CN109678993A - A kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection, preparation method and applications - Google Patents
A kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection, preparation method and applications Download PDFInfo
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Abstract
A kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection, preparation method and applications.The fluorescence probe is that internal standard signal element R is connected with weary oxygen detection unit platinum porphyrins by polyvinylpyrrolidone, and the Ratio-type namo fluorescence probe specific structure is as shown in formula I:Wherein, R is the fluorogen being not responding to oxygen；The polymerization degree n of polyvinylpyrrolidone is between 150-200.
Description
Technical field
The present invention relates to a kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection and its biology at
As application, belong to fluorescence probe field.
Background technique
Weary oxygen is a kind of histanoxia state, typically occurs in the pathogenic process of the diseases such as tumour, angiocarpy, apoplexy
In.Since the malignant proliferation of tumour causes new vessels abnormal and amount of blood supply deficiency, tumor hypoxia is considered as the one of solid tumor
Kind common trait (Nat.Rev.Cancer 2002,2,38).In clinical application, tumor hypoxia and poor prognosis are closely related,
And seriously hinder effective treatment (Nat.Rev.Cancer 2011,11,393) of tumour.Therefore, accurate, real-time monitoring swells
The weary oxygen degree of tumor is vital to the assessment of the diagnostic and therapeutic effects of cancer.
, Noninvasive highly sensitive by its and the advantages such as easy to operate, molecular optics imaging most develop as one kind
The visible technology of prospect is widely used in the weary oxygen detection of biosystem.However, most of existing literature report is weary
Oxygen fluorescence probe belongs to reacting activation fluorescence enhancement type (Org.Lett.2011,13,928；Angew.Chem.Int.Ed.2013,
52,13028；J.Am.Chem.Soc.2015,137,6407), be easy by the environment such as concentration and probe concentration, thickness of sample, temperature because
The influence of element, leads to that the accuracy of testing result is low, signal-to-noise ratio is high.Also, such fluorescence probe is undergone not in tumor hypoxia area
It is activated after reversible chemical reaction and generates fluorescence enhancement signal；When probe is transferred to the area Chang Yang, fluorescence signal by the area Fa Yang
Still in state of activation, it is easy to produce due to probe diffusion and weary oxygen zone-transfer bring false positive signal.In spite of few
Amount work reports the weary oxygen fluorescence probe of the Ratio-type based on fluorescence resonance energy transfer mechanism
(Angew.Chem.Int.Ed.2009,48,2741；Adv.Funct.Mater.2014,24,4823；Chem.Sci.2015,6,
1825), but they still suffer from and there are other interactions between donor and acceptor fluorescence group, such as proton and electronics turn
It moves, influences the accuracy of its testing result.In addition, most of small-molecule fluorescent probes show poor tumor-targeting, very
Its potential application in biosystem is limited in big degree.
Summary of the invention
For existing weary oxygen fluorescence probe the biosystem the problem of, the present invention provides a kind of internal standard Ratio-types
Namo fluorescence probe, which is characterized in that using, as internal standard signal element, oxygen is quick to the insensitive fluorogen of oxygen in the probe
The platinum porphyrins fluorogen of sense detects signal element as weary oxygen, and the two passes through water-soluble and biocompatibility polyvinylpyrrolidine
Alkanone is connected.Believe using the ratio of the fluorescence intensity of the fluorogen insensitive to oxygen and platinum porphyrins unit phosphorescence intensity as reading
Number, Ratio-type quantitative detection can be realized to tumor hypoxia.It is provided by the present invention based on oxygen to platinum porphyrins phosphorescence quenching mechanism
Namo fluorescence probe Reversible Cycle detection can be often carried out between oxygen region in weary oxygen-, realize that real-time tracing is weary in organism
Oxygen region and weary oxygen degree.Amphiphilic polymer can be self-assembly of the nano particle of stable homogeneous in water, have benefited from
High-permeability and retention effect (enhanced permeability and retention effect, EPR), so that the nanometer
Fluorescence probe has good tumor-targeting in biosystem.
