CN111704570B - Near-infrared reaction type fluorescent probe with heptamethine cyanine structure and preparation method and application thereof - Google Patents
Near-infrared reaction type fluorescent probe with heptamethine cyanine structure and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a near-infrared reaction type fluorescent probe with a heptamethine cyanine structure, a preparation method and application thereof, and belongs to the technical field of fluorescent probes. The near-infrared reaction type fluorescent probe with the heptamethine cyanine structure has a structural formula shown as a formula I, wherein R is1、R2Each independently selected from C2～C8And X is chlorine, iodine or bromine. In the compound shown in the formula I, the cyanine structure has the characteristic of near-infrared fluorescence emission, C2～C8The ether chain remarkably improves the water solubility of the heptamethine cyanine mother nucleus structure, the o-chlorobenzoyl group has specific chemical action on the cysteine, the selective recognition effect on the cysteine is realized, and meanwhile, the ether chain has good biological adaptability. The fluorescent probe provided by the invention is beneficial to improving the sensitivity and accuracy of cysteine fluorescent detection, and has an important role in expanding the application of the fluorescent probe technology in the field of analytical chemistry.
Description
Technical Field
The invention relates to a near-infrared reaction type fluorescent probe with a heptamethine cyanine structure, a preparation method and application thereof, and belongs to the technical field of fluorescent probes.
Background
The fluorescent probe technology has the advantages of convenient operation, no damage to samples, high selectivity, high sensitivity, capability of realizing in-situ detection and visual detection of biological samples and the like. At present, the analysis method based on the fluorescent probe technology is widely applied to important fields of analytical chemistry, environmental detection, biological imaging, disease diagnosis and the like.
Biological thiols, such as cysteine (Cys), homocysteine (Hcy) and Glutathione (GSH), are involved in many biological processes and play a key role in maintaining the proper redox state of biological systems. Cys and Hcy are important biomolecules required for maintaining immune competence and growth of aged cells and tissues, among others. Aberrant Cys expression levels are closely associated with slow growth, discoloration, liver damage, muscle and fat loss, skin damage, and cancer, among others. Homocysteine (Hcy) is involved in various types of vascular and renal diseases and is considered a risk factor for cardiovascular diseases and alzheimer's disease. Reduced Glutathione (GSH) is the most abundant intracellular non-protein thiol (1-10mM) and biomarker of oxidative stress. Studies have shown that GSH plays a key role in controlling oxidative stress to maintain redox homeostasis of cell growth and function. Abnormal levels of GSH are associated with various diseases, such as aids, cancer, liver damage, and neurodegenerative diseases, among others. Therefore, different biological thiols have different functions, so that the separation of other sulfhydryl-containing amino acids (homocysteine and glutathione) of cysteine fish is very important, and the biological thiol plays a very important role in researching the physiological and pathological processes of diseases and early diagnosis of the diseases.
Disclosure of Invention
The invention aims to provide a near-infrared reaction type fluorescent probe with a heptamethine cyanine structure, which has a selective recognition effect on cysteine.
The second object of the present invention is to provide a method for preparing the near-infrared reaction type fluorescent probe having a heptamethine cyanine structure.
The third purpose of the invention is to provide the application of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure in the detection of cysteine.
The technical scheme of the invention is as follows:
a near-infrared reaction type fluorescent probe with a heptamethine cyanine structure has a structural formula shown as a formula I:
wherein, R is1、R2Each independently selected from C2～C8And X is chlorine, iodine or bromine. It is understood that R3And X may be the same or different.
In the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure, the cyanine structure has the characteristic of near-infrared fluorescence emission, and the fluorescent probe designed based on the cyanine structure shows good application prospects in the fields of photodynamic/photothermal diagnosis and treatment, deep tissue response, subcellular organelle positioning and the like; the invention utilizes C2～C8The ether chain remarkably improves the water solubility of the heptamethine cyanine mother nucleus structure, realizes the selective recognition effect on the cysteine by utilizing the specific chemical action of the o-chlorobenzoyl group on the cysteine, and has good biological adaptability. The fluorescent probe provided by the invention is beneficial to improving the sensitivity and accuracy of cysteine fluorescent detection, and has an important role in expanding the application of the fluorescent probe technology in the field of analytical chemistry.
