CN105838355A - Micromolecule fluorescent probe and application thereof - Google Patents
Micromolecule fluorescent probe and application thereof Download PDFInfo
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Abstract
The invention discloses a micromolecule fluorescent probe and application thereof .The fluorescent probe is of the structure shown in the general formula I based on the TICT mechanism .Background fluorescence of the micromolecule fluorescent probe is weak in PBS buffer liquid, and the fluorescence intensity is obviously increased after the probe enters a hydrophobic cavity of human serum albumin HSA .The micromolecule fluorescent probe is good in selectivity and not disturbed by other common amino acid and protein molecules .In addition, response time is short after the probe interacts with HSA, and the probe can achieve specific binding of a FA1 locus of HSA .In the PBS buffer liquid, the fluorescence intensity and the HSA concentration show the good linear relation, and accurate detection can be achieved for human serum albumin HSA .In the real urine testing environment, the fluorescence intensity of the micromolecule fluorescent probe and albumin concentration in urine have the good linear relation, and therefore the micromolecule fluorescent probe has the good biological application prospect.
Description
Technical field
The present invention relates to field of fine chemical fluorescent probe, Preparation Method And The Use, particularly relate to a class based on
The small-molecule fluorescent probe of TICT mechanism, preparation method and the application in terms of albumin labelling and detection by quantitative thereof.
Background technology
Detection technique of fluorescence have highly sensitive, selectivity good, method is simple, in situ and the advantage such as detection in real time,
Become requisite detection means in modern biotechnology and life science.In protein labeling technology, glimmering
Cursor notation has obvious advantage relative to other traditional labeling methods, increasingly causes the concern of researcher.
Human serum albumin HSA is the protein that in serum, content is most, and its content in serum is at 30-50g/L
Left and right.Due to the Dialysis of kidney, in urine, albuminous content is at below 30mg/L.Albuminous physiology
Function includes maintaining the osmotic pressure of blood vessel, balanced nutritious, transport medicine and metabolite.Meanwhile, Microalbunin in urine
White content is relevant with kidney disease and diabetes, can be as kidney disease early treatment and intensification therapy of diabetes mellitus patients
Key index.Therefore its accurately detection in urine has very important biology and medical significance.
Imaging-PAM is the microanalysis skill that people provide that a kind of selectivity is good, highly sensitive, detection limit is low
Art, it has been widely used in the analysis detection of biomacromolecule such as protein, DNA and RNA.In document
The side's of having been reported that type dye, BODIPY class dyestuff and the albuminous detection in aqueous phase of polar sensitive probe.But send out
Now have good fluorescence character (such as high quantum production rate, long wavelength and stable in properties etc.), convieniently synthesized, can be low dense
The degree albuminous fluorescent probe of lower detection still faces the challenge.We have synthesized class HSA based on TICT mechanism
Fluorescent probe, it can realize the fluorescence response of OFF-ON in PBS.Such probe is convieniently synthesized,
Only need one-step or two-step reaction the most available, therefore there are good economic benefits.It addition, such probe has
TICT structure, while to HSA inner apolar environment sensitive, intramolecular freely rotatable can be empty by protein
Chamber is suppressed, and has relatively low detection limit and good sensitivity.In addition, by introducing rhodanine electron-withdrawing group
Group, it is achieved that probe is specific binding to HSA FA1 site, provides with protein bound mode for research dyestuff
New approach.
Summary of the invention
It is desirable to provide class small-molecule fluorescent probe based on TICT mechanism, can be used for human seralbumin in aqueous phase
The detection by quantitative of albumen HSA.
Present invention firstly provides small molecule fluorescent probe, there is following general structure I:
In formula I:
R1-R7It is each independently selected from H, C1-8Alkyl, replacement or unsubstituted phenyl；
Described substituted-phenyl is arbitrarily replaced by following group: CN, COOH, NH2、NO2、OH、SH、C1-6Alkane
Epoxide, C1-6Alkyl amino, C1-6Amide groups, halogen or C1-6Haloalkyl；
X is oxygen or sulfur；
N is selected from the integer of 1-8.
