CN106699734A - Fluorescent molecular probe and nanoprobe as well as preparation method and application thereof - Google Patents
Fluorescent molecular probe and nanoprobe as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN106699734A CN106699734A CN201611177239.1A CN201611177239A CN106699734A CN 106699734 A CN106699734 A CN 106699734A CN 201611177239 A CN201611177239 A CN 201611177239A CN 106699734 A CN106699734 A CN 106699734A
- Authority
- CN
- China
- Prior art keywords
- albumin
- probe
- fluorescent
- formula
- molecular probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
Abstract
The invention discloses a fluorescent molecular probe and a nanoprobe as well as a preparation method and application thereof. The fluorescent molecular probe contains a polyphenyl substituted imidazole ring and a quaternary ammonium salt structure, can be easily self-assembled into a non-fluorescent nanomaterial in an aqueous solution, and can be disassembled after being combined with albumin; probe molecules enter a hydrophobic cavity of albumin and have hydrogen bonds and hydrophobic effects with partial amino acid residues in the cavity, rotation of the probe molecules is limited, and fluorescence is produced; and the content of the albumin can be reflected by detecting the emitting light intensity of the fluorescence, so that the quantitative detection of the albumin is realized. The fluorescent molecular probe has the advantages of high sensitivity, strong resistance to interference and the like, is especially applicable to the quantitative detection of the albumin in a chemical solution system, a patient blood sample and the like, and has important practical values on detection of trace amount albumin, content detection of the albumin in patient blood and clinical medication guidance of the albumin.
Description
Technical field
It is more particularly to a kind of to there is water solubility and with poly- the present invention relates to a kind of fluorescent molecular probe and nano-probe
The fluorescent molecular probe and fluorescent nano probe of collection induction transmitting effect, and their preparation method, especially also relate to this glimmering
Optical molecule probe belongs to chemical analysis, bioanalysis detection technique field in the application of the context of detection of albumin.
Background technology
Human serum albumins (HSA) is a kind of main albumen in blood of human body, is maintaining plasma colloid osmotic pressure, transport
Endogenous and exogenous molecule include that many medicines play the effect of key.Research has shown that, concentration of the HSA in body fluid with it is many
Disease includes cancer, rheumatoid arthritis, myocardial ischemia (G.Fanali, A.di closely related with the progress of liver failure
Masi,V.Trezza,M.Marino,M.Fasano,P.Ascenzi,Mol Aspects Med.2012,33,209；
V.Arroyo,R.Garcia-Martinez,X.Salvatella,J.Hepatol.2014,61,396；C.-E.Ha,
N.V.Bhagavan,BBA-Gen.Subjects 2013,1830,5486；J.Rozga,T.Piatek,P.Malkowski,
Ann.Transpl.2013,18,205.)., used as a reliable health indicator, its normal level is about for the content of HSA in human body
It is 35-55g L-1(527-828μmolL-1).Low-level albumin and cirrhosis, chronic hepatitis and liver decline close phase in blood plasma
Close.Conversely, HSA excessive in urine is present, diagnosis diabetes, hypertension, angiocardiopathy and kidney trouble are acknowledged as
An index.Therefore, high selectivity, high sensitivity, the content for accurately detecting HSA in human body fluid, especially in serum
Content, has great importance in clinical diagnosis.
So far, including dyestuff combined with fluorescent analytic approach, based on antibody (SABC) method, and mass spectrum
Proteomic techniques, for the quantitative determination of albumin in blood sample.In numerous methods, the analysis based on fluorescence
Method has been widely used in not same due to its quick, sensitive, non-destructive and the excellent characteristics suitable for high flux detection
The detection of HAS contents in product.But, it has been reported that fluorescence probe have some limitation, for example, some probes are in the aqueous solution
Middle dissolubility is poor so that it be difficult in biosystem using (X.Fan, Q.He, S.Sun, H.Li, Y.Pei, Y.Xu,
Chem.Commun.2016,52,1178.).In addition, the probe in detecting albumin of some document reports, is by covalent bond and egg
White sulfydryl or amino combine to realize (P.Anees, S.Sreejith, A.Ajayaghosh,
J.Am.Chem.Soc.2014,136,13233；M.Cieplak,K.Szwabinska,M.Sosnowska,
B.K.C.Chandra,P.Borowicz,K.Noworyta,F.D’Souza,W.Kutner,
Biosens.Bioelectron.2015,74,960.).But, this irreversible combination may result in the denaturation of albumen.
Additionally, the fluorescence probe of most of detection albumin is by the use of traditional organic dye molecule as fluorescent emission group, this kind of point
Son in physiological buffer easily produce aggregation, however, aggregation after due to the excitation state nonradiative relaxation of aggregation cause it is glimmering
Optical quenching.Researchers are in order to overcome this defect, it has to used using extremely dilute probe solution and solubility probe high
In the detection of albumen.However, the problems such as probe of weak solution equally faces weak transmitting fluorescence, poor sensitivity.Even if additionally, dilute
Aggregation causes the influence being quenched cannot also to avoid completely in solution, because fluorescent probe molecule may be gathered in bioanalysis
On large biological molecule and the surface of the hydrophobic pocket of protein, cause fluorescent quenching.Therefore, design fluorescence probe has completely water-soluble
Property, and the defect that aggregation inducing is quenched can be overcome to detect human serum albumins be very crucial.
