CN107311977B - Indole ethylene compound and preparation method and application thereof - Google Patents

Indole ethylene compound and preparation method and application thereof Download PDF

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CN107311977B
CN107311977B CN201710505345.6A CN201710505345A CN107311977B CN 107311977 B CN107311977 B CN 107311977B CN 201710505345 A CN201710505345 A CN 201710505345A CN 107311977 B CN107311977 B CN 107311977B
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rna
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卢宇靖
王聪
阎雁
郑园园
蔡森源
龙威
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Abstract

The invention provides an indole ethylene compound with a structure shown in a formula (I), wherein indole-3-formaldehyde is introduced into a quinoline position of the compound, and a specific substituent is selected as a parent body to obtain a series of indole ethylene compounds with the structure shown in the formula (I), and the result shows that a fluorescence image marked by the fluorescence probe has obvious green light distribution in a cytoplasm region and a nucleolus region, and clearly prompts that the probe can specifically image RNA in cytoplasm and nucleolus in living cells.

Description

Indole ethylene compound and preparation method and application thereof
Technical Field
The invention relates to the field of fluorescent probes, in particular to an indole ethylene compound and a preparation method and application thereof.
Background
Small molecule probes are probes developed for a specific target biomolecule or bio-ion, which can specifically interact with a specific target molecule and can be detected by a specific detection technique. Compared with the common detection technology, the probe technology has the advantages of high sensitivity, strong specificity, rapidness, accuracy and the like, and is suitable for molecular imaging and real-time monitoring.
RNA (ribonucleic acid) has important regulation and control functions in the whole process of growth, development and apoptosis of organisms; in the process of occurrence of many diseases, RNA plays a key role, and for example, the occurrence of malignant tumor is closely related to the abnormal expression of RNA. However, the development and application of RNA analysis and detection technology is slow compared to DNA analysis and detection technology.
The RNA fluorescent probe has the advantage of high-selectivity and specificity recognition of RNA in a complex solution system or in a living cell environment, and has good application prospect in the field of RNA analysis and detection. Currently, only RNAselect available from Invitrogen is commercially available as an RNA fluorescent probe. The fluorescent probe has some defects in practical imaging application, such as the defects of slow response speed with RNA, high phototoxicity, poor light stability and the like, and the application value of the fluorescent probe is influenced by the defects. Therefore, the development of RNA small molecule fluorescent probes with excellent performance has strong market value.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide an indole ethylene compound, and a preparation method and an application thereof, wherein the indole ethylene compound provided by the present invention has low toxicity, fast response speed and strong light stability when used as an RNA fluorescent probe.
The invention provides an indole ethylene compound which has a structure shown in a formula (I),
Figure BDA0001333493040000021
wherein R is1、R3Independently selected from H, halogen, hydroxyl, alkoxy of C1-C6, imino of C2-C15 and alkyl of C1-C6;
R4alkyl selected from C1-C6;
R2selected from piperidinyl, morpholinyl, pyrrolyl or NR5R6；
R5、R6Independently selected from H, -CH3、-CH2CH3、-CH2CH2OH、-CH2CH2CH2OH, formula (R-1), formula (R-2), formula (R-3), formula (R-4) or formula (R-5),
Figure BDA0001333493040000022
Figure BDA0001333493040000023
and R is5、R6Not H at the same time.
Preferably, said R is1Selected from H, F, Cl, Br, -OH and-OCH3、-N(CH3)2Or C1-C6 alkyl.
Preferably, R3 is selected from H, F, Cl, Br, -OH, -OCH3、-N(CH3)2Or C1-C6 alkyl.
Preferably, said R is4Selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl or tert-butyl.
Preferably, the indoleethylene compound is a compound shown in formula (I-a), formula (I-b), formula (I-c), formula (I-d) or formula (I-e),
Figure BDA0001333493040000024
Figure BDA0001333493040000031
the invention also provides a preparation method of the indole ethylene compound, which comprises the following steps:
the compound of formula (II), the compound of formula (III) and R2Reacting with-H to obtain the indoleethylene compound with the structure of formula (I);
Figure BDA0001333493040000032
wherein R is1、R3Independently selected from H, halogen, hydroxyl, alkoxy of C1-C6, imino of C2-C15 and alkyl of C1-C6;
R4alkyl selected from C1-C6;
R2selected from piperidinyl, morpholinyl, pyrrolyl or NR5R6；
R5、R6Independently selected from H, -CH3、-CH2CH3、-CH2CH2OH、-CH2CH2CH2OH, formula (R-1), formula (R-2), formula (R-3), formula (R-4) or formula (R-5),
Figure BDA0001333493040000033
Figure BDA0001333493040000034
and R is5、R6Not H at the same time.
