Targeted near-infrared fluorescent compound, and preparation method therefor and use thereof

Abstract

The present invention relates to the technical field of organic fluorescent molecules, and provides a targeted near-infrared fluorescent compound, and a preparation method therefor and the use thereof. The targeted near-infrared fluorescent compound has a structure as represented by formula I. S0456 near-infrared small molecules are modified on a CXCR4 inhibitor by means of an organic total synthesis method, thereby obtaining the targeted near-infrared fluorescent compound represented by formula I. The targeted near-infrared fluorescent compound has a good active targeting effect during the identification of CXCR4-overexpressing tumors, such as triple-negative breast cancer and head and neck squamous cell carcinomas; and the targeted near-infrared fluorescent compound retains the water solubility of a dye and specificity to tumor cells, and has the advantages of good water solubility, high fluorescence quantum yield, etc.

Description

Targeted near-infrared fluorescent compound and preparation method and application thereof

[0001] Cross-reference to related applications

[0002] The present disclosure claims priority to the Chinese patent application No. 2024111251261, filed on August 16, 2024, entitled "Targeted near-infrared fluorescent compound and preparation method and application thereof", the entire content of which is incorporated herein by reference. TECHNICAL FIELD

[0003] The present disclosure relates to the technical field of near-infrared fluorescent molecules, in particular to a targeted near-infrared fluorescent compound and a preparation method and application thereof. BACKGROUND

[0004] In recent years, near-infrared imaging technology has provided an effective tool for precise tumor surgery. At present, the only clinically approved near-infrared imaging agent in China is indocyanine green (ICG), which provides tumor visualization capability for clinical surgery in early application. However, with the in-depth research, the tumor imaging effect of ICG is greatly affected by patient individual differences and operation time, and it is difficult to develop a standardized surgical protocol.

[0005] In addition, ICG also has the following problems: first, the aqueous solution has a low fluorescence quantum yield of <1%, which will greatly reduce the detection limit of the equipment. According to the Chinese expert consensus on the application of near-infrared fluorescence imaging of indocyanine green in laparoscopic radical gastrectomy (2019 version), the maximum injection dose of ICG in the human body should be less than 2 mg / kg, and under this dose, the concentration of ICG in the human tumor is reported in the literature to be 10-1000 nmol / L (reference: Wang Z, Ni K, Zhang X, Ai S, Guan W, Cai H, Wang Y, Lu Q, Lane LA (2018) Method for real-time tissue quantification of indocyanine green revealing optimal conditions for near infrared fluorescence guided surgery. Anal Chem 90:7922-7929.), which puts extremely high requirements on the detection equipment. Second, in a short period of time, the metabolic rate of ICG in the tumor and normal tissue is not much different, and only after more than 12 h can sufficient fluorescence contrast (tumor: normal tissue) be produced, increasing the burden on the hospital and the patient. Third, ICG is prone to photobleaching after aggregation, which greatly reduces its stability during imaging. Fourth, the enrichment of ICG in the tumor is through the EPR effect, lacking active targeting of tumor cells, which greatly reduces the accuracy of fluorescence tracer imaging. These factors greatly limit the clinical application of ICG, so it is necessary to further seek a tumor-specific near-infrared fluorescence tracer suitable for clinical surgical imaging.

[0006] In cancer, the CXCL12 / CXCR4 biological axis can stimulate the proliferation and metastasis of tumor cells, regulate the inflammatory state of the tumor, and activate the immune response within the tumor through autocrine and paracrine factors. CXCR4 is highly expressed in many solid tumors and hematological tumors, and is an effective target for tumor diagnosis and treatment. Small molecule compounds that act as CXCL12 / CXCR4 antagonists have great development and commercialization potential. So far, dozens of small molecule compounds have been studied and entered clinical trials.

[0007] Therefore, the present disclosure is proposed. SUMMARY

[0008] The purposes of the present disclosure include providing a targeted near-infrared fluorescent compound and a preparation method and application thereof. The targeted near-infrared fluorescent compound of the present disclosure is a novel NY-CXCR4 near-infrared fluorescent molecule targeting chemokine receptor type 4 receptor, which is obtained by connecting a near-infrared fluorescent molecule S0456 on a CXCR4 inhibitor. The targeted near-infrared fluorescent compound can specifically target triple-negative breast cancer and head and neck squamous cell carcinoma, and has the advantages of good water solubility and high fluorescence quantum yield.