It is an advantage of the invention to provide a kind of internal standard Ratio-type nanometer of reversible weary oxygen-often oxygen cycle detection is glimmering
Light probe, the probe are Formulas I compound represented:
Wherein, polymer compound as shown in Formula II is with compound shown in formula III through reversible addion-fragmentation chain transfer
It reacts to obtain with compound shown in formula IV after polymerization (RAFT) reaction；R is the fluorogen being not responding to oxygen；A is base shown in Formula V
Group (the curve place of marking is to replace position)；The polymerization degree n of polyvinylpyrrolidone is between 150-200.
Another object of the present invention is, provides a kind of method of compound shown in preparation formula I, the method includes walking as follows
It is rapid:
(1) in the absence of oxygen, in the presence of a free-radical initiator, will change shown in compound represented II and formula III
Conjunction object is in 65 DEG C~75 DEG C progress reversible addion-fragmentation chain transfer polymerization (RAFT) polymerization reactions (when the RAFT polymerization reaction
Between be 10 hours to 24 hours), obtain compound shown in Formula IV (its weight average molecular weight is between 15,000~40,000)；
(2) in the case where there is inert gas existence condition, compound shown in Formula IV is hydrolyzed 20 hours~24 in 40 DEG C of pure water
Hour, obtain compound shown in Formula VII；
(3) in the case where having inert gas and cesium carbonate existence condition, by compound shown in compound shown in Formula VII and formula IV,
In normal-temperature reaction 3 hours~5 hours, object (compound shown in Formulas I) is obtained.
Still a further object of the present invention is to provide above-mentioned internal standard Ratio-type namo fluorescence probe, and that weary oxygen is carried out in organism is glimmering
The application of light imaging.
Detailed description of the invention
Fig. 1 is (the 0.5mg mL of probe I -1-1, detailed in Example 1) dynamic light scattering (DLS) figure in water；
Wherein, abscissa is diameter (nm), and ordinate is intensity percent (%)
Fig. 2 is probe I -1, polymer VII-1, compound IV absorption spectrum comparison diagram；
Wherein, abscissa is wavelength (nm), and ordinate is absorbance.
Fig. 3 is (the 0.5mg mL of probe I -1-1) fluorescence emission spectrum under different oxygen in water；
Wherein, abscissa is wavelength (nm), and ordinate is fluorescence intensity (a.u.), excitation wavelength 405nm.
Fig. 4 is (the 0.5mg mL of probe I -1-1) in the ratio fluorescent value followed in ring filling nitrogen and oxygen water, wherein comparing
Rate value (ratio) is the ratio of fluorescence intensity and phosphorescence intensity at 675nm at 525nm.
Fig. 5 is (the 0.5mg mL of probe I -1-1) under aerobic and hypoxic condition with human lung carcinoma cell (A549 cell) be incubated for 1
Fluorescence imaging figure after hour.
Fig. 6 is the cell toxicity data figure of probe I -1；
Wherein, abscissa is concentration (mg mL-1), ordinate is versus cell survival rate (Relative cell
viability)。
Fig. 7 is (the 0.5mg mL of probe I -1-1) enter the living imaging figure after nude mice 12h by tail vein injection；
Wherein, left figure is the nude mice picture under light field, and right figure is the fluorescence imaging figure that 405nm light source irradiates lower nude mice.
Specific embodiment
In the present invention, the compound as shown in Formula II polymerize with compound shown in formula III through reversible addion-fragmentation chain transfer
(RAFT) polymer VI (presoma) weight average molecular weight obtained is preferably controlled 15,000~40,000.Such as dividing equally for presoma again
Son amount is too small, then its water solubility reduces, and is unfavorable for the application in biosystem.The weight average molecular weight of the right presoma is too
Greatly, then its nano particle diameter for being self-assembly of in aqueous solution is excessive, reduces its passive tumor-targeting, and it is swollen to be unfavorable for living body
The weary oxygen imaging of tumor.
