CN108342190A - A kind of namo fluorescence probe and its preparation method and application of ratio type detection HNO - Google Patents
A kind of namo fluorescence probe and its preparation method and application of ratio type detection HNO Download PDFInfo
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- CN108342190A CN108342190A CN201810020079.2A CN201810020079A CN108342190A CN 108342190 A CN108342190 A CN 108342190A CN 201810020079 A CN201810020079 A CN 201810020079A CN 108342190 A CN108342190 A CN 108342190A
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- 239000000523 sample Substances 0.000 title claims abstract description 83
- 238000001514 detection method Methods 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 150000001875 compounds Chemical class 0.000 claims abstract description 49
- 230000000694 effects Effects 0.000 claims abstract description 26
- 241000252212 Danio rerio Species 0.000 claims abstract description 16
- 229920000642 polymer Polymers 0.000 claims abstract description 16
- 125000001424 substituent group Chemical group 0.000 claims abstract description 16
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 13
- 150000003384 small molecules Chemical class 0.000 claims abstract description 10
- 239000000693 micelle Substances 0.000 claims abstract description 9
- 125000003368 amide group Chemical group 0.000 claims description 7
- 230000003834 intracellular effect Effects 0.000 claims description 7
- 230000032050 esterification Effects 0.000 claims description 6
- 238000005886 esterification reaction Methods 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 5
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 4
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 claims description 3
- -1 compound small molecule Chemical class 0.000 claims 1
- 230000005611 electricity Effects 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 210000001082 somatic cell Anatomy 0.000 abstract 1
- 239000007853 buffer solution Substances 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 5
- UYRPRYSDOVYCOU-UHFFFAOYSA-N 2-diphenylphosphanylbenzoic acid Chemical compound OC(=O)C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 UYRPRYSDOVYCOU-UHFFFAOYSA-N 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 238000002604 ultrasonography Methods 0.000 description 4
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- OAOSXODRWGDDCV-UHFFFAOYSA-N n,n-dimethylpyridin-4-amine;4-methylbenzenesulfonic acid Chemical compound CN(C)C1=CC=NC=C1.CC1=CC=C(S(O)(=O)=O)C=C1 OAOSXODRWGDDCV-UHFFFAOYSA-N 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000448255 Congiopodus peruvianus Species 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 229920002521 macromolecule Polymers 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N methylene chloride Substances ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical group C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000010148 water-pollination Effects 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 description 1
- LIQLLTGUOSHGKY-UHFFFAOYSA-N [B].[F] Chemical compound [B].[F] LIQLLTGUOSHGKY-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 229910052796 boron Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000011503 in vivo imaging Methods 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 150000003233 pyrroles Chemical class 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- YNQYZBDRJZVSJE-UHFFFAOYSA-M sodium;2,3-dihydroxypropyl 2,3-di(octadecanoyloxy)propyl phosphate Chemical compound [Na+].CCCCCCCCCCCCCCCCCC(=O)OCC(COP([O-])(=O)OCC(O)CO)OC(=O)CCCCCCCCCCCCCCCCC YNQYZBDRJZVSJE-UHFFFAOYSA-M 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
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- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
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Abstract
The present invention relates to the preparation method and application that a kind of ratio type detects the namo fluorescence probe of HNO.Namo fluorescence probe of the present invention is made by compound P BODIPY R through both sexes polymer packages；The compound P BODIPY R general structures are as follows：Wherein R1、R2For the substituent group of introducing, the electronic effect in fluorescent probe molecule is adjusted, ratio type variation occurs for fluorescence when it being made to respond HNO.The active small molecular HNO that namo fluorescence probe provided by the invention can be used in ratio type detection organism.In the present invention nano-micelle is prepared by introducing both sexes polymer and wrapping up fluorescence probe small molecule P BODIPY R, probe small molecule bio-compatibility is set to be greatly improved, the namo fluorescence probe is selectively preferable to HNO simultaneously, anti-interference ability is stronger, with higher sensitivity, real-time tracking detection can be carried out to the HNO in active somatic cell and in zebra fish body.