[0009] In order to achieve the above purposes of the present disclosure, the following technical solutions can be adopted:

[0010] In a first aspect, the present disclosure provides a targeted near-infrared fluorescent compound, which has a structure as shown in the following formula I:

[0011] In a second aspect, the present disclosure provides a preparation method of the targeted near-infrared fluorescent compound according to the first aspect, which comprises the following steps:

[0012] (1) reacting the compound S0456 and p-hydroxyphenylpropionate to obtain a compound A;

[0013] The structural formula of the compound A is as follows:

[0014] (2) reacting the compound A and dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate to obtain a compound B; the structural formula of the compound B is as follows:

[0015] (3) reacting the compound B and a CXCR4 inhibitor to obtain the targeted near-infrared fluorescent compound as shown in the formula I;

[0016] The structure of the compound CXCR4 inhibitor is as follows:

[0017] Optionally, in step (1), the molar ratio of the compound S0456 to p-hydroxyphenylpropionate is 1:(2-4).

[0018] Optionally, in step (1), the p-hydroxyphenylpropionate includes sodium p-hydroxyphenylpropionate.

[0019] Optionally, in step (1), the reaction is carried out in a solvent, and the solvent includes dimethyl sulfoxide.

[0020] Optionally, in step (1), the reaction is carried out in a protective gas atmosphere, and the protective gas includes nitrogen.

[0021] Optionally, in step (1), the reaction temperature is 60-70℃, and the reaction time is 5-7h.

[0022] Optionally, after the reaction in step (1), a step of post-treatment is further included:

[0023] The reaction solution obtained after the reaction in step (1) is added dropwise into a mixture of ethyl acetate and ethanol, and then filtered and dried to obtain compound A.

[0024] Optionally, in the mixture, the volume ratio of ethyl acetate to ethanol is 1:((0.5-2).

[0025] Optionally, in step (2), the molar ratio of compound A to dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate is 1:(1.2-2).

[0026] Optionally, in step (2), the reaction is carried out in the presence of a base, and the base includes N,N-diisopropylethylamine.

[0027] Optionally, in step (2), the molar ratio of compound A to the base is 1:(0.5-5).

[0028] Optionally, in step (2), the reaction is carried out in a solvent, and the solvent includes dimethyl sulfoxide.

[0029] Optionally, in step (2), the reaction temperature is 30-40°C, and the reaction time is 1-3h.

[0030] Optionally, after the reaction in step (2), a step of post-treatment is further included:

[0031] The reaction solution obtained after the reaction in step (2) is added dropwise into a solution of acetone, and then filtered and dried to obtain compound B.

[0032] Optionally, in step (3), the molar ratio of compound B to the CXCR4 inhibitor is 1:(0.5-2).

[0033] Optionally, in step (3), the reaction is carried out in the presence of a base, and the base includes N,N-diisopropylethylamine.

[0034] Optionally, in step (3), the molar ratio of compound B to the base is 1:(0.5-5).

[0035] Optionally, in step (3), the reaction is carried out in a solvent, and the solvent includes dimethyl sulfoxide.

[0036] Optionally, in step (3), the reaction temperature is 30-40°C, and the reaction time is 18-30h.

[0037] Optionally, after the reaction in step (3), a step of post-treatment is further included:

[0038] The reaction solution obtained after the reaction in step (3) is added dropwise into an acetone solution, filtered and dried to obtain the targeted near-infrared fluorescent compound shown in formula I.

[0039] In a third aspect, the present disclosure provides a use of the targeted near-infrared fluorescent compound or a racemate, stereoisomer, or pharmaceutically acceptable salt thereof according to the first aspect in the preparation of a reagent for tumor imaging.

[0040] Optionally, the tumor includes a triple-negative breast cancer tumor and / or a head and neck squamous carcinoma tumor.

[0041] In a fourth aspect, the present disclosure provides a use of the targeted near-infrared fluorescent compound or a racemate, stereoisomer, or pharmaceutically acceptable salt thereof according to the first aspect in the preparation of a reagent for identifying a CXCR4 overexpressing tumor.

[0042] Compared with the prior art, the present disclosure has the following beneficial effects:

[0043] (1) The present disclosure synthesizes the CXCR4 inhibitor modified S0456 to obtain the targeted near-infrared fluorescent compound NY-CXCR4 by using the method of organic total synthesis. The probe can actively target the chemokine receptor type 4 (CXCR4), can be rapidly cleared in normal tissues, and can be long-term retained in tumor sites, and has good active targeting effect in identifying triple-negative breast cancer and head and neck squamous carcinoma.