In order to further increase the water solubility of the compounds of formula I, preferably, R is1、R2Is composed of
Wherein
Indicates the attachment site to N.
A preparation method of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure comprises the following steps:
wherein, R is1、R2Each independently selected from C2～C8An ether chain of (a); x is chlorine, iodine or bromine;
(1) reacting the compound shown in the formula III with sodium acetate to obtain a compound shown in a formula II;
(2) the compound shown in the formula II reacts with o-chlorobenzoyl chloride under the action of triethylamine.
The preparation method of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure only needs two-step reaction to prepare the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure. The method has simple operation, mild condition and low toxicity.
It is understood that, in the step (1), after the reaction is completed, water is added to quench the reaction, and then the reaction solution is extracted with methylene chloride, and the obtained crude extract is subjected to silica gel column chromatography to obtain the compound represented by the formula II.
Preferably, in the step (1), the molar ratio of the compound shown in the formula III to sodium acetate is (0.9-1.1): (2.9-3.1). This amount of sodium acetate facilitates sufficient oxidation of the compound of formula III to the compound of formula II.
Preferably, in step (1), the solvent for the reaction is N, N-dimethylformamide. The N, N-dimethylformamide has certain alkalinity, which is beneficial to the reaction
Preferably, in step (1), the temperature of the reaction is 70 ℃; the reaction time is 5-7 h. The compound shown in the formula III can be promoted to react with sodium acetate by reasonably adjusting and optimizing the reaction temperature and time.
It will be appreciated that in step (2), for the reaction of the compound of formula II with the substituted benzoyl chloride, the substituted benzoyl chloride is added in the following manner: the substituted benzoyl chloride is dissolved in a solvent and then is dripped into a reaction system (a mixed system of a compound shown in a formula II, triethylamine and the solvent).
It is understood that, in step (2), after the reaction is completed, the compound represented by formula I can be obtained by evaporating the solvent, separating by silica gel column chromatography and anion exchange resin.
Preferably, in the step (2), the molar ratio of the compound shown in the formula II to the substituted benzoyl chloride is (1.8-2.2) to (2.8-3.2).
Preferably, in the step (2), the amount ratio of the compound represented by the formula II to triethylamine is (0.8-1.2) mol: (5-7) mL.
Preferably, in step (2), the reaction comprises: reacting for 25-35 min at 0 ℃, and then reacting for 10-15 h at 20-30 ℃. The substituted benzoyl chloride can be better dissolved in the solvent by two-stage reaction, which is beneficial to the full implementation of the subsequent reaction.
The near-infrared reaction type fluorescent probe with the heptamethine cyanine structure is applied to the detection of cysteine.
The near-infrared reaction type fluorescent probe with the heptamethine cyanine structure has the performance of visually detecting Cys at a cellular level and an animal level.
Preferably, the detection is performed at the cellular level and at the animal level
Preferably, the wavelength used for the detection is 635 nm. With 565nm as the excitation wavelength, Cys causes the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure to generate the fluorescence emission phenomenon at 635nm, and under the same condition, after the interaction with homocysteine (Hcy) or Glutathione (GSH) respectively, the probe only generates weak fluorescence emission at 635 nm. The near-infrared reaction type fluorescent probe with the heptamethine cyanine structure can be used as a fluorescent probe for selectively detecting Cys.