The content of the invention
For it is existing detection haemocyanin fluorescence probe performance exist deficiency, first purpose of the invention be
In a kind of good water solubility of offer, and there is the fluorescent molecular probe of aggregation inducing effect simultaneously.
It is to provide a kind of by good water solubility and the fluorescence with aggregation inducing effect point that second object of the present invention is
Sub- probe is self-assembly of fluorescent nano probe in aqueous.
It is that the fluorescence molecule for preparing that a kind of simple to operate, raw material of offer is easy to get is visited that third object of the present invention is
The method of pin.
It is to provide a kind of simple to operate, mild condition to prepare fluorescent nano probe that fourth object of the present invention is
Method.
It is to provide a kind of described fluorescent nano probe albumin in blood is detected that 5th purpose of the invention is
Application；Described fluorescent molecular probe is self-assembly of loose nanoparticle in aqueous, occurs after being acted on albumin
De- assembling, probe molecule limits the rotation of probe molecule into the hydrophobic cavity of albumen and launches fluorescence and realize to albumen
Detection, show sensitivity it is high, selectivity it is good the characteristics of.
In order to realize above-mentioned technical purpose, the invention provides a kind of with the glimmering of water-soluble and aggregation inducing transmitting effect
Optical molecule probe, with the structure of formula 1：
Wherein,
R1、R2And R3It is independently selected from hydrogen, halogen, alkyl, hydroxyl, alkoxy, nitro, carboxyl or amido；
R4Selected from C1~C13Aliphatic chain or C8~C13Alkane chain containing aryl.
Preferred scheme, R2And R3It is selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, hydroxyl, methoxyl group, nitro, carboxyl or diformazan
Amido；More preferably scheme, R2And R3It is selected from hydrogen.
Preferred scheme, R4Selected from C2~C8Alkane chain or 1,4- dimethylenebenzene groups.
Preferred scheme, R1Selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, hydroxyl, methoxyl group, nitro, carboxyl or dimethylamino,
More preferably scheme, R1Selected from hydrogen.
Present invention also offers a kind of fluorescent nano probe, it is self-assembly of by the fluorescent molecular probe.The structure of formula 1
Fluorescent molecular probe in aqueous by being self-assembly of state of aggregation nanoparticle, state of aggregation nanoparticle is main by fluorescence molecule
Probe monomer, oligomer and polymer are constituted.
The described fluorescence probe point with water-soluble and aggregation inducing transmitting effect is prepared present invention also offers a kind of
The method of son, the method is comprised the following steps：
1) 4- formylpyridines, the amino benzenes compounds of formula 2 and the benzil class compound of formula 3 are in acetic acid/ammonium acetate system
Cyclization is carried out, the intermediate of formula 4 is obtained；
2) intermediate of formula 4 carries out nucleophilic substitution with the bromo compound of formula 5 two, obtains the intermediate of formula 6；
3) with pyridine there is nucleophilic substitution in the intermediate of formula 6, obtain final product；
Wherein,
R1、R2And R3It is independently selected from hydrogen, halogen, alkyl, hydroxyl, alkoxy, nitro, carboxyl or amido；
R4Selected from C1~C13Aliphatic chain or C8~C13Alkane chain containing aryl.
Preferred scheme, 1) in course of reaction be：By the pyridine of 4- formoxyls and amino benzenes compounds in glacial acetic acid solvent
0.5~1.5h of stirring, adds benzil class compound and ammonium acetate, and 8~16h is reacted at a temperature of 110~130 DEG C.
Preferred scheme, 2) in reaction condition be：Using acetonitrile as solvent, reacted 6~10 hours at 80~95 DEG C.
Preferred scheme, 3) in reaction condition be：Using pyridine as solvent, reacted 6~10 hours at 80~95 DEG C.
Present invention also offers a kind of method for preparing described fluorescent nano probe, the method is by described fluorescence point
After sub- probe is dissolved in organic solvent, it is added in the aqueous solution, it is ultrasonically treated, obtain final product fluorescent nano probe.
Preferred scheme, organic solvent is selected from methyl alcohol, ethanol, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, acetonitrile
In at least one.
Preferred scheme, the aqueous solution is selected from pure water, physiological saline, phosphate buffer solution, 4- HEPESs and delays
Rush solution or three (methylol) aminomethane hydrochloride cushioning liquid.
Present invention also offers a kind of application of the fluorescent nano probe, fluorescent nano probe is applied to the white egg of serum
White detection.
Preferred scheme, white egg in chemical solution system, blood samples of patients or biological tissue is applied to by fluorescent nano probe
White quantitative fluorescence analysis detection.
Fluorescent probe molecule of the invention is a kind of typical bola types molecule, with many phenyl substituted imidazole rings and level Four
Ammonium salt structure, centre is connected by hydrophobic alkyl chain.Probe molecule has good water solubility, has been self-assembly of in water
Machine nanoparticle, and without fluorescence.After albumin identification, there is de- assembling, probe molecule enters the hydrophobic cavity of albumen,
There is hydrogen bond action and hydrophobic effect with the amino acid residue of albumen, ultimately result in the rotation of limitation probe molecule and produce glimmering
Light.Its Cleaning Principle such as Fig. 5.