The invention also provides application of the indole ethylene compound in preparation of a fluorescent probe for detecting RNA.
The invention also provides application of the indole ethylene compound in preparing a fluorescent probe for detecting RNA in aqueous solution.
The invention also provides application of the indole ethylene compound in preparing a fluorescent probe for detecting RNA in agarose gel or polyacrylamide gel.
The invention also provides application of the indole ethylene compound in preparation of a fluorescent probe for detecting RNA in cells.
Compared with the prior art, the indole ethylene compound has the structure shown in the formula (I), a series of indole ethylene compounds with the structure shown in the formula (I) are obtained by introducing indole-3-formaldehyde into quinoline positions and selecting specific substituent groups by taking the indole ethylene compounds as parent bodies, and experimental results show that fluorescence images marked by the fluorescent probe have obvious green light distribution in cytoplasm and nucleolus regions, clearly prompt that the probe can specifically image RNA in cytoplasm and nucleolus in living cells, and further prove that the fluorescent probe can be used for comparison tests with a traditional nucleolus fluorescent probe or cells subjected to RNA enzyme digestion experiments. Therefore, the fluorescent probe provided by the invention is a novel RNA selective recognition fluorescent probe molecule, and compared with a fluorescent probe with similar functions, the fluorescent probe provided by the invention has the characteristics of low biological toxicity, good membrane permeability, strong color development, good counterstain compatibility and strong light stability, indicates that the fluorescent probe has wide application as an RNA and nucleolus fluorescent probe, and is expected to be developed into a simple and visual biological detection reagent for RNA and nucleolus related physiological and pathological researches.
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FIG. 1 shows fluorescence spectra of a probe of a compound of formula (I-a) with five nucleic acids, da21, ds12, ds26 and RNA, at a concentration of 1: 1;
FIG. 2 is a curve fitted to the fluorescence data of probe titrations of st-DNA, ds26, RNA for compounds of formula (I-a);
FIG. 3 shows fluorescence spectra of probe-titrated RNA of a compound of formula (I-a);
FIG. 4 shows fluorescence spectra of compounds of formula (I-a) in which probe titrated RNA for C and (F-F)0)/F0A fitted curve;
FIG. 5 is a photograph showing the image of a cell obtained by counterstaining a probe of a compound of formula (I-a) with the dye DAPI on PC3 cells;
FIG. 6 shows cell staining experiments of the compound probe of formula (I-a) with RNase and DNase;
FIG. 7 shows the compound probe of formula (I-a) in gel electrophoresis with DNA and RNA.
Detailed Description
The invention provides an indole ethylene compound which has a structure shown in a formula (I),
Figure BDA0001333493040000051
wherein R is1、R3Independently selected from H, halogen, hydroxyl, alkoxy of C1-C6, imino of C2-C15 and alkyl of C1-C6;
R4alkyl selected from C1-C6;
R2selected from piperidinyl, morpholinyl, pyrrolyl or NR5R6；
R5、R6Independently selected from H, -CH3、-CH2CH3、-CH2CH2OH、-CH2CH2CH2OH, formula (R-1), formula (R-2), formula (R-3), formula (R-4) or formula (R-5),
Figure BDA0001333493040000052
Figure BDA0001333493040000053
and R is5、R6Not H at the same time.
According to the invention, said R1Preferably H, F, Cl, Br, -OH, -OCH3、-N(CH3)2Or C1-C6 alkyl, more preferably H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.
According to the invention, said R3Preferably H, F, Cl, Br, -OH, -OCH3、-N(CH3)2Or C1-C6 alkyl, more preferably H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.
According to the invention, said R4Preferably methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl or tert-butyl.
According to the invention, said R5、R6Preferably independently selected from-CH2CH2OH、-CH2CH2CH2OH, formula (R-1), formula (R-2), formula (R-3), formula (R-4) or formula (R-5),
Figure BDA0001333493040000054
wherein, in the structural formula:
Figure BDA0001333493040000055
the dotted line in (A) is the site of attachment to nitrogen.
