CN107118586A

CN107118586A - The purposes of the vinyl compound of nitrogen heterocycle substitution

Info

Publication number: CN107118586A
Application number: CN201610101494.1A
Authority: CN
Inventors: 张国庆; 陈彪; 徐程; 王浩; 杜嘉俊; 毕国强; 罗毅
Original assignee: University of Science and Technology of China USTC
Current assignee: University of Science and Technology of China USTC
Priority date: 2016-02-24
Filing date: 2016-02-24
Publication date: 2017-09-01

Abstract

The present invention relates to a series of new application of the vinyl compound of nitrogen heterocycles substitution, relate more specifically to be used for bio-imaging, pH value detection, biomacromolecule detection or the purposes of cell micro-environment detection, wherein Z, S by the compound or its salt that formula a is represented₁、S₂And R₁As defined herein.The diagnosticum detected by using such compound as the fluorescent dye of bio-imaging, the fluorescent pH probe of detection pH value and/or as tumour cell, the imaging effect of the present invention is excellent and shows the property of one pack system polychrome, available for the imaging of one pack system polychrome or conventional imaging etc., there is huge applications prospect in bio-imaging and lesion detection.

Description

The purposes of the vinyl compound of nitrogen heterocycle substitution

Invention field

The present invention relates to a series of new application of the vinyl compound of nitrogen heterocycles substitution.

Background technology

With the development of molecular biology, the research to organism has been introduced into molecule even single molecules level, still Traditional SABC and biochip technology can only be limited to experiment in vitro, and can not intuitively observe studied object, So can directly show the imaging method that tissue changes with intracellular molecules on a molecular scale in the urgent need to a kind of.Fluorescence molecule Imaging: Monitoring is sensitive, imaging is rapid, "dead" harm, and can observe polymolecular event simultaneously, therefore is widely used in molecule In the research of biology, especially in the research field of cranial nerve and tumor imaging.Imaging-PAM is especially more than nearly 20 Fast development is obtained over year, in the Nobel Prize in recent years, related to Imaging-PAM just has two, the promise of 2008 Bel's chemistry awards three scientists for giving and finding and transforming green fluorescent protein, and then, the Nobel chemistry Prize of 2014 Authorize again, the Nobel Prize is seldom issued to same field within the so short time and encouraged twice, it is seen that the weight of Imaging-PAM The property wanted.

Either any Imaging-PAM, one of most crucial key is all the selection of optical probe, especially Super-resolution fluorescence microtechnic, the requirement to optical probe is high.So-called optical probe, which is mainly, to be launched in the case where light is excited The dyestuff of fluorescence, and the resolution ratio and final effect being generally imaged are dependent on the property of dyestuff, therefore, optimization and exploitation have The fluorescent dye of excellent properties turns into the development priority of Imaging-PAM.Existing commercial dyes only have several moneys, the more commonly used Have AlexaFluor, ATTO dyes, Cy dyes and DyLight Fluor series, and various fluorescins.These are commercial Dyestuff is largely that, based on the modification to fluorescein, rhodamine, cumarin, BODIPY and dicyanin dyestuff, species is single, and Synthesis is complicated, expensive, and fluorescin volume is too big, influences the research of molecular level, therefore to the new of excellent The demand constraint of type fluorescent dye the development of imaging technique.

The part of compounds of the vinyl compound of nitrogen heterocycle substitution was once used in anti-nerve degenerative diseases field (WO 2010087306, which describe part azacyclo- substitution vinyl compound, is used for anti-nerve degenerative diseases agent, WO 2010087313 Describe stimulates medicament as neurite extension, and WO 2010087315 is described as anti-Alzheimer disease medicament, JP 2004250407), organic sulfur phosphorus Pesticides Testing (Tetrahedron Letters, 2010,51 (13)：1754-1757.), information Storage and copy material (JP 2009271202, JP 2004111011, JP 2002221790), the inspection of PrPC relevant disease Survey and remedy (WO 2004035522) field, the treatment conditioning agent (WO 2010068287, WO of HGF 2011019400, WO 2012047826), ACE2 activator (WO 2008066770), Terahertz generator (US 20080099698), zinc ion and mercury ion detecting (Bulletin of the Korean Chemical Society (2009), 30 (12), 3109-3112.), apolipoprotein A-1 expression stimulant (JP 2001131151) and detect solvent polarity (Chemical Reviews, 1994,94 (8)：2319-2358.).

Due to the importance of this kind of compound, therefore it is also the focus of synthetic methodology research, many seminars close to it Studied into method, obtained high yield simply synthetic method (The Journal of organic chemistry, 2011,76 (16)：6849-6855.).But, it is all the time to the vinyl compound luminosity that nitrogen heterocycle replaces Rest on theory stage (Dyes and pigments, 2007,72 (3)：357-362., Chemical Physics Letters 268 (1997) 434-438, Chemical Physics Letters 444 (2007) 71-75, Journal of Photochemistry and Photobiology A：Chemistry, 2011,218 (1)：76-86, Chemical Physics Letters 415 (2005) 217-222), seldom apply in actual bio-imaging or be imaged property to it Row carefully studies (CA 2129933 describes aminostyryl pyridiniujm as the fluorescence probe viscosity of viscosity).Without wide Be related to generally is used for bio-imaging using such compound as fluorescent dye, and this is due to the fluorescence imaging requirement of traditional concept Dyestuff is preferably insensitive to pH and cellular environment, is desirably to obtain narrow spectrum and polychrome imaging, but is found by our research, Especially histology can disclose many new phenomenons to the fluorescent dye of responsive type in being imaged in bio-imaging, show cell micro-loop The difference in border, this property can have important application in many aspects, such as determine intracellular microenvironment pH, distinguish pathologic group Knit (such as tumour) organelle different from normal structure, differentiation etc..

The dyestuff of fluorescence imaging is currently used for, such as based on modifications such as fluorescein, rhodamine, BODIPY and dicyanins Obtained commercial dyes, in practical application generally existing some shortcomings, first consist in synthesis complexity, purification difficult, obtained production Rate is low；And because dyestuff lights in water, very strong fluorescence background is produced during imaging, it is necessary to plus washing away unnecessary dyestuff, So that imaging operation is cumbersome, imaging time is lengthened, new uncontrollable factor is brought for research.

The probe of existing pH detections is also based on the modifications such as fluorescein, rhodamine, BODIPY and dicyanin mostly Obtain, there is the deficiency that these dyestuffs possess, document (TrAC Trends in Analytical Chemistry 2010, 29 (9), pp 1004-1013 and Chem.Rev., 2010,110 (5), pp 2709-2728) existing pH fluorescence is summarized well Probe deficiency and to the demand of new pH fluorescence probes, such as fluoresceins probe Stokes displacements are small, 8- hydroxyl pyrenes -1,3, 6- trisulfonic acid classes dyestuff can not enter cell, the easy photobleaching of cyanine class dyestuff, some dyestuff photostability difference or quantum yield It is low.

The content of the invention

By further investigation, inventors hereof have unexpectedly found that, compared to conventional fluorescent dyestuff, with following A series of vinyl compound of nitrogen heterocycles substitution of formula can be simply synthesized, image taking speed is fast and also water in entirely without Background, realizes disposable imaging, and can show one pack system polychrome imaging property, bio-imaging have it is huge it is potential should With.Meanwhile, the vinyl compound of nitrogen heterocycle substitution can delicately distinguish difference more than 0.2 with 0.2 difference sensitivity PH value, without background in water, improve pH detection signal to noise ratio.In addition, the vinyl compound of nitrogen heterocycle substitution Molecular weight is small, is easily accessible cell, and photoluminescence quantum yield is high and stably, and can characterize cell micro-environment feature.

In consideration of it, it is an object of the invention to provide a series of alkenes chemical combination of the substitution of nitrogen heterocycles with below general formula The new application of thing.

On the one hand, it is used for bio-imaging, pH value detection, life the invention provides the compound or its salt represented by formula a Thing macromolecular or the purposes of cell micro-environment detection,

Wherein

Z is represented optionally by a multiple group R₂Substituted pyrrole radicals, thiazolyl, isothiazolyl, oxazolyl, isoxazoles Base, pyrazolyl, imidazole radicals, 1H-1,2,3- triazolyls, pyridine radicals, pyridazinyl, pyrimidine radicals, pyrazinyl, 1,3,5-triazines base, Yin Diindyl base, purine radicals, carbazyl, quinolyl, isoquinolyl, pteridine radicals or quinoxalinyl；Preferably, in these groups, N is contained R on the heterocyclic group of atom₂Substituent is not on the N atoms；

S₁And S₂It is each independently selected from H, halogen, C1-C6 alkyl or C3-C6 cycloalkyl；

R₁Represent H, halogen, hydroxyl, amino, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1- C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl, C1-C6 alkoxies, C1- C6 alkyl aminos, hydroxyl C1-C6 alkyl aminos, C1-C6 alkylthio groups, C3-C7 Heterocyclylalkyls, C6-C10 aryl, heteroaryl, wherein The C6-C10 aryl and the heteroaryl are optionally selected from halogen, hydroxyl, amino, sulfydryl, sulfonyl, nitre by one or more Base, carbonyl, ester group, carboxyl, cyano group, amide groups, C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl, C1-C6 Alkoxy, C1-C6 alkyl aminos, hydroxyl C1-C6 alkyl aminos, the substituent of C1-C6 alkylthio groups and C3-C7 Heterocyclylalkyls take Generation；

R₂Represent H, hydroxyl, amino, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl or C1-C6 alcoxyls Base,

Term " heteroaryl " represent to refer to 4 to 8 single or multiple yuan of rings and containing 1,2 or 3 be individually selected from N, O and S hetero atom and at least one heterocycle therein are the groups of armaticity, and term " C3-C7 Heterocyclylalkyls " refers to contain There is a N atom as 3 to 7 yuan of saturated cyclic alkyl that ring memberses and other ring memberses are carbon atom；

In a preferred embodiment, the compound or its salt that is represented by formula a be by below general formula I, II, The compound or its salt that III or IV are represented：

Wherein each T independently represents C or N, U represent O, S or N, and R₁、R₂、S₁And S₂As defined above.

In a preferred embodiment, the compound or its salt represented by formula a is by following various expression Compound or its salt：

Wherein Z, R₁、R₂、S₁And S₂As defined above, T represents C or N, R₃Represent H, hydroxyl, amino, sulfydryl, sulfonyl, carboxylic Base, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl or C1-C6 Alkoxy.

Preferably, the compound or its salt is the compound or its salt by following various expression：

Wherein R₂、R₃、S₁、S₂With T as defined in claim 3, and R₆And R₇H, hydroxyl, ammonia are represented independently of one another Base, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl or C1-C6 alkoxies.

In a preferred embodiment, the compound or its salt with selected from alkanes, ethers, esters, alcohols or Solution in the solvent of ketone is used.

In a preferred embodiment, the solvent is n-hexane, dichloromethane, tetrahydrofuran, ethyl acetate, third Ketone, ethanol or methanol, and the concentration range of the solution is 0.1x10^-5M to 10x10^-5M。

In a preferred embodiment, under conditions of different solvents, difference pH, different cell micro-environments, describedization The solution of compound or its salt shows different colours in bioluminescence imaging.

In a preferred embodiment, when the compound or its salt is used to detect pH value, it can distinguish difference More than 0.2 pH value.

The invention provides a series of new application of the vinyl compound of nitrogen heterocycles substitution, such compound can be used Make the fluorescent dye of bio-imaging, can be used as detecting the fluorescent pH probe of pH value, the inspection available for large biological molecule such as protein Survey, and/or the diagnosticum detected as tumour cell, and the imaging effect of such compound is excellent and to show one pack system more The property of color, available for the imaging of one pack system polychrome or conventional imaging etc., has huge applications in bio-imaging and lesion detection Prospect.

Brief description of the drawings

Fig. 1：Ultraviolet-visible absorption spectroscopies of the molecule I-2060 in opposed polarity organic solvent), concentration is 1 × 10^-5M, The temperature of test is 298K.

Fig. 2：Steady-state fluorescence spectrum of the molecule I-2060 in opposed polarity organic solvent, concentration is 1 × 10^-5M, test Temperature is 298K.

Fig. 3：By different dyes, (blueness is I-2025, and rose is I-2064, and cyan is I-2061, and khaki is I- 2062, purple is I-2029, and livid purple color is I-2027, and red is I-2060, and cream-coloured is I-2045) concentration is made into respectively is The solution, is then adsorbed onto on filter paper by 60mmol/L ethanol solution, and the fluorescent effect figure under ultra violet lamp.

Fig. 4：I-2060 absorption lights not in filter paper bubble in the aqueous solution of different pH (pH is respectively 1-8 from left to right) Together.

Fig. 5：Dyestuff I-2060 is to pH quantitative response curves.

Fig. 6：The microphoto that (HeLa) living cells is imaged is drawn sea with molecule I-2060, wherein the excitation wavelength used For 488nm.

Fig. 7：The microphoto being imaged with molecule I-2060 to mouse neuron living cells, wherein the excitation light wave used is a length of 488nm。

Fig. 8：The microphoto being imaged with molecule I-2060 to fixed mouse neuronal cell.

Fig. 9：Ultraviolet-visible absorption spectroscopies of the molecule I-2064 in opposed polarity organic solvent, concentration is 1 × 10^-5M, The temperature of test is 298K.

Figure 10：Steady-state fluorescence spectrum of the molecule I-2064 in opposed polarity organic solvent, the concentration of solution is 1 × 10^- ⁵M, the temperature of test is 298K.

Figure 11：The filter paper being made of molecule I-2064 can distinguish the different solutions that pH value differs 0.2 precision.

Figure 12：The microphoto that living cells is imaged is drawn sea with molecule I-2064.

Figure 13：The microphoto being imaged with molecule I-2064 to fixed HeLa cell.

Figure 14：The microphoto being imaged with molecule I-2064 to neuron living cells.

Figure 15：The microphoto being imaged with molecule I-2064 to fixed neuronal cell.

Figure 16：The microphoto that same mouse brain frozen section is dyed and is imaged with molecule I-2064 and tradition Nissl, Wherein (a) is to be dyed with Nissl, and (b) is with molecule I-2064 dyeing.As can be seen from the figure the dye of the dyestuff from exploitation is utilized Color result can be seen that the difference between relatively sharp cell distribution and Different brain region becomes apparent.

Figure 17：The microphoto being imaged with molecule I-2064 to the gastric tissue for being derived from human body, (a) is normal structure, and (b) is Cancerous issue.

Figure 18：(BSA is ox to molecule I-2064 with different biological molecules or stearic acid in identical pH=7.4 PBS solution Seralbumin, FBS is hyclone, and Trypsin is trypsase, and Lipase is lipase, and Stearic Acid are tristearin Acid, stearic acid here is used as the lipid of analog cell film) combine after the obtained steady-state fluorescence spectrogram of derivative.

Figure 19：Ultraviolet-visible absorption spectroscopy figures of the molecule I-2067 in opposed polarity organic solvent, concentration is 1 × 10^- ⁵M, the temperature of test is 298K.

Figure 20：Steady-state fluorescence spectrograms of the molecule I-2067 in opposed polarity organic solvent, concentration is 1 × 10^-5M, is surveyed The temperature of examination is 298K.

Figure 21：The brain (a), trunk (b) and afterbody (c) of zebra fish living are coloured to respectively with molecule I-2067 as Microphoto, wherein dyeing time are 20 minutes.

Figure 22：The microphoto that (HeLa) living cells is imaged is drawn sea with molecule I-2084, wherein the excitation light wave used A length of 488nm.

Figure 23：With microphotos of the molecule I-2110 to vivo tumor imaging.