Description
Technical field
The invention belongs to biological chemical field, it is related to building a kind of novel ratio type can be used for detecting HNO in organism
Namo fluorescence probe, and in particular to a kind of preparation method and biologic applications of the namo fluorescence probe of detection HNO.
Background technology
BODIPY class compounds are that a kind of fluorescent dye haveing excellent performance passes through with higher fluorescence quantum yield
Boron bridged bond and methine key are fixed two pyrrole rings in one plane, so that molecule is had rigid coplanar structure, in exciting light
Under the action of can generate more strong fluorescence, and the rigid coplanar structure of its height makes it have higher Molar Extinction
Coefficient, preferable photochemistry and photophysics stability, excitation wavelength visible region, half-peak breadth is relatively narrow the advantages that, in addition its tie
Structure is easy to modify, and launch wavelength is adjusted near infrared region.These advantages make it in biologic applications（Especially as fluorescence probe
Signal detection group）Aspect is widely paid attention to.Synthesizing the research work of such dyestuff at present, there are still deficiencies, such as design
The poorly water-soluble of fluorescent probe molecule, detection range compares limitation, is mostly used in and detects exogenous small molecule etc..
Invention content
First purpose of the invention is to provide a kind of preferable namo fluorescence probe of water solubility of ratio type detection HNO.
Second purpose of the invention is to provide a kind of preparation method of the namo fluorescence probe of ratio type detection HNO.
The namo fluorescence probe that third purpose of the present invention is to provide a kind of ratio type detection HNO is detecting intracellular HNO
Application.
4th purpose of the invention is to provide a kind of namo fluorescence probe of ratio type detection HNO in detection zebra fish body
The application of HNO.
5th purpose of the invention is to provide a kind of compound of ratio type detection HNO.
6th purpose of the invention is to provide a kind of preparation method of the compound of ratio type detection HNO.
7th purpose of the invention is to provide a kind of compound of ratio type detection HNO in the application for detecting intracellular HNO.
8th purpose of the invention is to provide a kind of compound of ratio type detection HNO HNO in detection zebra fish body
Using.
Technical scheme is as follows：
A kind of ratio type namo fluorescence probe of detection HNO, which is characterized in that the namo fluorescence probe is by compound P-
BODIPY-R is made through both sexes polymer packages；
The micromolecular compound P-BODIPY-R general structures are as follows：
Wherein R1、R2For the substituent group of introducing（R1For the amido substituent group with electron, R2Do not have for generality and pushes away by force
The group of Draw electronic effect）, the electronic effect in fluorescent probe molecule is adjusted, ratio type occurs for fluorescence when it being made to respond HNO
Variation.
The compound HO-BODIPY-R has structure as follows：
Wherein R1、R2For the substituent group of introducing（R1For the amido substituent group with electron, R2Do not have for generality and pushes away by force
The group of Draw electronic effect）, the electronic effect in fluorescent probe molecule is adjusted, ratio type occurs for fluorescence when it being made to respond HNO
Variation.
The namo fluorescence probe that HNO is detected according to ratio type of the present invention is nano-micelle, and the nano-micelle outer layer is two
Property polymer, inside be compound P-BODIPY-R.
The namo fluorescence probe of HNO is detected according to ratio type of the present invention, the micromolecular compound identification base is three
Phenylphosphine formic acid ester structure.
The namo fluorescence probe of HNO is detected according to ratio type of the present invention, the polymer has hydrophily and oleophylic
Property.
The namo fluorescence probe of HNO, the R are detected according to ratio type of the present invention1、R2It is the substituent group of introducing, adjusts
Section R group (such as choose R1Group is the amido substituent group with electron, R2Do not have strong push-and-pull electronics effect for generality
The group answered) it can make fluorescence probe fluorescence when detecting HNO that ratio type variation occur.
The present invention also provides the preparation methods that a kind of ratio type detects the namo fluorescence probe of HNO, including walk as follows
Suddenly：
(1) using HO-BODIPY-R as raw material P-BODIPY-R is obtained through esterification（Adjust R1Group (such as choose R1Group
For the amido substituent group with electron), obtain the micromolecular compound that detection HNO is ratio type variation）；
(2) namo fluorescence probe that ratio type detects HNO is made through both sexes polymer packages in the compound P-BODIPY-R.