On the other hand, the present invention provides the preparation method of above-mentioned small-molecule fluorescent probe, is at toluene, ammonium acetate and ice
In the mixed system of acetic acid, compounds of formula II and compounds of formula III react gained:
Fluorescent probe of the present invention background fluorescence in PBS is more weak, at the hydrophobic cavity entering HSA
After, there is obvious Fluorescence Increasing, and its fluorescence intensity has good linear relationship with protein concentration, the most right
Other macro-molecular proteins and each amino acid have good selectivity, and are not done by internal common zwitterion
Disturb.It addition, its with HSA effect after response time shorter, and can tie with the FA1 locus specificity of HSA
Close.In PBS, its fluorescence intensity shows good linear relationship, to human serum with the concentration of HSA
Albumin HAS can receive orders detection.Under the test environment of true urine, its fluorescence intensity is dense with albumin in urine
Degree agriculture poplar has good linear relationship, therefore has good biologic applications prospect.In consideration of it, the present invention further side
The purpose in face is to provide the application in preparation Protein Detection compositions of described small-molecule fluorescent probe.And based on
This, it is provided that a kind of compositions for albumin detection, said composition includes that the small molecule fluorescent of the invention described above is visited
Pin.Described compositions is particularly suited for the inspection of the microalbumin in human serum albumin (HSA) and urine sample
Survey.
TICT mechanism small-molecule fluorescent probe of the present invention has a following significant feature:
(1) almost without fluorescence in PBS buffer solution, after entering protein cavity, there is obvious Fluorescence Increasing；
(2) probe fluorescence intensity in buffer and protein concentration present good linear relationship, can be used for white egg
White detection by quantitative；
(3) selectivity is good, to other protein and aminoacid almost without response；
(4) can be combined with albuminous FA1 locus specificity, the model of action for research dyestuff with protein carries
Supply powerful method；
(5) convieniently synthesized, product is easy to get.
Accompanying drawing explanation
Accompanying drawing 8 width of the present invention,
Fig. 1 be performance measurement experiment 1 in fluorescent probe compounds ES1, ES2, ES3 to human serum albumin (HSA)
Fluorescence titration experiment test figure.The concentration of probe molecule is 5 μMs, test system be in PBS buffer solution (pH=7.4,
10mM), the excitation wavelength of probe molecule ES1 be the excitation wavelength of 474nm, ES2 be 507nm, ES3 swash
Sending out wavelength is 430nm.
Fig. 2 be performance measurement experiment 2 in fluorescent probe compounds ES1 to human serum albumin (HSA) and difference
Aminoacid selectivity experiment block diagram.Probe molecule ES1 concentration is 5 μMs, and the concentration of HSA is 5 μMs, other
Amino acid whose concentration is 50 μMs.Excitation wavelength is 474nm, gathers the fluorescence intensity at 547nm.
Fig. 3 be performance measurement experiment 3 in fluorescent probe compounds ES1 to human serum albumin (HSA) and difference
The selectivity experiment block diagram of protein.Probe molecule ES1 concentration is 5 μMs, HSA and the concentration of other protein
For being 5 μMs.Excitation wavelength is 474nm, the fluorescence intensity at record 547nm.
Fig. 4 is that in performance measurement experiment 4, fluorescent probe compounds ES1 is combined ratio in fact with human serum albumin HSA
Test figure.The total concentration of fixing probe molecule ES1 Yu HSA is 10 μMs, constantly changes the concentration of probe and HSA
Ratio, respectively 1:9,2:8,3:7,4:6,5:5,6:4,7:3,8:2,9:1.Excitation wavelength is 474nm, record
Fluorescence intensity at 547nm.
Fig. 5 is fluorescent probe compounds ES1 and the positioning experiment of human serum albumin HSA in performance measurement experiment 5
Figure.Probe molecule concentration is 10 μMs, adds 2 μMs of HSA and makes itself and probe mixing 1h.Replace the concentration of medicine
It is respectively 5,10,15,20,30,40,60,80 μMs.
