CN106432334A - Mitochondria fluorescent probe with double fluorescence emission as well as preparation method and application thereof - Google Patents
Mitochondria fluorescent probe with double fluorescence emission as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN106432334A CN106432334A CN201610827552.9A CN201610827552A CN106432334A CN 106432334 A CN106432334 A CN 106432334A CN 201610827552 A CN201610827552 A CN 201610827552A CN 106432334 A CN106432334 A CN 106432334A
- Authority
- CN
- China
- Prior art keywords
- mitochondria
- fluorescent probe
- fluorescence probe
- fluorescent emission
- double fluorescent
- 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
- 210000003470 mitochondria Anatomy 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000007850 fluorescent dye Substances 0.000 title abstract description 11
- 210000004027 cell Anatomy 0.000 claims abstract description 24
- -1 phosphonate compound Chemical class 0.000 claims abstract description 18
- 150000001450 anions Chemical class 0.000 claims abstract description 5
- 125000000217 alkyl group Chemical group 0.000 claims abstract description 3
- 125000003118 aryl group Chemical group 0.000 claims abstract description 3
- 239000000523 sample Substances 0.000 claims description 29
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 claims description 21
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical group [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 claims description 20
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- XYFCBTPGUUZFHI-UHFFFAOYSA-O phosphonium Chemical compound [PH4+] XYFCBTPGUUZFHI-UHFFFAOYSA-O 0.000 claims description 17
- 238000006243 chemical reaction Methods 0.000 claims description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 12
- 150000001875 compounds Chemical class 0.000 claims description 12
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 claims description 12
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 235000019445 benzyl alcohol Nutrition 0.000 claims description 8
- 229960004217 benzyl alcohol Drugs 0.000 claims description 8
- 150000003003 phosphines Chemical class 0.000 claims description 7
- 150000004714 phosphonium salts Chemical class 0.000 claims description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 6
- PHSPJQZRQAJPPF-UHFFFAOYSA-N N-alpha-Methylhistamine Chemical compound CNCCC1=CN=CN1 PHSPJQZRQAJPPF-UHFFFAOYSA-N 0.000 claims description 6
- 230000002438 mitochondrial effect Effects 0.000 claims description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 3
- 229940006460 bromide ion Drugs 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 241000370738 Chlorion Species 0.000 claims description 2
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 2
- 229910052757 nitrogen Inorganic materials 0.000 claims description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 2
- 150000001412 amines Chemical class 0.000 claims 1
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 238000003786 synthesis reaction Methods 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 3
- 238000009826 distribution Methods 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 230000008859 change Effects 0.000 abstract description 2
- 239000008204 material by function Substances 0.000 abstract description 2
- 238000010923 batch production Methods 0.000 abstract 1
- 230000005284 excitation Effects 0.000 abstract 1
- 238000001215 fluorescent labelling Methods 0.000 abstract 1
- 229910052736 halogen Inorganic materials 0.000 abstract 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 5
- BBEAQIROQSPTKN-UHFFFAOYSA-N antipyrene Natural products C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 description 5
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthrene Natural products C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 description 5
- AGEZXYOZHKGVCM-UHFFFAOYSA-N benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 description 4
- IKEOZQLIVHGQLJ-UHFFFAOYSA-M mitoTracker Red Chemical compound [Cl-].C1=CC(CCl)=CC=C1C(C1=CC=2CCCN3CCCC(C=23)=C1O1)=C2C1=C(CCC1)C3=[N+]1CCCC3=C2 IKEOZQLIVHGQLJ-UHFFFAOYSA-M 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 238000003501 co-culture Methods 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- PMOIAJVKYNVHQE-UHFFFAOYSA-N phosphanium;bromide Chemical compound [PH4+].[Br-] PMOIAJVKYNVHQE-UHFFFAOYSA-N 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 238000000967 suction filtration Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXMKQUNVWSEMD-UHFFFAOYSA-N benzyl chloride Chemical compound ClCC1=CC=CC=C1 KCXMKQUNVWSEMD-UHFFFAOYSA-N 0.000 description 2