The preparation method of the namo fluorescence probe of the ratio type detection HNO, the polymer have hydrophily and oleophylic
Property.
The preparation method and synthetic route of the namo fluorescence probe of the ratio type detection HNO are as follows：
(a) DPTS, DIPC；(b) polymer.
The present invention is HO-BODIPY-R and 1,2-（Diphenylphosphino）Benzoic acid is obtained by processes such as esterification, packages
The namo fluorescence probe NanoP-BODIPY-R of HNO is detected to energy ratio type.
Originality of the present invention is to be based on BODIPY parents, by adjusting the R on precursor structure1, R2Group electronic effect, reaches
The purpose of HNO is detected to ratio type, while by introducing both sexes polymer packages, being prepared the form of nano-micelle, being obtained biology
The namo fluorescence probe NanoP-BODIPY-R of good compatibility, the clever lightness height of detection, strong antijamming capability.
The present invention also provides a kind of compounds detecting HNO for ratio type, which is characterized in that the compound P-
The general structure of BODIPY-R is as follows：
Wherein R1、R2For the substituent group of introducing（R1For the amido substituent group with electron, R2Do not have for generality and pushes away by force
The group of Draw electronic effect）, the electronic effect in fluorescent probe molecule is adjusted, ratio type occurs for fluorescence when it being made to respond HNO
Variation.
The present invention also provides a kind of preparation methods of the compound detecting HNO for ratio type：
Preparation process is as follows：
Using HO-BODIPY-R as raw material and 1,2-（Diphenylphosphino）Benzoic acid obtains P-BODIPY-R through esterification.
The present invention further provides a kind of ratio types to detect the namo fluorescence probe of HNO in the application for detecting intracellular HNO.
The present invention further provides a kind of ratio types to detect the namo fluorescence probe of HNO in detection zebra fish body
The application of HNO.
The present invention further provides a kind of ratio types to detect the compound of HNO in the application for detecting intracellular HNO.
The compound of the HNO HNO in detection zebra fish body is detected the present invention further provides a kind of ratio type to answer
With.
The detailed preparation method of the namo fluorescence probe of ratio type detection HNO provided by the invention：
According to the micromolecular compound P-BODIPY-R, preferably R for detecting HNO1For morpholine structure, R2It is for phenyl
Example illustrates.
Preparation process is as follows：
Using HO-BODIPY-N as raw material P-BODIPY-N is obtained through esterification.
The synthetic route of the compound P-BODIPY-N is as follows：
(1) synthesis of compound P-BODIPY-N
Take 1,2-（Diphenylphosphino）Benzoic acid is dissolved in dry CH2Cl2In, it is stirred at room temperature to being completely dissolved, is cooled to 0 DEG C
Afterwards, DPTS and DIPC is added, stirs 5min, adds HO-BODIPY-N, be slowly ramped to room temperature, washing is added after having reacted
It washs, merges organic phase, anhydrous sodium sulfate drying, column chromatography obtains Orange red solid.
(2) preparation of NanoP-BODIPY-R
Both sexes polymer is added ultrasound to clear shape, P-BODIPY-R ultrasounds then is added about by the ultra-pure water for taking 1ml
20min is extremely clarified, membrane filtration, you can the namo fluorescence probe for detecting HNO is made.
The present invention is provided to detect the compound P-BODIPY-R of HNO, general structure is as follows：
Wherein R1、R2It, can be by changing R for the substituent group of introducing1、R2Obtain the probe molecule P- with specific function
Then BODIPY-R improves water solubility using the method for both sexes polymer packages, the nanometer for detecting HNO is finally made
Fluorescence probe NanoP-BODIPY-N.
Term：
Absorption is absorption value.
Fluoresence intensity are fluorescence intensity.
BODIPY is two pyrroles's molecule of fluorine boron.
I568For the fluorescence intensity of 568nm.
I601For the fluorescence intensity of 601nm.