[0044] (2) The targeted near-infrared fluorescent compound NY-CXCR4 of the present disclosure has the advantages of water solubility and high fluorescence quantum yield of dye. It has great development potential in the fields of near-infrared fluorescent navigation tumor surgery and medical cell labeling. BRIEF DESCRIPTION OF DRAWINGS

[0045] In order to more clearly illustrate the technical solutions in the specific embodiments of the present disclosure or the prior art, the drawings needed in the specific embodiment or prior art description will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present disclosure, and those skilled in the art can also obtain other drawings according to these drawings without creative labor.

[0046] FIG. 1 is a high-performance liquid chromatography analysis diagram of NY-CXCR4 fluorescent molecules;

[0047] FIG. 2 is a mass spectrum diagram of NY-CXCR4 fluorescent molecules;

[0048] FIG. 3 is a fluorescent performance diagram of NY-CXCR4 fluorescent molecules;

[0049] FIG. 4 is a specific targeting effect of NY-CXCR4 on triple-negative breast cancer;

[0050] Figure 5 is the specific targeting effect of NY-CXCR4 on head and neck squamous cell carcinoma. DETAILED DESCRIPTION

[0051] Unless otherwise defined, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear; however, in the event of any latent ambiguity, definitions provided herein take precedent. In this application, the use of "or" means "and / or" unless otherwise stated. Moreover, the use of the "a", "an" or "the" are employed to describe a certain aspect intended to include one or more of them unless otherwise indicated.

[0052] It should be noted that specific details are set forth in the following description in order to provide a thorough understanding of the present disclosure. However, the present disclosure can be practiced without many of the details described in this description, which are not necessary to practice the present disclosure. In other instances, well known methods have not been described in detail in order not to unnecessarily obscure the present disclosure.

[0053] The technical solutions of the present disclosure will be described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of protection of the present disclosure.

[0054] In a first aspect, the present disclosure provides a targeted near-infrared fluorescent compound, which has the structure shown in the following formula I:

[0055] In the present disclosure, S0456 near-infrared small molecules are modified on CXCR4 inhibitors to obtain the targeted near-infrared fluorescent compound shown in the above formula I, which is referred to as NY-CXCR4 hereinafter. In the present disclosure, the CXCR4 inhibitor is selected from a structural analogue of plerixafor As a targeting group connected with a fluorescent group, the obtained NY-CXCR4 can be rapidly cleared in normal tissues, but can be retained for a long time at tumor sites. The connection with the fluorescent group further expands the application value of the drug, which is used for surgical navigation and tumor resection. The results show that the NY-CXCR4 obtained in the present disclosure has good active targeting effect in identifying triple-negative breast cancer and head and neck squamous cell carcinoma. In addition, NY-CXCR4 also has the advantages of water solubility of dye and high fluorescence quantum yield.

[0056] Plerixafor is a small molecule non-peptide molecule that specifically antagonizes the CXCR4 receptor. The primary structure of Plerixafor contains two cyclic polysaccharide rings connected by a phenyl linker. Under physiological conditions, the protonated nitrogen atoms on the ring interact with the carboxylic acid groups on CXCR4. This can inhibit the binding of CXCL12 to CXCR4, block downstream signaling and regulate various physiological activities. In the development of fluorescent probes with tumor specificity, anti-tumor drugs with tumor-specific killing effect have certain research value and can be used for the development of specific targeting fluorescent probes.

[0057] In a second aspect, the present disclosure provides a preparation method of the targeted near-infrared fluorescent compound according to the first aspect, the preparation method comprising the following steps:

[0058] (1) reacting compound S0456 and p-hydroxyphenylpropionate to obtain compound A;

[0059] The structural formula of compound A is as follows:

[0060] (2) reacting compound A and dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate to obtain compound B;

[0061] The structural formula of compound A is as follows:

[0062] (3) reacting compound B and a CXCR4 inhibitor to obtain the targeted near-infrared fluorescent compound shown in formula I;

[0063] The structure of the compound CXCR4 inhibitor is:

[0064] In some optional embodiments, the preparation method of the targeted near-infrared fluorescent compound can comprise:

[0065] (1) reacting compound S0456 and p-hydroxyphenylpropionate to obtain compound A; wherein the reaction formula is as follows:

[0066] (2) reacting compound A and dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate to obtain compound B; wherein the reaction formula is as follows:

[0067] (3) reacting compound B and a CXCR4 inhibitor to obtain the targeted near-infrared fluorescent compound shown in formula I; wherein the reaction formula is as follows:

[0068] In the present disclosure, a CXCR4 inhibitor is synthesized by a method of organic total synthesis The modified S0456 near-infrared small molecule targeting near-infrared fluorescent compound, NY-CXCR4, has good active targeting effect in identifying CXCR4 overexpressing tumor triple-negative breast cancer and head and neck squamous cell carcinoma. The targeting near-infrared fluorescent compound of formula I retains the water solubility and specificity of the dye to tumor cells, as a tracer, has the advantages of good water solubility and high fluorescence quantum yield, and has great development potential in the fields of fluorescent navigation tumor surgery and medical cell labeling.

