping reagent, terminated polymer, obtains the hyperbranched conjugated polymer two that end group is boric acid base group;
Preferably, the polyreaction in step (1) and (2) is Suzuki cross-coupling reaction, and the oil phase of reaction is toluene, and aqueous phase is Na
2cO
3solution, catalyzer is Pd (PPh
3)
4.
The end-capping reagent additionally added in step (2) is selected from one or more of following monomer: 9,9-di-n-hexyl-2,7-hypoboric acid fluorenes, 9,9-di-n-octyl-2,7-hypoboric acid fluorenes or
n-(n-hexyl)-3,6-hypoboric acid carbazole;
(3) adopt trivalent alcohol to carry out esterification to the boric acid of hyperbranched polymer end group, obtain boronic acid containing ester bond and have the hyperbranched conjugated polymer three of a large amount of terminal hydroxy group; Trivalent alcohol carries out the reaction of esterification to boric acid, adopts functional group's equimolar ratio reaction;
Preferably, trivalent alcohol can be selected from one or more of following monomer: glycerol, BT, 1,2; 5-penta triol, 1,2,6-hexanetriol, 1,2; 7-triol in heptan and the pungent triol of 1,2,8-etc., trivalent alcohol like any associated class all should drop in the protection domain of the application.
Preferably, the temperature of reaction in step (1), (2) and (3) is 111-125 DEG C, and the reaction times in step (1), (2) and (3) is respectively 1 to 5 day, 1 to 3 day and 4 little of 2 days.
Reaction solvent in step (1), (2) and (3) is toluene; The reaction product of step (2) and (3) with an organic solvent carries out aftertreatment, dry afterwards; Wherein, described organic solvent is selected from one or more of tetrahydrofuran (THF), anhydrous methanol, normal hexane and acetone; The siccative that dewaters that described drying process uses is for anhydrous magnesium sulfate.
(4) the described hyperbranched conjugated polymer three containing a large amount of terminal hydroxy group is reacted with poly glycol monomethyl ether p-toluenesulfonic esters (PEG-OTs) under highly basic exists, obtain the starlike conjugated polymers of boronic acid containing ester bond, i.e. described H
2o
2the fluorescence unit molecule conjugated polymers of response.
Preferably, in step (4), highly basic used is sodium hydride.Temperature of reaction in step (4) is 68 to 95 DEG C, and the reaction times is 1 to 3 day, and reaction solvent is tetrahydrofuran (THF), and PEG-OTs is the poly glycol monomethyl ether p-toluenesulfonic esters that molecular weight is greater than 1000.Reaction product with an organic solvent carries out aftertreatment, and organic solvent is selected from one or more of normal hexane, anhydrous methanol and ethanol.
Preferably, the starlike conjugated polymers of the described boronic acid containing ester bond prepared is prepared fluorescence unit molecule conjugated polymers micellar aqueous solution by the following method: dissolved in organic solvent by the starlike conjugated polymers of boronic acid containing ester bond, at room temperature instilled in ultrapure water also constantly to stir and carried out micella assembling, removing organic solvent, obtains fluorescence unit molecule conjugated polymers micellar aqueous solution.Preferably, described organic solvent be selected from tetrahydrofuran (THF) and
n,N'-dimethyl formamide.Preferably, the time of described assembling is 0.5 to 12 hour, and assembling temperature is 15 to 30 DEG C.In one embodiment of this invention, the molecular weight cut-off of the dialysis tubing that described removing organic solvent dialysis procedure uses is 3500, and water-dialyzing is ultrapure water.
The present invention also asks the hyperbranched conjugated polymer two protecting a kind of above-mentioned step (2) to prepare.
The present invention also asks the hyperbranched conjugated polymer three protecting a kind of above-mentioned step (3) to prepare.
The present invention also asks to protect a kind of above-mentioned fluorescence unit molecule conjugated polymers micellar aqueous solution prepared.
The present invention also asks to protect above-mentioned H
2o
2the fluorescence unit molecule conjugated polymers of response and micella thereof are at H
2o
2h in detection, cell
2o
2the application of imaging, tumour cell and Normocellular discriminating aspect.
