An organic compound biological probe and its preparation method and application

Abstract

The invention discloses an organic compound biological probe and a preparation method and application thereof, wherein the organic compound biological probe is In-AIE, and the method comprises: dissolving rutin in acetonitrile, and then adding 4-formyl pyridine and n-tributylphosphine for reflux reaction to obtain compound 1; dissolving 1,1,2-trimethyl-1H-benzo[e]indole and iodoethane in anhydrous acetonitrile for reflux to obtain compound 2; adding 2,4-dihydroxybenzaldehyde and anhydrous potassium carbonate to anhydrous acetonitrile and then dropping 1,4-dibromobutane for reflux reaction to obtain colorless oily compound 3; dissolving compound 2 and compound 3 in anhydrous ethanol for reflux reaction to obtain solid compound 4; dissolving compound 1 and compound 4 in anhydrous acetonitrile for reflux reaction and then purifying to obtain solid In-AIE. It can quickly enter cells within 1 min, has strong anti-photobleaching ability, and integrates multi-channel imaging, pH detection, mitochondrial targeting and photodynamic therapy.

Description

Technical Field

[0001] The present invention relates to the technical field of fluorescent probes, and in particular to an organic compound biological probe and a preparation method and application thereof. Background Art

[0002] The treatment of tumors has always been a difficult problem in scientific research and poses a great threat to human survival and development. Targeted therapy uses low-dose drugs to precisely act on the tumor site, avoiding toxic side effects on normal tissues and organs, and has the characteristics of high efficiency and low toxicity. Organelle targeted therapy can further improve the level of tumor treatment and reduce toxic side effects, so it is attracting more and more attention. Mitochondria, as the most special organelles in cells, not only provide chemical energy for cell life activities, but are also closely related to the occurrence, growth and spread of tumors. F16 is a small molecule with mitochondrial targeting function and selective inhibition of tumor growth screened by high-throughput computation, and plays an increasingly important role in the design and synthesis of mitochondrial targeted drugs.

[0003] Photodynamic therapy plays an important role in anti-cancer treatment. It has not only been used in clinical practice, but can also be used in combination with other therapies without affecting the subsequent use of other therapies. The main advantages of PDT are small side effects, less trauma and good selectivity. It consists of a photosensitizer, a light source and oxygen, where the light source depends on the spectral absorption range of the photosensitizer, so the photosensitizer is important. Given that mitochondria are the main site for aerobic respiration in living cells, mitochondrial-targeted photosensitizers help enhance phototoxicity and kill cancer cells. Many cyanine-derived dyes have been shown to have mitochondrial targeting and photodynamic therapy capabilities, and are easy to functionally design and synthesize, so they are ideal near-infrared (NIR) fluorophores for the preparation of biological diagnostic and therapeutic agents.

[0004] Organic fluorescent probes play an irreplaceable role in biological imaging, life activity detection, and qualitative or quantitative analysis of various biological substances. Compared with a large number of single-channel fluorescent imaging probes, multi-channel fluorescent probes are more easily applied to complex physiological environments, helping to obtain more comprehensive and multi-dimensional information, and greatly avoiding the possible error in the results caused by single-channel imaging.

[0005] Therefore, there is an urgent need to develop an organic compound bioprobe with multiple channels and its preparation method and application. Summary of the invention

[0006] The purpose of the present invention is to provide an organic compound biological probe and its preparation method and application. The organic compound biological probe of the present invention has aggregation-induced luminescence characteristics, pH sensitivity, multi-channel imaging, mitochondrial targeting, mitochondrial membrane potential sensitivity, strong anti-photobleaching ability and photodynamic therapy ability.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] In a first aspect of the present invention, an organic compound bioprobe is provided. The name of the organic compound bioprobe is In-AIE, and the structural formula is as follows:

[0009]

[0010] A second aspect of the present invention provides a method for preparing an organic compound biological probe, the method comprising:

[0011] Dissolve gramine in anhydrous acetonitrile, then add 4-formylpyridine and tributylphosphine to carry out reflux reaction, and then purify to obtain compound 1;