[0069] In an optional embodiment, in step (1), the molar ratio of compound S0456 and p-hydroxyphenyl propionate can be 1:(2-4), for example, it can be 1:2, 1:2.2, 1:2.4, 1:2.6, 1:2.8, 1:3, 1:3.2, 1:3.4, 1:3.6, 1:3.8 or 1:4, etc., or other values within the range of 1:(2-4).

[0070] In an optional embodiment, in step (1), the p-hydroxyphenyl propionate can include sodium p-hydroxyphenyl propionate.

[0071] In an optional embodiment, in step (1), the reaction is carried out in a solvent.

[0072] In an optional embodiment, in step (1), the solvent can include dimethyl sulfoxide.

[0073] In an optional embodiment, in step (1), the reaction is carried out under a protective gas atmosphere.

[0074] In an optional embodiment, in step (1), the protective gas can include nitrogen.

[0075] In an optional embodiment, in step (1), the temperature of the reaction can be 60-70℃, for example, it can be 60℃, 61℃, 62℃, 63℃, 64℃, 65℃, 66℃, 67℃, 68℃, 69℃ or 70℃, etc., or other values within the range of 60-70℃.

[0076] In an optional embodiment, in step (1), the reaction time can be 5-7h, for example, it can be 5h, 5.2h, 5.4h, 5.6h, 5.8h, 6h, 6.2h, 6.4h, 6.6h, 6.8h or 7h, etc., or other values within the range of 5-7h.

[0077] In an optional embodiment, in step (1), after the reaction is completed, a post-treatment step can be further included:

[0078] The reaction solution obtained after the reaction in step (1) is added dropwise into a mixture of ethyl acetate and ethanol, and after filtration and drying, compound A is obtained.

[0079] In an alternative embodiment, the volume ratio of ethyl acetate and ethanol in the mixture can be 1:(0.5-2), for example, it can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.85, 1:0.9, 1:0.95, 1:1, 1:1.05, 1:1.1, 1:1.15, 1:1.2, 1:1.4, 1:1.6, 1:1.8, or 1:2, etc., or other values within the range of 1:(0.5-2).

[0080] In an alternative embodiment, in step (2), the molar ratio of compound A and dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate can be 1:(1.2-2), for example, it can be 1:1.2, 1:1.25, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, or 1:2, etc., or other values within the range of 1:(1.2-2).

[0081] In an alternative embodiment, in step (2), the reaction is carried out in the presence of a base, which can include N,N-diisopropylethylamine.

[0082] In an alternative embodiment, in step (2), the molar ratio of compound A and the base can be 1:(0.5-5), for example, it can be 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, or 1:5, etc., or other values within the range of 1:(0.5-5).

[0083] In an alternative embodiment, in step (2), the reaction is carried out in a solvent.

[0084] In an alternative embodiment, in step (2), the solvent can include dimethyl sulfoxide.

[0085] In an alternative embodiment, in step (2), the temperature of the reaction can be 30-40℃, for example, it can be 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃, or 40℃, etc., or other values within the range of 30-40℃.

[0086] In an alternative embodiment, in step (2), the reaction can be performed for 1-3 hours, for example, 1 hour, 1.2 hours, 1.4 hours, 1.6 hours, 1.8 hours, 2 hours, 2.2 hours, 2.4 hours, 2.6 hours, 2.8 hours, or 3 hours, or other values within the range of 1-3 hours.

[0087] In an alternative embodiment, in step (2), after the reaction is completed, a post-treatment step can be further included.

[0088] The reaction solution obtained after the reaction in step (2) is added dropwise into an acetone solution, filtered and dried to obtain compound B.

[0089] In an alternative embodiment, in step (3), the molar ratio of compound B to the CXCR4 inhibitor can be 1:(0.5-2), for example, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.85, 1:0.9, 1:0.95, 1:1, 1:1.05, 1:1.1, 1:1.15, 1:1.2, 1:1.4, 1:1.6, 1:1.8, or 1:2, or other values within the range of 1:(0.5-2).