It is of the invention following (with R with water-soluble and aggregation inducing effect fluorescent molecular probe syntheti c route1、R2, and R3For
Hydrogen, R4=C2、C4、C6、C8、C10、C12It is specifically described as a example by alkane chain)：
The specific preparation method of the fluorescent molecular probe with water-soluble and aggregation inducing effect of the invention is by as follows
Step is realized：
A, 4- formylpyridines and aniline are dissolved in glacial acetic acid, then stirring at normal temperature 1 hour sequentially adds benzil
It is well mixed with ammonium acetate, continue to heat 120 DEG C of back flow reactions overnight, adjusted with sodium hydroxide solution after TLC detection reactions completely
PH has precipitation to produce to weakly acidic pH, and filtration under diminished pressure obtains filter cake, dries, and column chromatography separating purification obtains compound 1；
B, compound 1 is dissolved in acetonitrile, adds dibromo compound (1,2- Bromofume, Isosorbide-5-Nitrae-dibromobutane, 1,6-
Dibromo-hexane, 1,4- bis- (bromomethyl) benzene, the bromooctanes of 1,8- bis-, 1,10- dibromo-decanes or 1,12- dibromo-dodecanes)；System is mixed
Close it is uniform after, 90 DEG C are heated to reflux, and after reaction terminates, solvent is removed in vacuum distillation, column chromatography separating purification obtain compound 2,
Compound 4, compound 6, compound 8, compound 10, compound 12 or compound 14；
C, compound 2, compound 4, compound 6, compound 8, compound 10, compound 12 or compound 14 are dissolved in
In pyridine, system heats 90 DEG C of back flow reactions, and after reaction terminates, solvent is removed in vacuum distillation, and column chromatography separating purification obtains mesh
Mark product.
The preparation method of fluorescent nano probe of the invention is achieved by the steps of：
Target product is completely dissolved in mother liquor is prepared into organic solvent, then mother liquor is drawn with liquid-transfering gun, in ultrasound
Under the conditions of be added in a certain amount of aqueous solution, obtained final product after system stirring at normal temperature 30min detection the organic of seralbumin receive
The grain of rice.The particle diameter and pattern to form nanoparticle to observe using dynamic light scattering (DLS) and projection Electronic Speculum (TEM).
The method that technical scheme is based on fluorescent nano probe detection albumin is to take full advantage of fluorescence molecule
Probe is self-assembly of nanoparticle in aqueous, and the nanoparticle solution for being formed does not have fluorescence, and in the presence of albumin, hair
Raw de- assembling, probe molecule enters the hydrophobic cavity of albumen, and hydrogen bond action and hydrophobic work occur with the amino acid residue of albumen
With, ultimately result in limitation probe molecule rotation and produce fluorescence.The present invention be based on this and set up detection albumin side
Method, the anti-interference strong, sensitivity of the method is high, can be with wide popularization and application.
Fluorescent nano probe of the invention can be used in the detection of albumin in chemical simulation living things system；Also clinic can be used
The medically detection of Human Serum Albumin.
Compared with the prior art, the Advantageous Effects that technical scheme is brought：
1) fluorescent molecular probe of the invention has good water solubility, and non-blooming nanoparticle can be self-assembly of in water
The characteristics of, and there is aggregation inducing to launch effect for it, and de- assembling occurs when being acted on albumin, produce the fluorescence of 480nm, especially
Suitable for fluoroscopic examination.
2) preparation method of fluorescent probe molecule of the invention and fluorescent nano probe is simple, low cost, is conducive to big rule
Mould is produced.
3) fluorescent nano probe of the invention is used to detect the albumin in serum etc. that the probe to have fine to albumin
Selectivity, trypsase, papain, pepsin, immunoglobulin, glucose oxidase, histidine, half Guang ammonia
The detection to albumin such as acid, homocysteine, lysine, glutathione, ATP, glucose is not interfered with, the nano-probe
The fluorescence intensity of solution and the concentration of albumin in certain concentration range (0-15 μM of the concentration range of albumin) have good
Good linear relationship, shows quantitative determination characteristic, and the content inspection clinically to albumin in blood samples of patients can be met completely
The requirement of survey.
4) fluorescent nano probe of the invention is short to the response time of albumin (15s), and the sensitivity of measure is high, anti-interference
Ability is strong, simple to operate, with low cost so that it is adapted to promote the use.
Brief description of the drawings
【Fig. 1】The fluorescent nano probe dynamic light scattering diagram prepared in the embodiment of the present invention 1.
【Fig. 2】The projection electron microscope of the fluorescent nano probe prepared in the embodiment of the present invention 1.
【Fig. 3】The linear relationship of the fluorescence intensity of fluorescent nano probe and albumin concentration, horizontal seat in the embodiment of the present invention 1
Albumin concentration is designated as, ordinate is fluorescence intensity.
【Fig. 4】Selectivity of the fluorescent nano probe to albumin in the embodiment of the present invention 1.
【Fig. 5】It is used for the schematic diagram of albumin detection for fluorescent nano probe.
Specific embodiment
Implementation below is further intended to explaination and illustrates rather than limitation of the invention.
