CN108690009A - The luxuriant and rich with fragrance imidazole type mercaptoamino acid fluorescence probe of a kind of pair of transmitting and application - Google Patents
The luxuriant and rich with fragrance imidazole type mercaptoamino acid fluorescence probe of a kind of pair of transmitting and application Download PDFInfo
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- CN108690009A CN108690009A CN201810767806.1A CN201810767806A CN108690009A CN 108690009 A CN108690009 A CN 108690009A CN 201810767806 A CN201810767806 A CN 201810767806A CN 108690009 A CN108690009 A CN 108690009A
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- mercaptoamino acid
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- 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
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- 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
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- 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
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
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- 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
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- 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
- C09K2211/1048—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms with oxygen
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Abstract
The invention discloses a kind of fluorescence probes of detection mercaptoamino acid, belong to technical field of analytical chemistry.The probe is by 3-(1H- Fei Bings [9.10-d]Imidazoles -2- bases)-[1,1'Lian Ben ]- 4- alcohol and the chloro- 7- nitros benzo -2- oxa-s -1,3- diazole of 4- are prepared at room temperature in methylene chloride.The raw material for preparing of fluorescence probe of the present invention is easy to get, and synthetic method is simple, has many advantages, such as that highly selective, high sensitivity, low detection limit to mercaptoamino acid.Fluorescence probe of the present invention can be applied to the detection of the mercaptoamino acid in cell.
Description
Technical field
The present invention provides a kind of fluorescence probes of detection mercaptoamino acid, belong to fluorescent probe technique field.
Technical background
Cysteine(Cys), homocysteine(Hcy)And glutathione(GSH)It is very important three kinds in cell
Mercaptoamino acid, also referred to as biological thiol.Their effects in vivo are to participate in redox reaction process, adjust organism
Interior Cell Homeostasis and cell metabolism.First, during biological growth and development, cysteine(Cys)Shortage can cause
The loss of growth retardation, trichochromes depigmentation, liver.Secondly, in human plasma, homocysteine(Hcy)The increase of content
It is closely related with alzheimer disease, angiocardiopathy and osteoporosis diseases.Glutathione(GSH)Maintaining cell also
It is played an important role during former environment, including keeps the cysteine in protein(Cys)In reducing condition and
Be easy to damage the free radical of DNA and RNA by capture influences to protect cells from oxidative stress.Since biological thiol is important
Biological action, a kind of method of quick, sensitive, easy detection biological thiol of development is for academic research and potential disease
It diagnoses extremely important.
Currently, such as high performance liquid chromatography, mass spectrography, gas chromatography and electrochemical method may be used to biology
Mercaptan.However these methods have the shortcomings that operating procedure is complicated, instrument is expensive, the response time is long.In contrast, fluorescence
Probe have it is highly sensitive, easy to operate, detection time is short, inexpensive, and can realize to the real-time of cell and live body
The advantages of visualizing tracer can exactly make up the deficiency of traditional detection method.Therefore, it develops and can be used in detecting organism
The fluorescence probe of interior mercaptoamino acid contributes to the propulsion of the related fields such as environment or medicine.
Invention content
A kind of fluorescence probe of detection mercaptoamino acid of the present invention, which is characterized in that the change of the fluorescence probe
Learn structural formulaIt is shown:
The fluorescence probe of above-mentioned detection mercaptoamino acid is prepared in the following manner:
20mL dryings are added in the chloro- 7- nitros benzo -2- oxa-s -1,3- diazole of 4- and compound II under nitrogen atmosphere, ice bath
Triethylamine is added dropwise in dichloromethane.It is stirred overnight at room temperature, reaction terminates to be concentrated in vacuo, and column chromatography purifies to obtain yellow solid
Close object I.
It is detected while the mercaptoamino acid that fluorescence probe of the present invention can be used in aqueous solution.
It is further preferred that the fluorescence probe is used for the DMF in pH7.4 and the biological sulphur in PBS buffer solution mixed liquor
The quick detection of alcohol.By taking glutathione as an example, detection is limited to 1.34 × 10-7 mol/L。
Fluorescence probe of the present invention is for having mercaptoamino acid with PBS buffer solution mixed liquor in the DMF of pH7.4
Double transmitting response performances.The present invention passes through experimental verification, DMSO and PBS buffer solution mixed liquor of the fluorescence probe in pH7.4
In using the light source of 365nm wavelength as exciting light, solution hardly shine.With the addition of biological thiol, solution is in 365nm
Light excitation when, solution sends out strong blue-fluorescence at 480nm.But when being excited with the light of 470nm, solution exists
Strong green fluorescence is sent out at 550nm.The addition of other non-mercaptoamino acids does not influence the fluorescence of mercaptoamino acid.Cause
This, fluorescent probe compounds of the invention have very high selectivity to biological thiol.
The fluorescent probe compounds of the present invention in DMF and the PBS buffer solution mixed liquor of pH7.4, mercaptoamino acid it is dense
That spends therefore can be with the content of mercaptoamino acid in dosing solution with fluorescence intensity presentation linear relationship.
Fluorescence probe of the present invention can be used for the detection of mercaptoamino acid in cell.
Fluorescence probe of the present invention can be applied to the detection of intracellular mercaptoamino acid.Specifically detection method is:
Hela cells are cultivated 30 minutes in the solution of 10 μM of compound I at 37 DEG C, and cell does not have fluorescent emission.At 37 DEG C
After Hela cells are first cultivated 30 minutes in the solution of 10 μM of compound I, washed three times with PBS, then respectively with gluathione
Peptide, cysteine, homocysteine buffer solution are incubated 30 minutes, and cell sends out strong fluorescence.In fact, when using difference
Wavelength can send out the light of different colours when exciting.It is intracellular various that experiment shows that fluorescence probe of the present invention can be used in
The detection of biological thiol has potential application in biomedicine etc..