[0090] In an alternative embodiment, in step (3), the reaction is performed in the presence of a base.

[0091] In an alternative embodiment, in step (3), the base can include N,N-diisopropylethylamine.

[0092] In an alternative embodiment, in step (3), the molar ratio of compound B to the base can be 1:(0.5-2), for example, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8, 1:2, 1:2.2, 1:2.4, 1:2.6, 1:2.8, 1:3, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:6, 1:7, 1:8, 1:9, or 1:2, or other values within the range of 1:(0.5-2).

[0093] In an alternative embodiment, in step (3), the reaction is performed in a solvent, and the solvent can include dimethyl sulfoxide.

[0094] In an alternative embodiment, in step (3), the reaction can be performed at a temperature of 30-40°C, for example, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, or 40°C, or other values within the range of 30-40°C.

[0095] In an optional embodiment, in step (3), the reaction time can be 18-30 h, for example, it can be 18 h, 19 h, 20 h, 21 h, 22 h, 23 h, 24 h, 25 h, 26 h, 27 h, 28 h, 29 h or 30 h, etc., or other values in the range of 18-30 h.

[0096] In an optional embodiment, in step (3), after the reaction is completed, a post-treatment step can also be included:

[0097] The reaction solution obtained after the reaction in step (3) is added dropwise into an acetone solution, filtered and dried to obtain the targeted near-infrared fluorescent compound shown in formula I.

[0098] In a third aspect, the present disclosure provides a use of the targeted near-infrared fluorescent compound or a racemate, stereoisomer, or pharmaceutically acceptable salt thereof according to the first aspect in the preparation of a reagent for tumor imaging.

[0099] In an optional embodiment, the tumor can include a triple-negative breast cancer tumor and / or a head and neck squamous cell carcinoma tumor.

[0100] In a fourth aspect, the present disclosure provides a use of the targeted near-infrared fluorescent compound or a racemate, stereoisomer, or pharmaceutically acceptable salt thereof in the preparation of a reagent for identifying a CXCR4-overexpressing tumor.

[0101] In an optional embodiment, the use can be: using the targeted near-infrared fluorescent compound or a racemate, stereoisomer, or pharmaceutically acceptable salt thereof according to the first aspect to establish a tumor imaging model, and using an intraoperative fluorescence navigation device to diagnose the tumor effect.

[0102] In an optional embodiment, the present disclosure provides a triple-negative breast cancer tumor imaging model and a method for establishing the same, which can include the following steps: inoculating a CXCR4 high-expression cell line human triple-negative breast cancer on the back of a mouse, and injecting the targeted near-infrared fluorescent compound (NY-CXCR4) of the present disclosure through the tail vein.

[0103] In an optional embodiment, the present disclosure provides a triple-negative breast cancer tumor imaging model and a method for establishing the same, which can include the following steps: using a CXCR4 high-expression cell line human triple-negative breast cancer MDA-MB-231 to inoculate on the back of a Balb / c mouse, and injecting the targeted near-infrared fluorescent compound (NY-CXCR4) of the present disclosure through the tail vein.

[0104] After the above-mentioned triple negative breast cancer tumor imaging model is established, the small molecule can be used to diagnose the effect of triple negative breast cancer tumor by using intraoperative fluorescence navigation equipment; and the triple negative breast cancer can be removed intraoperatively, and it can be judged by pathological section that NY-CXCR4 can specifically target the triple negative breast cancer tumor.

[0105] In an alternative embodiment, the present disclosure provides a head and neck squamous cell carcinoma imaging model and a method for establishing the same, which can comprise the following steps: inoculating a CXCR4 high-expression cell line human head and neck squamous cell carcinoma on the back of a mouse, and injecting a targeted near-infrared fluorescent compound (NY-CXCR4) of the present disclosure through the tail vein.

[0106] In an alternative embodiment, the present disclosure provides a head and neck squamous cell carcinoma imaging model and a method for establishing the same, which can comprise the following steps: inoculating a CXCR4 high-expression cell line human head and neck squamous cell carcinoma FaDu on the back of a Balb / c mouse, and injecting a targeted near-infrared fluorescent compound (NY-CXCR4) of the present disclosure through the tail vein.

[0107] After the above-mentioned head and neck squamous cell carcinoma imaging model is established, the small molecule can be used to diagnose the effect of head and neck squamous cell carcinoma tumor by using intraoperative fluorescence navigation equipment. The head and neck squamous cell carcinoma can be removed intraoperatively, and it can be judged by pathological section that NY-CXCR4 can specifically target the head and neck squamous cell carcinoma tumor.