Embodiment

A series of vinyl compound small molecule organic dyestuff replaced based on nitrogen heterocycle of the present invention are as complete New fluorescent dye, shows excellent property in various bio-imagings, and photoluminescence quantum yield is high, image stabilization, without one As fluorescent dye photobleaching phenomenon, harmless to cell, without phototoxicity, it is a bit that imaging clearly, with conventional dyes compare performance It is not poor.This money dyestuff also has some excellent characteristics that other dyestuffs do not have simultaneously, for example, synthesize simple, most of dyestuff leads to Cross carbon-hydrogen bond activation and react a step with regard to product can be obtained, and yield is high；Post processing is simple, can be obtained by recrystallizing or crossing post The dyestuff of high-purity；Soon and stably, most of imaging time only needs to just be implemented as picture in several seconds image taking speed；Dyestuff is in water In do not light, therefore imaging need not be cleaned, and entirely without background, obtain the image of high-contrast；Dye property is stable, protects Deposit simple, and better tolerance in vivo, therefore imaging operation is simple, it is to avoid too many preparation is lost time, Excessive uncertain factor is brought to research, error is introduced, is delayed research aircraft meeting；Most of all, the dyestuff shows one pack system Polychrome imaging property, such dyestuff extreme sensitivity, after being combined with biological tissue, in different environments (such as pH, difference group Point, ion concentration etc.) in performance send different colours light, the imaging of one pack system polychrome is presented；Such dyestuff is also easy to be repaiied Decorations, with various antibody bindings, make specific fluorescence probe, or be modified into water miscible dyestuff etc..Therefore this contaminates Material has huge potential application in different kind organism imaging, internal pH detection and tumour cell detection.

Except with suitable with known fluorescent chemicals property, such dyestuff of the present invention also has excellent property Matter, the property that outstanding behaviours is imaged in one pack system polychrome, being imaged with regular growth, living cells imaging, various biological tissues cut Piece for example brain section imaging, the detection and imaging of pathological tissue, the imaging of such as cancerous tissue, imaging organs, various living animals into As in such as zebra fish imaging imaging, the dyestuff of a component is disclosed and detected with regard to that can show the property of multiple color Phenomenon and difference that some other dyestuffs of object can not be disclosed, these differences are probably microenvironment pH, all kinds of ion concentrations, oxygen Caused by change reducing condition, oxygen concentration, gas concentration lwevel, the pKa of institute's detection compound and other properties.This kind of dyestuff can The new window of a fan is opened for imaging and biological detection, many not detectable phenomenons and difference are disclosed.

Why very sensitive to environment the vinyl compound of nitrogen heterocycle substitution in the present invention is, there is one pack system polychrome Property is imaged, reason is in structure design while Intramolecular electron transfer (ICT) and pH responses are introduced, with molecule I-2064 Exemplified by, 4- [double (2- ethoxys) amino of N, N-] phenyl of alkenyl one end is an electron-donating group (Donor), the quinoline of the other end Quinoline base be by electronics group (Acceptor), by middle one conjugation of double bond formation to-by conjugated system (abbreviation D-A System), such system is separated due to electronic ground state with excitation state, the increase of excitation state dipole moment, steady by the molecule of surrounding environment Fixed degree is different so that energy extreme difference is different, and lighting also can be different, and the molecule of stable excitation state can be solvent molecule, also may be used To be the biomolecule combined with dyestuff, such as protein, lipid etc., surrounding molecules polarity are different so that dye molecule It is luminous also different, when dyestuff is combined from different biomolecule, realization be exactly different zones light it is different；While quinoline N atomic energy on base receives proton or loses proton, receives after proton, and the dipole moment of molecule further increases so that molecule pair Environment is more sensitive, and opposed polarity can not only be distinguished for molecule by macroscopically cashing, moreover it is possible to distinguish different pH；We are similar this When binding molecule in electric charge transfer (ICT) and pH response molecule be referred to as ICT-pH dyestuffs.The characteristics of this kind of dyestuff is nitrogen-containing hetero The vinyl compound of ring group substitution, it is different according to the pKa of N in nitrogen heterocyclic ring, the sensitivity to different pH scopes is shown, is adjusted simultaneously Substituent on whole molecule so that excite and the emission peak change of dyestuff, we design a whole system according to this theory The dyestuff of row.

Such dyestuff of the present invention can be used for regular growth imaging, living cells imaging, various biological tissue sections such as Brain section imaging, the detection and imaging of pathological tissue, the imaging of such as cancerous tissue, imaging organs, the imaging of various living animals are such as Zebra fish imaging etc., the present invention also includes the other application of synthesis, the application method of imaging and the such dyestuff of dyestuff, than Such as it is used for probe (pH) and display screen, different kind organism detection, art, design and commercial product.Type of service is included in various molten In liquid, solid, nano-particle, absorption is on all kinds of materials or is wrapped in matrix.It is specific the invention provides each of which Non-limiting examples.

Compound of the present invention can be with the biomolecule such as protein, nucleic acid, lipid, cellulose, chitosan well With reference to for bio-imaging and analysis, it is also possible to make the sensitivity detection reagent in biological characteristis.

The invention also discloses dyestuff in being imaged to various types of cells, can specifically with golgiosome, nucleus is various thin After birth and different organelles are combined and send fluorescence, and the micro-structural of these organelles can be finely showed exactly.

The invention also discloses confocal fluorescence microscope together, the various different fluorescence microscopes such as super-resolution fluorescence microscope The imaging data of combined use, with potential of the exploitation for commercial fluorescent dye.

Dyestuff of the present invention combines from different pKa protein and carries out electrophoresis detection, available for different proteinoid Detection.Dyestuff can also be used for the imaging technique of the specific markers such as Halo-tag, Snap-tag, antibody labeling by modifying, or For multicolor fluorescence imaging.

Compound of the present invention can be also used for doing the dyestuff of optical markings, and the optical markings can be albumen Matter, nucleic acid, oligomer, DNA, RNA, biological cell, lipid, single-, few-and polysaccharide, part, acceptor, polymer, medicine or poly- The mark of polymer beads, and the compound passes through functional group and the HO-, H of material as described above₂N-, HS- or HO₂C- functional groups Reaction, is used to determine protein, nucleic acid, oligomer, DNA, RNA, biological cell, lipid as dyestuff, single-, few-and polysaccharide, Part, acceptor, polymer, medicine or polymer beads.

Dyestuff of the present invention can also be adsorbed on filter paper, make the test paper of fluorescent pH response.Dye Adsorption in filter In the various matrix such as paper, glass plate, absorbent cotton, or combined with polymer matrix, or covalently combined with macromolecule, can be in different pH Under send different light.Also different fluorescence can be produced under all kinds of acid vapors such as acetic acid.

Dyestuff of the present invention also has pressure sensitive quality, rolling, the power such as shear, stretch in the presence of, with obvious Luminous red shift or blue shift, after a period of time recover.Available in mechanoluminescence and various detections application.

The invention discloses the synthesis of the compound, purifying and detection method.Disclose the various reasons of the compound Change the properties such as property, UV absorption, fluorescent emission.

Such compound of the present invention has good molten in the organic solvents such as ethanol, methanol, chloroform, ethyl acetate Solution property and strong fluorescence, with higher molar extinction coefficient.Majority of compounds only has less water solubility, and easily with Different kind organism molecule in water, which is combined, sends different fluorescence, plays the functions such as detection, analysis, imaging.All formulas can be with Optionally modification includes following solubilizing group, and it is selected from SO₃ ^-, PO₃ ^-, CO₂H, OH, NR₃ ⁺, and the ring of hydrophilic characteristics is provided Dextrin or sugar；The substituent is connected to chromophore optionally by aliphatic group or Heteroaliphatic groups or ring-like spacer base.Repair Compound after decorations solubilizing group has good aqueous solubility.

Such dyestuff of the present invention can be by the monochrome in ultraviolet (UV), visible or near-infrared (NIR) spectral region Light (laser/laser diode) or polychromatic light (white light source) simply excitation-emission fluorescence are high with high molar extinction coefficient Photostability and high storage stability, and entire series dyestuff can provide different lipophilic hydrophilies, for all kinds of detections, analysis With imaging etc. function.

It is one that the present invention, which is used to be implemented as in the middle of the vinyl compound that the dyestuff of picture replaces for nitrogen heterocycle, structural formula, Or multiple vinyl, the one end that is connected with vinyl is nitrogen heterocycle, and the other end is substituted benzene ring or other chemical groups, is used for The dyestuff of imaging includes the derivative and its salt for the vinyl compound that nitrogen heterocycle replaces.

The vinyl compound and its derivative of nitrogen heterocycle substitution be

The compound of following any formula：

Z is represented optionally by a multiple group R₂Substituted pyrrole radicals, thiazolyl, isothiazolyl, oxazolyl, isoxazoles Base, pyrazolyl, imidazole radicals, 1H-1,2,3- triazolyls, pyridine radicals, pyridazinyl, pyrimidine radicals, pyrazinyl, 1,3,5-triazines base, Yin Diindyl base, purine radicals, carbazyl, quinolyl, isoquinolyl, pteridine radicals, quinoxalinyl；

R₂Represent H, hydroxyl, amino, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl or C1-C6 alcoxyls Base；

Term " heteroaryl " represent to refer to 4 to 8 single or multiple yuan of rings and containing 1,2 or 3 be individually selected from N, O and S hetero atom and at least one heterocycle therein are the groups of armaticity, and term " C3-C7 Heterocyclylalkyls " refers to contain There is a N atom as 3 to 7 yuan of saturated cyclic alkyl that ring memberses and other ring memberses are carbon atom.

In the present invention, term " C1-C6 alkyl " represents such alkyl：It can be straight chain or side chain, carry Single or multiple branches, wherein the alkyl generally comprises 1 to 6 carbon atom, for example, methyl (Me), ethyl (Et), propyl group, Isopropyl (i- propyl group), normal-butyl, i- butyl (isobutyl group), 2- butyl (sec-butyl), t- butyl (tert-butyl group), isopentyl, 2- Ethyl-propyl, 1,2- Dimethyl-propyl etc..

Term " cyano group " refers to NC-.

Term " halogen " represents chlorine (Cl), iodine (I), fluorine (F) and bromine (Br).

Term " heteroaryl ", individually or with other moiety combinations, refer to such armaticity carbocylic radical, it has single 4 to 8 yuan of rings or comprising 6 to 14, particularly 6 to 10 annular atoms and containing 1,2 or 3 be individually selected from the miscellaneous of N, O and S At least one heterocycle is armaticity in multiple fused rings of atom, the group.The example of " heteroaryl " includes：Benzofuran Base, benzimidazolyl, 1H- benzimidazolyls, benzoxazinyl, benzoxazolyl, benzothiazine base, benzothiazolyl, benzo It is thienyl, BTA base, furyl, imidazole radicals, indazolyl, 1H- indazolyls, indyl, isoquinolyl, isothiazolyl, different Oxazolyl, oxazolyls, pyrazinyl, pyrazolyl, 1H- pyrazolyls, pyrazolo [1,5-a] pyridine radicals, pyridazinyl, pyridine radicals, pyrimidine Base, pyrrole radicals, quinolyl, tetrazole radical, thiazolyl, thienyl, triazolyl, 6,7- dihydros -5H- [1] benzazole base etc..Particularly " Heteroaryl " is quinolyl, isoquinolyl, pyridine radicals and pyrazinyl.Specifically " heteroaryl " is quinoline -2- bases, isoquinolin -1- first Base, pyridine -2- bases and pyrazine -2- bases.

Term " hydroxyl " expression-OH.

Term " amino " expression-NH₂。

Term " sulfydryl " expression-SH.

Term " sulfonyl " expression-S (O)₂-。

Term " carboxyl " expression-CO₂H。

Term " amide groups " expression-NHCOR, wherein R is C1-C6 alkyl.

Term " nitro " expression-NO₂。

Term " carbonyl " expression-CO-.

Term " ester group " expression-COOR, wherein R is C1-C6 alkyl.

In one embodiment, when Z group is different azacyclo-, below formula compound is formed：

Wherein T is carbon or nitrogen (T=C or N), U be oxygen element, element sulphur or nitrogen (T=O or S or N)；

In addition, the present invention also provides the compound of below formula：

Each substituent therein is as defined above.

In the present invention, all formulas can also optionally include following solubilizing group, ionization or ionizable substituents, It is selected from SO₃ ^-, PO₃ ^-, CO₂H, OH or NR₃ ⁺There is provided the cyclodextrin of hydrophilic characteristics or sugar or PEG chains；The substituent is optional Ground is connected to chromophore by aliphatic group or Heteroaliphatic groups or ring-like spacer base.

The salt of the vinyl compound of nitrogen heterocycle substitution of the present invention refers to the salt that the compound is formed with acid, The acid is, for example, strong inorganic acid, strong organic acid, saturation or undersaturated dicarboxylic acids or sulfonic acid, and the strong inorganic acid includes sulphur Acid, phosphoric acid or hydrochloric acid, the strong organic acid include acetic acid, and the saturation or undersaturated dicarboxylic acids include tartaric acid, lemon Acid, the sulfonic acid includes o-methyl-benzene sulfonic acid or p-methyl benzenesulfonic acid.

In one embodiment, the compound is formula I

And the non-limiting example of compounds of formula I is the compound with structural formula as shown in table 1 below：

Table 1

In one embodiment, the compound is formula II

And formula II non-limiting example is the compound with structural formula as shown in table 2 below：

Table 2

In one embodiment of the invention, the compound is general formula III

And the non-limiting example of general formula III is the compound with structural formula as shown in table 3 below：

Table 3

In embodiments of the invention, the compound is formula IV

And formula IV non-limiting example is the compound with structural formula as shown in Table 4 below：

Table 4

In addition, the present invention is also provided by binding antibody-dye techniques, above-mentioned dye composition is modified into energy and albumen The dyestuff of matter specific reaction, with following compound

Exemplified by, it can be modified to respectively by specifically being reacted with amino, sulfydryl, carbonyl, alkynyl or azido Compound with following structural formula：

The compound can also be combined with protein tag technology, make the fluorescence probe of specific marker target protein, If following two example is respectively that (Halo herein refers to a kind of protein tag technology to Halo-tag, passes through Halo parts Can be fluorescent dye and protein specific combine, detailed description is shown inACS Chem Biol.2008Jun 20；3(6)： 373-82.tag is the meaning of label) and SNAP-tag (what SNAP herein referred to is also a kind of protein tag technology, is passed through SNAP parts can be fluorescent dye and protein specific combine, detailed description is shown in Chemistry＆biology, 2008, 15(2)：128-136.tag refer to label) fluorescence probe

It will describe to be related in the present invention synthetic method of compound by way of example below.

A part of dyestuff in the present invention passes through 2- methylquinolines or 2- methyl-quinoxalines or 2- picolines or 2,6- bis- Picoline or 2- methylpyrazines and the molecule with carboxaldehyde radicals synthetic dyestuffs, root under the catalysis of para toluene sulfonamide or acetic anhydride Modification is further carried out according to needs.Also include first to being modified with the molecule of formaldehyde or other reactants, then be finally synthesizing Dyestuff.Same dyestuff is also not necessarily limited to a kind of synthetic method.It is any well known to a person skilled in the art chemical reaction and method all It can prepare and be not limited to but including people in the art with purifying the method for these compounds for carrying out synthesizing such compound The known method of member.

More specifically, the compounds of this invention is prepared by following general synthetic method：

The general synthesis step of the molecule 3：

Molecule 1 (being usually 2- methylquinolines) is mixed with equivalent molar ratio para toluene sulfonamide (TsNH2) and is dissolved in one In quantitative toluene, 120 DEG C are warming up to, electromagnetic agitation certain time (about 30 minutes), the molecule of equimolar ratio is added 2, reacted three to five days at 120 DEG C, with thin plate chromatography TLC monitoring reaction until molecule 1 reacts complete with molecule 2.Stop anti- It should heat and stir, be cooled to room temperature, place a period of time.For solid product, first carry out suction filtration and obtain solid, then use ethanol Or recrystallizing methanol, or directly purified with silica gel column chromatography (eluent substantially ethyl acetate: petroleum ether=1: 4-1: 2) product molecule 3, is obtained.It is dissolved in deuterated DMSO or deuterochloroform carrying out nucleus magnetic hydrogen spectrum sign, and carries out ESI mass spectrum tables Levy.