[0012] Dissolving 1,1,2-trimethyl-1H-benzo[e]indole and iodoethane in anhydrous acetonitrile to obtain a mixture, refluxing the mixture, cooling it to room temperature, performing solid-liquid separation, obtaining a solid, and purifying it to obtain compound 2;

[0013] 2,4-dihydroxybenzaldehyde and anhydrous potassium carbonate were added to anhydrous acetonitrile and heated under stirring, and then 1,4-dibromobutane was added dropwise for reflux reaction. After cooling to room temperature, solid-liquid separation was performed and the liquid was purified to obtain a colorless oily compound 3;

[0014] The compound 2 and the colorless oily compound 3 were dissolved in anhydrous ethanol, refluxed under argon protection, cooled to room temperature and purified to obtain a solid compound 4;

[0015] The compound 1 and the compound 4 are dissolved in anhydrous acetonitrile, and refluxed under argon protection. After cooling to room temperature, the mixture is purified to obtain a solid In-AIE, namely, the organic compound bioprobe.

[0016] The structural formulas of the compound 1, the compound 2, the colorless oily compound 3 and the compound 4 are shown below:

[0017]

[0018] Furthermore, the molar ratio of the gramine, the 4-formylpyridine and the n-tributylphosphine is 1:(1.1-1.5):(1.2-2).

[0019] Furthermore, the molar ratio of the 1,1,2-trimethyl-1H-benzo[e]indole to the ethyl iodide is 1:(1-5).

[0020] Furthermore, the molar ratio of the 2,4-dihydroxybenzaldehyde to the 1,4-dibromobutane is 1:(0.5-1).

[0021] Furthermore, the molar ratio of the compound 2 to the compound 3 is 1:(1-2).

[0022] Furthermore, the molar ratio of the compound 1 to the compound 4 is 1:(0.5-1).

[0023] In a third aspect of the present invention, use of the organic compound bioprobe in in vivo and in vitro multi-channel imaging is provided.

[0024] In a fourth aspect of the present invention, there is provided application of the organic compound biological probe in mitochondrial targeting and mitochondrial membrane potential detection.

[0025] In a fifth aspect of the present invention, application of the organic compound bioprobe in pH detection and photodynamic therapy is provided.

[0026] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0027] 1. The method for preparing the organic compound biological probe provided by the present invention has a simple synthesis and is conducive to commercial promotion and application.

[0028] 2. The organic compound biological probe In-AIE provided by the present invention has excellent application prospects, and has aggregation-induced emission characteristics, pH sensitivity, multi-channel imaging, mitochondrial targeting, mitochondrial membrane potential sensitivity, strong anti-photobleaching ability and photodynamic therapy ability. That is, In-AIE can quickly enter cells within 1 minute, has strong anti-photobleaching ability, and integrates multi-channel imaging, pH detection, mitochondrial targeting and photodynamic therapy; and can realize in vivo and in vitro near-infrared fluorescence imaging, which has very good application prospects for image-guided photodynamic therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description are some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0030] Figure 1 It is the synthesis of In-AIE;

[0031] Figure 2 is the H NMR spectrum of In-AIE;

[0032] Figure 3 is the mass spectrum of In-AIE;

[0033] Figure 4is the normalized UV absorption and solid fluorescence spectra of In-AIE;

[0034] Figure 5 This is the fluorescence spectra of In-AIE in different ratios of water and tetrahydrofuran;

[0035] Figure 6 This is the fluorescence spectra of In-AIE in solvents of different polarities;

[0036] Figure 7 is the fluorescence spectra of In-AIE in BF buffer solutions with different pH values;

[0037] Figure 8 The UV absorption graph of ABDA was used to detect the ability of In-AIE to produce singlet oxygen under white light irradiation;

[0038] Fig. 9 This is the MTT result graph of the phototoxicity and dark toxicity of In-AIE in different cell lines; Fig. 9 There are 4 columns at different concentrations, which are HeLa cells, COS-7 cells, HeLa cells irradiated with white light, and COS-7 cells irradiated with white light;

[0039] Fig.10 This is the intracellular multi-channel imaging of In-AIE;