Fig. 6 is fluorescent probe compounds ES1 and human serum albumin HSA compound system pair in performance measurement experiment 6
The response diagram of temperature.The concentration of probe molecule and HSA is 5 μMs, temperature chooses 20 respectively, 25,30,
35,40,45,50 degrees Celsius.
Fig. 7 is the fluorescent probe compounds ES1 time response to human serum albumin HSA in performance measurement experiment 7
Figure.The concentration of probe molecule is 5 μMs, and the concentration of the HSA of addition is 5 μMs.Abscissa is time (min),
Vertical coordinate is fluorescence intensity.
Fig. 8 is that in performance measurement experiment 8, fluorescent probe compounds ES1 is sensitive to the detection of human serum albumin HSA
Degree lab diagram.The concentration of probe molecule is 5 μMs.Figure a is to visit in (pH=7.4,10mM) in PBS buffer solution
Linear relationship between fluorescence intensity and the HSA concentration of pin molecule；Figure b is the fluorescence of true urine middle probe molecule
Linear relationship between intensity and HSA concentration.Excitation wavelength is 474nm, the fluorescence intensity at record 547nm.
Detailed description of the invention
Unless otherwise indicated, term used herein has following meanings.
Unless otherwise indicated, term used herein has following meanings.
Term used herein " alkyl " includes straight chained alkyl and branched alkyl.As mentioned by single alkyl such as " third
Base ", the most only refer in particular to straight chained alkyl, as mentioned by single branched alkyl such as " isopropyl ", the most only refer in particular to branched alkyl.Example
As, " C1-4Alkyl " include methyl, ethyl, n-pro-pyl, isopropyl, normal-butyl and the tert-butyl group etc..Similar rule
It is also applied in this specification other group used.
Small-molecule fluorescent probe of the present invention, has a following general structure I:
In formula I: R1-R7It is each independently selected from H, C1-8Alkyl, replacement or unsubstituted phenyl；
Described substituted-phenyl is arbitrarily replaced by following group: CN, COOH, NH2、NO2、OH、SH、C1-6Alkane
Epoxide, C1-6Alkyl amino, C1-6Amide groups, halogen or C1-6Haloalkyl；
In detailed description of the invention, described R1-R4It is each independently selected from H or C1-4Alkyl, preferably H or methyl,
Most preferably H.
In detailed description of the invention, described R5And R6It is each independently selected from C1-4Alkyl, preferably methyl or ethyl,
Most preferable.
In detailed description of the invention, described R7Selected from H or C1-4Alkyl, preferably H or methyl, most preferably H.
In formula I, described X is oxygen or sulfur, preferably S.
In formula I, described n selected from the integer of 1-8, the integer of preferably 1-3, particularly preferred 1 or 2, most preferably
1。
Each preferred feature described above can be mutually combined, and gained technical scheme should be included in this completely
Bright addressed scope.
The example of the combination of preferred feature, for specific embodiment of the present invention one of be, of the present invention little
Fluorescence probe, selected from following compound:
On the other hand, the present invention provides the preparation method of described small-molecule fluorescent probe, is at toluene, ammonium acetate and ice
In the mixed system of acetic acid, compounds of formula II and compounds of formula III react gained:
In detailed description of the invention, the molar ratio of described compound II and compound III is 0.1-1000:1, preferably
For 0.5-100:1, more preferably 0.5-10:1, further preferably 1-5:1, most preferably 1.5:1, there is condensation reaction, generate in the two
General structure is the target compound of I.
More than reaction can be carried out in aqueous organic solvent or anhydrous organic solvent.It is preferably anhydrous organic solvent.
Organic solvent includes but are not limited to toluene, benzene, dichloromethane, oxolane or acetonitrile etc., and preferably reaction dissolvent is
Toluene.Course of reaction judges the terminal of reaction by thin layer chromatography (TLC).
Above-mentioned reaction temperature is-10 DEG C-150 DEG C, preferably 111 DEG C-130 DEG C；React optional acidity or base catalyst is
Triethylamine, piperidines, ammonium acetate, sodium acetate and glacial acetic acid, preferred catalyst is ammonium acetate and glacial acetic acid mixed system.