Especially, described fluorescence unit molecule conjugated polymers micella being applied as in tumour cell and Normocellular discriminating, described fluorescence unit molecule conjugated polymers micella is to the H in tumour cell and normal cell
2o
2there is the fluorescence imaging of different colours.In one embodiment, the assembling micella of described fluorescence unit molecule conjugated polymers to the imaging of tumour cell display green fluorescence, and shows blue-fluorescence imaging to normal cell.
After described fluorescence unit molecule conjugated polymers micella enters tumour cell, high density H in tumour cell
2o
2under environment; the hydrophilic polymeric arms fracture of fluorescence unit molecule conjugated polymers; hydrophobic hyperbranched conjugated polymer core is assembled because losing the protection of polymeric arms; cause between super branched molecule and significantly strengthen with FRET in molecule; and with the change of fluorescence from blueness to green, to realize the imaging of tumour cell green fluorescence.
Described fluorescence unit molecule conjugated polymers micella utilizes H in tumour cell compared with normal cell
2o
2the difference that content is high, specifically only to H in tumour cell
2o
2respond and start FRET effect, presenting the green fluorescence of acceptor; And to H low in normal cell
2o
2content does not respond, and can not open FRET effect, can only be the blue-fluorescence of modern face.In view of conjugated polymers micella is to H in tumour cell and normal cell
2o
2have the fluorescence imaging that color is different, the present invention can realize differentiating tumour cell and Normocellular object.
Described tumour cell can be cervical cancer cell, breast cancer cell, melanoma cell, lung carcinoma cell, the one in kidney cancer cell, pancreatic cancer cell, lymphoma cell and myeloma cell.
H of the present invention
2o
2" ratio fluorescent " type unit molecule conjugated polymers micella of response, as probe, has following advantage:
1, due to the present invention's " ratio fluorescent " type H
2o
2the light stability of probe excellence, to the no dependence of concentration and probe concentration, environment and apparatus factor, can avoid traditional small molecules " fluorescence on-off " type H effectively
2o
2probe is because of serious photobleaching, concentration dependent and the apparatus factor (illumination stability) of probe itself and the data distortion caused and the technical matters such as sensitivity is low;
2, fluorescence unit molecule conjugated polymers of the present invention, comprises hydrophobic hyperbranched conjugated polymers core and hydrophilic polymeric arms, and by using H between core and arm
2o
2the boric acid ester bond of response connects, and in water, self-assembly forms nano-micelle, can be entered in cell by endocytosis soon; Based on the H of boric acid ester bond
2o
2h in responsiveness and tumour cell
2o
2h in (10 ~ 50 μMs) compared with normal cell
2o
2the feature that (0.05 ~ 0.7 μM) content is higher, fluorescence unit molecule conjugated polymers micella of the present invention, to H in tumour cell and normal cell
2o
2there is the fluorescence imaging of different colours, can specifically to H in tumour cell
2o
2response, cause hydrophilic polymeric arms fracture, hydrophobic hyperbranched conjugated polymer core is assembled because losing the protection of polymeric arms, causes FRET significantly to strengthen, with fluorescence color from blueness to the transformation of green; But, for normal cell, due to H
2o
2content is low, can not start FRET effect, finally presents blue-fluorescence imaging.Utilize this fluorescence unit molecule conjugated polymers micella to H in tumour cell and normal cell
2o
2there is the fluorescence imaging of different colours, can reach and differentiate tumour cell and Normocellular object;
3, fluorescence unit molecule conjugated polymers micella of the present invention is in tumour cell and normal cell discriminating, and contrast traditional discriminating means based on albumen and tumor markers identification, the present invention utilizes in tumour cell and normal cell first has different H
2o
2the microenvironment feature of content, utilize fluorescence unit molecule conjugated polymers micella to H
2o
2extremely sensitive advantage and utilize fluorescence unit molecule conjugated polymers micella normal cell not to be responded to the otherness of the imaging color in blue-fluorescence to tumor cell specific response in green fluorescence, very dexterously and intuitively by the difference of color, (blueness is normal cell, green is tumour cell), just can simply distinguish tumour cell and normal cell and differentiate, this technology be expected to be applied in the fields such as the front diagnosis of cancer and cell carcinogenesis research.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of the fluorescence unit molecule conjugated polymers that embodiment 1 is synthesized;
Fig. 2 is the fluorescence unit molecule conjugated polymers that synthesizes of embodiment 1 and intermediate thereof
1hNMR spectrogram;