In another optimal technical scheme of the invention, compound shown in Formula II (chain-transferring agent) and formula III shownization
The molar ratio of object (monomer) is closed between 100~400, preferred molar ratio is between 150~200；
In another optimal technical scheme of the invention, R is the fluorogen insensitive to oxygen, and optimal R is Formula IX
Shown compound；
A kind of side for the internal standard Ratio-type namo fluorescence probe preparing reversible weary oxygen-of the present invention often oxygen cycle detection
Method comprising following steps:
(1) compound, azodiisobutyronitrile and tetrahydrofuran shown in compound shown in Formula II and formula III are added to Shi Lai
In gram reaction tube, wherein the molar ratio of compound shown in compound, azodiisobutyronitrile shown in Formula II and formula III is 2:1:(100
~200), mixture is stirred to react 10 hours after freezing-pumping-dissolution recycles three times in liquid nitrogen at 65 DEG C.Reaction is completed
Afterwards, it is cooled to room temperature, is precipitated in petroleum ether, sediment is collected by filtration, and is dissolved in a small amount of methylene chloride, again in petroleum ether
Precipitating, is collected by filtration sediment, and vacuum drying obtains compound shown in Formula IV；
(2) compound shown in Formula IV and pure water are added in single port bottle, mixture under nitrogen protection, stirs at 40 DEG C
Reaction 20 hours, vacuum distillation remove water, product are dissolved in a small amount of methylene chloride, is precipitated in petroleum ether, precipitating is collected by filtration
Object, vacuum drying obtain compound shown in Formula VII；
(3) compound shown in compound, Formula V shown in Formula VII, cesium carbonate and acetonitrile are added in single port bottle, wherein formula
Compound shown in compound, Formula V shown in VII, cesium carbonate molar ratio be 1:2:10, under nitrogen protection, room temperature is anti-for mixture
It answers 5 hours.After reaction, vacuum distillation removes acetonitrile, and product is dissolved with a small amount of methylene chloride, precipitated in petroleum ether, takes out
Filter obtains sediment, and vacuum drying obtains compound shown in Formulas I.
Embodiment 1
The synthesis of compound shown in Formulas I -1 (being abbreviated as probe I -1):
Addition bromo- 1, the 8- naphthalic anhydride (2.0g, 7.22mmol) of 4- in 100mL single port bottle, ethanol amine (3.916g,
64.20mmol) and acetic acid (20mL), under nitrogen protection, oil bath back flow reaction 10 hours at 140 DEG C stop heating, naturally cold
But, it stands overnight, yellow solid is precipitated, filter, washing is dried in vacuo to obtain compound shown in Formula VIII；
1H NMR(400MHz,CDCl3, ppm) and δ=8.66 (dd, J=0.84,7.28Hz, 1H, phenyl-H), 8.58
(dd, J=0.96,8.48Hz, 1H, phenyl-H), 8.42 (d, J=7.88Hz, 1H, phenyl-H), 8.05 (d, J=
7.88Hz, 1H, phenyl-H), 7.85 (m, 1H, phenyl-H), 4.48 (t, J=4.9Hz, 2H ,-NCH2CH2OH),4.44(t,
J=5.0Hz, 2H ,-NCH2CH2OH).
In 100mL single port bottle, compound (1.75g, 5.47mmol) shown in addition Formula VIII, N methyl piperazine (3.15g,
31.26mmol) and glycol monoethyl ether (20mL), nitrogen protection, 140 DEG C back flow reaction 10 hours, be cooled to room temperature, constantly to
Water is added dropwise in reaction solution, until there is solid precipitation in reaction bottle wall, precipitation yellow solid overnight is filtered, and washing is dried in vacuo
Compound shown in Formula IX；
1H NMR(CDCl3, 400MHz, ppm) and δ=8.58 (dd, J=0.92,7.36Hz, 1H, phenyl-H), 8.51 (d,
J=8.08Hz, 1H, phenyl-H), 8.41 (dd, J=0.72,7.36Hz, 1H, phenyl-H), 7.69 (m, 1H, phenyl-
), H 7.21 (d, J=8.1.2Hz, 1H, phenyl-H), 4.45 (t, J=5.20Hz, 2H ,-NCH2CH2), OH 3.97 (t, J=
5.28Hz,2H,-NCH2CH2OH),3.32(s,4H,piperazineyl–H),2.75(s,4H,piperazineyl–H),2.45
(s,3H,methyl–H).