The method of purification of the small molecule fluorescent dyestuff of the present invention uses conventional method, is not particularly limited.Generally, instead
After should terminating, after directly filtering or boiling off solvent, utilize chromatographic column separating-purifying product.
Gained fluorescent dye can be reclaimed by separation well known in the art and purification technique, to reach the purity needed.
The various raw materials used in the present invention are all commercially available, or can be by the side of well known to a person skilled in the art
Method disclosed in method or prior art is prepared simply by raw material well known in the art.
It should be understood that the various ring substituents in the compounds of this invention have some can before above-mentioned steps is carried out or just
After completing, being introduced by the aromatics substitution reaction of standard or produced by conventional modified with functional group, this is included in
The method step aspect of the present invention.This reaction and modification include such as substituent group by the introducing of aromatics substitution reaction,
The reduction of substituent group, the alkylation of substituent group and the oxidation of substituent group.Reagent and reaction condition for these processes are
Known to chemical field.The instantiation of aromatics substitution reaction includes introducing nitro, with such as carboxylic acid halides and road with concentrated nitric acid
Lewis acid (such as aluminum chloride) introduces acyl group under the conditions of Friedel Crafts, with alkyl halide and lewis acid (such as trichlorine
Change aluminum) under the conditions of Friedel Crafts, introduce alkyl, and introduce halogen group.The instantiation modified includes passing through
Such as carry out catalytic hydrogenation with Raney nickel or carry out heat treated with ferrum in presence of hydrochloric acid, nitro is reduced ammonification
Base；Alkylthio group is oxidized to alkyl sulphinyl or alkyl sulphonyl.
Small-molecule fluorescent probe based on TICT mechanism as herein described has obvious fluorescence after entering albumin cavity
Recover, can be used for the detection by quantitative of microalbumin in urine.
Following non-limiting example can make those of ordinary skill in the art that the present invention be more fully understood, but not
Limit the present invention by any way.
Embodiment 1
The synthesis of fluorescent probe compounds ES1:
Dimethylamino benzaldehyde (1.49g, 10mmol) and rhodanine (1.33g, 10mmol) will be equipped with
In the round-bottomed flask of 10mL toluene, add 200mg ammonium acetate and 1mL glacial acetic acid as catalyst, be warmed up to
115 DEG C of back flow reaction.Reaction 24h, removes solvent, chromatographic column isolated red solid ES1 (78.25%) under reduced pressure.1H NMR (500MHz, DMSO-D6), δ: 3.04 (s, 6H), 4.27 (s, 1H), 6.82 (t, J=10Hz, 2H), 7.42
(t, J=10Hz, 2H), 7.53 (s, 1H)；13C NMR(500MHz,DMSO-D6),δ:112.172,117.313,
119.757,132.872,133.253,151.759,169.371,176.645,194.973,105.280ppm；TOF MS:m/z
calcd for[M+H]+:265.0469,found:265.0458.
Embodiment 2
The synthesis of fluorescent probe compounds ES2:
4-di methyl amino cinnamaldehyde (1.75g, 10mmol) is added dress with rhodanine (1.33g, 10mmol)
Have in the round-bottomed flask of 12mL toluene, add 200mg ammonium acetate and 1mL glacial acetic acid as catalyst, intensification
To 115 DEG C of back flow reaction.Reaction 24h, removes solvent, chromatographic column isolated dark brown solid ES2 (65.46%) under reduced pressure.1H NMR (500MHz, DMSO-D6), δ: 2.99 (s, 6H), 3.10 (q, J=25Hz, 1H), 6.72 (d, J=5Hz,
3H), 7.21 (q, J=45Hz, 2H), 7.51 (d, J=10Hz, 2H)；13C NMR(500MHz,DMSO-D6),δ:
45.75,111.89,112.10,118.48,123.31,129.41,129.73,130.97,145.41,151.45ppm；TOF
MS:m/z calcd for[M+H]+:291.0626,found:291.0618.