[0108] As mentioned in the present disclosure, the term "pharmaceutically acceptable salt" means that the salt is not only physiologically acceptable to the subject, but also can refer to a synthetic substance which is pharmaceutically useful, such as a salt formed as an intermediate when chiral resolution is performed, although such an intermediate salt cannot be directly administered to the subject, but the salt can play a role in obtaining the final product of the present disclosure.

[0109] As mentioned in the present disclosure, the pharmaceutically acceptable salt of the compound of formula I: a salt formed with a base or an alkali metal. The acid forming a pharmaceutically acceptable salt with the compound of formula I includes inorganic acid and organic acid. More specifically, the alkali metal forming a pharmaceutically acceptable salt with the compound of formula I includes but is not limited to lithium, sodium, potassium, magnesium, calcium, aluminum, zinc, etc.; the base forming a pharmaceutically acceptable salt with the compound of formula I but not limited to choline, diethanolamine, morpholine, etc.

[0110] The present disclosure is further illustrated by the following examples. Unless otherwise specified, the materials in the examples are prepared according to the existing methods or directly purchased from the market.

[0111] Example 1

[0112] This embodiment provides a targeted near-infrared fluorescent compound NY-CXCR4, which has the structure shown in Formula I:

[0113] The synthetic route for the targeted near-infrared fluorescent compound NY-CXCR4 is shown below:

[0114] The preparation method of the targeted near-infrared fluorescent compound NY-CXCR4 specifically includes the following steps:

[0115] (1) Dissolve SO456 (888 mg, 1 mmol) in 10 mL DMSO and stir until completely dissolved. Add sodium p-hydroxyphenylpropionate (424 mg, 2 mmol) to the flask, and react at 65 °C for 6 h under nitrogen protection. Add the resulting solution dropwise to a mixture of 200 mL ethyl acetate and ethanol (V 乙酸乙酯 :V 乙醇 =1:1), producing a dark green precipitate, which was filtered and dried under vacuum to give 1.05 g of solid (i.e., compound A), with a yield of 95%.

[0116] (2) Compound A (1g, 0.98mmol) was placed in a round-bottom flask and dissolved in 20mL of DMSO. Then, dipyrrolidinyl (N-succiniminooxy)carbomony hexafluorophosphate (483mg, 1.176mmol) and DIEA (151mg, 1.176mmol) were added in sequence. The mixture was stirred at 37℃ for 2h and precipitated in acetone solution. The precipitate was filtered and dried under vacuum to obtain 1.499g (i.e., compound B).

[0117] (3) Place compound B (1.15 g, 1 mmol) in a round-bottom flask, dissolve it in 20 mL of DMSO, and then add the following ingredients sequentially: (0.431 g, 1 mmol), DIEA (258 mg, 2 mmol), were stirred at 37 °C for 24 h, and precipitated in acetone solution. The precipitate was filtered, dried under vacuum, and 1.6 g was obtained. The product was purified by preparative liquid chromatography to obtain the targeted near-infrared fluorescent compound NY-CXCR4 shown in Formula I.

[0118] The structure was characterized by high performance liquid chromatography and mass spectrometry, and the structural determination results are as follows:

[0119] As shown in Figure 1, the purity of the targeted near-infrared fluorescent compound NY-CXCR4 is 96.1%.

[0120] As shown in Figure 2, LCMS(ESI):m / z:C 74 H 89 N7O 14S4, [M-3H] found 465.15, [M-2H] found 698.65.

[0121] As shown in Figure 3, the targeted near-infrared fluorescent compound NY-CXCR4 has a maximum excitation wavelength Ex max = 773 nm and a maximum fluorescent emission wavelength Em max = 795 nm, indicating that the molecule is a near-infrared fluorescent probe.

[0122] Application Example 1

[0123] The present application provides an application of a NY-CXCR4 near-infrared fluorescent molecule in specifically identifying triple-negative breast cancer.

[0124] A triple-negative breast cancer model was constructed using the MDA-MB-231 cell line: the human triple-negative breast cancer MDA-MB-231 cell line with high expression of CXCR4 was inoculated on the back of Balb / c mice, and 10 nmol of NY-CXCR4 molecules were injected into the tail vein of each mouse. The effect of the small molecule in diagnosing triple-negative breast cancer tumors was diagnosed using an intraoperative fluorescence navigation device. The triple-negative breast cancer was removed, and it was determined through pathological sectioning that NY-CXCR4 could specifically target triple-negative breast cancer tumors.