More specifically, the indoleethylene compound is shown as a formula (I-1), a formula (I-2), a formula (I-3), a formula (I-4) or a formula (I-5),
Figure BDA0001333493040000061
the invention also provides a preparation method of the indole ethylene compound, which comprises the following steps:
the compound of formula (II), the compound of formula (III) and R2Reacting with-H to obtain the indoleethylene compound with the structure of formula (I);
Figure BDA0001333493040000062
wherein R is1、R3Independently selected from H, halogen, hydroxyl, alkoxy of C1-C6, imino of C2-C15 and alkyl of C1-C6;
R4alkyl selected from C1-C6;
R2selected from piperidinyl, morpholinyl, pyrrolyl or NR5R6；
R5、R6Independently selected from H, -CH3、-CH2CH3、-CH2CH2OH、-CH2CH2CH2OH, formula (R-1), formula (R-2), formula (R-3), formula (R-4) or formula (R-5),
Figure BDA0001333493040000063
Figure BDA0001333493040000071
and R is5、R6Not H at the same time.
According to the invention, the compounds of the formula (II), the compounds of the formula (III), R2-H reaction to obtain an indoleethylene compound; wherein, the definition of each group is the same as that of the group in the compound, the method for reaction is not specially required in the invention, and the method for reaction can be used in the reaction which is known in the field.
Among them, in the present invention, the compound of formula (II) is preferably prepared by the following method,
reacting a compound of formula (IV) with R4I, reacting to obtain a compound with a structure shown in a formula (II);
Figure BDA0001333493040000072
wherein R is3Selected from H, halogen, hydroxyl, alkoxy of C1-C6, imino of C2-C15 and alkyl of C1-C6.
The reaction conditions in the present invention are not particularly limited, and any reaction conditions known in the art can be used for the reaction.
The invention also provides application of the indole ethylene compound in preparation of a fluorescent probe for detecting RNA.
The invention also provides application of the indole ethylene compound in preparing a fluorescent probe for detecting RNA in aqueous solution.
The invention also provides application of the indole ethylene compound in preparing a fluorescent probe for detecting RNA in agarose gel or polyacrylamide gel.
The invention also provides application of the indole ethylene compound in preparation of a fluorescent probe for detecting RNA in cells.
The invention provides an indole ethylene compound with a structure shown in a formula (I), wherein indole-3-formaldehyde is introduced into a quinoline position of the compound, and a specific substituent is selected as a parent body to obtain a series of indole ethylene compounds with the structure shown in the formula (I). Therefore, the fluorescent probe provided by the invention is a novel RNA selective recognition fluorescent probe molecule, and compared with a fluorescent probe with similar functions, the fluorescent probe provided by the invention has the characteristics of low biological toxicity, good membrane permeability, strong color development, good counterstain compatibility and strong light stability, indicates that the fluorescent probe has wide application as an RNA and nucleolus fluorescent probe, and is expected to be developed into a simple and visual biological detection reagent for RNA and nucleolus related physiological and pathological researches.
The following will clearly and completely describe the technical solutions of the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: synthesis of Compound 2 (4-chloro-1, 2-dimethylquinoline)
0.2g (1.1236mmol) of 4-chloro-2-methylquinoline is weighed in a 25ml round-bottom flask, methyl iodide and sulfolane are added, the mixture is heated to 40-60 ℃, after 15 hours of reaction, the mixture is cooled, anhydrous ether is added and then the mixture is shaken, the mixture is filtered, the solid is washed for a plurality of times, the mixture is weighed after vacuum drying, thin layer chromatography initially shows that no by-product is generated, and 0.345g of pure product 2 is obtained, wherein the yield is 95.8%.