Embodiment 3
The synthesis of fluorescent probe compounds ES3:
Will be to dimethylamino benzaldehyde (1.49g, 10mmol) and 2,4-thiazolidinedione (1.17g, 10mmol)
It is equipped with in the round-bottomed flask of 10mL toluene, adds 200mg ammonium acetate and 1mL glacial acetic acid as catalysis
Agent, is warmed up to 115 DEG C of back flow reaction.Reaction 24h, removes solvent, chromatographic column isolated yellow solid under reduced pressure
ES3 (85.60%).1H NMR (500MHz, DMSO-D6), δ: 3.01 (s, 6H), 6.83 (d, J=10Hz,
2H), 7.42 (d, J=5Hz, 2H), 7.67 (s, 1H), 12.31 (s, 1H)；13C NMR(500MHz,DMSO-D6),δ:
110.79,110.91,111.95,120.02,115.63,119.79,132.10,132.91,133.82,139.29,151.44,
167.48,168.11ppm；TOF MS:m/z calcd for[M-H]-:247.0541,found:247.0545.
Embodiment 4
Human serum albumin (HSA) fluorescence titration is tested by fluorescent probe compounds ES1, ES2 and ES3
5 μMs of small-molecule fluorescent probe ES1, ES2 and ES3 are joined (pH=7.4,10 in PBS buffer solution
MM), it is gradually added into the HSA of variable concentrations, as it is shown in figure 1,1a, 1b and 1c are respectively dyestuff ES1, ES2
With the ES3 fluorescence response figure to serum albumin HSA.The excitation wavelength of three example dyestuffs is respectively 474nm, 507nm
And 430nm.Along with the increase of HSA concentration, the fluorescence intensity at probe molecule maximum emission peak gradually strengthens.
Embodiment 5
The selectivity of human serum albumin (HSA) and different aminoacids is tested by fluorescent probe compounds ES1
The compound ES1 using above-mentioned synthesis evaluates the selectivity to different aminoacids: by 5 μMs of probe compounds
ES1 and 5 μMs of HSA and 50 μMs of variety classes aminoacid (glutamic acid, glycine, histidine, tryptophan,
Arginine, tyrosine, aspartic acid, agedoite, cysteine and lysine) join PBS
In (pH=7.4,10mM), the excitation wavelength of probe is 474nm, chooses a length of 547nm of transmitted wave, records phase
The fluorescence intensity change answered.Test result is as in figure 2 it is shown, after adding 5 μMs of human serum albumin HSAs, visit
The fluorescence intensity of pin increases 72 times, and the fluorescence that 50 μMs of amino acid whose additions of variety classes can't affect probe is strong
Degree.
Embodiment 6
The selectivity of human serum albumin (HSA) and different proteins is tested by fluorescent probe compounds ES1
The compound ES1 using above-mentioned synthesis evaluates the selectivity to different proteins: by 5 μMs of probe compounds
ES1 and 5 μMs of HSA and variety classes protein are (bovine serum albumin BSA, Chymotrypsin, protease, molten
Bacterium enzyme, chymotrypsinogen A, hemoglobin and histone) join (pH=7.4,10 in PBS
MM), the excitation wavelength of probe is 474nm, chooses a length of 547nm of transmitted wave, records corresponding fluorescence intensity and becomes
Change.Test result as it is shown on figure 3, owing to having similar structure and biological property, probe show HSA and
The high selectivity of BSA, when i.e. only adding HSA and BSA, probe compound just has obvious Fluorescence Increasing.
It addition, the fluorescence response different to HSA with BSA by comparing probe compound ES1, it has been found that add 5
After μM HSA, the fluorescence intensity of probe increases 60 times, and 5 μMs of BSA are only capable of making the fluorescence intensity of probe to increase
6 times, illustrate that probe compound ES1 has more preferable selectivity to HSA.