- 229940073608 benzyl chloride Drugs 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 238000012805 post-processing Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- TUQOTMZNTHZOKS-UHFFFAOYSA-N tributylphosphine Chemical compound CCCCP(CCCC)CCCC TUQOTMZNTHZOKS-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- GSNUFIFRDBKVIE-UHFFFAOYSA-N DMF Natural products CC1=CC=C(C)O1 GSNUFIFRDBKVIE-UHFFFAOYSA-N 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 230000004103 aerobic respiration Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- RPGWZZNNEUHDAQ-UHFFFAOYSA-N phenylphosphine Chemical compound PC1=CC=CC=C1 RPGWZZNNEUHDAQ-UHFFFAOYSA-N 0.000 description 1
- FAIAAWCVCHQXDN-UHFFFAOYSA-N phosphorus trichloride Chemical compound ClP(Cl)Cl FAIAAWCVCHQXDN-UHFFFAOYSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/54—Quaternary phosphonium compounds
- C07F9/5456—Arylalkanephosphonium compounds
-
- 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
- C09K9/00—Tenebrescent materials, i.e. materials for which the range of wavelengths for energy absorption is changed as a result of excitation by some form of energy
- C09K9/02—Organic tenebrescent 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
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1011—Condensed systems
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Biomedical Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a mitochondria fluorescent probe as well as a preparation method and application thereof, belonging to the field of functional materials. The mitochondria fluorescent probe has a double fluorescence emission function and is a quaternary phosphonate compound with a structural formula shown by formula I. In the formula I, R is alkyl or aryl, and the anion X is halogen ion. The fluorescent probe disclosed by the invention has very efficient mitochondria specificity recognition ability, and the fluorescent probe molecule presents a double fluorescence emission phenomenon under the excitation of different lights so as to realize precise double fluorescence labeling of mitochondria. In the invention, the fluorescent probe molecule also has excellent light stability, experiences relatively little influence of the charge change on the mitochondria surface, and can realize real-time tracking of the mitochondria distribution and form in a living cell. Moreover, the mitochondria fluorescent probe disclosed by the invention has the advantages of high water solubility, simple synthesis path, simple after-treatment, batch production, relatively low cost and the like and shows a broad market prospect.
Description
Technical field
The invention belongs to field of functional materials, particularly to a kind of mitochondria fluorescence probe with double fluorescent emission and its
Preparation method and application.
Technical background
Organelle plays extremely important effect in the functional expression and disease progression of normal cell.Line grain
Body is a kind of important eukaryotic device, is the important place that cell carries out aerobic respiration.Mitochondria is except to cell supplying energy
Outward, it also take part in cellular informatics transmission, cell differentiation, Apoptosis and the physiology mistake such as regulating cell growth and cell cycle
Journey.Therefore, development one class is come to mitochondria specificity fluorescent probe, regarding track mitochondria form in the cell and distribution, will help
Study some important cellular physiological processes in us.
Imaging-PAM has high room and time distinguishing because of it, has been widely used for various physiology courses at present
Monitoring, develop new mitochondria fluorescence probe also receive highest attention.Presently commercially available mitochondria fluorescence probe species is relatively
Few, costly, photostability need to improve further price.In recent years, there are some new mitochondria fluorescence probes in succession
It is reported, but the shortcomings of the mitochondria fluorescence probe still generally existing complex structure reported, synthesis difficulty.Additionally, having double
The mitochondria fluorescence probe of weight fluorescent emission performance just can carry out Dual positioning to mitochondria, reduces the interference of background fluorescence,
Selectivity such that it is able to make probe is higher, and positioning is more accurate.Presently commercially available and document report mitochondria fluorescence probe is general
For the molecular probe of substance fluorescent emission, thus, it is badly in need of the mitochondria fluorescence probe that exploitation one class has double fluorescent transmitting.