[0125] The test results are shown in Figure 4:

[0126] On the left is the effect of the small molecule in diagnosing triple-negative breast cancer tumors using an intraoperative fluorescence navigation device. When 10 nmol of NY-CXCR4 was injected, 8 h, 12 h, and 24 h later, the mouse's back only showed a high fluorescent signal in the tumor area, while the surrounding normal tissue showed a low fluorescent signal. This indicates that NY-CXCR4 has good specificity in identifying triple-negative breast cancer with overexpression of the CXCR4 receptor, and can achieve rapid clearance in normal tissue, while being retained in breast cancer tumor sites for a long time, thereby obtaining a higher tumor background signal ratio.

[0127] On the right is the fluorescent signal distribution of the isolated organs at the corresponding time. When 10 nmol of NY-CXCR4 was injected, the fluorescent intensity of the tumor tissue was significantly higher than that of the muscle tissue at each time point. This indicates that NY-CXCR4 can accumulate in the tumor, achieving intraoperative specific identification of breast cancer tumors.

[0128] Application Example 2

[0129] The present application provides an application of a NY-CXCR4 near-infrared fluorescent molecule in specifically identifying triple-negative breast cancer.

[0130] The CXCR4 high expression cell line human head and neck squamous carcinoma FaDu is inoculated on the back of Balb / c mice, 10 nmol NY-CXCR4 molecules are injected into the tail vein of each mouse, and the effect of the small molecule on the diagnosis of head and neck squamous carcinoma tumor is diagnosed by using the intraoperative fluorescence navigation device. The head and neck squamous carcinoma is removed, and it is judged by pathological section that NY-CXCR4 can specifically target the head and neck squamous carcinoma tumor.

[0131] The test results are shown in FIG. 5:

[0132] Among them, the left side is the effect of the small molecule on the diagnosis of head and neck squamous carcinoma tumor by using the intraoperative fluorescence navigation device, and 8h, 12h and 24h after injecting 10 nmol NY-CXCR4, only the tumor area of the mouse back shows a high fluorescence signal, while the surrounding normal tissue shows a low fluorescence. Therefore, it is proved that NY-CXCR4 has good specificity in identifying CXCR4 receptor overexpressing head and neck squamous carcinoma, can realize rapid clearance in normal tissue, and long-term retention in head and neck squamous carcinoma tumor site, thereby achieving the effect of in vivo diagnosis.

[0133] Among them, the right side is the fluorescence signal distribution of the isolated organs at the corresponding time, and after injecting 10 nmol NY-CXCR4, the fluorescence intensity of the tumor tissue is significantly higher than that of the muscle tissue at each time point. Therefore, it is proved that NY-CXCR4 can accumulate in the tumor, realizing the intraoperative specific recognition of head and neck squamous carcinoma tumor.

[0134] In summary, the purpose of the present disclosure includes providing a near-infrared fluorescent probe and its preparation method and application, which can actively target chemokine receptor 4 (CXCR4), rapidly clear in normal tissue, and long-term retention in tumor site, thereby achieving the effect of in vivo diagnosis. A synthesis method of a diagnostic metastatic lymph node near-infrared fluorescent tracer is disclosed, which is synthesized by using the method of organic total synthesis The modified S0456 near-infrared small molecule, NY-CXCR4, has good active targeting effect in identifying triple-negative breast cancer and head and neck squamous carcinoma. This near-infrared fluorescent dye retains the water solubility and specificity of the dye to tumor cells. The tracer has the advantages of good water solubility and high fluorescence quantum yield, and has great development potential in the fields of near-infrared fluorescence navigation tumor surgery and medical cell labeling.

[0135] The present disclosure provides a novel near-infrared fluorescent tracer targeting CXCR4 receptor for specific targeting of triple-negative breast cancer and head and neck squamous carcinoma. S0456 is used as the parent, S0456 and the hydroxyphenylpropionic acid are connected together, and finally NY-CXCR4 active targeting near-infrared fluorescent molecule is synthesized by condensation. It is proved by animal experiment that NY-CXCR4 can target tumor.

[0136] It should be finally pointed out that the above embodiments are only used to illustrate the technical solutions of the present disclosure, rather than limit them; although the present disclosure has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present disclosure. Industrial applicability

[0137] The targeting near-infrared fluorescent compound provided by the present disclosure has good active targeting effect in identifying CXCR4 overexpressing triple-negative breast cancer and head and neck squamous cell carcinoma; and the targeting near-infrared fluorescent compound retains the water solubility and specificity of the dye to tumor cells, and has the advantages of good water solubility and high fluorescence quantum yield.