Compound (3.218g, 9.49mmol) shown in addition Formula IX in 200mL single port bottle, triethylamine (0.96g,
9.49mmol) the CH removed water with molecular sieve2Cl22 bromo propionyl bromide (4.32g, 20.0mmol) is diluted in molecule and screened out by (80mL)
The CH of water2Cl2In (20mL), ice bath is protected from light down, is slowly added dropwise in single port bottle with constant pressure funnel, is added, nitrogen protection,
Stirring at normal temperature 10 hours, water washing (3 × 50mL) was spin-dried for solvent, column chromatography for separation purification, solvent (DCM:CH3OH=100:
1), ethyl alcohol recrystallization obtains compound shown in Formula X；
1H NMR(400MHz,CDCl3, ppm) and δ=8.58 (dd, J=0.88,7.24Hz, 1H, phenyl-H), 8.51 (d,
J=8.04Hz, 1H, phenyl-H), 8.35 (dd, J=0.80,8.26Hz, 1H, phenyl-H), 7.74-7.70 (m, 1H,
), phenyl-H 7.21 (d, J=8.04Hz, 1H, phenyl-H), 4.60-4.53 (m, 2H ,-NCH2CH2OH),4.52-4.45
(m,2H,-NCH2CH2), OH 4.32 (q, J=6.96Hz, 1H ,-CH (Br) CH3), 3.59 (t, J=4.68Hz, 4H,
piperazineyl–H),3.27(s,4H,piperazineyl–H),2.80(s,3H,-CH2N(CH2)CH3), 1.79 (d, J=
6.96Hz,3H,-CH(Br)CH3).
Compound (708mg, 1.49mmol) shown in addition Formula X in 100mL single port bottle, ehtyl potassium xanthate (1.194g,
7.50mmol) and acetone (50mL), stirring at normal temperature 24 hours under nitrogen protection, after the reaction was completed, vacuum distillation is spin-dried for solvent, column
Chromatography purification, solvent (DCM:CH3), OH=20:1 compound shown in Formula XI is obtained；
1H NMR(400MHz,CDCl3, ppm) and δ=8.58 (m, 1H, phenyl-H), 8.51 (d, J=8.04Hz, 1H,
), phenyl-H 8.35 (dd, J=0.80,8.26Hz, 1H, phenyl-H), 7.72-7.68 (m, 1H, phenyl-H), 7.22
(d, J=8.04Hz, 1H, phenyl-H), 4.59-4.43 (m, 6H ,-NCH2CH2OH,-NCH2CH2OH,-C(S)OCH2CH3),
4.32 (q, J=7.36Hz, 1H ,-CH (Br) CH3),3.31(s,4H,piperazineyl–H),2.75(s,4H,
piperazineyl–H),2.45(s,3H,-CH2N(CH2)CH3), 1.53 (d, J=7.36Hz, 3H ,-CH (Br) CH3),1.34
(t, J=7.12Hz, 2H ,-C (S) OCH2CH3).
Compound (30.0mg, 0.058mmol), N- ethenyl pyrrolidone shown in Formula XI are added in 25mL Shrek reaction tube
Ketone (1.938g, 17.46mmol), azodiisobutyronitrile (4.76mg, 0.028mmol) and tetrahydrofuran (2mL), it is cold in liquid nitrogen
Jelly-pumping-dissolving circulation removes three times removes oxygen, restores to room temperature, is placed in 65 DEG C of oil baths and reacts 24 hours, lead to big air quenching
It goes out reaction.CH is added2Cl2(5mL) is precipitated in 200mL petroleum ether, in triplicate, is filtered, and chemical combination shown in Formula XII is dried in vacuo to obtain
Object；
In 250mL single port bottle, compound (1.5g) and 100mL water shown in Formula XII is added, it is small to be stirred to react 20 at 40 DEG C
When, vacuum distillation removes water, a small amount of CH2Cl2(5mL) lysate precipitates in 200mL petroleum ether, filters, is dried in vacuo to obtain formula
Compound shown in VII-1.