Embodiment 1
The enhanced object 3 of aggregation inducing fluorescent emission of the invention (R in example1、R2、R3=H, R4=C2) synthesis,
Synthetic route is as follows：
The synthesis of compound 1：4- formylpyridines (1.06g, 10mmol) and aniline (0.91ml, 10mmol) are weighed respectively
It is dissolved in the middle of 100mL glacial acetic acid, is stirred at room temperature one hour.Benzil (2.1g, 10mmol) and ammonium acetate (3.55g,
46mmol) sequentially it is added in reaction system, compound reacts overnight under the conditions of 120 DEG C, reaction terminates rear reaction system and falls
Enter in 300mL frozen water, system pH to neutrality is reconciled with the sodium hydroxide solution of 0.1mmol/L, mixture filtering washes three with water
Time, silica gel column chromatography separating purification obtains product after vacuum drying.Yield is 42%.Nuclear magnetic resonance spectroscopy result is：1H NMR
(400MHz,CDCl3,δ):8.48-8.47 (d, J=2H), 7.58-7.60 (d, J=2H), 7.31-7.35 (m, 4H), 7.22-
7.29(m,7H),7.13-7.14(d,2H),7.09-7.10(d,2H)；13C NMR(100MHz,CDCl3,δ):149.71,
143.84,139.20,137.78,136.65,133.96,132.46,131.05,130.00,129.48,129.01,128.47,
128.34,128.30,128.27,127.32,126.97,122.32.HRMS(ESI)m/z:Calculated value C26H19N3,373.1579
[M]；It was found that 374.1651 [M+H]+.
The synthesis of compound 2：Weigh Compound 1 (0.19g, 0.50mmol) and compound glycol dibromide (93mg,
0.50mmol) it is dissolved in 15mL acetonitriles, room temperature is sufficiently stirred for, reaction system fully flows back 10 hours at 90 DEG C, the monitoring of TLC plates
After reaction completely, vacuum distillation removes solvent, the isolated product 2 of silica gel column chromatography.Yield is 45%.Nuclear magnetic resonance spectroscopy knot
It is really：1H NMR(400MHz,DMSO-d6,δ):9.09–9.11(d,2H),7.81-7.82(d,2H),7.51-7.55(m,7H),
7.25-7.42(m,8H),3.48-3.50(t,2H),3.40-3.43(t,2H)；13C NMR(100MHz,DMSO-d6,δ):
145.39,144.84,141.81,140.85,140.41,137.23,137.04,136.75,135.92,133.49,133.41,
131.35,130.79,129.16,128.96,127.06,123.72,123.48,60.74,32.08.
The synthesis of compound 3：Weigh Compound 2 (110mg, 0.20mmol) is dissolved in 10mL pyridine solutions, and room temperature is filled
Divide stirring, reaction system fully flows back 8 hours at 90 DEG C, after the monitoring of TLC plates is reacted completely, vacuum distillation removes solvent, silica gel
Column chromatography for separation obtains product 3.Yield is 53%.Nuclear magnetic resonance spectroscopy result is：1H NMR(400MHz,CDCl3,δ):9.14-
9.15(d,2H),8.94-8.96(d,2H),8.68-8.72(t,1H),8.19-8.23(t,2H)7.77-7.79(d,2H),
7.50-7.55(m,7H),7.27-7.37(m,8H),5.29-5.32(t,2H),5.20-5.23(t,2H),1.88-1.92(m,
4H),1.23-1.32(m,4H)；13C NMR(100MHz,DMSO-d6,δ):147.08,145.80,145.48,144.77,
140.57,140.43,136.80,135.84,133.45,131.37,130.66,130.47,130.11,129.78,129.15,
128.93,128.87,128.58,128.06,127.01,124.12,59.58,59.05.
Embodiment 2
The enhanced object 5 of aggregation inducing fluorescent emission of the invention (R in example1、R2、R3=H, R4=C4) synthesis,
Synthetic route is as follows：
The synthesis of compound 1：4- formylpyridines (1.06g, 10mmol) and aniline (0.91ml, 10mmol) are weighed respectively
It is dissolved in the middle of 100mL glacial acetic acid, is stirred at room temperature one hour.Benzil (2.1g, 10mmol) and ammonium acetate (3.55g,
46mmol) sequentially it is added in reaction system, compound reacts overnight under the conditions of 120 DEG C, reaction terminates rear reaction system and falls
Enter in 300mL frozen water, system pH to neutrality is reconciled with the sodium hydroxide solution of 0.1mmol/L, mixture filtering washes three with water
Time, silica gel column chromatography separating purification obtains product after vacuum drying.Yield is 42%.Nuclear magnetic resonance spectroscopy result is：1H NMR
(400MHz,CDCl3,δ):8.48-8.47 (d, J=2H), 7.58-7.60 (d, J=2H), 7.31-7.35 (m, 4H), 7.22-
7.29(m,7H),7.13-7.14(d,2H),7.09-7.10(d,2H)；13C NMR(100MHz,CDCl3,δ):149.71,
143.84,139.20,137.78,136.65,133.96,132.46,131.05,130.00,129.48,129.01,128.47,
128.34,128.30,128.27,127.32,126.97,122.32.HRMS(ESI)m/z:Calculated value C26H19N3,373.1579
[M]；It was found that 374.1651 [M+H]+.