Season, phosphonium salt structure was the very important Mitochondrially targeted site of a class, by having to having excellent fluorescence property
Fluorogen carries out the modification of season phosphonium salt structure, just can prepare efficiently, in bulk and a series of have excellent mitochondria recognition capability
Mitochondria fluorescence probe with excellent fluorescence property.
Content of the invention
It is an object of the invention to provide a kind of have mitochondria fluorescence probe of double fluorescent emission and preparation method thereof and
Application.Mitochondria fluorescence probe of the present invention is season phosphonium salt type molecular probe, and its molecular structure includes base fluorescence skeleton
With the season phosphonium salt group with mitochondria recognition performance, wherein fluorescence skeleton is connected by methylene group with season phosphonium salt group.
The present invention is achieved through the following technical solutions：
A kind of mitochondria fluorescence probe with double fluorescent emission, wherein play major function season phosphonium salt compound molecule knot
Structure is shown in formula I：
……………………………………（Formulas I）
Wherein, wherein R1, R2, R3Stand alone as alkyl or aryl；Anion X is halide ion.
Preferably, R in described structural formula1, R2, R3Stand alone as C1-C8Alkyl or phenyl.Further it is preferably R1=
R2=R3=butyl.
Preferably, the preferred bromide ion of anion X or chlorion, further preferred bromide ion in described structural formula.
One of most preferred configuration of mitochondria fluorescence probe with double fluorescent transmitting of the present invention is tributyl
（3- benzyl）Phosphonium bromide.
Present invention also offers a kind of preparation method of described season phosphonium salt type organic molecule piezochromic material, pass through
Benzylic halides and three replacement phosphine compounds react and are obtained, and it is as follows that it prepares reaction equation：
Specifically include following steps：
A, benzylalcohol and thionyl chloride or phosphorus tribromide are reacted under -10-80 C 0.5-24 hour generate corresponding benzyl
Halide；
B, by the benzylic halides of gained and three replacement phosphine compounds in a solvent, under room temperature to 120 C, reaction 2-24 is little
When can get target compound；
Preferably, in step a, benzylalcohol compound is 1 with the mol ratio of thionyl chloride or phosphorus tribromide:1~1:4；Used anti-
Answer solvent to be toluene, any one in oxolane, dichloromethane, chloroform, carbon tetrachloride.
Preferably, in step b, the mol ratio of benzylic halides and three replacement phosphine compounds is 1:0.5~1:3；Used
Reaction dissolvent is methyl alcohol, ethanol, acetone, ethyl acetate, DMF, dimethyl sulfoxide, any one in acetonitrile,
Ethyl acetate.
The preparation of mitochondria fluorescence probe of the present invention has the advantages of synthesis is simple and direct, and post processing is simple.
The present invention also provides above-mentioned mitochondria fluorescence probe that the mitochondrial mark in living cells and Mitochondrial Shape are regarded
The application of track.Described living cells is MG63 cell line, HUVEC cell line, HepG2 cell line and Hela cell line etc..
All features disclosed in this specification, or disclosed all methods or during step, except mutually exclusive
Feature and/or step beyond, all can combine by any way.
Beneficial effects of the present invention：
Heretofore described fluorescence probe has very efficient mitochondria specific recognition capability, and fluorescent probe molecule is not
Double fluorescent emission phenomena is presented under the exciting shared the same light, it is achieved thereby that mitochondria accurately double fluorescent mark.The present invention
Described in fluorescent probe molecule also there is excellent photostability, changed by mitochondrial surface electric charge affected less, can
Realize the distribution of living cells Mitochondria and form regards track in real time.Additionally, the serial mitochondria fluorescence probe that the present invention provides has
Good water solubility, synthetic route are simple and direct, post processing is simple, can produce in batches, the more low advantage of cost, show wide market
Prospect.
Brief description：
Fig. 1：Tributyl（3- benzyl）The hydrogen spectrogram of phosphonium bromide.
Fig. 2：MG63 cell and tributyl（3- benzyl）Phosphonium bromide（3.0 μM）Co-culture the confocal fluorescent photograph of 1 hour
Piece：（a）405 nm light excitated blue fluorescence；（b）488 nm light excite green fluorescence；（c）Photograph via bright field.