[0040] Fig.11 is the intracellular mitochondrial localization of In-AIE;

[0041] Fig.12 It is the intracellular mitochondrial membrane potential detection of In-AIE;

[0042] Fig.13 It is the anti-photobleaching experiment of In-AIE;

[0043] Fig.14 This is the intracellular fluorescence imaging of In-AIE at different pH;

[0044] Fig.15 It is a test of the level of intracellular reactive oxygen species after co-incubation of In-AIE and cells with or without illumination, including flow cytometry and confocal imaging. Fig.15 A is the flow cytometry diagram. Fig.15 B is the confocal imaging image;

[0045] Fig.16 This is a flow cytometric test diagram of cell apoptosis with or without illumination after In-AIE and HeLa cells were co-incubated for 24 hours;

[0046] Fig.17 These are the confocal images of In-AIE and HeLa cells after being co-incubated for different time periods with or without illumination;

[0047] Fig.18 This is the near-infrared fluorescence imaging of In-AIE in PC-3 tumor-bearing mice. DETAILED DESCRIPTION

[0048] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0049] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0050] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or obtained by existing methods.

[0051] The embodiment of the present invention provides an organic compound biological probe, and the overall idea is as follows:

[0052] According to a typical embodiment of the present invention, an organic compound bioprobe is provided. The name of the organic compound bioprobe is In-AIE, and the structural formula is as follows:

[0053]

[0054] According to a typical embodiment of the present invention, a method for preparing an organic compound biological probe is provided, wherein the reaction formula is as follows: Figure 1 As shown, the method includes:

[0055] Step S1, dissolving gramine in anhydrous acetonitrile, then adding 4-formylpyridine and n-tributylphosphine to carry out reflux reaction, and then purifying to obtain compound 1;

[0056] As an optional embodiment, in step S1, the molar ratio of the gramine, the 4-formylpyridine and the n-tributylphosphine is 1:(1.1-1.5):(1.2-2). This molar ratio range is conducive to complete reaction.

[0057] Step S2, dissolving 1,1,2-trimethyl-1H-benzo[e]indole and iodoethane in anhydrous acetonitrile to obtain a mixture, refluxing the mixture, cooling it to room temperature, performing solid-liquid separation, obtaining a solid, and purifying it to obtain compound 2;

[0058] The molar ratio of the 1,1,2-trimethyl-1H-benzo[e]indole to the ethyl iodide is 1:(1-5). This molar ratio range is conducive to complete reaction.

[0059] Step S3, adding 2,4-dihydroxybenzaldehyde and anhydrous potassium carbonate to anhydrous acetonitrile and heating under stirring, then adding 1,4-dibromobutane dropwise to carry out reflux reaction, cooling to room temperature, separating the solid and liquid, and purifying the liquid to obtain a colorless oily compound 3;

[0060] The molar ratio of the 2,4-dihydroxybenzaldehyde to the 1,4-dibromobutane is 1:(0.5-1). The molar ratio is in the range that is favorable for the reaction.

[0061] Step S4, dissolving the compound 2 and the colorless oily compound 3 in anhydrous ethanol, performing reflux reaction under argon protection, cooling to room temperature and then purifying to obtain a solid compound 4;

[0062] The molar ratio of the compound 2 to the compound 3 is 1:(1-2). This molar ratio range is conducive to complete reaction.

[0063] Step S5, dissolving the compound 1 and the compound 4 in anhydrous acetonitrile, performing reflux reaction under argon protection, cooling to room temperature and then purifying to obtain solid In-AIE, namely the organic compound bioprobe.

[0064] The molar ratio of the compound 1 to the compound 4 is 1:(0.5-1). This molar ratio range is conducive to complete reaction.

[0065] There is no strict order relationship among the steps S1-S5, and the order can be adjusted appropriately within the protection scope of the present invention.