The reaction equation of above-mentioned reaction is as follows:
In the three-necked flask of 500mL, be added parahydroxyben-zaldehyde (1.85mg, 15.2mmol), benzaldehyde (4.83g,
45.6mmol) and propionic acid (300mL), after being heated to 140 DEG C of reflux, with constant pressure funnel be added dropwise pyrroles (4.075g,
60.8mmol), continue 140 DEG C of back flow reactions one and a half hours, be cooled to room temperature, vacuum distillation removing propionic acid, column Chromatographic purification,
Solvent (DCM:PE=1:1) obtains compound shown in Formula XIII；
1H NMR(CDCl3, 400MHz, ppm): δ=8.88-8.84 (m, 8H, phenyl-H), 8.22-8.20 (m, 6H,
), pyrrolyl-H 8.06 (d, J=8.36Hz, 2H, pyrrolyl-H), 7.78-7.72 (m, 9H, phenyl-H), 7.18 (d, J
=8.36Hz, 2H, phenyl-H) 5.10 (s, 1H,-OH) -2.78 (s, 2H, pyrrolyl-H)
In 100mL single port bottle, be added Formula XIII shown in compound (700mg, 1.11mmol), triethylamine (169mg,
1.67mmol) and dry methylene chloride (30mL), weighing 2 bromo propionyl bromide (360mg, 1.67mmol) are dissolved in dry dichloro
In methane (10mL), ice bath is protected from light down, is slowly added dropwise with constant pressure funnel, drips off nitrogen protection, and stirring at normal temperature reaction 10 is small
When.After reaction, it being washed with water (3 × 30mL), collects organic phase, column chromatography for separation purifies, solvent (DCM:PE=2:
3) compound shown in Formula XIV, is obtained；
1H NMR(CDCl3, 400MHz, ppm): δ=8.78 (s, 8H, phenyl-H), 8.18-8.13 (m, 8H,
), pyrrolyl-H 7.73-7.65 (m, 9H, phenyl-H), 7.46 (d, J=8.40Hz, 2H, phenyl-H), 4.70 (q, J=
2.8Hz,1H,-CH(Br)CH3), 2.02 (d, J=2.8Hz, 3H ,-CH (Br) CH3-),-2.87(s,2H,pyrrolyl-H).
It is added platinous chloride (104mg, 0.39mmol) and benzonitrile (100mL) in 200mL single port bottle, under nitrogen protection,
180 DEG C of return stirrings of oil bath react 4 hours.After reaction, it is cooled to room temperature, compound shown in addition Formula XIV (100mg,
0.13mmol), 180 DEG C of return stirrings of oil bath react 2 hours.After reaction, vacuum distillation removes benzonitrile, column chromatography for separation
Purification, solvent (DCM:PE=3:2) obtain compound shown in formula IV；
1H NMR(400MHz,CDCl3, ppm) and δ=8.76 (m, 8H, phenyl-H), 8.16 (m, J=7.2Hz, 8H,
), pyrrolyl-H 7.73 (m, 9H, phenyl-H), 7.52 (m, 2H, phenyl-H), 4.77 (q, J=6.92Hz, 1H ,-CH
(Br)CH3), 2.10 (d, J=6.92Hz, 3H ,-CH (Br) CH3-).
Its structure is as follows:
The reaction equation of above-mentioned reaction is as follows:
In 100mL single port bottle, compound (1.368g) shown in addition Formula VII -1, compound shown in formula IV (50mg,
It 0.065mmol) with acetonitrile (20mL), is eventually adding cesium carbonate (137mg, 0.058mmol), stirring at normal temperature 5 hours, is evaporated under reduced pressure
Solvent is removed, is dissolved with a small amount of methylene chloride (5mL), is precipitated in 100mL petroleum ether, is removed small molecule in triplicate, obtain Formulas I -1
Shown compound.
Embodiment 2
Internal standard Ratio-type namo fluorescence probe I-1 diameter characterization
Embodiment 3 prepare fluorescence probe I-1 mesoporphyrin unit and polyvinylpyrrolidone be respectively provided with hydrophobicity and
Hydrophily can be self-assembly of nano particle in aqueous solution.As shown in Figure 1, dynamic light scattering test result shows fluorescence
Partial size of the probe I -1 in pure aquatic system is about 100nm, and polymer dispersity index (PDI) is 0.171, shows that fluorescence is visited
Needle I-1 has suitable partial size, is suitable for fluorescence imaging in organism.