Advantageous effects：
The namo fluorescence probe NanoP-BODIPY-R that the present invention provides a kind of ratio type detection HNO can be used for detecting cell and spot
Active small molecular HNO in horse fish body, the probe overcome some defects present in the relevant technologies, as such fluorescence probe utilizes
Triphenylphosphine structure is as recognition group, since excessive benzene ring structure so that such probe molecule is water-soluble universal poor.
In the present invention, the mode of probe small molecule is wrapped up by using amphipathic polymer, nano-micelle is prepared, obtains nano fluorescent
Probe NanoP-BODIPY-R solves such fluorescence probe to which the water-soluble of such probe molecule be greatly improved
The problem of macromolecule water-solubility difference；The fluorescence probe is selectively relatively good to HNO simultaneously, overcomes conventional fluorescent probe and is given birth in detection
The problem of being easy to be interfered by reducing substances in organism when HNO in object, the namo fluorescence probe anti-interference ability is stronger, tool
There is higher sensitivity.Meanwhile namo fluorescence probe of the present invention can be applied to HNO in cell and zebra fish body
Real-time tracking monitors.
The present invention provides the compound P-BODIPY-R of ratio type detection HNO a kind of, can be used for ratio type detection cell and
Zebra fish activity in vivo small molecule HNO, compound P-BODIPY-R is selectively preferable to HNO, and anti-interference ability is stronger, has more
High sensitivity.
Description of the drawings
Fig. 1 is the namo fluorescence probe NanoP-BODIPY-N characterization of size figures prepared；
Fig. 2 (A) is namo fluorescence probe NanoP-BODIPY-N in PBS buffer solutions（pH=7.4）Middle addition AS（HNO discharges
Agent）Front and back fluorescence emission spectrogram of compound；Fig. 2 (B) is P-BODIPY-N in PBS buffer solutions（pH=7.4）Before and after middle addition AS
Fluorescence emission spectrogram of compound；
Fig. 3 is namo fluorescence probe NanoP-BODIPY-N in PBS buffer solutions（PH=7.4) in disturbed test figure；
Fig. 4 is the fluorescence imaging design sketch of namo fluorescence probe NanoP-BODIPY-N HNO in detecting HepG2 cells；In figure
（A-D）It is blank control group, in figure（E-H）It is the effect after namo fluorescence probe NanoP-BODIPY-N and intracellular HNO effects
Fruit is schemed；
Fig. 5 is namo fluorescence probe NanoP-BODIPY-N in the fluorescence imaging design sketch for detecting the HNO in zebra fish body, figure
（A-D）It is blank control group, in figure（E-H）It is after namo fluorescence probe NanoP-BODIPY-N is acted on HNO in zebra fish body
Design sketch；
Fig. 6 is the hydrogen spectrum of compound P-BODIPY-N, deuterated reagent d6-DMSO；
Fig. 7 is the high resolution mass spec HRMS (m/z) of compound P-BODIPY-N.
Specific implementation mode
For a better understanding of the present invention, with reference to the embodiment content that the present invention is furture elucidated, but the present invention
Content is not limited solely to the following examples.
Embodiment：The synthesis of compound P-BODIPY-N
Take 1,2-（Diphenylphosphino）Benzoic acid is dissolved in dry CH2Cl2In, it is stirred at room temperature to being completely dissolved, is cooled to 0 DEG C
Afterwards, DPTS and DIPC is added, stirs 5min, adds HO-BODIPY-N, be slowly ramped to room temperature, washing is added after having reacted
It washs, merges organic phase, anhydrous sodium sulfate drying, column chromatography obtains Orange red solid.
The hydrogen of compound P-BODIPY-N is composed（Such as Fig. 6）, deuterated reagent d6-DMSO。
1H NMR (d6-DMSO, 600 MHz): δ 8.27-8.25 (q, 1H), δ 7.60-7.58 (t, 2H),
δ 7.54-7.53 (d, 2H), δ 7.42-7.38 (m, 10H), δ 7.22-7.19 (m,5H), δ 6.98-6.97
(d, 1H), δ 6.93-6.92 (d, 1H), δ 6.77-6.76 (d, 1H), δ 6.56-6.55 (m, 1H), δ
4.10 (s,1H), δ 3.82 (s, 1H).