Drawings
FIG. 1 shows the preparation of the compound of the formula II from example 41H NMR chart;
FIG. 2 shows the preparation of the compound of the formula Ia from example 41H NMR chart;
FIG. 3 shows the preparation of the compound of the formula Ia from example 413C NMR chart;
FIG. 4 is a HR-MS plot of the compound of formula Ia, prepared in example 4;
FIG. 5 shows UV spectroscopy experiments on compounds of formula Ia;
FIG. 6 shows fluorescence spectrum analysis of a compound represented by formula Ia;
FIG. 7 is a graph showing the selective fluorescence response spectrum of compound Ia;
FIG. 8 is a diagram of a confocal laser assay for selectively detecting Cys in HeLa cells with compound Ia; wherein (a) is HeLa cells stained by Hoechst 33342, (b) is HeLa cells and NEM incubated for 30min, (c) is HeLa cells and NEM incubated for 30min followed by incubation with Compound Ia (Probe, 2. mu.M) for 30min, (d) is cells without pre-scavenging thiol amino acids incubated with Probe (2. mu.M) for 30min, (e) is HeLa cells and NEM (1mM) incubated for 30min followed by incubation with glutathione GSH (100. mu.M) for 30min followed by incubation with Probe (2. mu.M) for 30min, (f) is HeLa cells and NEM incubated for 30min followed by incubation with homocysteine (Hcy) for 30min followed by incubation with Probe (10. mu.M) for 30min, (g) is HeLa cells and NEM followed by incubation with cysteine (Cys) for 30min followed by incubation with Probe (10. mu.M) for 30 min; (h) (i) relative fluorescence intensity statistics for purple channels (λ em: 780-820nm) and red channels (λ em: 580-670nm) from 1 to 7(a-g), respectively;
FIG. 9 is a graph of experiments showing that compound Ia selectively detects Cys in BALB/c nude mice.
Detailed Description
The present invention will be further described with reference to the following embodiments.
In the embodiment of the invention, the used instruments are of the following brands: bruker AV-400 nuclear magnetic resonance apparatus (Germany); hitachi U-2900 dual beam UV-visible spectrophotometer (japan); hitachi F-2500 fluorescence spectrophotometer (Japan); agilent 1100 series LC/MSD and AB SCIEX Triple TOFTM5600+ Mass Spectrometry (USA).
The specific embodiment of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure of the invention is as follows:
example 1
The structural formula of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure is shown as formula Ia.
Example 2
The structural formula of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure is shown as formula Ib.
Example 3
The structural formula of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure is shown as the formula ic.
Secondly, the specific embodiment of the preparation method of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure of the invention is as follows:
example 4
The preparation method of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure of the embodiment comprises the following steps:
(1) synthesis of Compound of formula III
Under the protection of nitrogen, the mixture is subjected to
Substituted hemicyanine (2.2mol) [ structural formula:
a solution of anhydrous acetic anhydride (10mL) and a solution of 2-chloro-1-formyl-3-hydroxymethylcyclohexene (172.6mg,1mmol) were added with sodium acetate (100mg, 1.2mmol) and reacted at 70 ℃ for 3 hours. The solution was cooled to room temperature and water (20mL) was added followed by CH2Cl2(30 mL. times.3). After removal of the solvent, the residue was purified by silica gel column Chromatography (CH)2 Cl 220/MeOH: 1) to give the compound of formula iii as a dark green powder (536.46mg, yield: 77.3%).
(2) Synthesis of Compound represented by formula II
To a solution of the compound represented by the formula III (314.52mg, 0.4mmol) in anhydrous N, N-dimethylformamide (15mL) was added sodium acetate (100mg, 1.2mmol) under nitrogen protection, and the mixture was reacted at 70 ℃ for 6 hours. The solution was cooled to room temperature and water (50mL) was added followed by CH2Cl2(30 mL. times.3). After removal of the solvent, the residue was purified by silica gel column Chromatography (CH)2 Cl 250 parts per MeOH: 1) to give the compound represented by formula ii as a dark red powder (135.21mg, yield: 68.4%).
Characterization of the compound of formula II gave the compound shown in FIG. 11H NMR chart, as is clear from FIG. 1,1H NMR(300MHz,CDCl3)δ8.35(d,J＝7.7Hz,1H),7.81–7.61(m,4H),7.41–7.27(m,3H),7.22–7.13(m,3H),6.34(d,J＝14.0Hz,2H),4.43(s,4H),3.92(s,4H),3.60(t,J＝4.3Hz,4H),3.45(dd,J＝5.7,3.0Hz,4H),3.28(s,6H),2.76(s,4H),1.85(s,15H).13C NMR(75MHz,CDCl3)δ173.04,149.99,144.04,142.55,140.84,128.70,127.78,125.22,122.02,111.49,102.34,71.36(d,J＝73.4Hz),68.02,58.71(d,J＝44.2Hz),49.32,28.20,18.42.HRMS(ESI)m/z:calcd for641.39490,found 641.39374。
(3) synthesis of Compounds of formula Ia
To a mixture of the compound represented by the formula II (128.08mg, 0.2mmol) and Et at 0 deg.C3N (0.3mL) in dichloromethane (20mL) was added dropwise o-chlorobenzoyl chloride (0.1mL, mixed with 10mL of dichloromethane). The reaction was stirred for 30 minutes under nitrogen, warmed to room temperature and stirred overnight. The reaction mixture was concentrated in vacuo to give the crude product, which was purified by silica gel column chromatography (DCM: MeOH ═ 20: 1) to give the compound of formula la as a green solid (33.26mg, 36.1%).