Fig. 3 is the fluorescence unit molecule conjugated polymers of embodiment 1 synthesis and the FTIR spectrogram of intermediate thereof;
Fig. 4 is transmission electron microscope (TEM) photo of fluorescence unit molecule conjugated polymers micella prepared by embodiment 1;
Fig. 5 is that the fluorescence unit molecule conjugated polymers micella of embodiment 1 preparation is to H
2o
2tEM photo after response;
Fig. 6 is that the fluorescence unit molecule conjugated polymers micella of embodiment 1 preparation is to H
2o
2the ultraviolet spectrogram of response;
Fig. 7 is that the fluorescence unit molecule conjugated polymers micella of embodiment 1 preparation is to H
2o
2the fluorescence spectrum figure of response;
Fig. 8 is that the fluorescence unit molecule conjugated polymers micella of embodiment 1 preparation is to H
2o
2the ratio fluorescent figure of response and solution fluorescence color camera;
Fig. 9 is that the fluorescence unit molecule conjugated polymers micella of embodiment 1 preparation is to tumour cell and normal cell fluorescent images;
Figure 10 is the schematic diagram of the fluorescence unit molecule conjugated polymers of the 2-in-1 one-tenth of embodiment;
Figure 11 is the schematic diagram of the fluorescence unit molecule conjugated polymers that embodiment 3 is synthesized;
Figure 12 is the schematic diagram of the fluorescence unit molecule conjugated polymers that embodiment 4 is synthesized;
Figure 13 is the schematic diagram of the fluorescence unit molecule conjugated polymers that embodiment 5 is synthesized;
Figure 14 is the schematic diagram of the fluorescence unit molecule conjugated polymers that embodiment 6 is synthesized.
Embodiment
The invention provides a kind of H
2o
2the fluorescence unit molecule conjugated polymers of response and micella and their preparation method and application, especially, can be used for differentiating tumour cell and normal cell.Fluorescence unit molecule conjugated polymers of the present invention, comprises hydrophobic hyperbranched conjugated polymers core and hydrophilic polymeric arms, by using hydrogen peroxide (H between described hydrophobic hyperbranched conjugated polymer core and hydrophilic polymeric arms
2o
2) the boric acid ester bond that responds connects.
Especially, during for differentiating tumour cell and normal cell, fluorescence unit molecule conjugated polymers micella of the present invention, can specifically only to H in tumour cell
2o
2response, cause the hydrophilic polymeric arms fracture of this fluorescence unit molecule conjugated polymers, its hydrophobic hyperbranched conjugated polymer core is assembled because losing the protection of polymeric arms, cause between super branched molecule and significantly strengthen with intramolecular fluorescence Resonance energy transfer (FRET), and with the change of fluorescence color before responding, present the green fluorescence of FRET acceptor; And to normal cell, due to H
2o
2the cause that content is low, can not start FRET effect, can only present the blue-fluorescence of FRET donor.In view of fluorescence unit molecule conjugated polymers micella of the present invention is to H in tumour cell and normal cell
2o
2there is the fluorescence imaging that color is different, can reach and differentiate tumour cell and Normocellular object.
Compared with prior art, the present invention utilizes first in tumour cell and normal cell and has different H
2o
2the microenvironment feature of content, utilize fluorescence unit molecule conjugated polymers micella to H
2o
2extremely sensitive advantage and utilize fluorescence unit molecule conjugated polymers micella normal cell not to be responded to the otherness of the imaging color in blue-fluorescence to tumor cell specific response in green fluorescence, very dexterously and intuitively by the difference of color, (blueness is normal cell, green is tumour cell), just can simply distinguish tumour cell and normal cell and differentiate, this technology be expected to be applied in the fields such as the front diagnosis of cancer and cell carcinogenesis research.
Below in conjunction with accompanying drawing, embodiments of the invention are elaborated.Specific embodiment is all implemented premised on technical scheme of the present invention, comprise detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
embodiment 1
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) preparation of intermediate:
As shown in Figure 1, by FRET donor monomer 9,9-di-n-hexyl-2,7-hypoboric acid fluorenes (1.05mmol), FRET donor monomer 9,9-di-n-hexyl-2,7-dibromo fluorenes (0.85mmol), FRET acceptor monomer 4, bromo-2,1, the 3-diazosulfides (0.1mmol) of 7-bis-, branched monomer 1,3,5-tribromo-benzene (0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days, obtain hyperbranched conjugated polymer one;
Continue additionally add under nitrogen atmosphere FRET donor monomer 9,9-di-n-hexyl-2,7-hypoboric acid fluorenes (
monomerA, 2.0mmol) and as end-capping reagent and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), in 500mL n-hexane/acetone (1:1), precipitation 3 times, obtains faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer two that end group is boric acid base group, be labeled as
hPFBT.