The high resolution mass spec HRMS (m/z) (such as Fig. 7) of compound P-BODIPY-N: Calcd for C43H35BF2N3O3P:
720.2519,Found: 720.2506 [M - H]-.
Embodiment：The preparation of namo fluorescence probe NanoP-BODIPY-N
MPEG-DSPE is added ultrasound to clear shape, P-BODIPY-N ultrasounds then is added about by the ultra-pure water for taking 1ml
20min is extremely clarified, membrane filtration, you can must detect the ratio type namo fluorescence probe of HNO.
Effect example：
It is namo fluorescence probe NanoP-BODIPY-N by characterizing the size measured with DLS and TEM with reference to figure 1.
It is namo fluorescence probe NanoP-BODIPY-N in PBS buffer solutions with reference to figure 2 (A)（0.01mol/L, pH=
7.4）In, the fluorescence emission spectrogram of compound before and after AS is added, Fig. 2 (B) is compound P-BODIPY-N in PBS buffer solutions
（0.01mol/L, pH=7.4）In, the fluorescence emission spectrogram of compound before and after AS is added；By being obtained in figure：Namo fluorescence probe
NanoP-BODIPY-N and compound P-BODIPY-N fluorescent emissions before and after AS is added have significant red shift, I601/I568's
Ratio values also have large change, illustrate that NanoP-BODIPY-N and compound P-BODIPY-N can be detected HNO.
It is namo fluorescence probe NanoP-BODIPY-N in PBS buffer solutions with reference to figure 3（PH=7.4) in disturbed test
Figure.NanoP-BODIPY-N is selectively preferable to HNO as seen from the figure.
It is namo fluorescence probe NanoP-BODIPY-N imaging effect figures in HepG2 cells with reference to figure 4.In figure（A-D）
It is NanoP-BODIPY-N cell imaging figures in the case where AS is not added, in figure（E-H）It is after AS is added, in probe and cell
The imaging effect figure of HNO effects.
It is namo fluorescence probe NanoP-BODIPY-N imaging effect figures in zebra fish with reference to figure 5, in figure（A-D）It is
NanoP-BODIPY-N zebra fish in-vivo imaging figures again in the case where AS is not added, in figure（E-H）For be added AS after, probe with
The imaging effect figure that HNO is acted in zebra fish body.
Present invention namo fluorescence probe claimed is the experiment proved that all have the work(described in similar said effect example
Can, i.e., the namo fluorescence probe and compound P-BODIPY-N of detection HNO provided by the invention；
Wherein example compound P-BODIPY-N structural formulas are as follows：
The namo fluorescence probe NanoP-BODIPY-R that the present invention provides a kind of ratio type detection HNO can be used for detecting cell and spot
Active small molecular HNO in horse fish body, the probe overcome some defects present in the relevant technologies, as such fluorescence probe utilizes
Triphenylphosphine structure is as recognition group, since excessive benzene ring structure so that such probe molecule is water-soluble universal poor.
In the present invention, the mode of probe small molecule is wrapped up by using amphipathic polymer, nano-micelle is prepared, obtains nano fluorescent
Probe NanoP-BODIPY-R solves such fluorescence probe to which the water-soluble of such probe molecule be greatly improved
The problem of macromolecule water-solubility difference；The fluorescence probe is selectively relatively good to HNO simultaneously, overcomes conventional fluorescent probe and is given birth in detection
The problem of being easy to be interfered by reducing substances in organism when HNO in object, the namo fluorescence probe anti-interference ability is stronger, tool
There is higher sensitivity.Meanwhile namo fluorescence probe of the present invention can be applied to HNO in cell and zebra fish body
Real-time tracking monitors.
The present invention provides the compound P-BODIPY-R of ratio type detection HNO a kind of, can be used for ratio type detection cell and
Zebra fish activity in vivo small molecule HNO, compound P-BODIPY-R is selectively relatively good to HNO, and anti-interference ability is stronger, has
Higher sensitivity.