Embodiment 3
Internal standard Ratio-type namo fluorescence probe I-1 absorption spectrum
Fluorescence probe I-1 prepared by Example 3 is dissolved in deionized water, is configured to 5mg mL-1Stock solution.Take 0.2mL
Above-mentioned solution is transferred in optical quartz cuvette (10 × 10mm), and deionized water is added and is diluted to 2mL, is tested it and is absorbed light
Spectrum.As shown in Fig. 2, fluorescence probe I-1 shows porphyrin fluorescence group characteristic absorption peak at 510nm；In addition, fluorescence probe I-1
Show a strong absworption peak at 405nm, and its half-peak breadth compared with porphyrin monomer the fluorescence peak width at 405nm, this is because
Naphthalimide unit has stronger wide band absorption in 350-470nm, shows that porphyrin and naphthalimide fluorogen are successfully connected to fluorescence
In probe I -1.
Embodiment 4
Internal standard Ratio-type namo fluorescence probe I-1 is to different oxygen spectral response
Deionized water is added in optical quartz cuvette in fluorescence probe I-1 stock solution 0.2mL prepared by Example 5
It is diluted to 2mL, after being passed through the oxygen of various concentration in solution, tests its emission spectrum.As seen from Figure 3, in dampening
Oxygen content reduces, and the 670nm emission peak intensity that platinum porphyrins unit generates gradually increases, but what naphthalimide part generated
525nm emission peak intensity is almost unchanged.The fluorescence signal that emission spectrum test result shows that naphthalimide unit generates can be made
For internal standard, the weary oxygen fluorescence probe of platinum porphyrins phosphorescent signal component ratio type with oxygen sensitive, realization quantifies oxygen content
Detection.
Embodiment 5
The often oxygen cycle detection of the reversible weary oxygen-of fluorescence probe I-1
Deionized water is added in optical quartz cuvette in fluorescence probe I-1 stock solution 0.2mL prepared by Example 5
It is diluted to 2mL, after being passed through oxygen or nitrogen in solution until being saturated, tests its emission spectrum in different phase respectively.
As shown in figure 4, platinum porphyrins unit phosphorescence is quenched in probe I -1, rate value (I in the case where being saturated Oxygen Condition525nm/I675nm) big
About 12.5；In the case where being saturated condition of nitrogen gas, phosphorescence restores at 675nm, and rate value is reduced to 0.5.By ten loop tests,
- 1 rate signal of probe I has preferable circulation repeatability, can be realized the often oxygen cycle detection of reversible weary oxygen-.
Embodiment 6
Fluorescence probe I-1 weary oxygen ratio imaging in living cells
The fluorescence probe I-1 stock solution 0.1mL of the preparation of Example 5 and three groups of human lung carcinoma cells (A549 cell) are in normal oxygen
Under the conditions of (21%O2) being incubated for 4 hours altogether, (probe ultimate density is 0.5mg mL-1), then three groups of A549 cells are respectively placed in
21%, 10%, it is further cultured for 1 hour under 1% oxygen concentration, above-mentioned A549 cell is carried out respectively by confocal microscope
Fluorescence imaging.As shown in figure 5, as oxygen concentration gradually decreases, platinum porphyrins unit 660-700nm wave band phosphorescent signal by
It is cumulative strong；Meanwhile naphthalimide unit is kept approximately constant in the fluorescence signal of 500-540nm wave band, therefore can be used as interior
Mark signal.It can as detection signal using the ratio of the fluorescence intensity of 500-540nm wave band and the phosphorescence intensity of 660-700nm wave band
To realize weary oxygen ratio imaging in living cells.In addition, cytotoxicity test results shows (Fig. 6), when -1 concentration of probe I reaches
6mg mL-1When, cell survival rate is about 90%, shows that probe I -1 has very low cytotoxicity.
Apply example 7
Tumor hypoxia imaging of the fluorescence probe I-1 in intravital mouse model
The fluorescence probe I-1 (0.5mg/mL, 100 μ L) that Example 5 is prepared passes through tail vein injection to tumor bearing nude mice 12h
After carry out living imaging.As shown in fig. 7, being only clearly observed very strong phosphorescent signal at mouse tumor position, show nanometer
Fluorescence probe I-1 can selectively targeted tumor tissues, and weary oxygen imaging is carried out to it.