By described hyperbranched conjugated polymer two
hPFBT, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT-OH(63.7%yield).
As shown in Figure 2, the nuclear-magnetism structure confirmation data of this polymkeric substance is as follows:
1hNMR (CDCl
3, ppm): 8.07,8.01,7.93,7.88-7.82,7.74-7.53,7.47-7.36,2.23-2.01,1.50-1.12,0.83-0.72.
As shown in Figure 3, the infrared structure of this polymkeric substance confirms that data are as follows: FITR (KBr, cm
-1): 3442,2955,2911,2849,1605,1455,1405,1255,1086,1017,880,817,724.
(2) preparation of fluorescence unit molecule conjugated polymers: by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT-
star-PEG
(81%yield).
As shown in Figure 2, the nuclear-magnetism structure confirmation data of this polymkeric substance is as follows:
1hNMR (CDCl
3, ppm): 8.06,8.02,7.94,7.88-7.81,7.75-7.51,7.46-7.32,3.81-3.30,2.21-2.01,1.51-1.12,0.83-0.71.
As shown in Figure 3, the infrared structure of this polymkeric substance confirms that data are as follows: FITR (KBr, cm
-1): 3423,2893,1640,1468,1347,1282,1260,1248,1116,958,827,814.
(3) preparation of fluorescence unit molecule conjugated polymers micella: by the starlike conjugated polymers HPFBT-of boronic acid containing ester bond
star-PEG(10mg) be dissolved in 2mL tetrahydrofuran (THF), at room temperature by its slowly instillation 3mL ultrapure water, and constantly stir 2 hours, removing organic solvent, obtains starlike conjugated polymers HPFBT-
star-PEG micellar aqueous solution.
to starlike conjugated polymers HPFBT-prepared by the present embodiment
star-PEG micellar aqueous solution carries out measuring and H
2
o
2
the example of response test is as follows:
Shown in the TEM image referring to Fig. 4, the above-mentioned starlike conjugated polymers HPFBT-prepared
star-PEG micellar size is about 62.9nm.
This starlike conjugated polymers HPFBT-
starthe H of-PEG micellar aqueous solution
2o
2response test one: at this HPFBT-
star0.1mMH is added in-PEG micellar solution
2o
2, constantly stir 60 minutes.Because boric acid ester bond meets H
2o
2fracture, cause hydrophilic PEG arm to come off, hydrophobic hyperbranched conjugated polymer core HPFBT assembles because losing the protection of PEG, and nano-particles size becomes large, and this is to H
2o
2the image of the nanoparticle after response is as shown in the TEM of Fig. 5, visible, H
2o
2after response, Nanoparticle Size is about 262.5nm.
This starlike conjugated polymers HPFBT-
starthe H of-PEG micellar aqueous solution
2o
2response test two: the HPFBT-preparing a series of 2 μ g/mL
star-PEG micella PBS(pH=7.4) solution, add the different concns H of 0 ~ 0.1mM respectively
2o
2solution, stirs 60 minutes.As shown in Figure 6, uv-visible absorption spectra demonstrates: along with H
2o
2the rising of concentration, maximum absorption band reduces gradually, and this is because boric acid ester bond meets H
2o
2fracture, hydrophobic HPFBT core because of lose PEG protection and caused by assembling.Same, as shown in Figure 7, in fluorescence emission spectrum, along with H
2o
2the rising of concentration, the fluorescence cancellation gradually of FRET donor, and the fluorescence of FRET acceptor significantly increases.Ratio (I is done to FRET acceptor/donor fluorescent emission peak
540/ I
425), and and H
2o
2concentration map, acquired results as shown in Figure 8: along with H
2o
2the rising of concentration, I
540/ I
425increase gradually, finally occur a platform, imply that boric acid ester bond is to H
2o
2response reaches balance.Meanwhile, the fluorescence color of solution also clearly illustrates, at H
2o
2after response, fluorescence becomes green from blueness.This to H
2o
2special response also with the characteristic of change in fluorescence, can perform well in because of H
2o
2the tumour cell that content is different and Normocellular discriminating.
starlike conjugated polymers HPFBT-prepared by the present embodiment
star-PEG micellar aqueous solution is used for tumour cell and Normocellular identification experiment example:
Adopt the cell imaging technology of laser confocal microscope.
This example adopts Cervical Cancer HeLa Cells, mouse normally becomes fiber L929 cell and uses 0.05mMH
2o
2the L929 cell of advanced processing is as cell model.First be that the circular lid slide of 8mm is placed in 24 orifice plates, then with every hole 1.0 × 10 by diameter
4the density bed board of individual cell, places incubator overnight incubation.After cell attachment, exhaust substratum, add the HPFBT-of 600 μ L
starthe serum-free DMEM solution of-PEG micella, cultivates after 4 hours, sucks supernatant liquor, wash 2 times with ice PBS, each 5 minutes.Add subsequently 4% paraformaldehyde fixing after, room temperature lucifuge places 30 minutes.Slightly dry a small amount of anti-cancellation mountant of rear dropping and cover sample, observe under laser confocal microscope after mounting.Excitation channel is 405nm, and reception channel is blue and green channel.
Result is as follows: as shown in Figure 9, and in (a), L929 normal cell presents blue-fluorescence, and in (b), HeLa tumour cell presents green fluorescence, and this is due to H in tumour cell compared with normal cell
2o
2content is higher, HPFBT-
star-PEG micella can be specific to H in tumour cell
2o
2imaging also display is different from Normocellular green.In order to simulate the H in tumour cell
2o
2environment, as shown in (c) in Fig. 9, uses 0.05mMH
2o
2the L929 cell of advanced processing also presents the green fluorescence consistent with HeLa tumour cell.This imaging to normal cell and tumour cell different colours, can effectively for the discriminating of normal cell and tumour cell.
In addition, the present embodiment tumour cell used can also be cervical cancer cell, breast cancer cell, melanoma cell, lung carcinoma cell, the one wherein such as kidney cancer cell, pancreatic cancer cell, lymphoma cell and myeloma cell; Normal cell used can also be the one wherein such as l cell, mouse placental fibroblasts, nephrocyte, pneumonocyte, medullary cell and lymphocyte.
embodiment 2
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) as shown in Figure 1, by FRET donor monomer
9,9-di-n-octyl-2,7-hypoboric acid fluorenes(1.05mmol), FRET donor monomer 9,9-di-n-octyl-2,7-dibromo fluorenes (0.85mmol), bromo-2,1, the 3-diazosulfides (0.1mmol) of FRET acceptor monomer 4,7-bis-, branched monomer 1,3,5-tribromo-benzene (0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days; Continue additionally to add under nitrogen atmosphere
9,9-di-n-octyl-2,7-hypoboric acid fluorenes(
monomerA, 2.0mmol) and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, at 35 DEG C, vacuum-drying 2 days, can obtain hyperbranched conjugated polymer two
hPFBT-2;
By described hyperbranched conjugated polymer two
hPFBT-2, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT2-OH, as shown in Figure 10.
(2) by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT2-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT2-
star-PEG
, as shown in Figure 10.
embodiment 3
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) as shown in Figure 1, by FRET donor monomer
9,9-di-n-hexyl-2-bromo-7-boric acid fluorenes(1.9mmol), bromo-2,1, the 3-diazosulfides (0.1mmol) of FRET acceptor monomer 4,7-bis-, branched monomer 1,3,5-tribromo-benzene (0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days; Continue additionally to add under nitrogen atmosphere
9,9-di-n-hexyl-2,7-hypoboric acid fluorenes(
monomerA, 2.0mmol) and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, at 35 DEG C, vacuum-drying 2 days, can obtain hyperbranched conjugated polymer two
hPFBT-3.
By described hyperbranched conjugated polymer two
hPFBT-3, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT3-OH, as shown in figure 11.
(2) by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT3-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT3-
star-PEG
, as shown in figure 11.
embodiment 4
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) as shown in Figure 1, by FRET donor monomer
9,9-di-n-octyl-2-bromo-7-boric acid fluorenes(1.9mmol), bromo-2,1, the 3-diazosulfides (0.1mmol) of FRET acceptor monomer 4,7-bis-, branched monomer 1,3,5-tribromo-benzene (0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days; Continue additionally to add under nitrogen atmosphere
9,9-di-n-octyl-2,7-hypoboric acid fluorenes(
monomerA, 2.0mmol) and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, at 35 DEG C, vacuum-drying 2 days, can obtain hyperbranched conjugated polymer two
hPFBT-4.
By described hyperbranched conjugated polymer two
hPFBT-4, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT4-OH, as shown in figure 12.
(2) by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT4-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT4-
star-PEG
, as shown in figure 12.
embodiment 5
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) as shown in Figure 1, by FRET donor monomer
9,9-di-n-hexyl-2-bromo-7-boric acid fluorenes(1.9mmol), bromo-2,1, the 3-diazosulfides (0.1mmol) of FRET acceptor monomer 4,7-bis-, branched monomer
1,3-bis-bromo-5-boric acid benzene(0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days; Continue additionally to add under nitrogen atmosphere
9,9-di-n-hexyl-2,7-hypoboric acid fluorenes(
monomerA, 2.0mmol) and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, at 35 DEG C, vacuum-drying 2 days, can obtain hyperbranched conjugated polymer two
hPFBT-5.
By described hyperbranched conjugated polymer two
hPFBT-5, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT5-OH, as shown in figure 13.
(2) by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT5-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT5-
star-PEG
, as shown in figure 13.
embodiment 6
The H of the present embodiment
2o
2the fluorescence unit molecule conjugated polymers of response and the preparation method of micella thereof comprise the steps:
(1) as shown in Figure 1, by FRET donor monomer
9,9-di-n-octyl-2-bromo-7-boric acid fluorenes(1.9mmol), bromo-2,1, the 3-diazosulfides (0.1mmol) of FRET acceptor monomer 4,7-bis-, branched monomer 1,3,5-tribromo-benzene (0.1mmol), Na
2cO
3(2M, 9.5mL, 19.0mmol) and 20mL toluene are added in reactor, under nitrogen atmosphere, add catalyst P d (PPh
3)
4(28.9mg, 0.025mmol), 120 DEG C of stirring and refluxing react 3 days; Continue additionally to add under nitrogen atmosphere
9,9-di-n-octyl-2,7-hypoboric acid fluorenes(
monomerA, 2.0mmol) and catalyst P d (PPh
3)
4(29mg), reaction mixture continues 120 DEG C of stirring and refluxing and reacts 2 days, terminates reaction, crosses and filters insolubles, except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, at 35 DEG C, vacuum-drying 2 days, can obtain hyperbranched conjugated polymer two
hPFBT-6.
By described hyperbranched conjugated polymer two
hPFBT-6, glycerol (5.0mmol) and 50mL toluene, join in 100mL reaction flask, under nitrogen atmosphere, adopt division box except the moisture in dereaction, 120 DEG C of stirring and refluxing react 12 hours, treat that solution cools, and except desolventizing, obtain faint yellow viscous liquid; By liquid dissolves in 10mL tetrahydrofuran (THF), precipitation 3 times in 500mL n-hexane/acetone (1:1), obtain faint yellow solid, vacuum-drying 2 days at 35 DEG C, can obtain the hyperbranched conjugated polymer three of boric acid base group end-blocking
hPFBT6-OH, as shown in figure 14.
(2) by the hyperbranched conjugated polymer three of described boric acid base group end-blocking
hPFBT6-OH, PEG-OTs(5mmol), NaH and 50mL tetrahydrofuran (THF), join in reaction flask, under nitrogen atmosphere, terminate reaction after 90 DEG C of return stirrings react 3 days, wash 3 times with deionized water, use anhydrous magnesium sulfate to carry out drying to solution afterwards to dewater, concentrated solution, precipitates three times, obtains faint yellow solid in 500mL anhydrous methanol, vacuum-drying 2 days at 35 DEG C, can obtain the starlike conjugated polymers of boronic acid containing ester bond
hPFBT6-
star-PEG
, as shown in figure 14.
The fluorescence unit molecule conjugated polymers micella providing a class novel of the present invention's innovation, connected to form by boric acid ester bond by hydrophobic hyperbranched conjugated polymers core and hydrophilic polymeric arms, fluorescence unit molecule conjugated polymers micella has H
2o
2responsiveness.The present invention utilizes in tumour cell and normal cell first has different H
2o
2the microenvironment feature of content, utilize fluorescence unit molecule conjugated polymers micella to H
2o
2extremely sensitive advantage and utilize fluorescence unit molecule conjugated polymers micella normal cell not to be responded to the otherness of the imaging color in blue-fluorescence to tumor cell specific response in green fluorescence, very dexterously and intuitively by the difference of color, (blueness is normal cell, green is tumour cell), just can simply distinguish tumour cell and normal cell and differentiate, this technology be expected to be applied in the fields such as the front diagnosis of cancer and cell carcinogenesis research.
Under the instruction of the present invention and above-described embodiment, those skilled in the art are easy to predict, cited or each raw material that exemplifies of the present invention or its equivalent alterations, each working method or its equivalent alterations can realize the present invention, and the parameter bound value of each raw material and working method, interval value can realize the present invention, do not enumerate embodiment at this.
Be only several specific embodiments of the application above, but the application is not limited thereto, the changes that any person skilled in the art can think of, all should drops in the protection domain of the application.
Claims (23)
1. a H
2o
2the fluorescence unit molecule conjugated polymers of response, is characterized in that, comprise hydrophobic hyperbranched conjugated polymer core and hydrophilic polymeric arms, pass through H between described hydrophobic hyperbranched conjugated polymer core and hydrophilic polymeric arms
2o
2the boric acid ester bond of response connects.
2. H as claimed in claim 1
2o
2the fluorescence unit molecule conjugated polymers of response, it is characterized in that, described hydrophobic hyperbranched conjugated polymer core prepares primarily of FRET donor monomer, FRET acceptor monomer and branched monomer.
3. H as claimed in claim 2
2o
2the fluorescence unit molecule conjugated polymers of response, it is characterized in that, described FRET donor monomer is the monomer of the bifunctional based on the brominated of fluorenes and/or boric acid base group, described FRET acceptor monomer is based on 2,1, the bifunctional monomer of the brominated and/or boric acid base group of 3-diazosulfide, described branched monomer is the monomer containing three or more bromines and/or boric acid base group functional group.
4. the H as described in claim 1 or 2 or 3
2o
2the fluorescence unit molecule conjugated polymers of response, it is characterized in that, described hydrophilic polymeric arms comprises hydrophilic polyoxyethylene glycol.
5. H as claimed in claim 1
2o
2the fluorescence unit molecule conjugated polymers of response, is characterized in that, described H
2o
2the fluorescence unit molecule conjugated polymers of response self-assembly can form unimolecular micelle in water medium, and described unimolecular micelle is assembled into fluorescence unit molecule conjugated polymers micella further, and this fluorescence unit molecule conjugated polymers micella can to H in water medium
2o
2respond specifically.
6. arbitrary described H in a claim 1-5
2o
2the fluorescence unit molecule conjugated polymers micella of response.
7. arbitrary described H in claim 1-5
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, is characterized in that, comprise the following steps:
(1) FRET donor monomer, FRET acceptor monomer and branched monomer are carried out Suzuki cross-coupling reaction under catalyst action, obtain hyperbranched conjugated polymer one;
(2) continue additionally to add catalyzer and the FRET donor monomer containing two boric acid base groups as end-capping reagent, terminated polymer, obtains the hyperbranched conjugated polymer two that end group is boric acid base group;
(3) adopt trivalent alcohol to carry out esterification to the boric acid of the end group of hyperbranched polymer, obtain boronic acid containing ester bond and have the hyperbranched conjugated polymer three of a large amount of terminal hydroxy group;
(4) the described hyperbranched conjugated polymer three containing a large amount of terminal hydroxy group is reacted with poly glycol monomethyl ether p-toluenesulfonic esters PEG-OTs under highly basic exists, obtain the starlike conjugated polymers of boronic acid containing ester bond, i.e. described H
2o
2the fluorescence unit molecule conjugated polymers of response.
8. H as claimed in claim 7
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, it is characterized in that, the starlike conjugated polymers of the described boronic acid containing ester bond prepared is prepared fluorescence unit molecule conjugated polymers micellar aqueous solution by the following method: dissolved in organic solvent by the starlike conjugated polymers of boronic acid containing ester bond, at room temperature instilled in ultrapure water or deionized water also constantly to stir and carried out micella assembling, remove organic solvent afterwards, obtain fluorescence unit molecule conjugated polymers micellar aqueous solution.
9. H as claimed in claim 7
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, it is characterized in that, described in step (1), the molar feed ratio of FRET acceptor monomer is: FRET acceptor monomer/(FRET donor monomer+FRET acceptor monomer)=5%; The charging capacity of branched monomer is: relative to the FRET donor monomer+FRET acceptor monomer of 2mmol, 0.1 ~ 0.5mmol.
10. H as claimed in claim 7
2o
2the preparation method of fluorescence unit molecule conjugated polymers of response, is characterized in that, the polyreaction in step (1) and (2) is Suzuki cross-coupling reaction, and the oil phase of reaction is toluene, and aqueous phase is Na
2cO
3solution, catalyzer is Pd (PPh
3)
4.
11. H as claimed in claim 7
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, it is characterized in that, temperature of reaction described in step (1), (2) and (3), refers to 111 to 125 DEG C.
12. H as claimed in claim 7
2o
2the preparation method of fluorescence unit molecule conjugated polymers of response, is characterized in that, the reaction times described in step (1), (2) and (3), refers to 1 to 5 day respectively, 1 to 3 day and 4 little of 2 days.
13. H as claimed in claim 7
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, it is characterized in that, highly basic used in step (4) is sodium hydride.
14. H as claimed in claim 7
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, it is characterized in that, temperature of reaction described in step (4), refers to 68 to 95 DEG C.
15. as the H as described in arbitrary in claim 7-11
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, is characterized in that, in the reaction times described in step (4), refer to 1 to 3 day.
16. as the H as described in arbitrary in claim 7-11
2o
2the preparation method of the fluorescence unit molecule conjugated polymers of response, is characterized in that, PEG-OTs described in step (4), refers to the poly glycol monomethyl ether p-toluenesulfonic esters that molecular weight is greater than 1000.
17. as claimed in claim 8 for differentiating tumour cell and Normocellular fluorescence unit molecule conjugated polymers micella, and it is characterized in that, the time of described assembling refers to 0.5 to 12 hour, and assembling temperature refers to 15 to 35 DEG C.
The hyperbranched conjugated polymer two that in 18. 1 kinds of claim 7-17, arbitrary described step (2) prepares.
The hyperbranched conjugated polymer three that in 19. 1 kinds of claim 7-17, arbitrary described step (3) prepares.
Arbitrary fluorescence unit molecule conjugated polymers micellar aqueous solution prepared in 20. 1 kinds of claim 7-17.
Arbitrary described H in 21. claim 1-6
2o
2the fluorescence unit molecule conjugated polymers of response and micella thereof are at H
2o
2h in detection, cell
2o
2the application of imaging, tumour cell and Normocellular discriminating aspect.
22. application of fluorescence unit molecule conjugated polymers micella as claimed in claim 21 in tumour cell and Normocellular discriminating, is characterized in that, described fluorescence unit molecule conjugated polymers micella is to the H in tumour cell and normal cell
2o
2there is the fluorescence imaging of different colours.
23. application of fluorescence unit molecule conjugated polymers micella as claimed in claim 21 in tumour cell and Normocellular discriminating, it is characterized in that, described fluorescence unit molecule conjugated polymers micella to the imaging of tumour cell display green fluorescence, and shows blue-fluorescence imaging to normal cell.
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| CN108559085A (en) * | 2018-03-02 | 2018-09-21 | 南京邮电大学 | A kind of preparation and application of the weary oxygen probe of double emitting conjugated polymers |
| CN109251743A (en) * | 2017-07-12 | 2019-01-22 | 上海交通大学医学院附属瑞金医院 | Hydrogen peroxide response ratiometric nanoprobe and application thereof |
| CN117018233A (en) * | 2023-08-14 | 2023-11-10 | 浙江大学 | Nanoparticle for dual-mode diagnosis of atherosclerosis and preparation method thereof |
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| CN103842472A (en) * | 2011-09-01 | 2014-06-04 | 香港科技大学 | Biocompatible nanoparticles with aggregation induced emission characteristics as fluorescent bioprobes and methods of using the same for in vitro and in vivo imaging |
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| CN117018233A (en) * | 2023-08-14 | 2023-11-10 | 浙江大学 | Nanoparticle for dual-mode diagnosis of atherosclerosis and preparation method thereof |
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