In one embodiment, the compounds of this invention is prepared by following general synthetic method：

The general synthesis step of the molecule 3 is：

Molecule 1 is mixed with the molecule 2 of equivalent molar ratio, add approximately twice as acetic anhydride, be warming up to backflow, reaction With thin plate chromatography TLC monitoring reaction until molecule 1 reacts complete with molecule 2.Vacuum spins off most of acetic anhydride and acetic acid, surplus Remaining reactant is added to the water, and is alkalized with 1M sodium hydroxides.Ethyl acetate or toluene are extracted three times, merge all organic Phase, is washed with saturated common salt, anhydrous sodium sulfate drying, and suction filtration is simultaneously spin-dried for organic solvent, methanol or ethyl alcohol recrystallization or straight Connect and purified with silica gel column chromatography, obtain molecule 3.It is dissolved in deuterated DMSO or deuterochloroform carrying out nucleus magnetic hydrogen spectrum sign, And carry out ESI mass spectral characteristis.

The general synthesis step of the molecule 3 is：

Molecule 1 is mixed with 4-butyl ammonium hydrogen sulfate, the sodium hydrate aqueous solution of 5 mol/Ls is added, is warming up to backflow, Add the molecule 2 with the equivalent molar ratio of molecule 1, back flow reaction.It is cooled to room temperature after the completion of reaction, suction filtration obtains crude product, then Recrystallization purification.

The general synthesis step of the molecule 3 is：

By molecule 1 (1mmol), molecule 2 (2mmol), trifluoromethanesulfonic acid calcium (5mol%) and tetrabutyl hexafluorophosphoric acid amine (2mol%) is dissolved in toluene, is heated to 130 degree of back flow reactions, with thin plate chromatography TLC monitoring reaction until molecule 1 and molecule 2 Reaction is complete.Stop reaction heating and stirring, be spin-dried for solvent.Crude product is dissolved in dichloromethane, is adsorbed in silica gel and passes through silicagel column Chromatography is purified.

In one embodiment, when the compound of synthesis carries hydroxyl, in order to protect hydroxyl in the reaction, two are added Hydrogen furans is protected, and compound is prepared by following general synthetic method：

The general synthesis step of the molecule 3 is：

Molecule 1 (1 equivalent), 3,4- dihydro -2H- pyrans (1.2 equivalent) and p-methyl benzenesulfonic acid (0.005 equivalent) are mixed It is dissolved in 10ml toluene, is warming up to 120 degree, molecule 2 (1 equivalent) and para toluene sulfonamide (1 equivalent) is added, at 120 DEG C Reaction one day, with thin plate chromatography TLC monitoring reaction until reaction is complete.Stop reaction, be cooled to room temperature, add 30ml water, 30ml tetrahydrofurans (THF) and 3ml acetic acid, place a period of time, separate out precipitation, and suction filtration obtains solid, and ethanol is washed, then uses ethanol Recrystallization, obtains target product.

In one embodiment, compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

1.44g 2- methyl-quinoxalines and 5mmol corresponding aldehyde are added and contain 0.215g methyl tricapryl ammonium chlorides In 70ml sodium hydroxide solutions (5M), heating stirring flows back 15 hours, cooling, precipitation filtering, and is washed with water, recrystallization or silicon Glue column chromatography purified product.

The general synthesis step of the molecule is：

1.3g guanidine hydrochlorides, 36ml sodium methoxides are added in 90ml absolute methanols, precipitation is filtered out, 1.5g is added into filtrate The pyridine of corresponding 2- Amino 3 cyanos -6- substitutions, is heated to reflux 18h, is cooled to room temperature, filter, precipitation is washed with methanol, Recrystallized in DMF.

The general synthesis step of the molecule is：

0.5mmol and 0.7mmol TES- alkene ether ketenes is dissolved in 2ml toluene, added in high-voltage tube, degasification 10min, adds 0.025mmol Pd (OAc) 2,0.05DtBPF, 0.7mmol CsF, 0.7mmol Bu3SnF, in argon atmosphere Under be heated to 120 DEG C of reaction 2-4h, be cooled to room temperature, add 10ml ether, gained slurries are filtered by diatomite, filtrate second Ether is extracted and washed with 1M sodium hydroxide solutions, concentration of organic layers, crude product purified by silica gel chromatography over CC (ethyl acetate: oil Ether=1: 18).

The general synthesis step of the molecule is：

1.88g4- methylpyrimidines and the corresponding aldehyde of 1.8mmol are added and contain 0.86g methyl tricapryl ammonium chlorides In 200ml sodium hydroxide solutions (5M), heating stirring backflow 2h, cooling, precipitation filtering, and be washed with water.Silica gel column chromatography is carried The purification of pure or means re-crystallization.

The general synthesis step of the molecule is：

Weigh s-triazine 1.8g (15mmol), corresponding aldehyde 2.23g (15mmol) and potassium hydroxide 0.34g and be dissolved in 60mL first In alcohol, return stirring reaction 14h. is filtrated to get faint yellow solid, with V (ethyl acetate): V (petroleum ether)=1: 2 mixture For eluant, eluent, pillar layer separation is carried out, target product is obtained.

The general synthesis step of the molecule is：

1.83ml 3- methyl pyridazine, the corresponding aldehyde of 4.07ml, 0.545g zinc chloride are mixed, stirred 2 hours at 150 DEG C. Room temperature is cooled to, adding sodium hydrate aqueous solution makes its split-phase, separates organic phase, uses saturated common salt water washing, anhydrous sodium sulfate Dry, be spin-dried for solvent, purify that (eluent is chloroform: methanol=20: 1) obtains target product with silica gel column chromatography

The general synthesis step of the molecule is：

Under argon atmosphere, 10mmol benzylalcohol, 10mmol triethylphosphine hydrogen are added into flame-dried microwave bottle Bromic acid, is sealed bottle with sealed membrane, and 8h is stirred at 100 DEG C.Diaphragm seal is removed, 4ml water and 11mol hydroxide sodium powder is added End, it is ultrasonic 5 minutes again after stirring 3 minutes.10mmol pyrrole-2-aldehyde is added, is shone after bottle capping at 100 DEG C in microwave 25min is penetrated, crude product uses anhydrous sodium sulfate drying, crude product purified by silica gel chromatography over CC after concentration after being extracted with dichloromethane (ethyl acetate: petroleum ether=1: 4)

The general synthesis step of the molecule is：

By 1.04gN, N- dimethyl thio ureas are dissolved in 50ml ethanol, then by 1.79g DMF two Dimethoxym ethane (DMF-DMA) is added thereto.Heating stirring 4h, rotates solvent, and surplus materials is dissolved in 50ml acetonitriles, added The corresponding Chinese cassia tree bromides of 15mmol, are heated to reflux 6h, are stirred for 30 minutes after adding 10ml triethylamines, are spin-dried for solvent, add 30ml water, (3 × 30mL) is extracted with dichloromethane, is associated with several layers of, anhydrous magnesium sulfate drying, crude product purified by silica gel column chromatography Method is purified.

The general synthesis step of the molecule is：

1.66g hydroxylamine hydrochlorides and 1.97ml pyridines and 1g A are added in 10ml ethanol, and flow back 6h, is cooled to room temperature, spins off Solvent, crude product imports and 1h is stirred in frozen water, precipitation filtering, dries, and recrystallized in the mixed liquor of methanol and dichloromethane Obtain target product.

The general synthesis step of the molecule is：

0.5g hydrazine hydrates, concentrated hydrochloric acid, 1.0g A are added in 10ml ethanol, flow back 6h, is cooled to room temperature, spins off solvent, Crude product imports and 1h is stirred in frozen water, precipitation filtering, dries, and be recrystallized to give mesh in the mixed liquor of methanol and dichloromethane Mark product.

It is can be seen that by above-mentioned synthetic method the present invention relates to the synthesis of a series of fluorescent dye compound simply, Most of dyestuff reacts a step with regard to that can obtain product by carbon-hydrogen bond activation, and yield is high, and post processing is simple, passes through recrystallization Or post excessively can obtain the dyestuff of high-purity.

Present invention below will be further described in conjunction with specific embodiments.

The synthesis (I-2025) of embodiment 1, (E) -2- (4- methoxyl-styrenes) quinoline

0.83g 2- methylquinolines, 1.00g para toluene sulfonamides (TsNH2) are mixed and are dissolved in 10ml toluene, is warming up to 120 DEG C, 0.79g P-methoxybenzal-dehyde is added, is reacted three days at 120 DEG C, with thin plate chromatography TLC monitoring reaction until anti- Should be complete.Stop reaction heating and stirring, be cooled to room temperature, suction filtration obtains solid, and ethanol is washed, then with ethyl alcohol recrystallization, obtains Target product is light yellow solid, yield 82%.

¹H NMR (400MHz, CDCl3) δ 8.12 (d, J=8.6Hz, 1H), 8.06 (d, J=8.5Hz, 1H), 7.85- 7.55 (m, 6H), 7.49 (t, J=7.5Hz, 1H), 7.32 (d, J=7.9Hz, 1H), 6.94 (d, J=8.7Hz, 2H), 3.86 (s, 3H)

The synthesis (I-2061) of embodiment 2, (E)-N- (4- (2- (quinoline -2- bases) vinyl) phenyl) acetamide

According to the identical process of embodiment 1, P-methoxybenzal-dehyde is simply replaced with to the acetylaminobenzene of equivalent Formaldehyde, obtained solid is washed with ethanol, then with ethyl alcohol recrystallization, thus obtains (E)-N- (4- (2- (quinoline -2- bases) vinyl) Phenyl) acetamide, light yellow solid, yield 89%.

¹H NMR (400MHz, CDCl3) δ 8.11 (d, J=8.6Hz, 1H), 8.07 (d, J=8.5Hz, 1H), 7.78 (d, J =8.0Hz, 1H), 7.73-7.45 (m, 8H), 7.33 (d, J=16.3Hz, 1H), 2.19 (s, 3H), 1.72 (s, 1H)

Embodiment 3, the program according to embodiment 1, simply replace with 2- methylquinolines 2, the 6- diformazans of equivalent respectively Base quinoline and the bromo- 2- methylquinolines of 6-, thus obtain (E)-N, N- dimethyl -4- (2- (6- methylquinoline -2- bases) vinyl) Aniline and (E) -4- (2- (6- bromoquinoline -2- bases) vinyl)-DMA.

(E)-N, N- dimethyl -4- (2- (6- methylquinoline -2- bases) vinyl) aniline (I-2063)

¹H NMR (300MHz, DMSO) δ 8.17 (d, J=8.6Hz, 1H), 7.83 (d, J=8.6Hz, 1H), 7.71 (dd, J =18.8,12.3Hz, 3H), 7.55 (d, J=8.6Hz, 3H), 7.17 (d, J=16.3Hz, 1H), 6.76 (d, J=8.8Hz, 2H), 2.97 (s, 6H), 2.48 (s, 3H)

Embodiment 4, (E) -4- (2- (6- bromoquinoline -2- bases) vinyl)-DMA (I-2069)

¹H NMR (400MHz, DMSO) δ 8.26 (d, J=8.7Hz, 1H), 8.19 (d, J=2.2Hz, 1H), 7.89-7.71 (m, 4H), 7.57 (d, J=8.8Hz, 2H), 7.19 (d, J=16.3Hz, 1H), 6.76 (d, J=8.9Hz, 2H), 2.95 (d, J =23.9Hz, 6H)

According to the program of embodiment 1, the various benzaldehyde-derivatives of equivalent will be simply replaced with to dimethylamino benzaldehyde Thing, obtains corresponding following compound.

Embodiment 5, (E) -4- (2- (quinoline -2- bases) vinyl) aniline (I-2062), yield 76%.

¹H NMR (400MHz, DMSO) δ 8.26 (d, J=8.6Hz, 1H), 7.91 (dd, J=13.5,8.2Hz, 2H), 7.77 (d, J=8.6Hz, 1H), 7.69 (dd, J=17.9,11.6Hz, 2H), 7.50 (t, J=7.4Hz, 1H), 7.42 (d, J= 8.4Hz, 2H), 7.13 (d, J=16.2Hz, 1H), 6.61 (d, J=8.4Hz, 2H), 5.54 (s, 2H)

The synthesis (I-2060) of embodiment 6, (E)-N, N- dimethyl -4- (2- (quinoline -2- bases) vinyl) aniline

¹H NMR (400MHz, CDCl3) δ 8.05 (dd, J=8.2,5.8Hz, 2H), 7.74 (d, J=7.9Hz, 1H), 7.71-7.57 (m, 3H), 7.54 (d, J=8.8Hz, 2H), 7.45 (t, J=7.4Hz, 1H), 7.22 (d, J=16.3Hz, 1H), 6.73 (d, J=8.8Hz, 2H), 3.01 (s, 6H)

Embodiment 7, (E) -2- (4- bromstyrols base) quinoline (I-2027), yield 94%.

¹H NMR (400MHz, CDCl₃) δ 8.14 (d, J=8.6Hz, 1H), 8.08 (d, J=8.5Hz, 1H), 7.79 (d, J =8.0Hz, 1H), 7.72 (t, J=7.7Hz, 1H), 7.68-7.60 (m, 2H), 7.56-7.47 (m, 5H), 7.38 (d, J= 16.3Hz, 1H)

Embodiment 8, (E) -2- (4- methyl styrenes base) quinoline (I-2028)

¹H NMR (400MHz, CDCl₃) δ 8.12 (d, J=8.6Hz, 1H), 8.07 (d, J=8.4Hz, 1H), 7.81-7.75 (m, 1H), 7.73-7.67 (m, 2H), 7.65 (d, J=7.7Hz, 1H), 7.54 (d, J=8.0Hz, 2H), 7.49 (dd, J= 11.0,3.9Hz, 1H), 7.37 (d, J=16.3Hz, 1H), 7.21 (d, J=7.9Hz, 2H), 2.39 (s, 3H)

Embodiment 9, (E)-methyl 4- (2- (quinoline -2- bases) vinyl) benzoic ether (I-2029), yield 79%.

¹H NMR (400MHz, acetone) δ 8.35 (d, J=8.5Hz, 1H), 8.07 (d, J=8.3Hz, 3H), 7.96 (d, J =5.0Hz, 1H), 7.93 (d, J=2.3Hz, 1H), 7.89 (dd, J=8.4,3.2Hz, 3H), 7.77 (dd, J=11.2, 4.1Hz, 1H), 7.64 (d, J=16.3Hz, 1H), 7.58 (t, J=7.5Hz, 1H), 3.93 (d, J=11.8Hz, 3H)

Embodiment 10, (E) -2,2 '-((4- (2- (quinoline -2- bases) vinyl) phenyl) azane diyl) diethanol (I- 2064), yield 81%.

¹H NMR (400MHz, DMSO) δ 8.26 (d, J=8.6Hz, 1H), 7.93 (d, J=8.3Hz, 1H), 7.89 (d, J =8.0Hz, 1H), 7.79 (d, J=8.7Hz, 1H), 7.75-7.66 (m, 2H), 7.51 (dd, J=15.8,8.0Hz, 3H), 7.17 (d, J=16.2Hz, 1H), 6.75 (d, J=8.8Hz, 2H), 4.82 (s, 2H), 3.58 (t, J=6.1Hz, 4H), 3.48 (t, J=6.1Hz, 4H)

Embodiment 11, the bromo- 4- of (E) -2,6- bis- (2- (quinoline -2- bases) vinyl) phenylacetic acid ester (I-2033), yield 86%.