The synthesis of compound 4：Weigh Compound 1 (0.19g, 0.50mmol) and compound 1,4- dibromobutanes (0.106g,
0.50mmol) it is dissolved in 15mL acetonitriles, room temperature is sufficiently stirred for, reaction system fully flows back 10 hours at 90 DEG C, the monitoring of TLC plates
After reaction completely, vacuum distillation removes solvent, the isolated product 4 of silica gel column chromatography.Yield is 51%.Nuclear magnetic resonance spectroscopy knot
It is really：1H NMR(400MHz,DMSO-d6,δ):8.90-8.92(d,2H),7.79-7.81(d,2H),7.48-7.53(m,7H),
7.26-7.36(m,8H),4.50-4.54(t,2H),3.54-3.57(t,2H),1.98-2.01(m,2H),1.79-1.82(m,
2H)；13C NMR(100MHz,DMSO-d6,δ):144.99,144.17,140.55,140.33,136.51,135.96,
133.54,131.38,130.63,129.71,129.30,129.12,128.98,128.91,127.99,127.01,124.12,
59.60,34.36,29.61,29.16.
The synthesis of compound 5：Weigh Compound 4 (120mg, 0.20mmol) is dissolved in 10mL pyridine solutions, and room temperature is filled
Divide stirring, reaction system fully flows back 8 hours at 90 DEG C, after the monitoring of TLC plates is reacted completely, vacuum distillation removes solvent, silica gel
Column chromatography for separation obtains product 5.Yield is 53%.Nuclear magnetic resonance spectroscopy result is：1H NMR(400MHz,DMSO-d6,δ):
9.29-9.30(d,2H),9.10-9.12(d,2H),8.61-8.65(t,1H),8.16-8.20(t,2H)7.77-7.79(d,
2H),7.49-7.53(m,7H),7.26-7.37(m,8H),4.76-4.80(t,2H),4.65-4.69(t,2H),1.91-1.98
(m,4H)；13C NMR(100MHz,DMSO-d6,δ):144.67,144.05,143.82,142.73,139.19,138.93,
135.12,134.58,132.18,130.01,129.29,129.07,128.35,127.94,127.76,127.54,127.21,
126.61,125.63,122.61,58.62,57.91,26.25,25.89.
Embodiment 3
The enhanced object 7 of aggregation inducing fluorescent emission of the invention (R in example1、R2、R3=H, R4=C6) synthesis,
Synthetic route is as follows：
The synthesis of compound 1：4- formylpyridines (1.06g, 10mmol) and aniline (0.91ml, 10mmol) are weighed respectively
It is dissolved in the middle of 100mL glacial acetic acid, is stirred at room temperature one hour.Benzil (2.1g, 10mmol) and ammonium acetate (3.55g,
46mmol) sequentially it is added in reaction system, compound reacts overnight under the conditions of 120 DEG C, reaction terminates rear reaction system and falls
Enter in 300mL frozen water, system pH to neutrality is reconciled with the sodium hydroxide solution of 0.1mmol/L, mixture filtering washes three with water
Time, silica gel column chromatography separating purification obtains product after vacuum drying.Yield is 42%.Nuclear magnetic resonance spectroscopy result is：1H NMR
(400MHz,CDCl3,δ):8.48-8.47 (d, J=2H), 7.58-7.60 (d, J=2H), 7.31-7.35 (m, 4H), 7.22-
7.29(m,7H),7.13-7.14(d,2H),7.09-7.10(d,2H)；13C NMR(100MHz,CDCl3,δ):149.71,
143.84,139.20,137.78,136.65,133.96,132.46,131.05,130.00,129.48,129.01,128.47,
128.34,128.30,128.27,127.32,126.97,122.32.HRMS(ESI)m/z:Calculated value C26H19N3,373.1579
[M]；It was found that 374.1651 [M+H]+.
The synthesis of compound 6：Weigh Compound 1 (0.19g, 0.50mmol) and compound 1,6- dibromo-hexanes (0.12g,
0.50mmol) it is dissolved in 15mL acetonitriles, room temperature is sufficiently stirred for, reaction system fully flows back 10 hours at 90 DEG C, the monitoring of TLC plates
After reaction completely, vacuum distillation removes solvent, the isolated product 6 of silica gel column chromatography.Yield is 54%.Nuclear magnetic resonance spectroscopy knot
It is really：1H NMR(500MHz,DMSO-d6,δ):8.91-8.93(d,2H),7.79-7.80(d,2H),7.49-7.54(m,7H),
7.28-7.37(m,8H),4.46-4.50(t,2H),3.51-3.54(t,2H),1.78-1.91(m,2H),1.78-1.81(m,
2H),1.24-1.43(m,4H)；13C NMR(125MHz,DMSO-d6,δ):143.36,142.41,138.93,138.61,
134.80,134.32,131.90,129.74,129.00,128.78,128.08,127.67,127.51,127.35,127.29,
126.35,125.36,122.40,58.74,33.83,30.66,30.14,25.66,23.30.