Embodiment 7
Fluorescent probe compounds ES1 is combined Ratio Experiments with human serum albumin HSA
Use the combination ratio of compound ES1 checking probe molecule and the human serum albumin of above-mentioned synthesis: Job ' s plot
Experiment is a kind of method for determining two kinds of material stoichiometric proportions.PBS (pH=7.4,10
MM), in test system, the total concentration of fixing probe molecule ES1 Yu HSA is 10 μMs, constantly changes probe
With the concentration ratio of HSA, respectively 1:9,2:8,3:7,4:6,5:5,6:4,7:3,8:2,9:1, test its phase
The change of the fluorescence spectrum answered.As it can be seen, when the concentration ratio of probe molecule and HSA is during for 5:5, fluorescence intensity
The strongest, this explanation probe molecule ES1 with HSA is to be combined with the ratio of 1:1.
Embodiment 8
Fluorescent probe compounds ES1 and the positioning experiment of human serum albumin HSA
Use the compound ES1 checking probe molecule of above-mentioned synthesis and the binding site of human serum albumin: select four
Planting different pharmaceutical, i.e. warfarin, ibuprofen, propofol and hemin and replace Experimental agents as medicine, they are respectively
Can be combined with FA7, FA3/4 and FA6 of human serum albumin HSA, FA3/4 and FA5 and FA1 site.
In the test system of PBS (pH=7.4,10mM), probe molecule concentration is 10 μMs, adds 2
μM HSA makes itself and probe mixing 1h, it is ensured that both effects completely.Add in above-mentioned supramolecular system subsequently
Four kinds of medicines of variable concentrations (5,10,15,20,30,40,60,80 μMs), the change of the fluorescence intensity of recording responses.As
Shown in Fig. 5, in the probe molecule mixed system with HSA, add the warfarin of variable concentrations, ibuprofen and the third pool
The fluorescence intensity of mixed system is had little to no effect by phenol, and hemin can the fluorescence of substantially cancellation system, probe is described
Compound ES1 is combined in FA1 site with human serum albumin HSA, for research dye molecule and protein
Interact and provide new method.
Embodiment 9
After fluorescent probe compounds ES1 and human serum albumin HSA effect, temperature-responsive is tested
The compound ES1 using above-mentioned synthesis evaluates after probe molecule acts on human serum albumin real to temperature-responsive
Test.According to the literature, the fluorescence intensity of the molecule with TICT character often shows the sensitivity to temperature.
In the test system of PBS (pH=7.4,10mM), probe molecule and the concentration of human serum albumin
It is 5 μMs, adjusts the temperature of test system the most from high to low and from low to high, the temperature of selection is respectively 20,
25,30,35,40,45 and 50 degrees Celsius.From Fig. 6 a, when temperature gradually rises, probe molecule with
The fluorescence intensity of protein complex system is gradually lowered, by Fig. 6 b, when temperature is gradually lowered, probe molecule and egg
The fluorescence intensity of white matter compound system gradually rises.
Embodiment 10
The response time of human serum albumin HSA is tested by fluorescent probe compounds ES1
The compound ES1 using above-mentioned synthesis evaluates the response time of probe molecule and HSA effect: buffer at PBS
In the test system of liquid (pH=7.4,10mM), probe molecule concentration is 5 μMs, and excitation wavelength is 474nm,
The change of fluorescence intensity at record 547nm.As shown in Figure 7, after adding 5 μMs of protein, probe molecule pair
The response of human serum albumin HSA is quite rapid, i.e. can reach balance in about 10s fluorescence intensity, and subsequently
At least 60min in keep constant.