¹H NMR (400MHz, CDCl₃) δ 8.17 (d, J=8.5Hz, 1H), 8.09 (d, J=8.4Hz, 1H), 7.81 (d, J =9.0Hz, 3H), 7.74 (dd, J=11.2,4.2Hz, 1H), 7.65-7.50 (m, 3H), 7.35 (d, J=16.2Hz, 1H), 2.42 (s, 3H)

Embodiment 12, (E) -2- (4- (amoxy) styryl) quinoline (I-2035), yield 94%.

¹H NMR (400MHz, CDCl₃) δ 8.12 (d, J=8.5Hz, 1H), 8.10-8.03 (m, 1H), 7.78 (d, J= 8.0Hz, 1H), 7.74-7.62 (m, 3H), 7.58 (d, J=8.6Hz, 2H), 7.50 (dd, J=13.3,6.0Hz, 1H), 7.31 (s, 1H), 6.93 (d, J=8.6Hz, 2H), 4.00 (t, J=6.6Hz, 2H), 1.86-1.74 (m, 2H), 1.51-1.35 (m, 4H), 0.94 (t, J=7.0Hz, 3H)

Embodiment 13, (E) -2- (4- (decyloxy) styryl) quinoline (I-2036), yield 93%.

¹H NMR (400MHz, CDCl₃) δ 8.11 (d, J=8.6Hz, 1H), 8.07 (d, J=8.4Hz, 1H), 7.78 (d, J =8.1Hz, 1H), 7.72-7.61 (m, 3H), 7.58 (d, J=8.7Hz, 2H), 7.48 (t, J=7.4Hz, 1H), 7.30 (d, J =7.1Hz, 1H), 6.93 (d, J=8.7Hz, 2H), 4.00 (dd, J=14.7,8.1Hz, 2H), 1.85-1.74 (m, 2H), (t, J=6.3Hz, the 3H) of 1.51-1.41 (m, 2H), 1.30 (d, J=18.5Hz, 12H), 0.89

Embodiment 14, (E) -2- (4- nitrostyrolenes base) quinoline (I-2044), yield 89%.

¹H NMR (400MHz, DMSO) δ 8.43 (d, J=8.6Hz, 1H), 8.29 (d, J=8.6Hz, 2H), 8.07-7.92 (m, 6H), 7.77 (dd, J=20.6,12.2Hz, 2H), 7.61 (t, J=7.4 Hz, 1H)

Embodiment 15, (E) -2- (4- (methylsulfonyl) styryl) quinoline (I-2045), yield 91%.

¹H NMR (400MHz, DMSO) δ 8.42 (d, J=8.6Hz, 1H), 8.07-7.89 (m, 8H), 7.79 (ddd, J= 8.4,6.9,1.4Hz, 1H), 7.71 (d, J=16.4Hz, 1H), 7.63-7.58 (m, 1H), 3.27 (s, 3H)

Embodiment 16, (E) -4- (2- (quinoline -2- bases) vinyl) benzonitrile (I-2046), yield 96%.

¹H NMR (400MHz, DMSO) δ 8.41 (d, J=8.5Hz, 1H), 8.05-7.86 (m, 8H), 7.78 (ddd, J= 8.4,6.9,1.4Hz, 1H), 7.69 (d, J=16.4Hz, 1H), 7.60 (ddd, J=8.0,6.9,1.1Hz, 1H)

Embodiment 17, N, N- dimethyl -4- ((1E, 3E) -4- (quinoline -2- bases) butyl- 1,3- diene -1- bases) aniline (I- 2067), yield 92%.

¹H NMR (400MHz, DMSO) δ 8.27 (d, J=8.6Hz, 1H), 7.91 (t, J=9.3Hz, 2H), 7.77-7.67 (m, 2H), 7.60 (dd, J=15.5,10.6Hz, 1H), 7.54-7.48 (m, 1H), 7.41 (d, J=8.8Hz, 2H), 6.98 (dd, J=15.3,10.7Hz, 1H), 6.86 (d, J=6.0Hz, 1H), 6.82 (d, J=6.1Hz, 1H), 6.72 (d, J= 8.9Hz, 2H), 2.09 (s, 6H)

Embodiment 18,2- ((1E, 3E) -4- (4- anisyls) butyl- 1,3- diene -1- bases) quinoline (I-2048), yield 81%.

¹H NMR (400MHz, DMSO) δ 8.30 (d, J=8.6Hz, 1H), 7.94 (dd, J=11.5,8.3Hz, 2H), 7.80-7.70 (m, 2H), 7.62 (dd, J=15.4,10.8Hz, 1H), 7.54 (dd, J=7.5,4.7Hz, 3H), 7.11 (dd, J =15.5,10.8Hz, 1H), 6.94 (dd, J=18.6,12.0Hz, 4H), 3.82-3.76 (m, 3H)

Embodiment 19, (E) -2- (4- chlorostyrenes base) quinoline (I-2049), yield 92%.

¹H NMR (400MHz, DMSO) δ 8.37 (d, J=8.6Hz, 1H), 8.00 (d, J=8.5Hz, 1H), 7.98-7.93 (m, 1H), 7.90-7.81 (m, 2H), 7.81-7.73 (m, 3H), 7.61-7.47 (m, 4H)

Embodiment 20, (E) -2- (2- (naphthalene -2- bases) vinyl) quinoline (I-2050), yield 91%.

¹H NMR (400MHz, DMSO) δ 8.39 (d, J=8.6Hz, 1H), 8.20 (s, 1H), 8.06-7.91 (m, 8H), 7.77 (ddd, J=8.4,6.9,1.4Hz, 1H), 7.65 (d, J=16.3Hz, 1H), 7.61-7.51 (m, 3H)

Embodiment 21, (E) -2- (2- (1H- indoles -5- bases) vinyl) quinoline (I-2051), yield 89%.

¹H NMR (400MHz, DMSO) δ 11.26 (s, 1H), 8.32 (d, J=8.6Hz, 1H), 8.01-7.84 (m, 5H), 7.73 (ddd, J=8.4,6.9,1.4Hz, 1H), 7.59-7.50 (m, 2H), 7.45 (d, J=8.5Hz, 1H), 7.38 (dd, J= 10.7,7.3Hz, 2H), 6.53-6.44 (m, 1H)

Embodiment 22, (E) -2- (methyl (4- (2- (quinoline -2- bases) vinyl) phenyl) amino) ethanol (I-2052), production Rate 88%.

¹H NMR (400MHz, DMSO) δ 8.27 (d, J=8.6Hz, 1H), 7.91 (dd, J=12.6,8.0Hz, 2H), 7.79 (d, J=8.7Hz, 1H), 7.76-7.68 (m, 2H), 7.58-7.46 (m, 3H), 7.18 (d, J=16.3Hz, 1H), 6.74 (d, J=8.9Hz, 2H), 4.73 (t, J=5.4Hz, 1H), 3.57 (q, J=6.0Hz, 2H), 3.45 (t, J=6.1Hz, 2H), 3.00 (s, 3H)

Embodiment 23, (E) -2- (2- (1H- indoles -4- bases) vinyl) quinoline (I-2053), yield 87%.

¹H NMR (400MHz, DMSO) δ 11.34 (s, 1H), 8.36 (d, J=8.6Hz, 1H), 8.19 (d, J=16.4Hz, 1H), 8.02 (dd, J=8.5,5.0Hz, 2H), 7.95 (d, J=8.0Hz, 1H), 7.75 (t, J=7.5Hz, 1H), 7.61- (s, the 1H) of 7.42 (m, 5H), 7.17 (t, J=7.7Hz, 1H), 6.97

Embodiment 24, (E) -2- (2- (pyrene -1- bases) vinyl) quinoline (I-2054), yield 96%.

¹H NMR (400MHz, DMSO) δ 8.98 (d, J=16.1Hz, 1H), 8.83 (d, J=9.4Hz, 1H), 8.66 (d, J =8.2Hz, 1H), 8.45 (d, J=8.6Hz, 1H), 8.40-8.31 (m, 4H), 8.25 (d, J=9.5Hz, 3H), 8.11 (dd, J =15.2,7.7Hz, 2H), 8.01 (d, J=7.6Hz, 1H), 7.84-7.76 (m, 2H), 7.64-7.57 (m, 1H)

The synthesis (I-2026) of embodiment 25, (E) -4- (2- (quinoline -2- bases) vinyl) phenol

By 0.78g parahydroxyben-zaldehydes (1 equivalent), 0.7 milliliter of 3,4- dihydro -2H- pyrans (1.2 equivalent) and 6.1mg pairs Toluene sulfonic acide (0.005 equivalent) mixing is dissolved in 10ml toluene, is warming up to 120 degree, is added 0.91g 2- methylquinolines (1 Equivalent) and 1.10g para toluene sulfonamides (1 equivalent), reacted one day at 120 DEG C, with thin plate chromatography TLC monitoring reaction until anti- Should be complete.Stop reaction, be cooled to room temperature, add 30ml water, 30ml tetrahydrofurans (THF) and 3ml acetic acid, separate out yellow and sink Form sediment, suction filtration obtains solid, and ethanol is washed, then with ethyl alcohol recrystallization, obtain target product for yellow solid, yield 76%.

¹H NMR (400MHz, acetone) δ 8.28 (d, J=8.6Hz, 1H), 8.02 (d, J=8.4Hz, 1H), 7.90 (d, J =8.1Hz, 1H), 7.85-7.76 (m, 2H), 7.76-7.70 (m, 1H), 7.60 (d, J=8.5Hz, 2H), 7.53 (dd, J= 11.0,4.0Hz, 1H), 7.31 (d, J=16.3Hz, 1H), 6.92 (d, J=8.6Hz, 2H)

The synthesis of embodiment 26, the bromo- 4- of (E) -2,6- bis- (2- (quinoline -2- bases) vinyl) phenol

By 2.00g 3,5- dibromine-4-hydroxy benzaldehydes are dissolved in 10ml acetic anhydrides, are heated to backflow, react two hours, Stop reaction, the acetic anhydride and acetic acid of residual are removed in rotation, then add 0.96ml 2- methylquinolines (1 equivalent) and 1.22g to toluene Sulfonamide (1 equivalent), is added in 10ml toluene, be warming up to 120 degree react three days, with thin plate chromatography TLC monitoring reaction until Reaction is complete.Stop reaction, be cooled to room temperature, be spin-dried for solvent, cross post, by obtained solid ethyl alcohol recrystallization, obtain product Q。

Take molecule Q 0.58g to be dissolved in 5ml ethanol, add sodium hydroxide 1.00g, 5ml water, heating reflux reaction is two small When, stop reaction, pH is adjusted to 7 with glacial acetic acid, obtain darkviolet precipitation, suction filtration goes out precipitation, washes, vacuum drying obtains product K。

(E)-bromo- 4- of 2,6- bis- (2- (quinoline -2- bases) vinyl) phenylacetic acid ester (I-2033), yield 76%.

(E)-bromo- 4- of 2,6- bis- (2- (quinoline -2- bases) vinyl) phenol (I-2031), yield 92%.

¹H NMR (400MHz, DMSO) δ 8.33 (d, J=8.6Hz, 1H), 7.95 (dd, J=9.8,6.6Hz, 4H), (d, J=16.2Hz, the 1H) of 7.80-7.65 (m, 3H), 7.55 (t, J=7.1Hz, 1H), 7.38

Embodiment 27, the program according to embodiment 5, simply replace with equivalent by 3,5- dibromine-4-hydroxy benzaldehydes The bromo- 4- hydroxy benzaldehydes of 3- or syringaldehyde or 3,4- 4-dihydroxy benzaldehyde or vanillic aldehyde or 3- hydroxyls -4-methoxybenzaldehyde, are obtained To corresponding following compound.

(E) the bromo- 4- of -2- (2- (quinoline -2- bases) vinyl) phenol (I-2032), yield 86%.

¹H NMR (400MHz, DMSO) δ 8.23 (d, J=8.6Hz, 1H), 7.89 (dd, J=12.3,8.1Hz, 2H), 7.70 (dt, J=8.6,7.8Hz, 3H), 7.62 (d, J=16.1Hz, 1H), 7.48 (t, J=7.4Hz, 1H), 7.36 (d, J= 8.5Hz, 1H), 7.03 (d, J=16.1Hz, 1H), 6.69 (d, J=8.0Hz, 1H)

Embodiment 28, (E) -2,6- dimethoxy-4 's-(2- (quinoline -2- bases) vinyl) phenol (I-2037), yield 93%.

¹H NMR (400MHz, DMSO) δ 8.74 (s, 1H), 8.32 (d, J=8.6Hz, 1H), 7.94 (t, J=8.8Hz, 2H), 7.82 (d, J=8.6Hz, 1H), 7.75 (dd, J=15.6,5.5Hz, 2H), 7.54 (t, J=7.1Hz, 1H), 7.37 (d, J=16.3Hz, 1H), 7.05 (s, 2H), 3.85 (s, 6H)

Embodiment 29, (E) -4- (2- (quinoline -2- bases) vinyl) benzene -1,2- diphenol (I-2039), yield 84%.

¹HNMR (400MHz, DMSO) δ 9.34 (s, 1H), 9.10 (s, 1H), 8.30 (d, J=8.6Hz, 1H), 7.94 (dd, J=15.2,8.1Hz, 2H), 7.83 (d, J=8.6Hz, 1H), 7.76-7.63 (m, 2H), 7.53 (t, J=7.4Hz, 1H), (d, J=8.1Hz, the 1H) of 7.14 (dd, J=12.4,9.1Hz, 2H), 7.02 (dd, J=8.2,1.8Hz, 1H), 6.79

Embodiment 30, (E) -2- methoxyl groups -4- (2- (quinoline -2- bases) vinyl) phenylacetic acid ester (I-2041), yield 89%.

¹H NMR (300MHz, DMSO) δ 8.37 (d, J=8.6Hz, 1H), 8.03-7.93 (m, 2H), 7.86 (dd, J= 12.5,10.1Hz, 2H), 7.80-7.72 (m, 1H), 7.61-7.49 (m, 3H), 7.32 (dd, J=8.2,1.7Hz, 1H), 7.14 (d, J=8.1Hz, 1H), 3.89 (s, 3H), 2.28 (s, 3H)

Embodiment 31, (E) -2- methoxyl groups -4- (2- (quinoline -2- bases) vinyl) phenol (I-2040), yield 86%.

¹H NMR (400MHz, DMSO) δ 8.31 (d, J=8.6Hz, 1H), 7.94 (dd, J=12.8,8.2Hz, 2H), 7.82 (d, J=8.6Hz, 1H), 7.78-7.69 (m, 2H), 7.53 (t, J=7.0Hz, 1H), 7.33 (dd, J=14.3, 9.0Hz, 2H), 7.13 (dd, J=8.2,1.7Hz, 1H), 6.83 (d, J=8.1Hz, 1H), 3.87 (s, 3H)

Embodiment 32, (E) -2- methoxyl groups -5- (2- (quinoline -2- bases) vinyl) phenylacetic acid ester (I-2042), yield 85%.

¹H NMR (400MHz, DMSO) δ 8.37 (d, J=8.6Hz, 1H), 7.98 (dd, J=16.3,7.9Hz, 2H), 7.91-7.81 (m, 2H), 7.76 (ddd, J=8.4,6.9,1.4Hz, 1H), 7.61-7.50 (m, 3H), 7.32 (dd, J=8.2, 1.8Hz, 1H), 7.16 (dd, J=7.6,4.3Hz, 1H), 3.89 (s, 3H), 2.29 (d, J=6.8Hz, 3H)

Embodiment 33, (E) -2- methoxyl groups -5- (2- (quinoline -2- bases) vinyl) phenol (I-2043), yield 79%.