Characterization of the Compound of formula Ia gave the compound shown in FIG. 21H NMR chart as shown in FIG. 313C NMR chart, as shown in HR-MS chart of FIG. 4, can be understood from FIGS. 2 to 4,1H NMR(300MHz,CDCl3)δ8.35(dd,J＝7.8,1.7Hz,1H),7.81–7.68(m,4H),7.61(dd,J＝7.8,1.7Hz,1H),7.35(dd,J＝7.0,1.5Hz,2H),7.30(d,J＝3.5Hz,2H),7.23(d,J＝1.5Hz,3H),7.20(d,J＝7.2Hz,2H),6.31(d,J＝14.1Hz,2H),4.41(t,J＝5.4Hz,4H),3.93(d,J＝5.2Hz,4H),3.65–3.57(m,4H),3.48–3.41(m,4H),3.28(s,6H),2.75(t,J＝6.1Hz,4H),2.41(s,5H),2.10–1.97(m,2H),1.45(s,13H).13C NMR(75MHz，CDCl3) δ 197.93,141.32,140.61,129.93,129.41,128.17,123.24,122.00,115.91,71.43,70.27,67.05,58.60,54.74,49.96,23.03,22.92,16.61 hrms (esi) m/z: calculated 779.38214, found 779.38263.
In another example of the method for producing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure, the compound of formula Ib in example 2 can be obtained by exchanging the compound of formula Ia obtained in example 4 with an anion resin. In addition, the chloride ion can also be replaced by iodide ion by anion resin exchange (compound of formula Ic in example 3).
Third, related test example
Test example 1
The main structures of the compounds Ib and ic are basically the same as that of Ia, but the contained anions are different, and the properties of the main structures are not influenced by the anions, so that the properties of the three compounds are similar, and the test example only takes Ia as an example to explain the application of the near-infrared reaction type fluorescent probe in cysteine detection.
(1) Ultraviolet and fluorescence spectrum experiments for selective detection of cysteine (Cys) by compound Ia
The compound ia obtained in example 4 was mixed with DMSO/PBS buffer (2:8, v/v, 10mM, pH 7.4) at 25 ℃ at ambient temperature to form a Probe solution (Probe), which was then subjected to uv spectroscopy and fluorescence spectroscopy. The result shows that the maximum absorption peak is at 780 nm; when the probe is excited at 780nm wavelength, the fluorescence emission of the probe is at 780-830nm, and the near-infrared emission characteristic is shown.
When GSH (glutathione) (Probe + GSH), Cys (cysteine) (Probe + Cys) and Hcy (homocysteine) (Probe + Hcy) are respectively added into the Probe solution, only Cys enables the maximum absorption peak of the Probe at 780nm to be reduced, and a new absorption peak appears at 565 nm; while the absorption peak of the probe at 780nm hardly changed when Hcy and GSH were added, respectively (as shown in FIG. 5).
Fluorescence spectrum tests were performed on Probe, Probe + GSH, Probe + Cys, and Probe + Hcy, respectively, using 565nm as the excitation wavelength, and the test results are shown in FIG. 6. Cys alone enhanced the fluorescence emission of the probe at 635nm (fluorescence intensity increased by about 35-fold), whereas Hcy and GSH were added under the same conditions, respectively, and the fluorescence emission intensity of the probe at 635nm was weaker. Fluorescence spectrum tests were performed on Probe, Probe + GSH, Probe + Cys, and Probe + Hcy, respectively, using 575nm as an excitation wavelength. The results show that Cys alone enhanced the fluorescence emission of the probe at 635nm (fluorescence intensity increased by about 78-fold). Under the same conditions, when Hcy and GSH were added separately, the fluorescence emission of the probe at 635nm was weak.