Embodiment 11
The fluorescent probe compounds ES1 detection sensitivity to human serum albumin HSA
The compound ES1 using above-mentioned synthesis evaluates the probe molecule detection sensitivity to HSA: first delay at PBS
Rushing in the test system of liquid (pH=7.4,10mM), the concentration of dyestuff is 10 μMs, is separately added into above-mentioned system
Concentration is 0.002,0.004,0.006,0.008,0.013,0.018,0.023,0.028 and 0.033mg/mL,
Excitation wavelength is 474nm, the fluorescence intensity change at record 547nm.As shown in Figure 8 a, the fluorescence intensity of probe
Along with the increase of HSA concentration gradually strengthens, and its fluorescence intensity and human serum albumin HSA are at 0-0.033
There is in the range of mg/mL good linear relationship (R2=0.997).Calculate detection be limited to by 3 σ/k simultaneously
0.132mg/L, therefore probe molecule ES1 can be used for the detection by quantitative of low concentration HSA.It addition, we also investigate
Compound ES1 is in response condition to HSA of true urine test system.We select healthy male volunteers to carry
The urine specimen of confession, using clinical method to record the content of microalbumin in this urine specimen is 0.0035
Mg/mL, and as benchmark, in above-mentioned system, add 10 μMs of probe molecule ES1, be subsequently added variable concentrations
The HSA of (0.005,0.01,0.015,0.025,0.035,0.045 and 0.055mg/mL), excitation wavelength is
474nm, the fluorescence intensity change at record 547nm.As shown in Figure 8 b, in the test system of true urine
Side, the fluorescence intensity of probe molecule ES1 has good linear relationship as the concentration of human serum albumin HSA
(R2=0.999), illustrate that probe compound ES1 has good biologic applications.
Claims (9)
1. small molecule fluorescent probe, has a following general structure I:
In formula I:
R1-R7It is each independently selected from H, C1-8Alkyl, replacement or unsubstituted phenyl；
Described substituted-phenyl is arbitrarily replaced by following group: CN, COOH, NH2、NO2、OH、SH、C1-6Alkane
Epoxide, C1-6Alkyl amino, C1-6Amide groups, halogen or C1-6Haloalkyl；
X is oxygen or sulfur；
N is selected from the integer of 1-8.
Small-molecule fluorescent probe the most according to claim 1, it is characterised in that described R1-R7The most independent
Ground is selected from H or C1-4Alkyl.
Small-molecule fluorescent probe the most according to claim 2, it is characterised in that described R1-R4、R7Each
Independently selected from H or methyl.
Small-molecule fluorescent probe the most according to claim 2, it is characterised in that described R5And R6The most solely
On the spot selected from C1-4Alkyl.
Small-molecule fluorescent probe the most according to claim 1, it is characterised in that described n is whole selected from 1-3's
Number.
Small-molecule fluorescent probe the most according to claim 1, selected from following compound:
7. the preparation method of the small-molecule fluorescent probe described in claim 1, it is characterised in that be at toluene, acetic acid
In the mixed system of ammonium and glacial acetic acid, compounds of formula II and compounds of formula III react gained:
8. the application in preparation Protein Detection compositions of the small-molecule fluorescent probe described in claim 1.
9. for the compositions of albumin detection, including the small-molecule fluorescent probe described in claim 1.
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| CN110372681A (en) * | 2019-07-29 | 2019-10-25 | 天津医科大学 | A kind of application of the self-assembled nanometer fluorescence probe for selective enumeration method human serum albumins |
| CN110386931A (en) * | 2018-04-20 | 2019-10-29 | 南京大学 | A kind of human albumin's fluorescence probe and its preparation method and purposes |
| CN112823278A (en) * | 2018-08-27 | 2021-05-18 | 香港科技大学 | Method for detecting human serum albumin in biological liquid |
| WO2021175203A1 (en) * | 2020-03-02 | 2021-09-10 | The Hong Kong University Of Science And Technology | Aie probe and application thereof |
| CN114940686A (en) * | 2022-05-13 | 2022-08-26 | 西北工业大学深圳研究院 | Photodynamic physiotherapy micromolecule and preparation method and use method thereof |
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| WO2021175203A1 (en) * | 2020-03-02 | 2021-09-10 | The Hong Kong University Of Science And Technology | Aie probe and application thereof |
| CN114940686A (en) * | 2022-05-13 | 2022-08-26 | 西北工业大学深圳研究院 | Photodynamic physiotherapy micromolecule and preparation method and use method thereof |
| CN114940686B (en) * | 2022-05-13 | 2023-11-28 | 西北工业大学深圳研究院 | Photodynamic physiotherapy small molecule and preparation method and use method thereof |
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