¹H NMR (400MHz, DMSO) δ 9.37 (s, 1H), 8.31 (d, J=8.6Hz, 1H), 7.94 (dd, J=12.5, 7.9Hz, 2H), 7.82 (d, J=8.6Hz, 1H), 7.73 (ddd, J=8.4,5.9,3.2Hz, 2H), 7.56-7.50 (m, 1H), 7.34 (dd, J=14.6,9.1Hz, 2H), 7.14 (dd, J=8.2,1.9Hz, 1H), 6.82 (d, J=8.1Hz, 1H), 3.87 (s, 3H)

Embodiment 34, according to embodiment 5 Part II similar program, 0.5g molecules H is dissolved in 5ml ethanol, then Sodium hydroxide 1.00g, 5ml water is added, two hours of heating reflux reaction, stops reaction, adjusts pH to 7 with glacial acetic acid, is sunk Form sediment, suction filtration goes out precipitation, wash, vacuum drying obtains product I.

(E) -4- (2- (quinoline -2- bases) vinyl) benzoic acid (I-2030), yield 75%.

¹H NMR (400MHz, DMSO) δ 13.03 (s, 1H), 8.40 (d, J=8.5Hz, 1H), 8.05-7.84 (m, 8H), 7.77 (t, J=7.3Hz, 1H), 7.61 (dd, J=21.4,11.9Hz, 2H)

The synthesis (I-2059) of embodiment 35, (E) -2- (4- (azetidine -1- bases) styryl) quinoline

By 1.24g 4-Fluorobenzaldehydes (0.01mol), 0.57g heterocyclics butane (0.01mol) and 1.38g potassium carbonate (0.01mol) is well mixed, and adds 10ml dimethyl sulfoxide (DMSO)s, and ultrasound 30 minutes, pour into 400ml water, use 100ml second under water-bath Ether is extracted three times, merges ether phase, and saturated common salt washing, anhydrous sodium sulfate drying is spin-dried for, input next step reaction.

According to the program of embodiment 1, simply by dimethylamino benzaldehyde is replaced with equivalent to azete piperidinyl benzene first Aldehyde, obtains (E) -2- (4- (azetidine -1- bases) styryl) quinoline, yield 54%.

¹H NMR (400MHz, CD₃OD_SPE) δ 8.23 (d, J=8.6Hz, 1H), 7.95 (d, J=8.5Hz, 1H), 7.85 (d, J=8.5Hz, 2H), 7.71 (t, J=7.2Hz, 1H), 7.63 (d, J=16.3Hz, 1H), 7.51 (t, J=7.9Hz, 3H), 7.18 (d, J=16.4Hz, 1H), 6.48 (d, J=8.4Hz, 2H), 3.93 (t, J=7.2Hz, 4H), 2.46-2.33 (m, 2H).

Embodiment 36,2,2 '-((4- ((1E, 3E) -4- (quinoline -2- bases) butyl- 1,3- diene -1- bases) phenyl) azane two Base) diethanol synthesis (I-2068)

0.89g 4- (two (2- hydroxyethyls) amino) benzaldehyde is dissolved in 10ml tetrahydrofurans, 2.58g (1,3- bis- is added Oxygen cyclopenta -2- methyl) triphenylphosphinebromide, 0.24g sodium hydrides and the ether of 0.13g 18- crown-s 6.React 30 minutes, rise under ice bath To room temperature reaction 24 hours, add 4ml concentrated hydrochloric acids and be quenched, potassium carbonate is neutralized, ethyl acetate extraction, sodium sulphate is dried, was spin-dried for Column chromatography.

According to the program of embodiment 1, (E) -3- (4- (two (2- hydroxyls of equivalent are simply replaced with to dimethylamino benzaldehyde Base ethyl) amino) phenyl) acryloyl group aldehyde, reacts and obtains 2,2 '-((4- ((1E, 3E) -4- (quinoline -2- bases) butyl- 1,3- bis- Alkene -1- bases) phenyl) azane diyl) diethanol, yield 33%.

¹H NMR (400MHz, DMSO) δ 8.26 (d, J=8.6Hz, 1H), 7.91 (dd, J=13.8,8.3Hz, 2H), 7.78 (d, J=8.6Hz, 1H), 7.71 (dd, J=16.2,8.3Hz, 2H), 7.51 (dd, J=16.3,7.9Hz, 3H), 7.17 (d, J=16.2Hz, 1H), 6.72 (dd, J=17.1,8.7Hz, 2H), 4.80 (t, J=5.1Hz, 2H), 3.57 (dd, J= 11.6,5.8Hz, 4H), 3.48 (t, J=6.2Hz, 4H)

The synthesis (I-2003) of embodiment 37, (E) -1,2- bis- (quinoline -2- bases) ethene

4ml acetic anhydrides will be added in 0.78g 2- formaldehyde quinoline and 0.71g 2- methylquinolines, 150 DEG C of back flow reactions 24 are small When, remaining acetic acid acid anhydride and acetic acid are removed in rotation, add 30ml water, adjust neutral with sodium hydroxide, ethyl acetate extraction, organic phase saturation Brine It, anhydrous sodium sulfate drying crosses post, obtains (E) -1,2- bis- (quinoline -2- bases) ethene, yield 51% after being spin-dried for.

¹H NMR (200MHz, CDCl₃)-δppm：7.51 (dd, J=7.6,7.2Hz, 2H, H-3), 7.71 (dd, J=7.9, 7.1Hz, 2H, H-7), 7.81 (d, J=9.6,4H, H-3,5), 7.87 (s, 2H, H-1 '), 8.08 (d, J=8.4Hz, 2H, H- 8), 8.19 (d, J=8.6Hz, 2H, H-4)；

The synthesis (II-2070) of embodiment 38, (E)-N, N- dimethyl -4- (2- (pyrazine -2- bases) vinyl) aniline

2- methylpyrazines 0.47g (5mmol) is mixed with 0.17g 4-butyl ammonium hydrogen sulfates (0.5mmol), 5 moles are added Every liter of sodium hydrate aqueous solution 25ml, is warming up to backflow, adds 5mmol to dimethylamino benzaldehyde, back flow reaction.React It is cooled to room temperature after, suction filtration obtains crude product, then obtains product, yield 56% with ethyl alcohol recrystallization purification.

¹H NMR (400MHz, CDCl₃) δ 8.81 (s, 1H), 7.84 (d, J=8.1Hz, 2H), 7.76 (d, J=8.5Hz, 2H), (s, the 6H) of 7.29 (d, J=8.0Hz, 2H), 6.66 (d, J=8.8Hz, 2H), 3.09

Embodiment 39,4,4 '-((1E, 1 ' E)-pyridine -2,6- diyl two (ethene -2,1- diyl)) two (N, N- dimethyl Aniline) synthesis (II-2071)

By 2,6- lutidines (1mmol), to dimethylamino benzaldehyde (2mmol), trifluoromethanesulfonic acid calcium (5mol%) It is dissolved in tetrabutyl hexafluorophosphoric acid amine (2mol%) in toluene, is heated to 130 degree of back flow reactions, monitors anti-with thin plate chromatography TLC With molecule 2 it should react complete up to molecule 1.Stop reaction heating and stirring, be spin-dried for solvent.Crude product is dissolved in dichloromethane, inhales Invest silica gel to purify by silica gel column chromatography, yield 51%.

¹H NMR(DMSO-d₆)：δ 3.02 (12H, s), 6.78 (4H, d, J=8.8Hz), 7.32 (2H, d,³J= 16.0Hz), 7.65-7.71 (6H, m), 8.08 (2H, d, J=8.1Hz), 8.00 (1H, t J=8.1Hz)

The synthesis (I-2098) of embodiment 40, (E) -4- (2- (isoquinolyl-1) vinyl)-DMA

According to the program of embodiment 1,2- methylquinolines are simply replaced with to the methylisoquinolinium of equivalent, (E) -4- is obtained (2- (isoquinolyl-1) vinyl)-DMA, yield 83%.

¹H NMR (400MHz, CDCl₃) δ 8.52 (d, J=5.6Hz, 1H), 8.39 (d, J=8.4Hz, 1H), 7.97 (d, J =15.4Hz, 1H), 7.86-7.77 (m, 2H), 7.67 (t, J=7.4Hz, 1H), 7.62 (d, J=8.5Hz, 3H), 7.50 (d, J =5.6Hz, 1H), 6.75 (d, J=8.6Hz, 2H), 3.03 (s, 6H)

Embodiment 41, the program according to embodiment 11, simply will replace with 4- (two (2- hydroxyls to dimethylamino benzaldehyde Ethyl) amino) benzaldehyde, obtain (E) -2,2 '-((4- (2- (isoquinolyl-1) vinyl) phenyl) azane diyl) diethanols (I-2099), yield 80%.

¹H NMR (400MHz, DMSO) δ 8.65 (d, J=8.5Hz, 1H), 8.46 (d, J=5.5Hz, 1H), 8.00-7.85 (m, 3H), 7.76 (t, J=7.5Hz, 1H), 7.71-7.60 (m, 4H), 6.75 (d, J=8.4Hz, 2H), 4.81 (t, J= 5.3Hz, 2H), 3.67-3.53 (m, 4H), 3.49 (d, J=5.9Hz, 4H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 42, (E) -2- styryls quinoxaline (I-2100)

Silica gel column chromatography purification (eluent substantially ethyl acetate: petroleum ether=1: 50), it is white to obtain target product Solid.¹H NMR (400MHz, CDCl3) δ 9.03 (s, 1H), 8.07 (d, J=8.0Hz, 2H), 7.87 (d, J=16.0Hz, 1H), 7.77-7.69 (m, 2H), 7.66 (d, J=7.2Hz, 2H), 7.44-7.34 (m, 4H)

Embodiment 43, (E) -2- [2- (4- methyl mercaptos phenyl)-vinyl]-quinoxaline (I-2101)

Silica gel column chromatography purification (eluent substantially dichloromethane: ethyl acetate=8: it is pale red 2) to obtain target product Color solid, yield 24%.¹H NMR (500MHz, CDCl3) δ 2.53 (s, 3H), 7.27 (d, 2H, J=8Hz), 7.33 (d, 1H, J =16Hz), 7.58 (d, 2H, J=8Hz), 7.72-7.68 (m, 1H), 7.77-7.74 (m, 1H), 7.83 (d, 1H, J=16Hz), 8.08-8.05 (m, 2H), 9.02 (s, 1H)

Embodiment 44, (E) -2- [2- (the chloro- phenyl of 4-)-vinyl]-quinoxaline (I-2102)

Silica gel column chromatography purification (eluent substantially dichloromethane: ethyl acetate=8: it is pale red 2) to obtain target product Color solid, yield 56%.¹H NMR (500MHz, CDCl3) δ 7.35 (d, 1H, J=16Hz), 7.40 (d, 2H, J=8Hz), 7.60 (d, 2H, J=8Hz), 7.74-7.71 (m, 1H), 7.79-7.76 (m, 1H), 7.84 (d, 1H, J=16Hz), 8.10- 8.06 (m, 2H), 9.03 (s, 1H)

Embodiment 45,2- [2- (4-N, N- dimethyl-amino-phenyl)-vinyl]-quinoxaline (I-2103)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for light yellow solid, yield 78%.¹H NMR (500MHz, CDCl3) δ 3.04 (s, 6H), 6.74 (d, 2H, J=8Hz), 7.18 (d, 1H, J=16Hz), 7.56 (d, 2H, J =8Hz), 7.67-7.63 (m, 1H), 7.74-7.71 (m, 1H), 7.81 (d, 1H, J=16Hz), 8.04-8.01 (m, 2H), 9.01 (s, 1H)

Embodiment 46,2- [2- (4- piperidin-1-yls-phenyl)-vinyl]-quinoxaline (I-2104)

Silica gel column chromatography purification (eluent substantially ethyl acetate: petroleum ether=5: 5) obtains target product solid for yellow Body, yield 50%.¹H NMR (500MHz, CDCl3) δ 1.64-1.60 (m, 2H), 1.71-1.70 (m, 4H), 3.30-3.28 (m, 4H), 6.93 (d, 2H, J=8Hz), 7.20 (d, 1H, J=16Hz), 7.55 (d, 2H, J=8Hz), 7.68-7.65 (m, 1H), 7.75-7.71 (m, 1H), 7.80 (d, 1H, J=16Hz), 8.05-8.02 (m, 2H), 9.01 (s, 1H)

Embodiment 47,2- [2- (4-N, N- diphenyl amino-phenyl)-vinyl]-quinoxaline (I-2105)

Silica gel column chromatography purification (eluent substantially ethyl acetate: petroleum ether=6: 2) obtains target product for crocus Solid, yield 45%.¹H NMR (500MHz, CDCl3) δ 7.11-7.07 (m, 4H), 7.15 (dd, 4H, J1=8.5Hz, J2= 1.5Hz), 7.27 (d, 1H, J=16Hz), 7.30 (dd, 4H, J1=8Hz, J2=8.5Hz), 7.52 (d, 2H, J=8Hz), 7.70-7.67 (m, 1H), 7.76-7.73 (m, 1H), 7.81 (d, 1H, J=16Hz), 8.05-8.03 (m, 2H), 9.02 (s, 1H)。

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 48,2.4 diaminourea -7- styryls pteridine (I-2106)

Yield 89%.Relative molecular weight：264mp(from DMF)303℃.

Embodiment 49,2,4- diaminourea -7- (3,4- dichlorostyrene base) pteridine (I-2107)

Yield 79%.Relative molecular weight：333mp(from DMF)338℃.

Embodiment 50,2,4- diaminourea -7- (3,4- methylene-dioxy styryl) pteridine (I-2108)

Yield 91%.Relative molecular weight：308.mp(from DMF)335℃.

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 51, (E) -- 2- styryl -1H- indoles (IV-2042)

White solid, yield：72%.¹H NMR (400MHz, CDCl3)：δ 8.23 (bs, 1H), 7.57 (d, J=7.8Hz, 1H), 7.49 (d, J=7.8Hz, 2H), 7.42-7.30 (m, 3H), 7.26 (d, J=7.4Hz, 1H), 7.18 (t, J=7.48Hz, 1H), 7.14-7.04 (m, 2H), 6.89 (d, J=16.0Hz, 1H), 6.60 (s, 1H).

Embodiment 52, (E) -- 6- methoxyl group -2- styryl -1H- indoles (IV-2043)

White solid, yield：74%.¹H NMR (400MHz, CDCl3)：δ 8.12 (bs, 1H), 7.55-7.40 (m, 3H), 7.34 (t, J=7.8Hz, 2H), 7.27-7.23 (m, 2H), 7.06 (d, J=16.5Hz, 1H), 6.86-6.80 (m, 1H), 6.79-6.72 (m, 1H), 6.52 (s, 1H), 3.85 (s, 3H).

Embodiment 53, (E) -6- (tert-butyl group) -2- styryl -1H- indoles (IV-2044)

White solid, yield：73%.¹H NMR (400MHz, CDCl3)：δ 8.15 (bs, 1H), 7.54-7.43 (m, 3H), 7.40-7.31 (m, 3H), 7.27-7.21 (m, 1H), 7.17 (dd, J=8.3Hz, 1.5Hz, 1H), 7.10 (d, J=16.5Hz, 1H), 6.84 (d, J=16.5Hz, 1H), 6.55 (s, 1H), 1.37 (s, 9H).

Embodiment 54, (E) -2- styryls -6- (trifluoromethyl) -1H- indoles (IV-2045)

White solid, yield：68%.¹H NMR (400MHz, CDCl3)：δ 8.42 (bs, 1H), 7.72-7.57 (m, 2H), 7.50 (d, J=7.5Hz, 2H), 7.44-7.26 (m, 4H), 7.11 (d, J=16.5Hz, 1H), 6.98 (d, J=16.5Hz, 1H), 6.65 (s, 1H).