Fig. 3：MG63 cell and tributyl（3- benzyl）Phosphonium bromide（3.0 μM）With commercially available mitochondria fluorescence probe Mito-
The confocal fluorescent photo that Tracker Red FM contaminates altogether：（a）Tributyl（3- benzyl）Phosphonium bromide（3.0 μM, 1 h）, 405
Nm light excitated blue fluorescence；（b）Tributyl（3- benzyl）Phosphonium bromide（3.0 μM, 1 h）, 488 nm light excite green fluorescence；
（c）Mito-Tracker Red FM, the excitated red fluorescence of 552 nm light；（d）Stacking chart's fluorescence of figure a and figure c is in purple；（e）
Stacking chart's fluorescence of figure b and figure c is in yellow green；（f）Send out photograph via bright field.
Specific embodiment：
Specific embodiment by the following examples is described in further detail to the above of the present invention again.Should manage
Solution, instantiation described herein only in order to explain the present invention, is not intended to limit the present invention.Without departing from the present invention's
Any modification made within spirit and principle, and the equivalent made according to ordinary skill knowledge and customary means
Or improve, all should include within the scope of the present invention.
Embodiment 1：The synthesis of intermediate benzyl bromine
At room temperature, by phosphorus tribromide（0.49 g）Add benzylalcohol（0.42 g）Carbon tetrachloride（50 mL）In solution.Heat up
To flowing back, reaction stopped reaction after 2 hours.After removing carbon tetrachloride, gained solid is through methyl alcohol（40 mL）Washing, suction filtration and true
Sky obtains orange object benzyl bromine after being dried（0.37 g, 73%）.
Embodiment 2：The synthesis of intermediate benzyl chloride
At room temperature, by thionyl chloride（0.4 mL）Add benzylalcohol（The toluene of 0.3 g（9 mL）In solution.Continue reaction 24
After hour, stop reaction.Decompression removes excessive thionyl chloride, and residue obtains target after water washing, suction filtration and vacuum drying
Thing is light yellow solid（0.31 g, 97%）.
Embodiment 3：Object tributyl（3- benzyl）The synthesis of phosphonium bromide
Under nitrogen, by benzyl bromine（0.69 g）, tributylphosphine（0.75 mL）And ethyl acetate（15 mL）Add in reaction bulb.
It is warming up to reaction under 80 C and stop reaction after 8 hours.Suction filtration, solid with ethyl acetate washs to obtain target compound tributyl（3-
Pyrene benzyl）Phosphonium bromide is yellow solid（0.90 g, 82%）.
Embodiment 4：Object tributyl（3- benzyl）The synthesis of phosphorus chloride
In embodiment 3 benzyl bromine is changed into benzyl chloride and successfully prepare tributyl（3- benzyl）Phosphorus chloride.
Embodiment 5：
In embodiment 1-4, benzylalcohol is changed into pyrene benzylalcohol and tributyl is obtained respectively（3- pyrene benzyl）Phosphonium bromide and tributyl（3-
Pyrene benzyl）Phosphorus chloride.
Embodiment 6：Object tributyl（3- benzyl）Phosphonium bromide and the co-cultivation of MG63 cell
First, MG63 cell is cultivated 24 hours under 37 C.Subsequently, remove culture medium, add compound tributyl（3-
Benzyl）Phosphonium bromide（3 μM）DMEM solution, co-culture 1 hour.Washed after three times with DMEM, seen with confocal microscope
Examine and obtain Fig. 2.
Exciting light in Fig. 2 a is 405 nanometers；Exciting light in Fig. 2 b is 488 nanometers. compound three as can be seen from Figure 2
Butyl（3- benzyl）Phosphonium bromide can be successfully entered cell and be mainly distributed in cytoplasm.Additionally, Fig. 2 also clearly shows that,
Compound tributyl（3- benzyl）Phosphonium bromide presents double fluorescent emission phenomena in the cell.