The above display describes the main application of the basic principles and main features and the present invention of the present invention.The skill of the industry
Art personnel it should be appreciated that the present invention is not limited to the above embodiments, the above embodiments and description only describe
The principle of the present invention, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these
Changes and improvements all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and
Its equivalent defines.
Claims (9)
1. a kind of namo fluorescence probe of ratio type detection HNO, by fluorescence probe small molecule P-BODIPY-R through both sexes polymer
Package is made；
The fluorescence probe small molecule P-BODIPY-R general structures are as follows：
Wherein R1、R2For the electronic effect in the adjustable fluorescent probe molecule that is introduced into and when it being made to respond HNO, ratio occurs for fluorescence
The substituent group of formula variation.
2. the namo fluorescence probe of ratio type detection HNO according to claim 1, which is characterized in that the nano fluorescent is visited
Needle is nano-micelle, and the outer layer of the nano-micelle is both sexes polymer, and inside is micromolecular compound P-BODIPY-R.
3. the namo fluorescence probe of ratio type detection HNO according to claim 1, which is characterized in that the R1To have to electricity
Amido substituent group, the R of son2For the general group for not having strong push-and-pull electronic effect.
4. the namo fluorescence probe of ratio type detection HNO according to claim 1, which is characterized in that R1For morpholine base, R2
For phenyl.
5. the preparation method of the namo fluorescence probe of ratio type detection HNO, includes the following steps described in a kind of claim 1：
(1) P-BODIPY-R fluorescence probe small molecules are obtained through esterification using HO-BODIPY-R as raw material,
The compound HO-BODIPY-R has structure as follows：
Wherein R1、R2For the electronic effect in the adjustable fluorescent probe molecule that is introduced into and when it being made to respond HNO, ratio occurs for fluorescence
The substituent group of formula variation；
(2) nano-micelle is made through both sexes polymer packages in the compound small molecule P-BODIPY-R, and then obtains ratio
Formula detects the namo fluorescence probe of HNO.
6. a kind of compound for detecting HNO, which is characterized in that the compound P-BODIPY-R, general structure are as follows：
Wherein R1、R2For the electronic effect in the adjustable fluorescent probe molecule that is introduced into and when it being made to respond HNO, ratio occurs for fluorescence
The substituent group of formula variation.
7. a kind of preparation method of the compound described in claim 6 for detecting HNO, includes the following steps：
(1) P-BODIPY-R compounds are obtained through esterification using HO-BODIPY-R as raw material,
The compound HO-BODIPY-R has structure as follows：
Wherein R1For the amido substituent group with electron, R2For the general group for not having strong push-and-pull electronic effect.
8. the ratio type namo fluorescence probe for detecting HNO described in a kind of claim 1 is detecting in intracellular HNO or zebra fish body
The application of HNO.
9. detecting answering for HNO in intracellular HNO or zebra fish body for detecting the compound of HNO described in a kind of claim 6
With.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110885675A (en) * | 2019-11-22 | 2020-03-17 | 山东师范大学 | Nano fluorescent probe, preparation method and application thereof in detection of HNO in Golgi apparatus |
| CN111830000A (en) * | 2020-07-23 | 2020-10-27 | 南开大学 | Method for loading probe on nanoparticles to act on zebra fish embryo/juvenile fish |
| CN114957290A (en) * | 2021-12-30 | 2022-08-30 | 九江学院 | EPR probe for detecting HNO, and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110885675A (en) * | 2019-11-22 | 2020-03-17 | 山东师范大学 | Nano fluorescent probe, preparation method and application thereof in detection of HNO in Golgi apparatus |
| CN110885675B (en) * | 2019-11-22 | 2022-07-12 | 山东师范大学 | Nano fluorescent probe, preparation method and application thereof in detection of HNO in Golgi apparatus |
| CN111830000A (en) * | 2020-07-23 | 2020-10-27 | 南开大学 | Method for loading probe on nanoparticles to act on zebra fish embryo/juvenile fish |
| CN114957290A (en) * | 2021-12-30 | 2022-08-30 | 九江学院 | EPR probe for detecting HNO, and preparation method and application thereof |
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