Embodiment 55, (E) -5- nitro -2- styryl -1H- indoles (IV-2046)

Crocus solid, yield：68%.¹H NMR (400MHz, CDCl3)：δ 8.76 (bs, 1H), 8.30 (s, 1H), 8.00 (dt, J=8.7Hz, 1.7Hz, 1H), 7.52 (d, J=7.8Hz, 2H), 7.45-7.37 (m, 2H), 7.33-7.30 (m, 1H), 7.25 (d, J=1.4Hz, 1H), 7.10 (s, 2H), 6.70 (s, 1H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 56,4- [2- (the chloro- phenyl of 4-)-vinyl]-pyrimidine (II-2072)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for pale red solid, yield 77%.¹H NMR (500MHz, CDCl3) δ 7.00 (d, 1H, J=16Hz), 7.28 (d, 1H, J=5Hz), 7.35 (d, 2H, J=8Hz), 7.50 (d, 2H, J=8Hz), 7.83 (d, 1H, J=16Hz), 8.66 (d, 1H, J=5Hz), 9.15 (s, 1H)

Embodiment 57,4- [2- (the bromo- phenyl of 4-)-vinyl]-pyrimidine (II-2073)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for pale red solid, yield 78%.¹H NMR (500MHz, CDCl3) δ 7.02 (d, 1H, J=16Hz), 7.28 (d, 1H, J=5Hz), 7.44 (d, 2H, J=8Hz), 7.52 (d, 2H, J=8Hz), 7.81 (d, 1H, J=16Hz), 8.67 (d, 1H, J=5Hz), 9.16 (s, 1H)

Embodiment 58,4- [2- (4- methyl mercaptos phenyl)-vinyl]-pyrimidine (II-2074)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for cream solid, yield 59%.¹H NMR (500MHz, CDCl3) δ 2.51 (s, 3H), 7.00 (d, 1H, J=16Hz), 7.26 (d, 1H, J=5Hz), 7.27 (d, 2H, J =8Hz), 7.50 (d, 2H, J=8Hz), 7.83 (d, 1H, J=16Hz), 8.65 (d, 1H, J=5Hz), 9.14 (s, 1H)

Embodiment 59,4- [2- (4-N, N- dimethyl-amino-phenyl)-vinyl]-pyrimidine (II-2075)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for yellowish red color solid, yield 78%.¹H NMR (500MHz, CDCl3) δ 3.03 (s, 6H), 6.71 (d, 2H, J=8Hz), 6.84 (d, 1H, J=16Hz), 7.24 (d, 1H, J =5Hz), 7.50 (d, 2H, J=8Hz), 7.81 (d, 1H, J=16Hz), 8.58 (d, 1H, J=5Hz), 9.09 (s, 1H)

Embodiment 60,4- [2- (4- piperidin-1-yls-phenyl)-vinyl]-pyrimidine (II-2076)

Target product is recrystallized to give in the mixed liquor of dichloromethane and heptane for yellowish red color solid, yield 67%.¹H NMR (500MHz, CDCl3) δ 1.64-1.62 (m, 2H), 1.72-1.67 (m, 4H), 3.29-3.27 (m, 4H), 6.87 (d, 1H, J =16Hz), 6.90 (d, 2H, J=8 Hz), 7.27 (d, 1H, J=5Hz), 7.49 (d, 2H, J=8Hz), 7.80 (d, 1H, J= 16Hz), 8.59 (d, 1H, J=5Hz), 9.10 (s, 1H)

Embodiment 61,4- [2- (4-N, N- diphenyl amino-phenyl)-vinyl]-pyrimidine (II-2077)

Silica gel column chromatography purification (eluent substantially ethyl acetate: dichloromethane=3: 7) obtains target product for yellow Solid, yield 46%.¹H NMR (500MHz, CDCl3) δ 6.91 (d, 1H, J=16Hz), 7.04 (d, 2H, J=8Hz), 7.08 (t, 2H, J=7Hz), 7.13 (d, 4H, J=7.5Hz), 7.30-7.26 (m, 5H), 7.45 (d, 2H, J=8Hz), 7.82 (d, 1H, J=16Hz), 8.62 (d, 1H, J=5Hz), 9.13 (s, 1H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 62,2,4- dimethyl -6- (4 '-N, N- dimethylaminostyryl) -1,3,5- s-triazine (II-2078)

Faint yellow solid, yield 36%.¹H NMR (CDCl3,600MHz) δ：8.16 (d, J=15.6Hz, 1H), 7.55 (d, J=8.40Hz, 2H), 6.86 (d, J=15.6Hz, 1H), 6.72 (d, J=8.40Hz, 2H), 3.04 (s, 6H), 2.62 (s, 6H)

Embodiment 63,2- styryl -4,6- dimethyl -1,3,5- s-triazine (II-2079)

White solid, yield 35%.¹H NMR (CDCl3,600MHz) δ：8.21 (d, J=15.6Hz, 1H), 7.64 (d, J =7.2Hz, 2H), 7.38~7.43 (m, 3H), 7.07 (d, J=15.6Hz, 1H), 2.65 (s, 6H)

Embodiment 64,2,4- dimethyl -6- (pN, N- dimethylaminostyryl)-s- triazines (II-2080)

Yellow solid, yield：59%.¹H NMR(CDCl3)δ：8.13 (d, 1H, J=15.60Hz), 7.50 (d, 2H, J= 9.26Hz), 6.80 (d, 1H, J=15.60Hz), 6.65 (d, 2H, J=9.26Hz), 3.41 (q, 4H, J=6.99Hz), 2.59 (s, 6H), 1.20 (t, 6H, 6.83Hz)

Embodiment 65,2,4- dimethyl -6- (m hydroxy styrenes base)-s- triazines (II-2081)

Yellow solid, yield：30%.¹H NMR (400MHz, DMSO)：8.47 (d, J=16Hz, 1H), 7.70 (d, J= 7.6,1H), 7.28 (dd, J=7.6, J=7.2,1H), 7.19 (d, J=16,1H), 6.87 (dd, J=7.6, J=7.2,1H), 2.56 (s, 6H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 66,3- [2- (4-N, N- dimethyl-amino-phenyl)-vinyl]-pyridazine (II-2082)

Yield 49%.¹H NMR (500MHz, CDCl3) δ 3.02 (s, 6H), 6.72 (d, 2H, J=8Hz), 7.15 (d, 1H, J=16Hz), 7.37 (dd, 1H, J1=8Hz, J2=5Hz), 7.50 (d, 2H, J=8Hz), 7.57 (dd, 1H, J1=8Hz, J2 =2Hz), 7.60 (d, 1H, J=16Hz), 8.97 (dd, 1H, J1=5Hz, J2=2Hz)

Embodiment 67,3- (2- phenyl vinyls) pyridazine (II-2083)

Yield：81%.¹H NMR (400MHz, CDCl3) d 7.32-7.47 (5H, m), 7.59-7.73 (4H, m), 9.06 (1H, dd, J=4.8,1.6Hz)

Embodiment 68,1.3- (2- p-methylphenyls-vinyl) pyridazine (II-2084)

Yield：60%.¹H NMR (CDCl3) d 9.03 (dd, 1H, J 1Z 4.8Hz and J 2Z1.6Hz), 7.72- 7.58 (m, 2H), 7.51 (s, 1H), 7.48 (s, 1H), 7.44 (dd, 1H, J 1Z 8.6Hz, J 2Z 4.9Hz), 7.31 (d, 1H, J Z 10.7Hz), 7.19 (s, 1H), 7.14 (s, 1H), 2.38 (s, 3H)

Embodiment 69,3- (2- biphenyl -4- bases-phenyl) pyridazine (II-2085)

Yield 50%.¹H NMR (CDCl3) d 9.05 (dd, 1H, J 1Z 4.8Hz and J 2Z 1.5Hz), 7.74 (d, 1H, J Z 16.5Hz), 7.61-7.66 (m, 7H), 7.36-7.49 (m, 4H), 7.13 (d, 1H, J Z 16.5Hz)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 70, (E-2- phenyl vinyls -1 (H)-pyrroles (III-2069)

Yield：98%.¹H NMR (200MHz, CDCl3)：D 6.33 (s, 1H), 6.43 (s, 1H), 6.69 (d, JHH= 16.5Hz, 1H), 6.83 (s, 1H), 7.02 (d, JHH=16.5Hz, 1H), 7.38 (m, 5H), 8.31 (s, 1H)

Embodiment 71, (E) -2- (4 '-methoxyphenyl) vinyl -1- (H)-pyrroles (III-2070)

Yield：83%.¹H NMR (200MHz, CDCl3)：D 3.82 (s, 3H), 6.26 (m, 2H), 6.62 (d, JHH= 16.4Hz, 1H), 6.85 (m, 4H), 7.34 (m, 2H), 8.3 (s, 1H)

Embodiment 72, (E) -2- (4 '-aminomethyl phenyl) vinyl -1- (H)-pyrroles (III-2071)

Yield：93%.¹H NMR (200MHz, CDCl3)：D 2.36 (s, 3H), 6.26 (t, JHH=5.6Hz, 1H), 6.35 (m, 1H), 6.65 (d, JHH=16.4Hz, 1H), 6.82 (m, 1H), 6.95 (d, JHH=16.4Hz, 1H), 7.15 (d, JHH=8.0Hz, 2H), 7.34 (d, JHH=8.2Hz, 2H), 8.34 (s, 1H)

Embodiment 73, (E) -2- (2 '-bromophenyl) vinyl -1- (H)-pyrroles (III-2072)

Yield：92%.¹H NMR (CDCl3,600MHz)：D 6.26 (d, JHH=6.0Hz, 1H), 6.39 (s, 1H), 6.86 (s, 1H), 6.93 (d, JHH=16.2Hz, 1H), 6.99 (d, JHH=16.8Hz, 1H), 7.07 (t, JHH=7.2Hz, 1H), 7.28 (t, JHH=7.8Hz, 1H), 7.56 (d, JHH=8.4Hz, 1H), 7.60 (d, JHH=7.8Hz, 1H), 8.44 (s, 1H)

Embodiment 74, (E) -2- (2 '-fluorophenyl) vinyl -1- (H)-pyrroles (III-2073)

Yield：95%.¹H NMR (CDCl3,200MHz)：D 6.29 (d, JHH=8.4Hz, 1H), 6.77 (m, 1H), 6.82 (d, JHH=16.6Hz, 2H), 7.16 (m, 4H), 7.54 (t, JHH=7.8Hz, 1H), 8.43 (s, 1H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

By 1.04gN, N- dimethyl thio ureas are dissolved in 50ml ethanol, then by 1.79g DMF two Dimethoxym ethane (DMF-DMA) is added thereto.Heating stirring 4h, rotates solvent, and surplus materials is dissolved in 50ml acetonitriles, added The corresponding Chinese cassia tree bromides of 15mmol, are heated to reflux 6h, are stirred for 30 minutes after adding 10ml triethylamines, are spin-dried for solvent, add 30ml water, (3 × 30mL) is extracted with dichloromethane, is associated with several layers of, anhydrous magnesium sulfate drying, crude product purified by silica gel column chromatography Purification.

Embodiment 75,2- dimethyl -5- [(E) -2- phenyl vinyls] thiazole (III-2074)

Eluent：Petroleum ether: ethyl acetate=1: 1, yield 50%.¹H NMR (CDCl3,300.10MHz)：δ=3.13 (s, 6H, NMe2), 6.47 (d, J=15.9Hz, 1H), 7.08 (d, J=15.9Hz, 1H), 7.16 (s, 1H, H4), 7.17-7.21 (m, 1H), 7.27-7.33 (m, 2H), 7.37-7.40 (m, 2H) ppm

Embodiment 76,2- dimethyl -5- [(E) -2- (4- nitrobenzophenones) vinyl] thiazole (III-2075)

Eluent：Petroleum ether: ethyl acetate=1: 1, yield 80%.¹H NMR (CDCl3,400.91MHz)：δ=3.17 (s, 6H, NMe2), 6.45 (d, J=15.8Hz, 1H), 7.24-7.28 (m, 2H), 7.48 (d, J=8.8Hz, 2H), 8.16 (d, J =8.8Hz, 2H) ppm.

Embodiment 77,2- dimethyl -5- [(E) -2- (4- methoxyphenyls) vinyl] thiazole (III-2076)

Eluent：Petroleum ether: ethyl acetate=1: 1, yield 13%.¹H NMR (CDCl3,400.91MHz)：δ=3.13 (s, 6H, NMe2), 3.81 (s, 3H, OMe), 6.44 (d, J=15.9Hz, 1H), 6.86 (d, J=8.8Hz, 2H), 6.94 (d, J =15.9Hz, 1H), 7.12 (s, 1H, H4), 7.32 (d, J=8.8Hz, 2H) ppm

Embodiment 78, (E) -5- (4- chlorostyrenes base) -4- methylthiazols (III-2077)

Eluent：Petroleum ether: ethyl acetate=1: 1, yield 43%..¹H NMR (400MHz, CDCl3) δ 8.57 (s, 1H), 7.39 (d, J=8.5Hz, 2H), 7.31 (d, J=8.5Hz, 2H), 7.14 (d, J=15.9Hz, 1H), 6.75 (d, J= 16.0Hz, 1H), 2.54 (s, 3H).

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 79, (E) -4- (2- (methyl isoxazole -5- bases) vinyl) phenol (III-2078)

Light tan solid, yield：61%.¹H NMR (400MHz, CDCl3) d：7.42 (m, 2H), 7.39 (d, 1H, J= 16Hz), 6.84 (d, 1H, J=16Hz), 6.86 (m, 2H), 6.0 (s, 1H), 5.6 (s, 1H), 2.3 (s, 3H)

Embodiment 80, (E) -5- (3,4- dimethoxy-styryl) -3- methyl-isoxazoles (III-2079)

Half white solid, yield 61%.¹H NMR (400MHz, CDCl3) d：7.26 (s, 1H), 7.06 (m, 1H), 6.88 (d, 1H), 7.2 (d, 1H, J=16Hz), 6.76 (d, 1H, J=16Hz), 6.0 (s, 1H), 3.93 (s, 6H), 2.31 (s, 3H)

Embodiment 81, (E) -5- (4- methoxyl-styrenes) -3- methyl-isoxazoles (III-2080)

Half white solid, yield 61%.¹H NMR (400MHz, CDCl3) d：7.45 (m, 2H), 7.22 (d, 1H, J= 16Hz), 6.75 (d, 1H, J=16Hz), 6.9 (m, 2H), 6.0 (s, 1H), 3.8 (s, 3H), 2.3 (s, 3H)

Following compound is prepared by following general synthetic method：

The general synthesis step of the molecule is：

Embodiment 82, (E) -4- (2- (3- methyl isophthalic acid H- pyrazoles -5- bases) vinyl) benzene -1,2- glycol (III-2081)

Light tan solid, yield 90%.¹H NMR (400MHz, DMSO) d；12.38 (s, 1H), 9.0 (s, 2H), 6.9 (m, 2H), 6.7 (m, 3H), 6.2 (s, 1H), 2.1 (s, 3H)

Embodiment 83, (E) -4- (2- (3- methyl isophthalic acid H- pyrazoles -5- bases) vinyl) phenol (III-2082)

Light tan solid, yield 79%.¹H NMR (400MHz, DMSO-d 6) d ppm：12.37 (s, 1H), 9.5 (s, 1H), 7.3 (d, 2H), 6.8 (d, 1H), 6.7 (m, 1H), 6.6 (d, 2H), 6.1 (s, 1H), 2.2 (s, 3H)

Embodiment 84, (E) -5- (4- methoxyl groups) -3- methyl isophthalic acid H- pyrazoles (III-2083)

Light tan solid, yield 61%.¹H NMR (400MHz, DMSO-d6) dppm 2.53 (s, 1H), 7.47 (d, 2H), 6.9 (m, 4H), 6.2 (s, 1H), 3.7 (s, 3H), 2.2 (s, 3H)

Embodiment 85, (III-2084)

By 0.3mmol nitroolefins, 0.45mmol NaN3 are added in 3mlDMF, and 0.15mmol is added after stirring to first Benzene sulfonic acid, a hour is stirred at 60 DEG C, is cooled to room temperature, is added water quenching and is gone out reaction, (3 × 10ml) is extracted with ethyl acetate, Merge organic layer anhydrous sodium sulfate drying, be spin-dried for solvent, with silica gel column chromatography purified product (eluent is ethyl acetate: Petroleum ether=1: 5) yield：74%.1HNMR (DMSO-d6,400MHz), 1drop TFA：δ 8.09 (s, 1H), 7.57 (s, 2H), (7.35-7.26 m, 5H)

Embodiment 86,2,4- dimethyl -5- [(E)-styryl] oxazoles (III-2085)

Yield：89%.¹H NMR (400MHz, CDCl3) 2.21 (s, 3H), 2.45 (s, 3H), 6.81 (d, J=16.5Hz, 1H), 6.90 (d, J=16.5Hz, 1H), 7.22-7.26 (m, 1H), 7.32-7.36 (m, 2H), 7.45 (d, J=7.3Hz, 2H)

Embodiment 87, (E) -2- (methyl (4- (2- (quinoline -2- bases) vinyl) phenyl) amino) ethanol-Halo labels (Halo herein refers to a kind of protein tag technology, can be that fluorescent dye and protein are special by Halo parts for synthesis Combine different in naturely, earliest detailed description is shown inACS Chem Biol.2008Jun 20；3(6)：373-82.)