Embodiment 7：Object tributyl（3- benzyl）Phosphonium bromide and commercially available mitochondrial dye Mitotracker Red
FM common location is analyzed
First, MG63 cell is cultivated 24 hours under 37 C.Subsequently, remove culture medium, add compound tributyl（3-
Benzyl）Phosphonium bromide（3 μM）DMEM solution, co-culture 1 hour.After being washed with DMEM, add 0.5 μM of commercially available mitochondria dye
Material Mitotracker Red FM, continues culture 0.5 hour.Washed after three times with DMEM, observed with confocal microscope
Obtain Fig. 3.
Exciting light in Fig. 3 a is 405 nanometers；Exciting light in Fig. 3 b is 488 nanometers；Exciting light in Fig. 3 c is received for 522
Rice.Can be seen that compound tributyl from stacking chart 3d and 3e（3- benzyl）The pigmented section of phosphonium bromide and commercially available mitochondria dye
The pigmented section of material Mitotracker Red FM is basically identical, and compound tributyl is described（3- benzyl）Phosphonium bromide has non-
Often good mitochondria regards track ability.
Embodiment 8：
In preparation method described in embodiment 1-7, it is respectively adopted C1-C8Alkylphosphines or Phenylphosphine replace tributylphosphine.Success
A series of mitochondria fluorescence probes with double fluorescent transmitting have been obtained.Wherein comprehensive with the product of preparation in embodiment 3
Can be best.
Embodiment 9：
In preparation method described in embodiment 1 or 4, adjustment relevant parameter carries out serial experiment：
Select -10 DEG C, 0 DEG C, 40 DEG C, 60 DEG C, 80 DEG C of replacement room temperature reactions respectively in step a, the reaction time controls in 0.5-
24 hours；Benzylalcohol compound is controlled to be 1 with the mol ratio of phosphorus tribromide respectively:1、1:2 and 1:4；It is respectively adopted toluene, dichloro
Methane, chloroform, carbon tetrachloride replace oxolane.
20 DEG C, 50 DEG C, 120 DEG C are selected to replace 80 DEG C of reactions in stepb respectively, the reaction time controls in 2-24 hour；
The mol ratio of control Benzyl halides compound and three replacement phosphine compounds is 1 respectively:0.5、1:1 and 1:3；Be respectively adopted methyl alcohol,
Ethanol, acetone, N,N-dimethylformamide, dimethyl sulfoxide, acetonitrile replace ethyl acetate.
By experiment parameter screening find above-mentioned each under the conditions of all can successfully prepare the mitochondria with double fluorescent transmitting
Fluorescence probe, but still be optimal with the design parameter described in embodiment 3.
Comparative example 1：
With reference to embodiment 6 and 7 methods describeds, it is respectively adopted embodiment 5 and prepares tributyl（3- pyrene benzyl）Phosphonium bromide and tributyl
（3- pyrene benzyl）Phosphorus chloride replaces tributyl（3- benzyl）Phosphonium bromide is marked experiment, gained in result display embodiment 5
Although product also shows that certain double fluorescent emission effects, the resultant effect such as its luminous intensity is substantially not so good as institute in the present invention
State the season phosphine salt type compound using benzyl as skeleton.
The foregoing is only the preferred embodiments of the present invention, for the purpose of the present invention, be merely illustrative, and non-limiting
's；Those of ordinary skill in the art understand, in patent requirements limited range of the present invention, it can be carried out many changes,
Modification, or even equivalent change, but fall within protection scope of the present invention.
Claims (10)
1. a kind of mitochondria fluorescence probe with double fluorescent emission it is characterised in that containing molecular structure season shown in formula I
Phosphine salt compounds：
……………………………………（Formulas I）
Wherein R1, R2, R3Stand alone as alkyl or aryl；Anion X is halide ion.
2. the mitochondria fluorescence probe with double fluorescent emission according to claim 1 is it is characterised in that described structural formula
Middle R1, R2, R3Stand alone as C1-C8Alkyl or phenyl.