[0066] The present invention provides an organic compound biological probe and a preparation method thereof, which breaks the conventional thinking of the prior art and adopts a simple four-step method to synthesize the organic compound biological probe In-AIE. In-AIE can quickly enter cells within 1 minute, has strong resistance to photobleaching, and integrates multi-channel imaging, pH detection, mitochondrial targeting and photodynamic therapy. It is understood that In-AIE is the first organic biological probe that can achieve multi-channel imaging of green, yellow, red, and NIR fluorescence in cells, and can achieve near-infrared imaging both in vivo and in vitro. In addition, the probe can not only locate the mitochondria in cells well, but also has the ability of photodynamic therapy. Under white light irradiation, singlet oxygen ( 1 O2), increasing the level of cellular reactive oxygen species (ROS) and inducing cell apoptosis.

[0067] The following will describe in detail an organic compound biological probe and its preparation method and application in combination with examples and experimental data.

[0068] Example 1: Organic synthesis of In-AIE

[0069] like Figure 1 As shown, 2 mmol of gramine was dissolved in 15 mL of acetonitrile, and 2.5 mmol of 4-formylpyridine and 3 mmol of n-tributylphosphine were added at room temperature. After reflux reaction for 22 hours, the temperature was lowered and rotary evaporation was performed. Compound 1 was obtained by purification using a silica gel column with a yield of 49%.

[0070] 5 mmol of 1,1,2-trimethyl-1H-benzo[e]indole and 10 mmol of iodoethane were dissolved in 10 mL of anhydrous acetonitrile. The mixture was refluxed for 12 h. After cooling to room temperature, the solid was filtered out and purified using a silica gel column to obtain compound 2 with a yield of 85%.

[0071] 2 mmol 2,4-dihydroxybenzaldehyde and 2.1 mmol anhydrous potassium carbonate were added to 25 mL anhydrous acetonitrile and heated under stirring. 2 mmol 1,4-dibromobutane was added dropwise and refluxed for 5 h. After cooling to room temperature, the filtrate was filtered and the solvent was evaporated by rotary evaporator. The colorless oily compound 3 was separated and purified by silica gel column with a yield of 67%.

[0072] 1 mmol of compound 2 and 1 mmol of compound 3 were dissolved in 20 mL of anhydrous ethanol and refluxed for 12 h under argon protection. After the system was cooled to room temperature, the solvent was evaporated under reduced pressure and passed through a silica gel column to obtain solid compound 4 with a yield of 86%.

[0073] 0.52 mmol of compound 1 and 0.5 mmol of compound 4 were dissolved in 15 mL of anhydrous acetonitrile and refluxed for 24 h under argon protection. After the system was cooled to room temperature, the solvent was evaporated under reduced pressure and purified by silica gel column to obtain solid In-AIE with a yield of 46%. The H NMR spectrum is Figure 2 , the mass spectrum is Figure 3 .

[0074] Example 2: Characterization of In-AIE

[0075] 1. Take 10 μM In-AIE in Example 1 and perform UV spectrum test and solid fluorescence test. The results are as follows: Figure 4 shown by Figure 4 It can be seen that In-AIE has a wide visible absorption range and NIR solid luminescence ability; the proportion of tetrahydrofuran in water is continuously increased, and the fluorescence intensity change of 10μM probe is detected. The results are as follows Figure 5 shown by Figure 5 It can be seen that In-AIE has induced luminescence characteristics;

[0076] 2. Detect the fluorescence properties of the probe in solvents of different polarities. The results are as follows: Figure 6 shown by Figure 6 It can be seen that the polarity of the solvent does not cause a significant shift in the fluorescence peak of In-AIE, and the fluorescence emission range of In-AIE in organic solvents is concentrated in the visible region;

[0077] 3. Detect the change of fluorescence intensity of the probe at different pH values. Figure 7 shown by Figure 7 It can be seen that within the physiological pH range, the fluorescence intensity of In-AIE increases significantly with the increase of pH;

[0078] 4. 9,10-Anthracenediyl-bis(methylene)dimalonic acid (ABDA) was used to detect the ability of 10 μM probe to generate singlet oxygen under white light irradiation. The results are as follows: Figure 8 shown by Figure 8 It can be seen that In-AIE has the ability to generate singlet oxygen under white light irradiation.