The synthesis of molecule 1,2,3,4

By 2.1g molecules 1 (20mmol), 2- (2- amino ethoxies) ethanol) it is dissolved in 50ml ethanol, it is cooled to 0 degree, plus Enter 4.36g di-tert-butyl dicarbonates, react 5 hours at room temperature, ethanol is removed in rotation, add 20ml dichloromethane and 20ml water, then use Dichloromethane is extracted twice, and merges organic phase, and saturated common salt is washed, and anhydrous sodium sulfate drying, suction filtration is spin-dried for, after silicagel column Obtain molecule 2, yield 90%.

The nuclear magnetic data of molecule 2：¹H NMR (400MHz, CDCl₃) δ 3.77-3.72 (m, 2H), 3.61-3.52 (m, 4H), (s, the 9H) of 3.33 (t, J=5.1Hz, 2H), 1.53 (s, 1H), 1.45

2.15g molecules 2 (10.5mmol) are dissolved in 20ml tetrahydrofurans and 10ml DMF, are cooled to 0 degree, sodium hydride is added (14mmol), reacts half an hour, adds the chloro- 1- iodohexanes (16mmol) of 6-, 0 degree of reaction half an hour, reacts at room temperature one day, plus Enter saturated ammonium chloride and reaction is quenched, ethyl acetate extraction merges organic phase, and saturated common salt washing, anhydrous sodium sulfate drying is taken out Filter, is spin-dried for, molecule 3, yield 65% is obtained after silicagel column.

The nuclear magnetic data of molecule 3：¹H NMR (400MHz, CDCl₃) δ 3.63-3.59 (m, 2H), 3.59-3.51 (m, 6H), 3.47 (t, J=6.7Hz, 2H), 3.32 (t, J=5.1Hz, 2H), 1.77 (dd, J=14.6,6.8Hz, 2H), 1.66-1.56 (m, 2H), 1.50-1.42 (m, 11H), 1.38 (ddd, J=9.1,5.8,2.8Hz, 2H)

1.2g molecules 3 (4.1mmol) are dissolved in 30ml dichloromethane, are cooled to 0 degree, 5ml trifluoroacetic acids are added, reaction 4 is small When, rotation removes solvent and unnecessary trifluoroacetic acid, adds 30ml methanol, is cooled to 5 degree, adds 1.65g potassium carbonate, reacts half an hour, Suction filtration, is spin-dried for, and adds 20ml water, and ethyl acetate extraction merges organic phase, and saturated common salt washing, anhydrous sodium sulfate drying is taken out Filter, is spin-dried for, molecule 4, yield 72% is obtained after silicagel column.

The nuclear magnetic data of molecule 4：¹H NMR (400MHz, CDCl₃) δ 6.50 (s, 3H), 3.80-3.61 (m, 4H), 3.61- 3.50 (m, 4H), 3.46 (dd, J=8.6,4.8Hz, 2H), 3.02 (t, J=4.7Hz, 2H), 1.84-1.71 (m, 2H), 1.65- 1.54 (m, 2H), 1.50-1.30 (m, 4H)

0.3g molecules 5 are dissolved in 5ml and dry DMF, are cooled to 0 degree, add 40mg sodium hydrides, react 15 minutes, add monovalent The 3- bromo-propionic acid tert-butyl esters, react half an hour, be warming up to 50 degree and react 24 hours, DMF is removed in rotation, add 20ml water, ethyl acetate Extraction, merges organic phase, and saturated common salt washing, anhydrous sodium sulfate drying, suction filtration is spin-dried for, and is obtained molecule 6 after silicagel column, is produced Rate 45%.

The nuclear magnetic data of molecule 6¹H NMR (400MHz, DMSO) δ 8.27 (d, J=8.6Hz, 1H), 7.91 (dd, J=12.9, 8.2Hz, 2H), 7.80 (d, J=8.6Hz, 1H), 7.75-7.68 (m, 2H), 7.55 (d, J=8.8Hz, 2H), 7.50 (t, J= 7.5Hz, 1H), 7.19 (d, J=16.3Hz, 1H), 6.76 (d, J=8.8Hz, 2H), 3.63 (d, J=4.7Hz, 2H), 3.59 (d, J=4.8Hz, 2H), 3.00 (s, 3H), 1.42-1.38 (m, 9H), 1.36 (d, J=6.0Hz, 2H), 0.91 (t, J= 7.4Hz, 2H)

0.1g molecules 6 are dissolved in 2ml ethanol, add monovalent molecule 4, back flow reaction 8 hours is spin-dried for, obtained after silicagel column To molecule 7, yield 67%.

The nuclear magnetic data of molecule 7¹H NMR (400MHz, DMSO) δ 8.27 (d, J=8.6Hz, 1H), 7.91 (dd, J=12.9, 8.2Hz, 2H), 7.80 (d, J=8.6Hz, 1H), 7.75-7.68 (m, 2H), 7.55 (d, J=8.8Hz, 2H), 7.50 (t, J= 7.5Hz, 1H), 7.19 (d, J=16.3Hz, 1H), 6.76 (d, J=8.8Hz, 2H), 6.50 (s, 3H), 3.80-3.61 (m, 4H), 3.61-3.50 (m, 4H), 3.59 (d, J=4.8Hz, 2H), 3.46 (dd, J=8.6,4.8Hz, 2H), 3.02 (t, J= 4.7Hz, 2H), 3.00 (s, 3H), 1.36 (d, J=6.0Hz, 2H), 1.84-1.71 (m, 2H), 1.65-1.54 (m, 2H), 1.50-1.30 (m, 4H), 0.91 (t, J=7.4Hz, 2H)

The compound explained below for utilizing the present invention is used for the application aspect of bio-imaging purposes as dyestuff, first describes Once the step of different imaging operations, convenient no longer to repeat write operation step every time in embodiment.

Application aspect 1：As dyestuff, the living cells in HeLa cell is imaged the compound of the present invention

Imaging method 1：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Cell dyeing and image-forming step are as follows：

1. by the HeLa cell of passage (be derived from Hai Ruita lachs (Henrietta Lacks) cervical cancer cell it is thin Born of the same parents are, are wide variety of cells in biomedical research.) cultivate in the 0.17um culture dish of sheet glass is covered with.Culture The component of base is：2.75 milliliters of water, 2.5 milliliters of F12 solution, 20 milliliters of DMEM solution, 0.625 milliliter of 1M HEPEs buffer solutions, 2.5 milliliters of growth factor 0.5ml, serum)

2. taking the storing liquid of 10ul dyestuffs, it is added to pipettor in the Tissue Culture Dish containing 1ml cell culture mediums.

3. cell is placed in 37 degree, 5% CO2 incubator, 20min is dyed.

4. being moved to sheet glass in the culture dish containing extracellular fluid with tweezers, the specific component of extracellular fluid is：150mM NaCl, 3mM KCl, 3mM CaCl2,10mM HEPEs buffer solutions, 5mM glucose.

5. being imaged with Zeiss (zeiss) 710 Laser Scanning Confocal Microscope, image-forming objective lens are 20 times of hydroscopes, the numerical aperture of the object lens Footpath is 1.0.

6. being imaged using the scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 462nm-538nm, 490nm-535nm and 555nm-675nm, scanning range about 150um x150um；Sweep time is 0.5- 1min obtains cell imaging image per frame.

Application aspect 2：Dyestuff fixes cell imaging in HeLa cell

Imaging method 2：Dyestuff with 0.06mmol/L concentration to be dissolved in ethanol solution, room temperature storage.

Cell dyeing and image-forming step

1. fixing the HeLa cell of passage ten minutes with 3% paraformaldehyde, cell is placed in 37 degree, 5% CO2 culture In case, 20min is dyed.

2. being moved to sheet glass in the culture dish containing extracellular fluid with tweezers, the specific component of extracellular fluid is：150mM NaCl, 3mM KCl, 3mM CaCl2,10mM HEPEs buffer solutions, 5mM glucose.

3. being imaged with Zeiss (zeiss) 710 Laser Scanning Confocal Microscope, image-forming objective lens are 20 times of hydroscopes, the numerical aperture of the object lens Footpath is 1.0.

4. being imaged using scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 462nm-538nm, 490nm-535nm and 555nm-675nm, scanning range about 150umXl50um.Sweep time is 0.5-1min Per frame, cell imaging image is obtained.

Application aspect 3：Dyestuff carries out living cells imaging in neuronal cell.

It is imaged imaging method 3：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage Deposit.

Cell dyeing and image-forming step are as follows：

1. the pregnant mouse being pregnant 18 days is lethal using the method for cervical dislocation, take out the cultured hippocampal neuron of tire mouse head In the 0.17um culture dish of sheet glass is covered with.The component of culture medium is：2.75 milliliters of water, 2.5 milliliters of F12 solution, DMEM 20 milliliters of solution, 0.625 milliliter of 1M HEPEs buffer solutions, growth factor 0.5ml, 2.5 milliliters of serum), it will be trained outside neuronal soma Support 14-18 days

2. taking the storing liquid of 10ul dyestuffs, the culture containing 1ml culture mediums is added to pipettor neuron culture dish In.

3. cell is placed in 37 degree, 5% CO2 incubator, 20min is dyed.

6. being imaged using the scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 490nm-535nm and 555nm-675nm, scanning range about 150um x150um；Sweep time is the every frames of 0.5-1min, obtains thin Born of the same parents' image.

Application aspect 4：Cell imaging is fixed in dyestuff in neuronal cell

Imaging method 4：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Cell dyeing and image-forming step are as follows：

2. fixing neuron ten minutes with 3% paraformaldehyde, cell is placed in 37 degree, 5% CO2 incubator, contaminated Color 20min.

3. being moved to sheet glass in the culture dish containing extracellular fluid with tweezers, the specific component of extracellular fluid is：150mM NaCl, 3mM KCl, 3mM CaCl2,10mM HEPEs buffer solutions, 5mM glucose.

4. being imaged with Zeiss (zeiss) 710 Laser Scanning Confocal Microscope, image-forming objective lens are 20 times of hydroscopes, the numerical aperture of the object lens Footpath is 1.0.

5. being imaged using scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 490nm-535nm and 555nm-675nm, scanning range about 150umX150um.Sweep time is the every frames of 0.5-1min, obtains thin Born of the same parents' image.

Application aspect 5：Dyestuff is dyed with being imaged in Histological section

Imaging method 5：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Imaging method and step

1. rat head sample sections are obtained into rat cerebral tissue's section of 20um thickness using freezing-microtome, and paste In on slide.

2. 0.06mmol/L dyestuff storing liquid is mixed in equal volume with phosphate buffer (PBS).

3. taking 30ul mixed liquors to add to 10mmX10mm histological sample, 10min is dyed at normal temperatures

4. mixed liquor is removed, 1ml PBS are added dropwise and observe for 710 time on sheet glass, being placed in Zeiss (zeiss).

5. being imaged with Zeiss (zeiss) 710 Laser Scanning Confocal Microscope, image-forming objective lens are 20 times of hydroscopes, the numerical aperture of the object lens Footpath 1.0.

6. being imaged using scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 490nm-535nm and 555nm-675nm, scanning range about 0.8mmX0.8mm.Sweep time is the every frames of 0.5-1min, is obtained into As image.

Application aspect 6：Dyestuff is in zebra fish visitain and imaging

Imaging method 6：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Imaging method and step

1. taking the storing liquid of 200ul 0.06mmol/L dyestuff, being added dropwise to support has in the 20ml solution of zebra fish.

2. dye 20min-120min under normal temperature.

3. with 16.8ug/ml ms-222 anesthesia zebra fish, about 3min.

4. in the PBS that zebra fish is embedded in 1.5% agarose, about 10min to agarose solidifies.

6. being imaged using scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 490nm-535nm and 555nm-675nm, scanning range about 3mmX3mm.Sweep time is 2min per frame.

Application aspect 7：Dyestuff is to staining pathologic section and imaging

Imaging method 7：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Imaging method and step

1. pathological tissue to be cut to 5um frozen section using freezing microtome, it is placed on slide.

2. fix section, 5min with 10% formalin.

3. 0.06mmol/L dyestuff storing liquid is mixed in equal volume with phosphate buffer (PBS).

4. the slide for posting pathological section is placed in mixed liquor, 3-5s is dyed.

5. being imaged with Zeiss (zeiss) 710 Laser Scanning Confocal Microscope, image-forming objective lens are 20 times of hydroscopes, numerical aperture 1.0.

6. being imaged using scanning cofocal microscope, a length of 488nm of excitation light wave, the transmitting spectral coverage of receiving is respectively 490nm-535nm and 555nm-675nm, scanning range about 0.8mmX0.8mm.Sweep time is 0.5-1min per frame.

Application aspect 8：Tumor imaging of the dyestuff to living animal

Imaging method 8：Dye composition is dissolved in ethanol solution with 0.06mmol/L concentration, room temperature storage.

Imaging method and step

1. G1 breast tumor cells are mixed with matrigel according to 3: 1,

2. mixed liquor is injected near the mammary gland of NOD/SCID mouse of 5-8 weeks

3. culture injection has mouse 1-2 weeks of tumour

4. from yellow Jackets of the abdominal cavity to mouse plastic injection quality proportion 80mg/kg, anesthetized mice 20 minutes

5. cutting the skin of mouse with scalpel, tumor tissues and part normal structure is set to be exposed in air

6. dyestuff F2 is mixed with PBS using 1: 1,1ml mixed liquors are taken to drop in the tumour and normal structure of mouse exposure, Dyeing 2-5 minutes

7. observing the fluorescence signal of intravital mouse using the MVX10 Stereo microscopes of Olympus companies, visual field is 2cmX1.5cm。

The photoluminescent property of the compounds of this invention explained below and imaging applications example, imaging method therein is with being described above Imaging method 1-8 refer to, description can be made simple.

In the application example being imaged below for the dyestuff of different structure using above-mentioned different imaging methods, the present invention Dyestuff meet ICT-pH structures dyestuff can show following Examples realization property, different molecular difference be exciting light and Launch the difference of light, the response range to pH is different, part non-limiting examples is only enumerated below, by non-limiting to these Example is specific and describes in detail, and to the disclosure of dye property, dyestuff is described using distinct methods to the imaging of different samples And the analysis to image, show the one pack system polychrome that the excellent imaging property and dyestuff of dyestuff in the present invention are shown It is imaged property.

Application examples 1：Molecule I-2060

Photoluminescent properties of the molecule I-2060 in different solvents is shown in Fig. 1 (ultraviolet absorption spectrum) and Fig. 2 (fluorescence emission spectrum), by The two collection of illustrative plates can be seen that molecule I-2060 is very sensitive to environment polarity, as solvent polarity increases (n-hexane, dichloromethane Alkane, tetrahydrofuran, ethyl acetate, acetone, ethanol, methanol), spectrum peak red shift, this is due to the distinctive confessions of molecule I-2060 Electronics base-conjugation-electron-withdrawing group structure has the property of Intramolecular electron transfer, the stabilization of different polarity to excited electronic state Property it is different, polar solvent stabilizing power is stronger so that can extreme difference it is smaller, so spectral red shift.