3. the mitochondria fluorescence probe with double fluorescent emission according to claim 1 is it is characterised in that described structural formula
Middle R1=R2=R3=butyl.
4. the mitochondria fluorescence probe with double fluorescent emission according to claim 1 is it is characterised in that described structural formula
Middle anion X is bromide ion or chlorion.
5. the mitochondria fluorescence probe with double fluorescent emission according to claim 1 is it is characterised in that described season phosphonium salt
Class compound is specially tributyl（3- benzyl）Phosphonium bromide.
6. the mitochondria fluorescence probe with double fluorescent emission described in a kind of claim 1 preparation method it is characterised in that
Specifically include following steps：
A, benzylalcohol and thionyl chloride or phosphorus tribromide are reacted under -10-80 C 0.5-24 hour generate corresponding benzyl
Halide；
B, by the benzylic halides of gained and three replacement phosphine compounds in a solvent, under room temperature to 120 C, reaction 2-24 is little
When can get target compound.
7. preparation method according to claim 6 is it is characterised in that in step a, benzylalcohol compound and thionyl chloride or
The mol ratio of phosphorus tribromide is 1:1~1:4；Reaction dissolvent used is toluene, oxolane, dichloromethane, chloroform, carbon tetrachloride
In any one.
8. preparation method according to claim 6 is it is characterised in that in step b, benzylic halides and three replacement phosphines
The mol ratio of compound is 1:0.5~1:3；Reaction dissolvent used is methyl alcohol, ethanol, acetone, ethyl acetate, N, N- dimethyl formyl
Any one in amine, dimethyl sulfoxide, acetonitrile.
9. the mitochondria fluorescence probe with double fluorescent emission described in a kind of claim 1 application it is characterised in that by its
For track is regarded to the mitochondrial mark in living cells and Mitochondrial Shape.
10. according to claim 9 application it is characterised in that described living cells be MG63 cell line, HUVEC cell line,
At least one in HepG2 cell line and Hela cell line.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610827552.9A CN106432334B (en) | 2016-09-18 | 2016-09-18 | Mitochondria fluorescence probe with double fluorescent emissions and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610827552.9A CN106432334B (en) | 2016-09-18 | 2016-09-18 | Mitochondria fluorescence probe with double fluorescent emissions and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106432334A true CN106432334A (en) | 2017-02-22 |
| CN106432334B CN106432334B (en) | 2018-05-22 |
Family
ID=58168077
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610827552.9A Expired - Fee Related CN106432334B (en) | 2016-09-18 | 2016-09-18 | Mitochondria fluorescence probe with double fluorescent emissions and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106432334B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108444962A (en) * | 2018-03-05 | 2018-08-24 | 北京化工大学 | It is a kind of based on formaldehyde colorimetric probe and formaldehyde fluorescent test paper, preparation method and application method |
| CN109627264A (en) * | 2018-08-21 | 2019-04-16 | 河北大学 | Targetted mitochondria acid anhydride fluorescence probe and its synthetic method and application |
| CN111956610A (en) * | 2020-07-21 | 2020-11-20 | 四川大学 | Drug-loading system for diagnosis and treatment of atherosclerosis and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105017298A (en) * | 2014-04-28 | 2015-11-04 | 中国科学院烟台海岸带研究所 | Bodipy compound and application thereof |
-
2016
- 2016-09-18 CN CN201610827552.9A patent/CN106432334B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105017298A (en) * | 2014-04-28 | 2015-11-04 | 中国科学院烟台海岸带研究所 | Bodipy compound and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 姜娜等: ""线粒体荧光探针最新研究进展"", 《化工学报》 * |
| 杨志广等: ""有机双光子线粒体内活性小分子荧光探针研究进展"", 《分子测试学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108444962A (en) * | 2018-03-05 | 2018-08-24 | 北京化工大学 | It is a kind of based on formaldehyde colorimetric probe and formaldehyde fluorescent test paper, preparation method and application method |