1H NMR(400MHz，DMSO)8.56(d，J＝8.4Hz，1H)，8.46(d，J＝8.3Hz，1H)，8.22(t，J＝8.1Hz，1H)，8.01(t，J＝7.9Hz，1H)，7.55(s，J＝7.4Hz，1H)，4.20(s，3H)，3.74(s，1H)，2.68(s，3H)。
Example 2: synthesis of Compound (I-a)
Weighing 0.319g (0.001mol) of compound 2, adding the weighed compound into a round-bottom flask containing 10ml of ethanol, adding 0.217g, namely 1.5 times of mol of indole-3-formaldehyde, stirring for 5 minutes at room temperature, adding 1ml of piperidine, reacting for 5 hours at 80 ℃, cooling to room temperature, adding 10ml of ethyl acetate into a solution after the reaction, performing suction filtration after shaking, washing precipitates with a small amount of ethanol, and performing vacuum drying to obtain 0.409g of compound (I-a) as a tan solid, wherein the structure is as follows, and the yield is 85.1%: 1H NMR (400MHz, DMSO)11.82(s, 1H), 8.27(d, J ═ 8.8Hz, 1H), 8.11(d, J ═ 8.3Hz, 1H), 8.05-7.98(m, 2H), 7.74(t, J ═ 7.6Hz, 1H), 7.63(dd, J ═ 11.8, 9.1Hz, 2H), 7.51-7.43(m, 2H), 7.25(t, J ═ 7.6Hz, 1H), 7.06(t, J ═ 7.5Hz, 1H), 6.98(s, 1H), 4.23(s, 3H), 3.76(s, 4H), 1.84(s, 4H), 1.78(s, 2H).
Figure BDA0001333493040000091
Example 3: synthesis of Compound (I-b)
The preparation method of this example is the same as that of example two except that morpholine is used instead of piperidine,the product is a reddish-brown solid, namely the compound (I-b), and the structural formula is as follows, and the yield is 78.2 percent:1H NMR(400MHz，DMSO)12.03(s，1H)，8.30(dd，J＝12.3，3.2Hz，2H)，8.18(t，J＝5.2Hz，3H)，8.02(t，J＝7.5Hz，1H)，7.73(t，J＝7.6Hz，1H)，7.58-7.51(m，2H)，7.43(d，J＝15.7Hz，1H)，7.32-7.24(m，2H)，4.27(s，3H)，3.97-3.88(m，4H)，3.72(d，J＝4.2Hz，4H)。
Figure BDA0001333493040000092
example 4: synthesis of Compound (I-c)
This example was carried out in the same manner as example II except that pyrolidine was used in place of piperidine, and the product was a yellowish brown solid, i.e., compound (I-c), which had the following structural formula and a yield of 76.8%:1H NMR(400MHz，DMSO)11.85(s，1H)，8.49(d，J＝8.5Hz，1H)，8.17-8.09(m，2H)，8.01(dt，J＝15.8，5.3Hz，3H)，7.65(t，J＝7.7Hz，1H)，7.51(d，J＝7.7Hz，1H)，7.33(d，J＝15.7Hz，1H)，7.28-7.20(m，2H)，7.00(s，1H)，4.11(s，3H)，4.01(s，4H)，2.06(s，4H)。
Figure BDA0001333493040000101
example 5: synthesis of Compound (I-d)
The procedure of this example is the same as in example two except that piperidine is replaced with 1- (2-aminoethyl) piperidinane, and the product is a tan solid, compound (I-d), having the following structural formula, in 78.2% yield:1H NMR(400MHz，DMSO)11.91(s，1H)，8.74(s，1H)，8.49(d，J＝8.1Hz，1H)，8.18(d，J＝8.8Hz，1H)，8.10(dd，J＝14.9，11.3Hz，3H)，8.03-7.97(m，1H)，7.73(t，J＝7.7Hz，1H)，7.53(d，J＝7.1Hz，1H)，7.38(d，J＝15.8Hz，1H)，7.30-7.21(m，2H)，7.15(s，1H)，4.13(d，J＝11.7Hz，3H)，3.78(s，2H)，2.66(d，J＝21.8Hz，2H)，1.50(s，4H)，1.38(d，J＝4.2Hz，2H)。
Figure BDA0001333493040000102
example 6: synthesis of Compound (I-e)
The procedure of this example is the same as in example two except that 3-amino-1-propanol is used in place of piperidine, and the product is a dark yellow solid which is compound (I-e) having the following formula in 81.2% yield:1H NMR(400MHz，DMSO)11.90(s，1H)，8.87(s，1H)，8.50(d，J＝8.2Hz，1H)，8.18(d，J＝8.8Hz，1H)，8.14-8.11(m，1H)，8.11-8.02(m，2H)，7.99(t，J＝7.7Hz，1H)，7.72(t，J＝7.6Hz，1H)，7.55-7.49(m，1H)，7.37(d，J＝15.8Hz，1H)，7.29-7.22(m，2H)，7.15(s，1H)，4.78(s，1H)，4.14(s，3H)，3.71(s，2H)，3.61(d，J＝4.4Hz，2H)，2.00-1.86(m，2H)。
Figure BDA0001333493040000111
example 7: fluorescence spectrum experiment of fluorescent probe of compound of formula (I-a) for different nucleic acid selectivity
The fluorescence intensity of 5mM stock solution of the compound of formula (I-a) was diluted to 5uM, and then different kinds of nucleic acids were added to measure the respective fluorescence intensities by a fluorescence spectrophotometer (slit width 10nm, scanning speed 400nm/min, excitation wavelength 455nm), and the results are shown in FIG. 1, in which FIG. 1 shows the fluorescence spectra of the probe of the compound of formula (I-a) and five kinds of nucleic acids, da21, ds12, ds26 and RNA, at a concentration of 1: 1; as can be seen from the figure, the fluorescent probe of the compound of formula (I-a) has the strongest fluorescence intensity after binding to RNA, but weaker fluorescence intensity for other double-stranded, single-stranded and G-quadruplex DNAs.