Beneficial effects of the present invention:
A kind of fluorescence probe of detection biological thiol according to the present invention, has the advantage that:1, the probe can be with biological sulphur
Alcohol reacts, and fluorescent probe compounds is caused to have stronger Fluorescence intensity emission in two wave bands;2, the probe is detecting
There is higher selectivity during biological thiol;3, detection of the probe class for the various biological thiols in living cells.
Description of the drawings
Fig. 1 is compound I1H NMR spectras.
Fig. 2 is the selective fluorescence spectra of compound I.
Fig. 3 is that the lower compound I of 365nm excitations changes fluorescence spectra with glutathione concentrations.
Fig. 4 is that the lower compound I of 470nm excitations changes fluorescence spectra with glutathione concentrations.
Fig. 5 is the cell imaging figure of the compound I and glutathione under 405nm laser excitations.
Fig. 6 is the bio-imaging figure of the compound I and glutathione under 470nm laser excitations.
Specific implementation mode
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings, but not limited to this.It is each in embodiment
Kind raw material is bought both from market.
The synthesis of 1 compound I of embodiment
Under nitrogen protection, ice bath, by 0.25 g of compound II and the chloro- 7- nitros benzo -2- oxa-s -1,3- of 0.24 gram of 4- bis-
Azoles is dissolved in dry dichloromethane, and triethylamine is being added, is being stirred overnight at room temperature, and reaction terminates, and vacuum concentration is slightly produced
Object obtains compound I, yield 35% through column chromatography.Fig. 1 is the nuclear-magnetism figure of the probe.
The selectivity of 2 probe compound I of embodiment is analyzed
Disposably it is added to a concentration of 5 μM of compound in the DMF/PBS buffer solutions of the mercaptoamino acid containing 10mM
In IDMF/PBS buffer solutions, the variation of solution fluorescence is detected.As a result such as Fig. 2, as a length of 365nm of excitation light wave, probe chemical combination
Object I goes out to have strong blue-fluorescence to glutathione, cysteine, homocysteine in 480nm, and probe compound I
Other non-mercaptoamino acids are not obvious responsed to, when being excited with the light of 480nm, probe compound I sends out green fluorescence,
He is non-, and mercaptoamino acid does not influence.It is excellent to illustrate that probe compound I has glutathione, cysteine, homocysteine
Different selectivity, and can double transmitting responses.
3 probe compound I of embodiment changes response analysis to sulfhydryl amino acid concentration
By the glutathione containing various concentration, cysteine, homocysteine buffer solution in be added 10 μM of probe chemical combination
In the buffer solution of object I, the fluorescence intensity at 480nm, 550nm is with glutathione, cysteine, homocysteine amount
Increase in regularity change.Such as Fig. 3 and Fig. 4, illustrate probe compound I to the Concentration Testing ranges of various mercaptoamino acids compared with
Wide and sensitivity is higher.
4 probe compound I of embodiment detects mercaptoamino acid in living cells
It is added in the culture solution of Hela cells in 10 μM of compound I, then cultivates at 37 DEG C, be then separately added into again
It is imaged after glutathione, cysteine, homocysteine.The imaging results of glutathione such as Fig. 5 and Fig. 6.Experiment shows to visit
Mercaptoamino acid in needle compound I on cell realizes double light imagings, can be used for the detection of intracellular mercaptoamino acid.
Claims (2)
1. a kind of fluorescence probe to mercaptoamino acid specificly-response, which is characterized in that the chemical constitution of the fluorescence probe
Formula is as shown in I。
2. according to a kind of fluorescence probe of detection mercaptoamino acid described in claim 1, which is characterized in that the fluorescence is visited
Needle can be applied to the detection of the mercaptoamino acid in cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810767806.1A CN108690009A (en) | 2018-07-13 | 2018-07-13 | The luxuriant and rich with fragrance imidazole type mercaptoamino acid fluorescence probe of a kind of pair of transmitting and application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810767806.1A CN108690009A (en) | 2018-07-13 | 2018-07-13 | The luxuriant and rich with fragrance imidazole type mercaptoamino acid fluorescence probe of a kind of pair of transmitting and application |
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| CN108690009A true CN108690009A (en) | 2018-10-23 |
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| CN201810767806.1A Pending CN108690009A (en) | 2018-07-13 | 2018-07-13 | The luxuriant and rich with fragrance imidazole type mercaptoamino acid fluorescence probe of a kind of pair of transmitting and application |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830312A (en) * | 2015-04-07 | 2015-08-12 | 济南大学 | Fluorescence-enhanced probe compound preparation method and trivalent chromium ion detection method |
| CN107602502A (en) * | 2017-09-29 | 2018-01-19 | 安徽农业大学 | A kind of ESIPT types fluorescence probe and application for biological thiol detection |
-
2018
- 2018-07-13 CN CN201810767806.1A patent/CN108690009A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104830312A (en) * | 2015-04-07 | 2015-08-12 | 济南大学 | Fluorescence-enhanced probe compound preparation method and trivalent chromium ion detection method |
| CN107602502A (en) * | 2017-09-29 | 2018-01-19 | 安徽农业大学 | A kind of ESIPT types fluorescence probe and application for biological thiol detection |
Non-Patent Citations (1)
| Title |
|---|
| ZHENTAO ZHU等: "《New NBD-based fluorescent probes for biological thiols》", 《TETRAHEDRON LETTERS》 * |
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Application publication date: 20181023 |