The synthesis of compound 7：Weigh Compound 6 (110mg, 0.21mmol) is dissolved in 10mL pyridine solutions, and room temperature is filled
Divide stirring, reaction system fully flows back 8 hours at 90 DEG C, after the monitoring of TLC plates is reacted completely, vacuum distillation removes solvent, silica gel
Column chromatography for separation obtains product 3.Yield is 53%.Nuclear magnetic resonance spectroscopy result is：1H NMR(400MHz,DMSO-d6,δ):
9.29-9.30(d,2H),9.10-9.12(d,2H),8.61-8.65(t,1H),8.16-8.20(t,2H)7.77-7.79(d,
2H),7.49-7.53(m,7H),7.26-7.37(m,8H),4.76-4.80(t,2H),4.65-4.69(t,2H),1.91-1.98
(m,4H)；13C NMR(100MHz,DMSO-d6,δ):145.13,144.56,144.34,143.21,139.78,139.43,
135.62,135.15,132.74,130.58,129.85,129.62,128.91,128.52,128.33,128.19,128.11,
127.71,127.17,126.18,123.15,59.97,59.24,29.97,29.65,24.24,24.20.
Embodiment 4
The preparation of fluorescence organic nano probe：
The dimethyl sulfoxide solution of 20 μ L compounds 7 (1mM) is taken with liquid-transfering gun, under ultrasonic bar in be added to 2mL phosphoric acid delay
Rush make in solution (10mM, pH=7.4) compound 3 ultimate density be 10 μM.30min systems are stirred under room temperature condition opalescence
Produce, in order to verify its nanometer of Assembling Behavior, dynamic light scattering experiment determines its average grain diameter for 101nm, as shown in Figure 1.
Embodiment 5
Projection electron microscope (TEM) test of the fluorescent nano probe of preparation：
Prepared solution in the drop embodiment 4 of absorption one, drips on copper mesh, with filter suck dry moisture, dries naturally, is placed in throwing
Observed in radio mirror.Projection electron microscopic picture is as shown in Figure 2.
Embodiment 6
The fluorescence intensity of fluorescent nano probe and the linear relationship of albumin concentration：
To the albumin solution liquid that 10 μ L are separately added into system prepared in embodiment 4, make the final dense of albumin
Degree is respectively reached, 1.5 μM, 3.0 μM, 4.5 μM, 6.0 μM, 7.5 μM, 9.0 μM, 10.5 μM, 12.0 μM, 13.5 μM, 15.0 μM,
18.0μM、21μM.After the completion of all test solutions are prepared, it is well mixed using vortex instrument.Measured after incubation at room temperature 1min
Its fluorescent emission intensity at 480nm.Acquired results as shown in Figure 3, the fluorescence intensity of 480nm and albumin in system
Concentration has good linear relationship between 0-15 μM.
Embodiment 7
The selectivity of fluorescent nano probe dialogue Protein Detection：
Using solution prepared in embodiment 4 as fluorescent nano probe, and evaluate selection of the probe to albumin
Property, the excitation wavelength of the nano-probe is 380nm.The concentration of chemical combination 3 is 10 μM in the system, adds white egg to the system respectively
Make its concentration in vain for 15 μM, and 5 times of trypsase of molar equivalent, papain, pepsin, immunoglobulin, Portugals
After grape carbohydrate oxidase, histidine, cysteine, homocysteine, lysine, glutathione, ATP, glucose mixing fully,
After incubation at room temperature 1min, survey its fluorescence emission spectrum and record the intensity of fluorescent emission at 480nm.Result as shown in Figure 5, only
Have in the system for adding albumin and produce strong blue-green fluorescent, and add be barely perceivable in the solution of other species it is bluish-green
Color fluorescence.The above results show that fluorescence organic nano grain detection albumin has good selectivity and practical application.
Finally, it is accordingly required in particular to explanation, above citing is only some specific embodiments of the invention.The present invention is obvious not
It is limited to above example, all changes that the person skilled of this area can directly be derived from present disclosure or associated
Shape, is considered as protection scope of the present invention.
Claims (10)
1. a kind of fluorescent molecular probe for launching effect with water-soluble and aggregation inducing, it is characterised in that：With the structure of formula 1：
Wherein,
R1、R2And R3It is independently selected from hydrogen, halogen, alkyl, hydroxyl, alkoxy, nitro, carboxyl or amido；
R4Selected from C1~C13Aliphatic chain or C8~C13Alkyl chain containing aryl.
2. the fluorescent molecular probe for launching effect with water-soluble and aggregation inducing according to claim 1, its feature exists
In：R2And R3It is selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, hydroxyl, methoxyl group, nitro, carboxyl or dimethylamino；R4Selected from C2~C8
Alkane chain or 1,4- dimethylenebenzene groups.
3. the fluorescent molecular probe for launching effect with water-soluble and aggregation inducing according to claim 1, its feature exists
In：R1Selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, hydroxyl, methoxyl group, nitro, carboxyl or dimethylamino.
4. a kind of fluorescent nano probe, it is characterised in that：The fluorescent molecular probe self assembly as described in any one of claims 1 to 3
Formed.
5. the fluorescent molecular probe for launching effect with water-soluble and aggregation inducing described in any one of claims 1 to 3 is prepared
Method, it is characterised in that：Comprise the following steps：
1) 4- formylpyridines, the amino benzenes compounds of formula 2 and the benzil class compound of formula 3 are carried out in acetic acid/ammonium acetate system
Cyclization, obtains the intermediate of formula 4；
2) with the bromo compound of formula 5 two there is nucleophilic substitution in the intermediate of formula 4, obtain the intermediate of formula 6；
3) with pyridine there is nucleophilic substitution in the intermediate of formula 6, obtain final product；
Wherein,
R1、R2And R3It is independently selected from hydrogen, halogen, alkyl, hydroxyl, alkoxy, nitro, carboxyl or amido；
R4Selected from C1~C13Aliphatic chain or C8~C13Alkane chain containing aryl.