[0079] Example 3: Phototoxicity and dark toxicity of In-AIE

[0080] HeLa cells and COS-7 cells were used as research objects. Different concentrations of In-AIE in Example 1 were incubated with cells for 24 hours for cytotoxicity MTT test. The group treated with white light for 0.5 hours after adding the probe for 2 hours was used as the phototoxicity test group, and the group not treated with white light was used as the dark toxicity test group. The MTT results are shown in Figure 2. Fig. 9 shown.

[0081] Depend on Fig. 9 It can be seen that the phototoxicity of In-AIE in HeLa cells is much stronger than that in COS-7 cells, and the dark toxicity increases with the increase of concentration.

[0082] Example 4: Cell imaging test of In-AIE

[0083] 5 μM of In-AIE in Example 1 was taken and incubated with HeLa cells for 2 h, and then fluorescence confocal imaging was performed. Fig.10 This indicates that In-AIE has green, yellow, red and near-infrared fluorescence imaging capabilities. Mitochondrial green dye was added and incubated for 15 min. Fig.11 This shows that In-AIE has good mitochondrial targeting ability.

[0084] HeLa cells were mixed with 1 μL PBS, 10 ng mL -1 After oligomycin and 20 μM CCCP were co-incubated for 0.5 h, they were co-incubated with 1 μM In-AIE in Example 1 for 0.5 h. The fluorescence confocal imaging results are shown in FIG. Fig.12As shown, it shows that changes in mitochondrial membrane potential will affect the fluorescence intensity of In-AIE in cells, and In-AIE has the ability to detect mitochondrial membrane potential.

[0085] 5 μM of In-AIE in Example 1 was taken and incubated with HeLa cells for 2 h, and then laser scanning was performed continuously for 300 times. The fluorescence confocal images and signal intensity changes of different imaging channels were collected and analyzed. The results are as follows: Fig.13 As shown, it indicates that In-AIE has anti-photobleaching ability.

[0086] Example 5: Intracellular pH detection by In-AIE

[0087] After 1 μM In-AIE in Example 1 was incubated with HeLa cells for 2 h, the culture medium was replaced with BF buffer solutions with pH values ​​of 5, 6, 7, and 8, respectively. A laser confocal imager was used to detect the effect of pH on the green, red, and near-infrared multi-channel imaging of the probe in cells and to analyze the fluorescence intensity. The results are shown in FIG. Fig.14 shown.

[0088] Depend on Fig.14 It can be seen that changing the cell pH will affect the fluorescence intensity of In-AIE in the cell, and In-AIE has the ability to detect changes in intracellular pH by multi-channel imaging.

[0089] Example 6: Study on the intracellular phototoxicity mechanism of In-AIE

[0090] Different concentrations of In-AIE in Example 1 were co-incubated with HeLa cells for 2 h, then irradiated with white light for 0.5 h, 1 μM dihydroethidium (DHE) reactive oxygen species detection probe was added and incubated for another 0.5 h. After the cells were collected, flow cytometry and confocal imaging were performed. The results are shown in FIG. Fig.15 shown by Fig.15 It can be seen that the combined treatment of In-AIE and light leads to an increase in the level of reactive oxygen species in cells;

[0091] After co-incubation of HeLa cells with different concentrations of In-AIE in Example 1 for 24 hours, the cell status was detected using the Annexin V-FITC / PI cell apoptosis detection kit. The "In-AIE with white light" group was incubated with the probe and cells for 2 hours and then irradiated with white light for 0.5 hours, while the "In-AIE without white light" group was not treated with any white light. The test results are shown in Figure 2. Fig.16 shown by Fig.16 It can be seen that the combined treatment of In-AIE and light induces cell apoptosis;

[0092] After 5 μM In-AIE in Example 1 was co-incubated with HeLa cells for 2 h or 24 h, the group that was irradiated with white light for 0.5 h after the incubation time reached 2 h was the "+white light" group. After co-incubation with 1 mg / mL Hoechst 33342 for 5 min, the multi-channel imaging of the cells was observed using a laser confocal microscope. The results are as follows: Fig.17 shown by Fig.17 It can be seen that as the incubation time of In-AIE and cells increases from 2h to 24h, the multi-channel fluorescence intensity in the cells does not decrease significantly, while the cells shrink slightly. The combined treatment of In-AIE and light causes the cells to shrink and deform further, and the cell nucleus breaks into fragments, representing cell apoptosis.