| CN108444962B (en) * | 2018-03-05 | 2021-01-12 | 北京化工大学 | Perylene-based formaldehyde colorimetric probe and formaldehyde fluorescent test paper, and preparation method and use method thereof |
| CN109627264A (en) * | 2018-08-21 | 2019-04-16 | 河北大学 | Targetted mitochondria acid anhydride fluorescence probe and its synthetic method and application |
| CN109627264B (en) * | 2018-08-21 | 2021-01-05 | 河北大学 | Target mitochondrial perylene anhydride fluorescent probe and synthetic method and application thereof |
| CN111956610A (en) * | 2020-07-21 | 2020-11-20 | 四川大学 | Drug-loading system for diagnosis and treatment of atherosclerosis and preparation method thereof |
| CN111956610B (en) * | 2020-07-21 | 2021-11-30 | 四川大学 | Drug-loading system for treating atherosclerosis and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106432334B (en) | 2018-05-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108440475B (en) | Ratio type fluorescent probe for distinguishing lipid droplets with different polarities and preparation method and application thereof | |
| Chakraborty et al. | Phenoxo bridged luminescent dinuclear zinc (II) and cadmium (II) complexes of 2-[[[2-(2-pyridyl) ethyl] imino] methyl] phenol: Crystal structure, photophysical and thermal studies | |
| CN106432334A (en) | Mitochondria fluorescent probe with double fluorescence emission as well as preparation method and application thereof | |
| Amatori et al. | Modulating the sensor response to halide using NBD-based azamacrocycles | |
| CN102807591B (en) | Double benzimidazole ligand based metal complex and preparation method and application thereof | |
| CN103524539B (en) | Luminescent material Zn (phen) (HL) 2and preparation method thereof | |
| Song et al. | Synthesis, crystal structure and luminescence properties of lanthanide complexes with a new semirigid bridging furfurylsalicylamide ligand | |
| CN105541876B (en) | N-heterocyclic carbine metal complex of anthraquinone bridging and preparation method and application | |
| US8134017B1 (en) | Compound and use thereof | |
| CN104744453B (en) | Hemicyanine compound for detecting polarity of mitochondria | |
| CN102993763A (en) | Single charge boron fluroride complexing dipyrrole methenyl fluorochrome and application thereof | |
| CN106632063A (en) | Compound I and compound II based on phenanthroimidazole, and preparation method and applications thereof | |
| CN104151867B (en) | Temperature response type cyclodextrin probe double with pH and preparation method thereof | |
| Huang et al. | Employing heterometallic trinuclear cluster as building subunit to construct a new coordination polymer with rare 3D inorganic–Cd–O–Ba–connectivity | |
| CN103773060B (en) | Organic fluorescent dye molecule and synthetic method thereof and application | |
| CN103450234B (en) | A kind of cadmium coordination polymer and preparation method thereof | |
| Li et al. | Synthesis and characterization of two self-catenated networks and one case of pcu topology based on the mixed ligands | |
| WO2021244345A1 (en) | Cyanine derivative compound, preparation method therefor, and application thereof | |
| Zhao et al. | Luminescent 3D homochiral coordination polymers of europium (III) and terbium (III) based on amino acid-functionalized isophthalic acid | |
| Yang et al. | Syntheses, structures, magnetic and fluorescence properties of coordination polymers constructed with the reduced Schiff base carboxylic acid ligand | |
| CN112778258B (en) | Amide derivative neutral mitochondrial fluorescent marker and preparation method and application thereof | |
| CN104610218A (en) | Xanthene-1,8-diketone derivatives as well as preparation method and application thereof | |
| CN114773875A (en) | Azaindole-squarylium cyanine dye, and synthesis method and application thereof | |
| CN103709197A (en) | Substituted salicylaldehyde-TCF (Tricresyl Phosphate) derivative as well as preparation method and application thereof | |
| CN103275107B (en) | Annular metal complex and preparation method thereof and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180522 Termination date: 20190918 |