Nucleic acids for use in selective assays
Figure BDA0001333493040000112
Example 8: fluorescence titration experiment of fluorescent probe of compound of formula (I-a) on different nucleic acids
A5 mM stock solution of the compound was diluted to a concentration of 5. mu.M, placed in a fluorescence spectrophotometer, and the concentration of different nucleic acids in the solution was gradually increased, and fluorescence intensity measurement was performed. The measurement conditions were: the width of the slit is 10nm, the scanning speed is 400nm/min, and the excitation wavelength is 455 nm; the results are shown in FIG. 2, FIG. 2 is a curve fitted to the fluorescence data of probe titrations of st-DNA, ds26, RNA for compounds of formula (I-a); as can be seen from the figure, the fluorescence intensity is gradually increased along with the increase of the concentration of the nucleic acid, and the fluorescence intensity of the fluorescent probe combined with the RNA is stronger, which indicates that the fluorescent probe has obvious selectivity on the RNA.
Nucleic acids for use in fluorescence titration experiments
Figure BDA0001333493040000121
Example 9: determination of the detection Limit for RNA by the fluorescent Probe of the Compound of formula (I-a)
Diluting a compound stock solution with 5mM to a concentration of 5 mu M, scanning by a fluorescence spectrophotometer (slit width is 10nm, scanning speed is 200nm/min, excitation wavelength is 455nm), slowly adding RNA into the compound stock solution to saturate the compound stock solution, and detecting results are shown in figure 3, wherein figure 3 is a fluorescence spectrum of titrating RNA by a compound probe with the formula (I-a), wherein in the figure, curves of increasing the added RNA amount are sequentially formed from bottom to top, and as can be seen from figure 3, the fluorescence intensity is continuously increased along with the increase of RNA, which indicates that the compound can be combined with the RNA and generate stronger fluorescence.
Calculation formula of detection limit
LOD＝K×Sb/m
LOD (binding constant of Compound), m is the slope of the line drawn between concentration C and (F-F0)/F0, SbFor standard deviation of multiple measurements with instrument blanks, the value of K is usually taken to be 3 according to the International Union of pure and applied chemistry recommendations, where C is compared with (F-F)0)/F0The fitted curves are shown in FIG. 4, and FIG. 4 shows the fluorescence spectra of RNA titrated with the compound probe of formula (I-a) for C and (F-F)0)/F0A fitted curve; according to calculation, the LOD of the fluorescent probe of the compound shown in the formula (I-a) for RNA detection is 8 mug/L.
Example 10: cell imaging assay with fluorescent probes for Compounds of formula (I-a)
Cells were seeded in 6-well plates to a cell density of about 5000 cells/mL and then incubated at 37 ℃ with 5% CO2Culturing for 70h in the environment, then discarding the cell culture solution in the 6-well plate in the previous step, washing with precooled 1 × PBS for 3 times, then adding precooled pure methanol 1.5mL, placing for 2min at normal temperature in a dark place, finally discarding pure methanol and washing with precooled 1 × PBS for 3 times, adding 1mL of 5 mu M compound and placing for 20min, discarding the compound solution in the 6-well plate in the previous step, washing with precooled 1 × PBS for 3 times, adding 1mL of 1 mu M DAPI solution into the 6-well plate, placing for 2min at 37 ℃, then washing with precooled 1 × PBS for 6 times, soaking for 5min each time, and observing the cell staining condition under an inverted fluorescence microscope.