Claims

1. A near-infrared fluorescent targeting compound, characterized in that, The targeting near-infrared fluorescent compound has a structure shown in the following formula I:

2. A method of preparing a targeted near-infrared fluorescent compound according to claim 1, characterized in that, The preparation method comprises the following steps: (1) reacting compound S0456 and p-hydroxyphenyl propionate to obtain compound A; The structural formula of the compound A is as follows: (2) reacting compound A and dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate to obtain compound B; The structural formula of the compound B is as follows: (3) reacting compound B and a CXCR4 inhibitor to obtain the targeted near-infrared fluorescent compound shown in formula I; The structure of the compound CXCR4 inhibitor is:

3. The method for preparing the targeted near-infrared fluorescent compound according to claim 2, characterized in that, In step (1), the molar ratio of compound S0456 to p-hydroxyphenyl propionate is 1:(2-4).

4. The method of producing a targeted near-infrared fluorescent compound according to claim 2 or 3, characterized by, In step (1), the p-hydroxyphenyl propionate comprises sodium p-hydroxyphenyl propionate.

5. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 4, characterized in that, In step (1), the reaction is carried out in a solvent, and the solvent comprises dimethyl sulfoxide.

6. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 5, characterized in that, In step (1), the reaction is carried out in a protective gas atmosphere, and the protective gas comprises nitrogen.

7. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 6, characterized in that, In step (1), the temperature of the reaction is 60-70°C, and the reaction time is 5-7 hours.

8. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 7, characterized in that, In step (1), after the reaction is completed, a post-treatment step is further included: The reaction liquid obtained after the reaction in step (1) is added dropwise into a mixed solution of ethyl acetate and ethanol, and then filtered and dried to obtain the compound A.

9. The method for preparing the targeted near-infrared fluorescent compound according to claim 8, characterized in that, In the mixed solution, the volume ratio of ethyl acetate to ethanol is 1:(0.5-2).

10. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 9, characterized in that, In step (2), the molar ratio of compound A to dipyrrolidinyl (N-succinimidoxyl) carbonium hexafluorophosphate is 1:(1.2-2).

11. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 10, characterized in that, In step (2), the reaction is carried out in the presence of a base, and the base comprises N,N-diisopropyl ethylamine.

12. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 11, characterized in that, In step (2), the molar ratio of compound A to the base is 1:(0.5-5).

13. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 12, characterized in that, In step (2), the reaction is carried out in a solvent, and the solvent comprises dimethyl sulfoxide.

14. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 13, characterized in that, In step (2), the temperature of the reaction is 30-40°C, and the reaction time is 1-3 hours.

15. The method of preparing a targeted near-infrared fluorescent compound according to any one of claims 2 to 14, wherein, In step (2), after the reaction is completed, a post-treatment step is further included: The reaction liquid obtained after the reaction in step (2) is added dropwise into a solution of acetone, and then filtered and dried to obtain the compound B.

16. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 15, characterized in that, In step (3), the molar ratio of compound B to the CXCR4 inhibitor is 1:(0.5-2).

17. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 16, characterized in that, In step (3), the reaction is carried out in the presence of a base, and the base comprises N,N-diisopropyl ethylamine.

18. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 17, characterized in that, In step (3), the molar ratio of compound B to the base is 1:(0.5-5).

19. The method of producing a targeted near-infrared fluorescent compound according to any one of claims 2 to 18, characterized in that, In step (3), the reaction is carried out in a solvent, and the solvent comprises dimethyl sulfoxide.

20. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 19, characterized in that, In step (3), the temperature of the reaction is 30-40°C, and the reaction time is 18-30 hours.

21. The method for preparing the targeted near-infrared fluorescent compound according to any one of claims 2 to 20, characterized in that, In step (3), after the reaction is completed, a post-treatment step is further included: The reaction liquid obtained after the reaction in step (3) is added dropwise into a solution of acetone, and then filtered and dried to obtain the targeted near-infrared fluorescent compound shown in formula I.

22. Use of the targeted near-infrared fluorescent compound or a racemate, stereoisomer, pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a reagent for tumor imaging.

23. The use according to claim 22, characterized in that, The tumor comprises a triple-negative breast cancer tumor and / or a head and neck squamous carcinoma tumor.

24. Use of a targeted near-infrared fluorescent compound of claim 1, or a racemate, stereoisomer, pharmaceutically acceptable salt thereof, in the manufacture of a reagent for identifying CXCR4 overexpressing tumors.

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