6. preparation according to claim 5 has the fluorescence organic nano probe of water-soluble and aggregation inducing transmitting effect
Method, it is characterised in that：
1) course of reaction in is：Terephthalaldehyde and amino benzenes compounds are stirred into 0.5~1.5h in glacial acetic acid solvent, then
Benzil class compound and ammonium acetate are added, 8~16h is reacted at a temperature of 110~130 DEG C；
2) reaction condition in is：Using acetonitrile as solvent, reacted 6~10 hours at 80~95 DEG C；
3) reaction condition in is：Using pyridine as solvent, reacted 6~10 hours at 80~95 DEG C.
7. a kind of method of the fluorescent nano probe prepared described in claim 4, it is characterised in that：Claims 1 to 3 is any
After fluorescent probe molecule described in is dissolved in organic solvent, it is added in the aqueous solution, it is ultrasonically treated, obtain final product fluorescent nano probe.
8. the method for preparing fluorescent nano probe according to claim 7, it is characterised in that：Described organic solvent is selected from
At least one in methyl alcohol, ethanol, dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, acetonitrile；The described aqueous solution is selected from pure
Water, physiological saline, phosphate buffer solution, 4- HEPESs cushioning liquid or three (methylol) aminomethane hydrochlorides
Cushioning liquid.
9. the application of the fluorescent nano probe described in claim 4, it is characterised in that：It is applied to the detection of albumin.
10. the application of fluorescent nano probe according to claim 9, it is characterised in that：It is applied to chemical solution system, suffers from
The quantitative fluorescence analysis detection of albumin in person's blood or biological tissue.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611177239.1A CN106699734B (en) | 2016-12-19 | 2016-12-19 | Fluorescent molecular probe and nano probe as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611177239.1A CN106699734B (en) | 2016-12-19 | 2016-12-19 | Fluorescent molecular probe and nano probe as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106699734A true CN106699734A (en) | 2017-05-24 |
| CN106699734B CN106699734B (en) | 2020-03-10 |
Family
ID=58938601
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611177239.1A Active CN106699734B (en) | 2016-12-19 | 2016-12-19 | Fluorescent molecular probe and nano probe as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106699734B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110642839A (en) * | 2019-08-14 | 2020-01-03 | 深圳大学 | Nano probe and preparation method and application thereof |
| WO2020007210A1 (en) * | 2018-07-05 | 2020-01-09 | 湖南超亟检测技术有限责任公司 | Water-soluble fluorescent probe and nanoparticles capable of being used for ovarian cancer and having aggregation induced emission effect, and preparation methods and applications of water-soluble fluorescent probe and nanoparticles |
| CN111610168A (en) * | 2019-02-26 | 2020-09-01 | 香港科技大学 | AIE molecule for detecting potential bloodstains and application thereof |
| CN111795960A (en) * | 2020-08-10 | 2020-10-20 | 齐齐哈尔大学 | Molecular platform for detecting different forms of iodine by spectrometry and colorimetry, and preparation method and application thereof |
| CN113620884A (en) * | 2020-05-09 | 2021-11-09 | 中国科学院大连化学物理研究所 | Compound, preparation method thereof and application of compound as fluorescent probe |
| CN114015022A (en) * | 2021-11-16 | 2022-02-08 | 西安交通大学 | Cationic conjugated polymer, ratiometric fluorescent probe based on cationic conjugated polymer, preparation method and application |
-
2016
- 2016-12-19 CN CN201611177239.1A patent/CN106699734B/en active Active
Non-Patent Citations (4)
| Title |
|---|
| ELI G. HVASTKOVS ET AL.: "Electrochemical Detection of DNA Hybridization via Bis-Intercalation of a Naphthylimide-Functionalized Viologen Dimer", 《ANALYTICAL CHEMISTRY》 * |
| FILIP ZEMEK ET AL.: "Acetylcholinesterase Reactivators (HI-6, Obidoxime,Trimedoxime, K027, K075, K127, K203, K282):Structural Evaluation of Human Serum Albumin Binding and Absorption Kinetics", 《INT. J. MOL. SCI.》 * |
| SHAHI IMAM REJA ET AL.: "A TICT based NIR-fluorescent probe for human serum albumin: a pre-clinical diagnosis in blood serum", 《CHEM. COMMUN.》 * |
| YUANYUAN YUE ET AL.: "Synthesis of imidazole derivatives and the spectral characterization of the binding properties towards human serum albumin", 《SPECTROCHIMICA ACTA PART A: MOLECULAR AND BIOMOLECULAR SPECTROSCOPY》 * |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210029793A (en) * | 2018-07-05 | 2021-03-16 | 후난 타겟팅 디텍션 테크놀로지 컴퍼니 리미티드 | Water-soluble fluorescent probe and nanoparticles with agglutination-induced emission effect for use in ovarian cancer, and their manufacturing method and application |
| WO2020007210A1 (en) * | 2018-07-05 | 2020-01-09 | 湖南超亟检测技术有限责任公司 | Water-soluble fluorescent probe and nanoparticles capable of being used for ovarian cancer and having aggregation induced emission effect, and preparation methods and applications of water-soluble fluorescent probe and nanoparticles |