Claims (8)
1. a kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection, which is characterized in that internal standard signal list
First R is connected with weary oxygen detection unit platinum porphyrins by polyvinylpyrrolidone, and the Ratio-type namo fluorescence probe is specifically tied
Structure is as shown in formula I:
Wherein, R is the fluorogen being not responding to oxygen；The polymerization degree n of polyvinylpyrrolidone is between 150-200.
2. the internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-of one kind according to claim 1 often oxygen cycle detection,
It is characterized in that, compound shown in structural formula I polymer compound as shown in Formula II and formula III is through reversible addition-fracture chain
Transfer polymerization (RAFT) reacts to obtain with compound shown in formula IV afterwards, and structural formula is as follows:
Wherein, the A in formula IV is group shown in Formula V.
3. the internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-of one kind according to claim 1 often oxygen cycle detection,
It is characterized in that, the weight average molecular weight of the structural formula I polymer is more than or equal to 15000.
4. the internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-of one kind according to claim 2 often oxygen cycle detection,
It is characterized in that, the molar ratio of compound shown in compound and formula III shown in Formula II is between 100~400.
5. the internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-of one kind according to claim 4 often oxygen cycle detection, special
Sign is that the molar ratio of compound shown in compound and formula III shown in Formula II is between 150~200.
6. the internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-of one kind according to claim 1 often oxygen cycle detection,
It is characterized in that, the R group is compound shown in Formula IX.
7. a kind of preparation method of the internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection, which is characterized in that
The internal standard Ratio-type namo fluorescence probe of the reversible weary oxygen-often oxygen cycle detection is made by following method: in the condition of anaerobic
Under, it is compound shown in Formula IV compound represented II and formula III in tetrahydrofuran by R group, it is poly- by RAFT in 65 DEG C
The polymer hydrolysis obtained after closing reaction 10 hours, the polymer after hydrolysis is again with compound shown in formula IV in acetonitrile, Yu Chang
Temperature reaction obtains object in 5 hours；
Structural formula is as follows:
Wherein, the A in formula IV is group shown in Formula V.
8. a kind of internal standard Ratio-type namo fluorescence probe of reversible weary oxygen-often oxygen cycle detection as described in claim 1 is in life
The application of weary oxygen fluorescence imaging in objects system.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112745303A (en) * | 2019-10-30 | 2021-05-04 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN113984729A (en) * | 2021-10-29 | 2022-01-28 | 中国科学院自动化研究所 | Synthesis method and application of hypoxic response type ratiometric probe |
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| CN106634965A (en) * | 2016-12-08 | 2017-05-10 | 南方科技大学 | Extracellular ratio-type oxygen sensing probe, and preparation method and applications thereof |
| CN106905389A (en) * | 2017-01-24 | 2017-06-30 | 华东理工大学 | A kind of beta galactosidase fluorescence probe with Intracellular retention ability |
| CN108333162A (en) * | 2018-02-07 | 2018-07-27 | 南方科技大学 | A kind of fluorescence oxygen probe and its preparation method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US6726925B1 (en) * | 1998-06-18 | 2004-04-27 | Duke University | Temperature-sensitive liposomal formulation |
| CN106634965A (en) * | 2016-12-08 | 2017-05-10 | 南方科技大学 | Extracellular ratio-type oxygen sensing probe, and preparation method and applications thereof |
| CN106905389A (en) * | 2017-01-24 | 2017-06-30 | 华东理工大学 | A kind of beta galactosidase fluorescence probe with Intracellular retention ability |
| CN108333162A (en) * | 2018-02-07 | 2018-07-27 | 南方科技大学 | A kind of fluorescence oxygen probe and its preparation method and application |
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| CN112745303A (en) * | 2019-10-30 | 2021-05-04 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN112745303B (en) * | 2019-10-30 | 2022-04-22 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN113984729A (en) * | 2021-10-29 | 2022-01-28 | 中国科学院自动化研究所 | Synthesis method and application of hypoxic response type ratiometric probe |
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