Molecule I-2025, I-2061, I-2063, I-2069, I-2062, I-2027, I-2036, I-2044, I-2045, I- 2046、I-2048、I-2051、I-2052、I-2026、I-2030、I-2059、I-2098、I-2099、I-2103、I-2104、I- 2106；II-2070、II-2074、II-2075、II-2076、II-2078、II-2079、II-2080、II-2081、II-2082、 II-2083、II-2084、II-2085；III-2069、III-2070、III-2071、III-2074、III-2075、III- 2076、III-2077、III-2078、III-2080、III-2082、III-2083、III-2084、III-2085；IV-2042、 IV-2043, IV-2044, IV-2045, IV-2046 etc. can show similar property.

This dye disclosed by the invention all carries Intramolecular electron transfer property, to showing to environment polarity very Sensitive the characteristics of, this feature can be found out from following more examples.

Molecule I-2060 is made into about 60mmol/L concentration, is adsorbed onto on filter paper, fluorescence paper, specific effect is obtained See that (blueness is I-2025 to Fig. 3, and rose is I-2064, and cyan is I-2061, and khaki is I-2062, and purple is I-2029, blue or green Purple is I-2027, and red is I-2060, and cream-coloured is I-2045), this kind of filter paper lighted in the different pH aqueous solution it is different (see Fig. 4), the response to pH is shown, its emission spectra is quantitatively determined and finds that it has good response (see Fig. 5) to pH, under Face can it is further seen that such dyestuff can distinguish pH value differ 0.2 precision different solutions, it is very sensitive.

HeLa cell imaging is carried out with molecule I-2060, method is the method for dye image method 1, it is figure to obtain image 6, under 488nm exciting light, the transmitting spectral coverage of receiving is respectively 490nm-535nm (the picture left above), 555nm-675nm (upper rights Figure) and three different wave lengths of 462nm-538nm (lower-left figure) are interval can be collected into transmitting light, show one as smart as a new pin The imaging of multicomponent polychrome picture (bottom-right graph), temporarily what region of the luminous correspondence cell of unclear different colours or Molecule, this needs further to study, but such dyestuff obviously discloses many new phenomenons in cell, be it is biological into As opening the new gate of a fan.

Neuron living cells imaging is carried out with molecule I-2060, method is the method for dye image method 3, and obtaining image is Fig. 7, under 488nm exciting light, the transmitting spectral coverage of receiving is respectively 490nm-535nm (the picture left above), 555nm-675nm (right Upper figure) and three different wave lengths of 462nm-538nm (lower-left figure) interval can be collected into transmitting light, show one and float very much The picture (bottom-right graph) of bright multicomponent polychrome imaging, it is seen that dyestuff different types of cell can realize one pack system polychrome into Picture.

Neuronal cell imaging is fixed with molecule I-2060, method is the method for dye image method 4, obtains image For Fig. 8, hence it is evident that it can be seen that the difference that the nucleus of neuronal cell lights with neuron dendron, comparison diagram 7 and Fig. 8, are entering When row living cells is imaged, nucleus is not almost dyed, but fixed cell dyes obvious yellow, can significantly distinguish work Cell and fixed cell.

By be can be seen that to molecule I-2060 in the imaging of different type cell as a kind of brand-new fluorescent dye, Excellent property is shown in various bio-imagings disclosed by the invention, photoluminescence quantum yield is high, and image stabilization is not observed To photobleaching phenomenon and phototoxicity, it is tally in every detail that imaging clearly, with conventional dyes compare performance.This money dyestuff also has it simultaneously Some excellent characteristics that his dyestuff does not have：Image taking speed soon and stably, most of imaging time only need to several seconds it is just real Ready-made picture；Dyestuff is not lighted in water, therefore imaging need not be cleaned, and entirely without background, obtains the image of high-contrast； Dye property is stable, preserves simple, and better tolerance in vivo, therefore imaging operation is simple, it is to avoid too many preparation Work is lost time, and also brings excessive uncertain factor to research, introduces error, is delayed research aircraft meeting；It is most important, the dyestuff Show one pack system polychrome imaging property, such dyestuff extreme sensitivity, after being combined with biological tissue, in different environments Performance sends different colours light in (such as pH, different component, ion concentration etc.), and the imaging of one pack system polychrome is presented；Therefore this Dyestuff has huge potential application in different kind organism imaging, internal pH detection and tumour cell detection.

Molecule I-2025, I-2061, I-2063, I-2069, I-2062, I-2027, I-2036, I-2044, I-2045, I-2046、I-2048、I-2051、I-2052、I-2026、I-2030、I-2059、I-2098、I-2099、I-2I03、I-2104、 I-2106；II-2070、II-2074、II-2075、II-2076、II-2078、II-2079、II-2080、II-2081、II- 2082、II-2083、II-2084、II-2085；III-2069、III-2070、III-2071、III-2074、III-2075、 III-2076、III-2077、III-2078、III-2080、III-2082、III-2083、III-2084、III-2085；IV- 2042nd, IV-2043, IV-2044, IV-2045, IV-2046, which can become, reveals the imaging and pH response similar to molecule I-2060 Property, difference is that glow color is different, and pH response range is different.

Application examples 2：Molecule I-2064

Photoluminescent properties of the molecule I-2064 in different solvents is shown in Fig. 9 (ultraviolet absorption spectrum) and Figure 10 (fluorescence emission spectrum), It can be seen that molecule I-2064 is same very sensitive to environment polarity by the two collection of illustrative plates, keep the superiority of this dye Matter, relative to molecule I-2060, molecule I-2064 luminous more red shift, the penetration power to tissue is mutually better, and also explanation can be with By adjusting molecule substituent, this kind of dyestuff is made the dyestuff of the luminous whole visible spectrum of covering, with very big commercial valency Value.

Molecule I-2064 is made into about 60mmol/L concentration, is adsorbed onto on filter paper, fluorescence paper, specific effect is obtained See that (blueness is I-2025 to Fig. 3, and rose is I-2064, and cyan is I-2061, and khaki is I-2062, and purple is I-2029, blue or green Purple is I-2027, and red is I-2060, and cream-coloured is I-2045), the filter paper that molecule I-2064 is made can distinguish pH value difference 0.2 The different solutions of precision, it is very sensitive, see Figure 11, the figure is shown as pH increases from 4.4 peak values for increasing to 7.2,500nm, The different pH solution of 0.2 precision can be distinguished..

HeLa cell imaging is carried out with molecule I-2064, method is the method for dye image method 1, it is figure to obtain image 12, the cell image obtained in the case of a kind of molecules of only I-2064 shows blueness and green in two regions respectively, shows Intracellular different material can be distinguished by showing.

HeLa cell imaging is fixed with molecule I-2064, method is the method for dye image method 2, and obtaining image is Figure 13, the imaging of contrast living cells and fixed cell imaging, it can be clearly seen that cell can be dyed to nucleus after fixed, and And cell imaging color red shift after fixing, present the cellular change of cell after death.

Neuron living cells imaging is carried out with molecule I-2064, method is the method for dye image method 3, and obtaining image is Figure 14, image shows that the aixs cylinder and dendron of neuron are dyed to green, and cell space is yellow, and nucleus is red, shows this Different cellular environment at three, in addition it is also necessary to further study this difference at three.

Neuronal cell imaging is fixed with molecule I-2064, method is the method for dye image method 4, obtains image For Figure 15, the fixed neuronal cell dendron of image display and aixs cylinder are yellow, and nucleus is red, and comparison diagram 14 illustrates solid Cellular environment there occurs that change causes dyeing change after fixed, illustrate that I-2064 can distinguish neuronal cell living and dead god Through first cell.

Histological section's imaging is carried out with molecule I-2064, method is the method for dye image method 5, it is figure to obtain image 16 ((a) is Nissl dyeing, and (b) dyes for I-2064).Histological section used in the figure is mouse brain frozen section, existing Imaging be Nissl dyeing method, the present invention carries out being coloured to picture and Nissl imaging contrasts with molecule I-2064, can be with Find out the details that can show of Nissl imagings, molecule I-2064 can show, but molecule I-2064 obtain be one pack system polychrome into The coloured picture of picture, also presents many details not seen before, indicates the heterogeneity that may be present of mouse brain different zones, These properties are likely to relevant with brain function subregion.

Staining pathologic section and imaging of the dyestuff to people are carried out with molecule I-2064, method is the side of dye image method 7 Method, obtains image for Figure 17, and the gastric tissue for being derived from human body is imaged with molecule I-2064, and (a) is normal structure, and (b) is canceration Tissue, Figure 17 shows that the dyeing of normal structure is partially green partially yellow, and the dyeing of cancerous tissue is partially red because cancerous tissue with just Often tissue is compared, and physiological environment difference is very big, and cancerous tissue pH is lower than normal structure, and this dyestuff is to environment especially pH It is very sensitive, therefore among detection of this dyestuff available for cancerous tissue and cancer cell, with very big practical application meaning Justice.

Further molecule I-2064 is combined with different biological molecules and protein to investigate luminosity, Figure 18 shows point (BSA is bovine serum albumin(BSA) to sub- I-2064, and FBS is tire ox with different biological molecules or stearic acid in same pH PBS solution Serum, Trypsin is trypsase, and Lipase is lipase, and Stearic Acid are stearic acid, and stearic acid is used for analog cell The lipid of film) the luminous difference that combines, the microenvironment of protein determines the luminous of dyestuff, illustrates that the dyestuff can be used for different eggs The detection and differentiation of white matter and biomolecule.

Application examples 3：Molecule I-2067

Photoluminescent properties of the molecule I-2067 in different solvents is shown in Figure 19 (ultraviolet absorption spectrum) and Figure 20 (fluorescence emission spectrum), The more red shift of molecule I-2067 average molecular I-2060 spectrum is can be seen that by the two collection of illustrative plates, this is due to add one pair Key, caused by conjugation becomes big.

With molecule I-2067 in zebra fish visitain and imaging, method is the method for dye image method 6, obtains image 21, real-time one pack system polychrome imaging of the zebra fish in live body is obtained, Figure 21 is respectively the head of zebra fish from top to bottom ((a)), trunk ((b)) and afterbody ((c)), as can be seen from Figure a kind of single molecule I-2067 zebra fish different parts are contaminated respectively Into yellow green, yellow and partially red.

Application examples 4：Molecule I-2084

HeLa cell imaging is carried out with molecule I-2084, method is the method for dye image method 1, it is figure to obtain image 22, under 488nm exciting light, the transmitting spectral coverage of receiving is respectively 490nm-535nm (the picture left above), 555nm-675nm (upper rights Figure) and three different wave lengths of 462nm-538nm (lower-left figure) are interval can be collected into transmitting light, show one as smart as a new pin The imaging of multicomponent polychrome picture (bottom-right graph), the cell image obtained in the case of a kind of molecules of only I-2084 shows Blueness, green and red multicolor image.

Molecule I-2069, I-2052, I-2055, I-2058, I-2059, I-2070, I-2071, I-2098, I-2099, I- 2109th, II-2070, I-2110 etc. have the property similar to above-mentioned molecule I-2084.

Application examples 5：Molecule I-2110

The tumor imaging of living animal (NOD/SCID mouse) is carried out with molecule I-2110, method is dye image method 8 Method, obtain image for Figure 23, image show, kind has the mouse of tumour by after the dyeing, and tumor group is woven with bright green Color fluorescence, and normal structure does not have fluorescence or there was only very low background fluorescence.

It is described above, it is only the illustrative embodiments of the present invention, but protection scope of the present invention is not limited thereto, and appoints Why not pass through the change or replacement that creative work is expected, should all be included within the scope of the present invention.Therefore, it is of the invention Protection domain should be determined by the scope of protection defined in the claims.

Claims

1. bio-imaging, pH value detection, biomacromolecule detection or cell micro-loop are used for by the formula a compound or its salts represented The purposes of border detection,

Wherein

Z is represented optionally by a multiple group R₂Substituted pyrrole radicals, thiazolyl, isothiazolyl, oxazolyl, isoxazolyls, pyrrole Oxazolyl, imidazole radicals, 1H-1,2,3- triazolyls, pyridine radicals, pyridazinyl, pyrimidine radicals, pyrazinyl, 1,3,5-triazines base, indyl, Purine radicals, carbazyl, quinolyl, isoquinolyl, pteridine radicals or quinoxalinyl；

R₁Represent H, halogen, hydroxyl, amino, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1-C6 alkane Base, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl, C1-C6 alkoxies, C1-C6 alkane Base amino, hydroxyl C1-C6 alkyl aminos, C1-C6 alkylthio groups, C3-C7 Heterocyclylalkyls, C6-C10 aryl, heteroaryl are wherein described C6-C10 aryl and the heteroaryl optionally by it is one or more selected from halogen, hydroxyl, amino, sulfydryl, sulfonyl, nitro, Carbonyl, ester group, carboxyl, cyano group, amide groups, C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl, C1-C6 alkane Substituent substitution in epoxide, C1-C6 alkyl aminos, hydroxyl C1-C6 alkyl aminos and C1-C6 alkylthio groups；

R₂Represent H, hydroxyl, amino, sulfydryl, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl or C1-C6 alkoxies；

Term " heteroaryl " represent to refer to 4 to 8 single or multiple yuan of rings and containing 1,2 or 3 be individually selected from N, O and S hetero atom and at least one heterocycle therein are the groups of armaticity, and term " C3-C7 Heterocyclylalkyls " refers to containing one N atoms are used as 3 to 7 yuan of saturated cyclic alkyl that ring memberses and other ring memberses are carbon atom.

2. purposes according to claim 1, it is characterised in that it is described by the formula a compound or its salts represented be by with The compound or its salt that lower formula I, II, III or IV are represented：

Wherein each T independently represents C or N, U represent O, S or N, and R₁、R₂、S₁And S₂As defined in claim 1.

3. purposes according to claim 1, it is characterised in that it is described by the formula a compound or its salts represented be by with Under various expression compound or its salt：

Wherein Z, R₁、R₂、S₁And S₂As defined in claim 1, T represents C or N, R₃Represent H, hydroxyl, amino, sulfydryl, sulphonyl Base, carboxyl, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 cycloalkyl or C1-C6 alkoxies.

4. purposes according to claim 3, it is characterised in that the compound or its salt is the change by following various expression Compound or its salt：

Wherein R₂、R₃、S₁、S₂With T as defined in claim 3, and R₆And R₇H, hydroxyl, amino, mercapto are represented independently of one another Base, sulfonyl, carboxyl, cyano group, amide groups, C1-C6 alkyl, hydroxyl C1-C6 alkyl, C2-C6 alkenyls, C2-C6 alkynyls, C3-C6 Cycloalkyl or C1-C6 alkoxies.

5. purposes according to claim 1, it is characterised in that the compound or its salt is the change with following structural formula Compound or its salt：

6. purposes according to claim 1, it is characterised in that the compound or its salt is the change with following structural formula Compound or its salt：

7. the purposes according to any one of claim 1-6, it is characterised in that the compound or its salt is with selected from alkane Solution in hydro carbons, ethers, esters, the solvent of alcohols or ketone is used.

8. purposes according to claim 7, it is characterised in that the solvent be n-hexane, dichloromethane, tetrahydrofuran, Ethyl acetate, acetone, ethanol or methanol, and the concentration range of the solution is 0.1x10^-5M to 10x10^-5M。

9. the purposes according to any one of claim 1-6, it is characterised in that thin in different solvents, difference pH or different Under conditions of born of the same parents' microenvironment, the solution of the compound or its salt shows different colours in bioluminescence imaging.

10. purposes according to claim 9, it is characterised in that when the compound or its salt is used to detect pH value, its The pH value of difference more than 0.2 can be distinguished.