[0093] Example 7: In-AIE near-infrared imaging of tumor-bearing mice

[0094] PC-3 tumor-bearing mice were used as animal models for in vivo imaging tests. 5 mg / mL of In-AIE in Example 1 was injected intratumorally. Near-infrared imaging tests were performed at different time points using a small animal in vivo imaging system. Near-infrared fluorescence signals at 710 nm were collected under 561 nm excitation light. The results are shown in Figure 2. Fig.18 shown.

[0095] Depend on Fig.18 It can be seen that In-AIE can realize in vivo near-infrared fluorescence imaging. The fluorescence intensity reaches the maximum value 8 hours after injection, and the near-infrared fluorescence signal basically disappears after 72 hours.

[0096] In summary, the organic compound bioprobe of the present invention has aggregation-induced luminescence properties, pH sensitivity, multi-channel imaging, mitochondrial targeting, mitochondrial membrane potential sensitivity, strong anti-photobleaching ability and photodynamic therapy ability.

[0097] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0098] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0099] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. An organic compound biological probe, characterized in that: The name of the organic compound bioprobe is In-AIE, and the structural formula is as follows: 。 2. A method for preparing an organic compound biological probe according to claim 1, characterized in that: The method comprises: Dissolve gramine in anhydrous acetonitrile, then add 4-formylpyridine and n-tributylphosphine to carry out reflux reaction, and then purify to obtain compound 1; Dissolving 1,1,2-trimethyl-1H-benzo[e]indole and iodoethane in anhydrous acetonitrile to obtain a mixture, refluxing the mixture, cooling it to room temperature, performing solid-liquid separation, obtaining a solid, and purifying it to obtain compound 2; 2,4-dihydroxybenzaldehyde and anhydrous potassium carbonate were added to anhydrous acetonitrile and heated under stirring, and then 1,4-dibromobutane was added dropwise for reflux reaction. After cooling to room temperature, solid-liquid separation was performed and the liquid was purified to obtain a colorless oily compound 3; The compound 2 and the colorless oily compound 3 were dissolved in anhydrous ethanol, and refluxed under argon protection, and then cooled to room temperature and purified to obtain a solid compound 4; The compound 1 and the compound 4 are dissolved in anhydrous acetonitrile, refluxed under argon protection, cooled to room temperature and purified to obtain solid In-AIE, i.e., the organic compound bioprobe; The structural formulas of the compound 1, the compound 2, the colorless oily compound 3 and the compound 4 are shown below: ； ； ； 。 3. The method for preparing an organic compound biological probe according to claim 2, characterized in that: The molar ratio of the gramine, the 4-formylpyridine and the n-tributylphosphine is 1:(1.1-1.5):(1.2-2).

4. The method for preparing an organic compound biological probe according to claim 2, characterized in that: The molar ratio of the 1,1,2-trimethyl-1H-benzo[e]indole to the ethyl iodide is 1:(1-5).

5. The method for preparing an organic compound biological probe according to claim 2, characterized in that: The molar ratio of the 2,4-dihydroxybenzaldehyde to the 1,4-dibromobutane is 1:(0.5-1).

6. The method for preparing an organic compound biological probe according to claim 2, characterized in that: The molar ratio of the compound 2 to the compound 3 is 1:(1-2).

7. The method for preparing an organic compound biological probe according to claim 2, characterized in that: The molar ratio of the compound 1 to the compound 4 is 1:(0.5-1).

8. Use of the organic compound bioprobe according to claim 1 in the preparation of products for in vivo and in vitro multi-channel imaging.

9. Use of the organic compound biological probe according to claim 1 in the preparation of products for mitochondrial targeting and mitochondrial membrane potential detection.

10. Use of the organic compound bioprobe according to claim 1 in preparing products for pH detection and photodynamic therapy.

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