The results are shown in FIG. 5, FIG. 5 is a photograph of an image of a cell counterstained PC3 with the compound probe of formula (I-a) and the dye DAPI; as can be seen from the figure, the fluorescent probe of the invention acts on RNA in nucleus and cytoplasm, and simultaneously proves that the series of fluorescent ligands can image and detect RNA in a cell system.
Example 11: RNase and DNase cell imaging experiment of compound fluorescent probe of formula (I-a)
Cells were seeded in 6-well plates to a cell density of about 5000 cells/mL and then incubated at 37 ℃ with 5% CO2Culturing for 72h in the environment, discarding the cell culture solution in the 6-well plate, washing with pre-cooled 1 × PBS 3 times, adding pre-cooled pure methanol 1.5mL, standing at room temperature in dark for 2min, discarding pure methanol, washing with pre-cooled 1 × PBS 3 times, adding 1mL of 5uM compound, standing for 20min, discarding the compound solution in the 6-well plate, washing with pre-cooled 1 × PBS 3 times, adding 1mL of DNase, DNase-Free RNase solution into six-well plate, and mixing with 5% CO at 37 deg.C2Culturing for 3h, discarding the enzyme solution in the 6-well plate, washing with precooled 1 × PBS for 3 times, soaking for 5min each time, observing the cell staining condition under an inverted fluorescence microscope, and obtaining the result shown in figure 6, wherein figure 6 is a cell staining experiment of a compound probe of formula (I-a) and RNase and DNase, wherein, a is a fluorescence image without enzyme, b is fluorescence imaging after DNA enzyme is added, c is fluorescence imaging after RNA enzyme is added, the fluorescence intensity has no obvious change after DNA enzyme is added, and RN is performed after RNA enzyme is addedA is hydrolyzed, and the fluorescence almost completely disappears, which indicates that the fluorescent probe selectively binds to RNA in the cell and generates fluorescence.
Example 12: nucleic acid gel electrophoresis experiment
Preparing 1000ml of 1 XTAE electrophoresis buffer solution, respectively weighing 0.2g of BIOWESTAGROSE (Spanish agarose) and 20ml of electrophoresis buffer solution into a conical flask, heating and boiling by a microwave oven, putting the boiled gel into a water bath kettle at 65 ℃, adding 5mM of compound 2uL for shaking up when the gel is cooled to 65 ℃, pouring the gel, taking out a comb and a partition plate after the gel is cooled, putting the comb and the partition plate into an electrophoresis tank until the buffer solution is submerged for 1-2mM, preparing 5M DNA (sample loading buffer solution in mixed solution is 1 x), preparing 5mg/L RNA, respectively adding 10uL into a gel point sample hole, switching on the whole electrophoresis apparatus, running at 100V for 15min, taking out a gel block, and placing the gel block on the gel electrophoresis apparatus for observation.
The results are shown in FIG. 7, and FIG. 7 is a gel electrophoresis chart of the probe of the compound of formula (I-a) with DNA and RNA; as can be seen from the figure, the compound of formula (I-a) binds to RNA with a stronger fluorescence, whereas for double-stranded ds26, it fluoresces less strongly, indicating a better selectivity.
The above description of the embodiments is only intended to facilitate the understanding of the method of the invention and its core idea. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.

Claims (6)

1. An indoleethylene compound has a structure shown in a formula (I-a) or a formula (I-c),
Figure 801264DEST_PATH_IMAGE001
Figure 469006DEST_PATH_IMAGE002
formula (I-a) and formula (I-c).
2. A preparation method of indole ethylene compounds comprises the following steps:
the compound of formula (II), the compound of formula (III) and R2Reacting with-H to obtain the indoleethylene compound with the structure of formula (I);
Figure 137885DEST_PATH_IMAGE003
a compound of the formula (II),
Figure 114937DEST_PATH_IMAGE004
a compound of the formula (III),
Figure 869266DEST_PATH_IMAGE005
a compound of the formula (I),
wherein R is1、R3Independently selected from H;
R4is selected from methyl; r2Selected from piperidyl or pyrrolyl.
3. An application of the indole ethylene compound of claim 1 in preparing a fluorescent probe for detecting RNA.
4. An application of the indole ethylene compound of claim 1 in preparing a fluorescent probe for detecting RNA in an aqueous solution.
5. The use of the indoleethylene compound of claim 1 in the preparation of a fluorescent probe for detecting RNA in agarose gel or polyacrylamide gel.
6. An application of the indoleethylene compound of claim 1 in preparing a fluorescent probe for detecting RNA in a cell.
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