(2) Selective response experiment of compound Ia to Cys
Compound ia was incubated in DMSO/PBS buffer (2:8, v/v, 10mM, pH 7.4) with 40-fold equivalents of each amino acid or bioactive sulfur species for 120 minutes at 25 ℃ ambient temperature. The change in fluorescence intensity of compound Ia in the corresponding 22 solutions is shown in FIG. 7. As can be seen from FIG. 7, for different amino acids, for example, cysteine (Hcy), Glutathione (GSH), glutamic acid (Glu), leucine (Leu), glycine (Gly), isoleucine (Ile), phenylalanine (Phe), alanine (Ala), threonine (Thr), glutamine (Gln), asparagine (Asn), methionine (Met), serine (Ser), proline (Pro), tryptophan (Trp), lysine (Lys), arginine (Arg), histidine (His) and active sulfur (NaHS, S)2O3 2-) The compound Ia has obvious selective response characteristics to Cys, which shows that the compound Ia can be used as a novel fluorescent probe for selectively detecting Cys, and the experimental result is shown in figure 7, wherein in figure 7, the amino acids corresponding to each icon are as follows: (1) free; (2) cys; (3) hcy; (4) a GSH; (5) glu; (6) leu; (7) gly; (8) ile; (9) phe; (10) ala; (11) thr; (12) gln; (13) asn; (14) met; (15) ser; (16) pro; (17) try; (18) lys; (19) arg; (20) his; (21) NaHS; (22) s2O3 2-.(λex＝575nm，λem＝630nm)。
(3) Cys fluorescence imaging detection experiment of compound Ia in living cells and animals
The growth inhibition of HeLa cells by compound Ia was first tested using a cytotoxicity assay at a concentration of 5. mu.M. After the HeLa cells and the compound Ia are incubated together for 24 hours at the temperature of 37 ℃, the cell survival rate is more than 90 percent, and the result shows that the compound Ia does not generate obvious toxicity to the HeLa cells, so the compound Ia can be applied to a living cell experiment.
The ability of compound ia to detect cysteine (Cys) in living cells was then examined using a confocal laser imaging assay. As shown in FIG. 8, the cells were incubated with thiol scavenging reagent (NEM) for 30min to eliminate endogenous thiol amino acids produced in living cells, and then incubated with compound Ia for 30min (see FIGS. 8a-b), and the cells were moved to a confocal laser microscope to observe fluorescence emission. The results show that the Red Channel (Red Channel) has no fluorescence emission phenomenon, while the Purple Channel (Purple Channel) has stronger Purple fluorescence emission (as shown in figure 8c), which shows that only the autofluorescence emission of the compound Ia is generated in the cell, and the Red fluorescence emission phenomenon after the probe and cysteine (Cys) react does not occur. In cells without the pre-cleared thiol amino acids, after incubation with compound ia for 30 minutes, only weak fluorescence is observed in the purple Channel, and strong fluorescence emission occurs in the Red Channel (as shown in fig. 8d), which indicates that compound ia can react with thiol amino acids in living cells, and then Red fluorescence emission occurs. Meanwhile, after the cells in which endogenous thiol amino acids are removed in advance (co-incubated with NEM) are co-incubated with Glutathione (GSH), homocysteine (Hcy) and cysteine (Cys), respectively, the cells only incubated with cysteine (Cys) show a strong red fluorescence emission phenomenon in a red channel (as shown in FIGS. 8e-g), and the red fluorescence emission phenomena are consistent (as shown in FIGS. 8h-i, statistical information of purple fluorescence intensity and red fluorescence intensity of 1-7(a-g), respectively), which indicates that the red fluorescence emission phenomenon of the compound Ia in living cells is caused by cysteine (Cys). Therefore, the series of experiments prove that the compound Ia can realize the imaging detection of cysteine (Cys) in living cells.
This test example further examined the ability of compound Ia to selectively detect cysteine (Cys) in animals in a BALB/c nude mouse model (FIG. 9). A thiol scavenger (NEM) is intraperitoneally injected into the left lower part of BALB/c nude mouse to eliminate thiol amino acid generated in vivo, and then a cysteine (Cys) solution and a compound Ia solution are intraperitoneally injected into the same part. The results of the experiment show that strong red fluorescence emission appears after the probe injection, and the intensity of fluorescence emission decreases with time (40 min), indicating that part of cysteine (Cys) is cleared by NEM, and thus the amount of interaction with compound Ia decreases, resulting in a decrease in fluorescence intensity (NEM + in FIG. 9). While the red fluorescence intensity in control mice without prior injection of NEM (i.e., NEM-in FIG. 9) increased over time. This experiment further demonstrates the ability of compound Ia to detect cysteine (Cys) in nude mice. The area selected by the curve in fig. 9 is the lower left part of the mouse injected in the abdominal cavity, and the explanatory text therein expresses the areas injected in the abdominal cavity and imaged by fluorescence, and the displayed definition is not high due to the resolution problem of the original picture.
The results show that the compound shown in the formula I subjected to structural modification based on heptamethine cyanine is a selective cysteine near-infrared fluorescent probe with a novel structure.
Claims (10)
1. A near-infrared reaction type fluorescent probe with a heptamethine cyanine structure is characterized in that the structural formula of the near-infrared reaction type fluorescent probe with the heptamethine cyanine structure is shown as a formula I:
wherein, R is1、R2Each independently selected from C2～C8And X is chlorine, iodine or bromine.
2. The near-infrared reaction type fluorescent probe having a heptamethine cyanine structure of claim 1, wherein R is1、R2Is composed of
3. A method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as claimed in claim 1 or 2, comprising the steps of:
wherein, R is1、R2Each independently selected from C2～C8The ether chain of (a), wherein X is chlorine, iodine or bromine;
(1) reacting the compound shown in the formula III with sodium acetate to obtain a compound shown in a formula II;
(2) the compound shown in the formula II reacts with o-chlorobenzoyl chloride under the action of triethylamine.
4. The method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as in claim 3, wherein in the step (1), the molar ratio of the compound represented by the formula III to sodium acetate is (0.9-1.1): (2.9-3.1).
5. The method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as claimed in claim 3, wherein in the step (1), the reaction temperature is 70 ℃; the reaction time is 5-7 h.
6. The method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as in any one of claims 3 to 5, wherein in the step (2), the molar ratio of the compound represented by the formula II to the o-chlorobenzoyl chloride is (1.8-2.2): (2.8-3.2).
7. The method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as claimed in any one of claims 3 to 5, wherein in step (2), the amount ratio of the compound represented by formula II to triethylamine is (0.8 to 1.2) mol: (5-7) mL.
8. The method for preparing a near-infrared reaction type fluorescent probe having a heptamethine cyanine structure as in any one of claims 3 to 5, wherein in the step (2), the reaction comprises: reacting for 25-35 min at 0 ℃, and then reacting for 10-15 h at 20-30 ℃.
9. The use of the near-infrared reaction type fluorescent probe having a heptamethine cyanine structure according to claim 1 or 2 for detecting cysteine.
10. The use according to claim 9, said detection being carried out at the cellular level and at the animal level.
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| CN108641710A (en) * | 2018-05-22 | 2018-10-12 | 中国人民解放军第二军医大学 | A kind of fluorescence probe and its preparation method and application of detection protein sulphur sulfhydrylation |
| CN109370247A (en) * | 2018-09-05 | 2019-02-22 | 西安电子科技大学 | Conjugated chain functionalization benzindole heptamethine cyanine and application |
| CN109796779A (en) * | 2017-11-17 | 2019-05-24 | 中国科学院宁波材料技术与工程研究所 | A kind of preparation method of seven methines benzindole cyanine dye |
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| CN109796779A (en) * | 2017-11-17 | 2019-05-24 | 中国科学院宁波材料技术与工程研究所 | A kind of preparation method of seven methines benzindole cyanine dye |
| CN108641710A (en) * | 2018-05-22 | 2018-10-12 | 中国人民解放军第二军医大学 | A kind of fluorescence probe and its preparation method and application of detection protein sulphur sulfhydrylation |
| CN109370247A (en) * | 2018-09-05 | 2019-02-22 | 西安电子科技大学 | Conjugated chain functionalization benzindole heptamethine cyanine and application |
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