| CN110684014A (en) * | 2018-07-05 | 2020-01-14 | 湖南超亟检测技术有限责任公司 | Water-soluble fluorescent probe and nanoparticle with aggregation-induced emission effect for ovarian cancer and preparation method and application thereof |
| KR102556554B1 (en) * | 2018-07-05 | 2023-07-17 | 후난 타겟팅 디텍션 테크놀로지 컴퍼니 리미티드 | Water-soluble fluorescent probes and nanoparticles with aggregation-induced release effects that can be used for ovarian cancer and their preparation methods and applications |
| CN110684014B (en) * | 2018-07-05 | 2023-05-16 | 湖南超亟检测技术有限责任公司 | Water-soluble fluorescent probe and nanoparticle with aggregation-induced emission effect and preparation methods and application thereof |
| CN111610168B (en) * | 2019-02-26 | 2023-05-05 | 香港科技大学 | AIE molecule for detecting potential blood stain and application thereof |
| CN111610168A (en) * | 2019-02-26 | 2020-09-01 | 香港科技大学 | AIE molecule for detecting potential bloodstains and application thereof |
| CN110372681B (en) * | 2019-07-29 | 2022-06-07 | 天津医科大学 | Application of self-assembled nano fluorescent probe for selectively detecting human serum albumin |
| 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 |
| CN110642839B (en) * | 2019-08-14 | 2022-04-01 | 深圳大学 | Nano probe and preparation method and application thereof |
| CN110642839A (en) * | 2019-08-14 | 2020-01-03 | 深圳大学 | Nano probe and preparation method and application thereof |
| CN113620884A (en) * | 2020-05-09 | 2021-11-09 | 中国科学院大连化学物理研究所 | Compound, preparation method thereof and application of compound as fluorescent probe |
| CN111795960A (en) * | 2020-08-10 | 2020-10-20 | 齐齐哈尔大学 | Molecular platform for detecting different forms of iodine by spectrometry and colorimetry, and preparation method and application thereof |
| CN114015022B (en) * | 2021-11-16 | 2022-08-09 | 西安交通大学 | Cationic conjugated polymer, ratio-type fluorescent probe based on cationic conjugated polymer, preparation method and application |
| CN114015022A (en) * | 2021-11-16 | 2022-02-08 | 西安交通大学 | Cationic conjugated polymer, ratiometric fluorescent probe based on cationic conjugated polymer, preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106699734B (en) | 2020-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106699734A (en) | Fluorescent molecular probe and nanoprobe as well as preparation method and application thereof | |
| JP5313249B2 (en) | Fluorescence resonance energy transfer detection using nanoparticles | |
| JP5817838B2 (en) | Fluorescence immunoassay method using polypeptide complex containing fluorescently labeled antibody variable region | |
| US20040171827A1 (en) | Phthalocyanine dyes | |
| JP2015172199A (en) | Fluorescent compounds | |
| CN105572095B (en) | A kind of detection reagent and quantitative detecting method of human serum albumins | |
| CN105928920B (en) | A kind of detection method based on aggregation-induced emission and aptamer | |
| Li et al. | A visible and near-infrared dual-fluorescent probe for discrimination between Cys/Hcy and GSH and its application in bioimaging | |
| JP2008502681A (en) | Substituted azaporphine as a fluorescent label | |
| CA2630046A1 (en) | Fluorescent membrane intercalating probes and methods for their use | |
| WO2009006443A1 (en) | Large stoke shift nir dyes | |
| BRPI0711844A2 (en) | method and system for determining sample matrix effects, and disposable cartridge | |
| CN109633171A (en) | The quickly fluorescence immune chromatography test paper bar and its preparation and application of detection GFAP | |
| JP2022543618A (en) | Chemiluminescent compounds for multiplexing | |
| CN110201191A (en) | A kind of functional protein and the compound of cyanine dye molecule and its preparation method and application | |
| CN104198740B (en) | A kind of to glucose and the synchronous nano biological sensor detecting of cholesterol | |
| Warerkar et al. | A hemicyanine based fluorescence turn-on sensor for amyloid fibril detection in the far-red region | |
| US10761025B2 (en) | Method for detecting biomaterial using linear upconversion fluorescent property | |
| CN102967708A (en) | Homogeneous immunoassay based method for synchronous fluorescence detection of multiple disease markers | |
| CN106632305B (en) | A kind of fluorescence probe and nanoparticle and preparation method and application with water-soluble and aggregation inducing transmitting effect | |
| CN111499555B (en) | Two-photon fluorescence labeling probe, synthetic method thereof and application of new coronavirus diagnosis | |
| CN105682689B (en) | For detecting the diagnostic device of disease correlation target structure | |
| CN110183376B (en) | Fluorescent probe for detecting human serum albumin and synthetic method and application thereof | |
| Dhiman et al. | “Turn-On” monopodal and dipodal nanoprobes for serum albumins–a case of shift in selectivity towards BSA and a Z-to U-like conformational change | |
| JP2003525915A (en) | Fluorescent membrane intercalating probe and method of using the same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |