Near-infrared fluorescent probe molecules for kidney clearance as well as preparation method and application of near-infrared fluorescent probe molecules

Abstract

The invention discloses a near-infrared fluorescent probe molecule for kidney clearance as well as a preparation method and application of the near-infrared fluorescent probe molecule. The chemical structural formula of the near-infrared fluorescent probe molecule is # imgabs0 #, wherein X is S or Se. The molecule adopts a D-pi-A-pi-D structural design, and a triphenylamine group is used as an electron donor and a rotor unit, so that a molecular skeleton is obviously distorted, and the molecule is endowed with an aggregation-induced emission (AIE) characteristic. The introduction of the ethylene glycol fragment can be used for regulating and controlling the hydrophilicity of the probe. The strong electron acceptor has excellent electron withdrawing capability, can enhance the intramolecular charge transfer effect, reduces the molecular band gap, and extends the absorption / emission wavelength to a near-infrared region. The kidney clearance near-infrared fluorescent probe molecule provided by the invention has kidney clearance capability, is simple and convenient in synthesis route, low in cost and excellent in biocompatibility, and is suitable for realizing early diagnosis of acute kidney injury caused by various causes through near-infrared two-region fluorescence imaging.

Description

Technical Field

[0001] The present invention relates to the technical field of fluorescent probe molecules, and in particular to a class of renal clearance near-infrared fluorescent probe molecules, their preparation methods and applications. Background Art

[0002] Acute Kidney Injury (AKI) is a severe clinical syndrome characterized by a rapid decline in renal function, mainly manifested as an increase in serum creatinine levels and a decrease in urine output. AKI has the characteristics of high incidence and high mortality. More than 50% of patients need to be transferred to the intensive care unit during hospitalization, bringing a heavy economic burden to patients and society. The causes of AKI are complex and diverse, and can be classified into three categories according to semi-anatomical classification: prerenal, renal, and postrenal. Currently, the clinical diagnosis of AKI mainly relies on serum biomarkers such as serum creatinine (sCr) and blood urea nitrogen (BUN). However, these indicators have a lag in reflecting renal function damage and are difficult to achieve early and accurate diagnosis. In addition, although renal biopsy is considered the gold standard for AKI diagnosis, its invasive operation may cause tissue bleeding and damage, limiting its wide application in clinical practice.

[0003] Traditional imaging techniques such as ultrasound (US), computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET) have their respective application values in the monitoring of kidney diseases, but there are also many limitations, including insufficient tissue contrast, high cost, long time consumption, radiation exposure, potential nephrotoxicity, and contrast agent allergic reactions. In contrast, the optical imaging technique mediated by near-infrared organic fluorescent molecules has the advantages of high sensitivity, real-time non-invasiveness, high spatio-temporal resolution, etc., and especially shows better tissue penetration ability in the near-infrared second window (NIR-II, 1000 - 1700 nm). However, currently, most NIR-II probes have strong hydrophobicity. After encapsulation with amphiphilic polymers, their sizes are relatively large (tens to hundreds of nanometers), and they are easily retained in the liver or spleen and difficult to be metabolized and cleared by the kidneys. Summary of the Invention

[0004] The technical problem to be solved by the present invention is that most of the existing NIR-II fluorescent probes are polymers with relatively large sizes and cannot be metabolized by the kidneys, making it difficult to conduct pharmacokinetic studies and achieve the diagnosis of kidney diseases; in addition, the development of water-soluble near-infrared fluorescent probe molecules is still in its infancy, and there is a lack of a general synthesis method. The present invention aims to develop a new method for preparing water-soluble NIR-II fluorescent probes and expand new applications.

[0005] The technical solution adopted by the present invention to solve this technical problem is: by introducing an ethylene glycol fragment, a novel water-soluble NIR-II fluorescent small molecule probe with renal clearance ability is constructed, and its specific chemical structural formula is:

[0006] Among them, X is S or Se.

[0007] The present invention also provides a method for preparing a renal clearance near-infrared fluorescent probe molecule, and the method includes the following steps:

[0008] Add compound III, an S-containing receptor, Pd(PPh3)4, and a carbonate into a mixed solvent of toluene and water, and heat and stir the reaction under the protection of an inert gas. After the reaction is completed, cool to room temperature, and successively perform dilution, extraction, and drying treatments to obtain a renal clearance near-infrared fluorescent probe molecule;

[0009] Or add compound III, a Se-containing receptor, Pd(PPh3)4, and a carbonate into a mixed solvent of toluene and water, and heat and stir the reaction under the protection of an inert gas. After the reaction is completed, cool to room temperature, and successively perform dilution, extraction, and drying treatments to obtain a renal clearance near-infrared fluorescent probe molecule;

[0010] The structural formula of the compound III is:

[0011] Optionally, in the method for preparing the renal clearance near-infrared fluorescent probe molecule, the molar ratio of the compound III, the S-containing receptor, Pd(PPh3)4, and the carbonate is 30:10:1:300.

[0012] Optionally, in the method for preparing the renal clearance near-infrared fluorescent probe molecule, the molar ratio of the compound III, the Se-containing receptor, Pd(PPh3)4, and the carbonate is 30:10:1:300.

[0013] Optionally, in the method for preparing the renal clearance near-infrared fluorescent probe molecule, the temperature of the heating and stirring is 90-110°C, and the time of the heating and stirring is 12-24 h.

[0014] Optionally, in the method for preparing the renal clearance near-infrared fluorescent probe molecule, the S-containing receptor is 4,8-dibromo-6-(2-ethylhexyl)-[1,2,5]thiadiazolo[3,4-f]benzotriazole; the carbonate is selected from one of potassium carbonate, sodium carbonate, and cesium carbonate.

[0015] Optionally, in the method for preparing the renal clearance near-infrared fluorescent probe molecule, the Se-containing receptor is 4,8-dibromo-6-(2-ethylhexyl)-[1,2,5]selenadiazolo[3,4-f]benzotriazole.

[0016] Second aspect, an application of the renal-clearing near-infrared fluorescent probe molecule as described above in the early diagnosis of acute kidney injury models.

[0017] Optionally, an application of the renal-clearing near-infrared fluorescent probe molecule in the preparation of early diagnosis reagents for acute kidney injury, wherein the acute kidney injury models include: unilateral renal ischemia-reperfusion models, cisplatin-induced acute kidney injury models, and unilateral ureteral ligation models.

[0018] Beneficial effects: The novel water-soluble NIR-II fluorescent probe small molecule provided by the present invention has a D-π-A-π-D (D is an electron donor, A is an electron acceptor) structure, and the introduction of polyethylene glycol fragments endows the material with adjustable hydrophilicity. The obtained NIR-II fluorescent probe small molecule can self-assemble into nanoparticles in the aqueous phase, and it can be observed through NIR-II fluorescence imaging that this material can achieve efficient renal clearance. Description of the Drawings

[0019] Figure 1 is the synthetic route diagram of the novel water-soluble NIR-II organic small molecule probe provided in Example 1 of the present invention;

[0020] Figure 2 is the 1H nuclear magnetic resonance spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0021] Figure 3 is the 13C nuclear magnetic resonance spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0022] Figure 4 is the 1H nuclear magnetic resonance spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0023] Figure 5 is the 13C nuclear magnetic resonance spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0024] Figure 6 is the HRMS mass spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention;

[0025] Figure 7 is the HRMS mass spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention;

[0026] Figure 8It is the ultraviolet absorption and fluorescence emission spectrogram of the PEG-TBS and PEG-TBSe materials prepared in Example 1 of the present invention, indicating that the prepared compounds have near-infrared absorption / emission effects.

[0027] Figure 9 It is the fluorescence signal change diagram of different concentrations of the PEG-TBS and PEG-TBSe materials prepared in Example 1 of the present invention. It can be seen that the synthesized materials have the property of aggregation-induced emission.

[0028] Figure 10 It is the clearance rate diagram of the PEG-TBSe material prepared in Example 1 of the present invention over time. From Figure 10 it can be seen that PEG-TBSe has excellent renal clearance characteristics.

[0029] Figure 11 It is the cell survival rate diagram of the PEG-TBSe material prepared in Example 1 of the present invention co-incubated with HK-2 cells (human renal tubular epithelial cells) and SV-HUC-1 cells (human ureteral epithelial immortalized cells) for 72 hours. From Figure 11 it can be seen that PEG-TBSe has excellent cell compatibility.

[0030] Figure 12 It is the fluorescence imaging diagram after injecting the PEG-TBSe material prepared in Example 1 of the present invention into a unilateral renal ischemia-reperfusion mouse model. From Figure 12 it can be seen that PEG-TBSe can accurately identify the damaged left kidney and can be used as a reagent for the early diagnosis of pre-renal acute kidney injury.

[0031] Figure 13 It is the fluorescence imaging diagram after injecting the PEG-TBSe material prepared in Example 1 of the present invention into a cisplatin-induced acute kidney injury mouse model. From Figure 13 it can be seen that PEG-TBSe can accurately identify the damaged bilateral kidneys and can be used as a reagent for the early diagnosis of renal acute kidney injury.

[0032] Figure 14 It is the fluorescence imaging diagram after injecting the PEG-TBSe material prepared in Example 1 of the present invention into a unilateral ureteral ligation mouse model. From Figure 14 it can be seen that PEG-TBSe can accurately identify the damaged bilateral kidneys and can be used as a reagent for the early diagnosis of post-renal acute kidney injury. Detailed implementation manner

[0033] The present invention provides a method for preparing a water-soluble NIR-II fluorescent probe molecule with renal clearance ability and its application. To make the objectives, technical solutions and advantages of the present invention clearer and more definite, the present invention will be further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. The experimental methods without specific conditions noted in the following examples are carried out according to conventional methods and conditions, or are selected according to the product specifications. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

[0034] Currently, most NIR-II diagnostic and therapeutic probes encapsulated by amphiphilic polymers are easily retained in the liver or spleen due to their relatively large sizes (tens to hundreds of nm). Only those probes smaller than the renal filtration threshold (about 6 nm) can be cleared by renal metabolism. Therefore, designing NIR-II diagnostic and therapeutic probes with renal clearance ability is of great significance for achieving non-invasive in-situ diagnosis of AKI.

[0035] The embodiments of the present invention provide a class of water-soluble near-infrared fluorescent probe molecules with renal clearance ability, and their chemical structural formula is:

[0036]

[0037] This molecule adopts a D-π-A-π-D structure design. The triphenylamine group serves as an electron donor and rotor unit, causing the molecular skeleton to exhibit significant distortion, thereby endowing it with aggregation-induced emission (AIE) characteristics. The introduction of the ethylene glycol fragment can be used to regulate the hydrophilicity of the probe. The strong electron acceptor has excellent electron-withdrawing ability, which can enhance the intramolecular charge transfer effect, reduce the molecular bandgap, and extend the absorption / emission wavelength to the near-infrared region. The size of this organic small molecule probe is 5.0 - 5.5 nm and can be cleared by renal metabolism.

[0038] The following specific preparation examples are used to further explain and illustrate the above technical solutions provided by the present invention.

[0039] Example 1

[0040] As Figure 1 shown, the synthesis of compounds PEG-TBS and PEG-TBSe includes the following steps:

[0041] Step 1: Synthesis of compound 1

[0042] 4-Bromo-N,N-bis(4-methoxyphenyl)aniline (10 g, 25.8 mmol) was added to a solution of DCM (50 mL), and boron tribromide (18.829 g, 76 mmol) was slowly added thereto at 0 °C. The resulting mixture was stirred overnight, and water was added to cool the reaction. After extraction with DCM three times, it was dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain a green solid (8.5 g, 93%). 1 H NMR (600 MHz, DMSO-d6) δ 9.39 (s, 2H), 7.28–7.25 (m, 2H), 6.97–6.93 (m, 4H), 6.77–6.74 (m, 4H), 6.60–6.58 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 154.92, 148.86, 138.50, 131.93, 127.92, 119.81, 116.79, 110.18.

[0043] Step 2: Synthesis of Compound 2

[0044] In a 50 mL two-necked round-bottom flask, 1 (0.1775 g, 1 mmol) and MeCN (20 mL) were mixed, and then mPEG420-OTs (0.4951 g, 3 mmol) and K2CO3 (0.2070 g, 1.5 mmol) were added. The mixture was heated to 85 °C and stirred overnight. After completion of the reaction, the solvent was removed by rotary evaporation. Chromatographic purification (PE:DCM = 10:1) gave a colorless transparent oily liquid (0.245 g, 94%). 1 H NMR (500 MHz, Chloroform-d) δ 7.18–7.14 (m, 2H), 6.94–6.90 (m, 4H), 6.77–6.74 (m, 4H), 6.71 (d, J = 8.9 Hz, 2H), 4.03 (dd, J = 5.7, 4.0 Hz, 4H), 3.77 (dd, J = 5.6, 4.1 Hz, 4H), 3.66–3.64 (m, 4H), 3.59–3.57 (m, 56H), 3.48 (dd, J = 3.9, 2.0 Hz, 4H), 3.31 (d, J = 1.8 Hz, 6H). 13 C NMR (126 MHz, Chloroform-d) δ 155.22, 147.87, 140.69, 131.76, 126.43, 122.14, 115.52, 112.44, 71.94, 70.82, 70.64, 70.58, 70.52, 69.76, 67.69, 59.04.

[0045] Step 3: Synthesis of Compound 3

[0046] Dissolve Compound 2 (0.5256 g, 1 mmol) in ultradry THF (12 mL), and add n-BuLi (0.44 mL, 1.1 mmol) at -78 °C. After stirring for 1 h, add tributyltin chloride (1.1 mmol). After stirring overnight at room temperature, quench the reaction with KF solution. Extract the mixture three times with EA, combine the organic phases and dry over Na2SO4. After removing the solvent, use directly without further purification.

[0047] Step 4: Synthesis of Compound PEG-TBS

[0048] Mix Compound 3 (366 mg, 0.3 mmol), the S-containing acceptor (45 mg, 0.1 mmol), and toluene / water (25 mL, v:v = 4:1), then add Pd(PPh3)4 (11.5 mg, 0.01 mmol) and K2CO3 (138 mg, 3 mmol). Heat to 120 °C under inert gas protection and stir overnight to obtain the crude product. After cooling to room temperature, dilute the crude product with water successively, extract three times with DCM, and dry over anhydrous sodium sulfate. Purify the obtained product by column chromatography to obtain a blue solid product (92 mg, 37%), with a particle size of 5.5 nm. 1 H NMR (600 MHz, THF-d8) δ 8.45 (d, J = 9.0 Hz, 4H), 7.15–7.13 (m, 8H), 7.02 (d, J = 9.0 Hz, 4H), 6.93–6.91 (m, 8H), 4.83 (d, J = 6.9 Hz, 2H), 4.10 (m, 8H), 3.80 (m, 8H), 3.64 (d, J = 5.1 Hz, 8H), 3.42–3.57 (m, 120H), 3.42 (d, J = 4.7 Hz, 8H), 3.26 (s, 12H), 1.41–1.38 (m, 5H), 1.29–1.26 (m, 4H), 0.95 (t, J = 7.5 Hz, 3H), 0.85 (t, J = 7.3 Hz, 3H). 1313C NMR(151MHz,THF-d8)δ156.90,152.41,149.54,145.24,141.44,133.15,127.97,127.14,119.58,118.06,116.29,72.91,71.69,71.52–71.60(multiple peaks of the PEG chains),71.36,70.60,68.71,41.57,31.50,29.30,23.85,14.42,10.85.HRMS(ESI)calculated for C 126 H 200 N7O 40 S[M + 3H] 3+ [m / z]828.1190; Found: 828.1183.

[0049] Synthesis of Compound PEG-TBSe

[0050] Compound 3(366 mg, 0.3 mmol), Se-containing acceptor(45 mg, 0.1 mmol) and toluene / water(25 mL, v:v = 4:1) were mixed, and then Pd(PPh3)4(11.5 mg, 0.01 mmol) and K2CO3(138 mg, 3 mmol) were added. The mixture was heated to 120 °C under inert gas protection and stirred overnight to obtain the crude product. After cooling to room temperature, the crude product was diluted with water successively, extracted three times with DCM, and dried over anhydrous sodium sulfate. The obtained product was purified by column chromatography to obtain a green solid product(132 mg, 52%), with a particle size of 5.5 nm. 1 1H NMR(600 MHz, THF-d8)δ8.35(d, J = 8.4 Hz, 4H), 7.15–7.12(m, 8H), 7.00(d, J = 8.4 Hz, 4H), 6.92–6.90(m, 8H), 4.76(d, J = 6.9 Hz, 2H), 4.10(t, J = 5.0 Hz, 8H), 3.80(t, J = 4.9 Hz, 8H), 3.65–3.63(m, 8H), 3.42–3.53(m, 120H), 3.43(d, J = 4.5 Hz, 8H), 3.27(s, 12H), 1.42–1.38(m, 5H), 1.30(d, J = 9.9 Hz, 4H), 0.95(t, J = 7.4 Hz, 3H), 0.86(t, J = 7.3 Hz, 3H). 1313C NMR (151 MHz, THF-d8) δ 158.89, 156.77, 149.18, 145.85, 141.57, 133.59, 129.06, 127.84, 119.62, 118.19, 116.26, 72.90, 71.68, 71.54 (multiple peaks of the PEG chains), 71.35, 70.59, 68.70, 61.76, 58.95, 41.45, 31.49, 29.28, 23.85, 14.44, 10.88. HRMS (ESI) calculated for C 126 H 200 N7O 40 Se [M+3H] 3+ [m / z] 844.1005; Found: 844.1000.

[0051] Example 2

[0052] The preparation methods of the compounds PEG-TBS and PEG-TBSe specifically include the following steps:

[0053] As Figure 1 shown: Figure 1 is the synthetic route diagram of the novel water-soluble NIR-II organic small molecule probe provided in Example 1 of the present invention;

[0054] As Figure 2 shown: Figure 2 is the 1H NMR spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0055] As Figure 3 shown: Figure 3 is the 13C NMR spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0056] As Figure 4 shown: Figure 4 is the 1H NMR spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0057] As Figure 5 shown: Figure 5 is the 13C NMR spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention in deuterated tetrahydrofuran;

[0058] As Figure 6 shown: Figure 6It is the HRMS mass spectrum of the water-soluble near-infrared organic small molecule probe PEG-TBS prepared in Example 1 of the present invention;

[0059] As Figure 7 shown: Figure 7 It is the HRMS mass spectrum of the water-soluble NIR-II organic small molecule probe PEG-TBSe prepared in Example 1 of the present invention;

[0060] As Figure 8 shown: Figure 8 It is the ultraviolet absorption and fluorescence emission spectra of the PEG-TBS and PEG-TBSe materials prepared in Example 1 of the present invention, indicating that the prepared compounds have near-infrared absorption / emission effects.

[0061] As Figure 9 shown: Figure 9 It is the fluorescence signal change diagram of the PEG-TBS and PEG-TBSe materials with different concentrations prepared in Example 1 of the present invention. It can be seen that the synthesized materials have the property of aggregation-induced emission.

[0062] As Figure 10 shown: Figure 10 It is the clearance rate diagram of the PEG-TBSe material prepared in Example 1 of the present invention over time. From Figure 10 it can be seen that PEG-TBSe has excellent renal clearance characteristics.

[0063] As Figure 11 shown: Figure 11 It is the cell survival rate diagram of the PEG-TBSe material prepared in Example 1 of the present invention co-incubated with HK-2 cells (human renal tubular epithelial cells) and SV-HUC-1 cells (human ureteral epithelial immortalized cells) for 72 hours. From Figure 11 it can be seen that PEG-TBSe has excellent cell compatibility.

[0064] As Figure 12 shown: Figure 12 It is the fluorescence imaging diagram of the PEG-TBSe material prepared in Example 1 of the present invention injected into a unilateral renal ischemia-reperfusion mouse model. From Figure 12 it can be seen that PEG-TBSe can accurately identify the damaged left kidney and can be used for the early diagnosis of pre-renal acute kidney injury.

[0065] As Figure 13 shown: Figure 13 It is the fluorescence imaging diagram of the PEG-TBSe material prepared in Example 1 of the present invention injected into a cisplatin-induced acute kidney injury mouse model. From Figure 13 it can be seen that PEG-TBSe can accurately identify the damaged bilateral kidneys and can be used for the early diagnosis of renal acute kidney injury.

[0066] As shown in Figure 14 the following figure: Figure 14 This is the fluorescence imaging diagram after injecting the PEG-TBSe material prepared in Example 1 of the present invention into a unilateral ureteral ligation mouse model. It can be seen from Figure 14 this that PEG-TBSe can accurately identify the damaged bilateral kidneys and can be used for the early diagnosis of post-renal acute kidney injury.

[0067] To sum up: The embodiments of the present invention provide a class of novel water-soluble NIR-II organic small molecule probes, and their chemical structural formula is:

[0068]

[0069] wherein X is S or Se. This molecule adopts a D-π-A-π-D structure design. The triphenylamine group serves as an electron donor and a rotor unit, making the molecular backbone show a significant twist, thereby endowing it with aggregation-induced emission (AIE) characteristics. The introduction of the ethylene glycol fragment can be used to regulate the hydrophilicity of the probe. The strong electron acceptor has excellent electron-withdrawing ability, which can enhance the intramolecular charge transfer effect, reduce the molecular band gap, and extend the absorption / emission wavelength to the near-infrared region. The water-soluble near-infrared fluorescent probe molecule proposed by the present invention has renal clearance ability. Its synthesis route is simple, the cost is low, and the biocompatibility is excellent. It is suitable for realizing the early diagnosis of acute kidney injury caused by various etiologies through fluorescence imaging in the second near-infrared region.

[0070] It should be understood that the application of the present invention is not limited to the above examples. For those of ordinary skill in the art, improvements or transformations can be made according to the above description, and all such improvements and transformations should fall within the protection scope of the appended claims of the present invention.

Claims

1. A class of renal clearance near-infrared fluorescent probe molecules, characterized in that, Its chemical structural formula is: Wherein, X is S or Se.

2. A method for preparing a renal clearance near-infrared fluorescence probe molecule as described in claim 1, characterized in that, Including: Compound III, S-containing receptor, Pd(PPh3)4 and carbonate are added to a mixture of toluene and water, and heated and stirred under the protection of an inert gas to obtain a first crude product; After cooling the temperature of the first crude product to room temperature, it is diluted, extracted and dried in sequence to obtain a renal clearance near-infrared fluorescent probe molecule; The structural formula of Compound III is:

3. The preparation method of the renal clearance near-infrared fluorescence probe molecule according to claim 2, wherein, Compound III, Se-containing receptor, Pd(PPh3)4 and carbonate are added to a mixture of toluene and water, and heated and stirred under the protection of an inert gas to obtain a second crude product; After cooling the temperature of the second crude product to room temperature, it is diluted, extracted and dried in sequence to obtain a renal clearance near-infrared fluorescent probe molecule; The structural formula of Compound III is:

4. The preparation method of the renal clearance near-infrared fluorescent probe molecule according to claim 2, wherein, The molar ratio of Compound III, S-containing receptor, Pd(PPh3)4 and carbonate is 30:10:1:

300.

5. The preparation method of the renal clearance near-infrared fluorescence probe molecule according to claim 3, wherein The molar ratio of Compound III, Se-containing receptor, Pd(PPh3)4 and carbonate is 30:10:1:

300.

6. The preparation method of the renal clearance near-infrared fluorescence probe molecule according to claim 2 or 3, characterized in that, The temperature of the heating and stirring is 90 - 110 °C, and the time of the heating and stirring is 12 - 24 h.

7. The preparation method of the renal clearance near-infrared fluorescence probe molecule according to claim 2, wherein, The S-containing receptor is 4,8-dibromo-6-(2-ethylhexyl)-[1,2,5]thiadiazolo[3,4-f]benzotriazole; the carbonate is selected from one of potassium carbonate, sodium carbonate and cesium carbonate.

8. The preparation method of the near-infrared fluorescent probe molecule for renal clearance according to claim 3, wherein The Se-containing receptor is 4,8-dibromo-6-(2-ethylhexyl)-[1,2,5]selenadiazolo[3,4-f]benzotriazole.

9. Use of a renal clearance near-infrared fluorescent probe molecule as described in claim 1 in the early diagnosis of an acute kidney injury model.

10. Use of the near-infrared fluorescence probe molecule for renal clearance according to claim 9 in the preparation of a reagent for early diagnosis of acute kidney injury, characterized in that, The acute kidney injury model includes: a unilateral renal ischemia-reperfusion model, a cisplatin-induced acute kidney injury model, and a unilateral ureteral ligation model.

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