Compound capable of being used as glioma PBCT / BNCT diagnosis and treatment agent as well as preparation method and application thereof

Abstract

The invention discloses a compound capable of being used as a glioma PBCT / BNCT diagnosis and treatment agent as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The structural formula of the compound is as follows: # imgabs0 #, wherein BG is selected from any one of alkyl boric acid, alkyl boric acid ester and alkyl trifluoroborate; the B atom comprises 10B and / or 11B; n = 0, 1, 2 or 3. The heptamethine cyanine compound with the boric acid derivative unit at the tail end of the alkyl side chain contains boron, can penetrate through a blood brain barrier, and can be used as a boron delivery agent for PBCT or BNCT treatment of glioma. The compound can emit fluorescence under the irradiation of near-infrared light, and can be used as a near-infrared fluorescent probe for glioma. Therefore, the compound can be used as a glioma PBCT or BNCT diagnosis and treatment integrated reagent.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to a class of compounds that can be used as glioma PBCT / BNCT diagnostic and therapeutic agents, and a preparation method and application thereof. Background Art

[0002] Central nervous system tumors are one of the main diseases that endanger public health. The most common central nervous system tumors include primary brain gliomas, primary brain lymphomas, metastatic tumors, etc., as well as some intracranial and extracranial tumors of meningeal or neural origin (such as meningioma, hemangiopericytoma, schwannoma, etc.). Glioma, primary brain lymphoma, and metastatic tumors account for nearly 90% of intracranial malignant tumors. Among them, brain gliomas are characterized by high mortality and disability rates. They are the primary cause of cancer death in men under 40 years old and women under 20 years old, and the incidence rate is gradually increasing in the elderly population. Brain gliomas are located in the brain, the most important tissue organ in the human body. They often grow in an infiltrative manner and have no obvious boundaries with the surrounding normal brain tissue. Traditional surgical and radiotherapy methods are difficult to completely and effectively remove tumor lesions; in addition, the existence of the blood-brain barrier also limits the effective distribution of therapeutic drugs in the brain. Almost all macromolecular drugs, such as monoclonal antibodies and peptides, cannot penetrate the blood-brain barrier. Even most small molecule drugs cannot penetrate the blood-brain barrier. How to deliver drugs across the blood-brain barrier to target brain tumors has become a major difficulty in treating gliomas.

[0003] Boron neutron capture therapy (BNCT) is the use of biocompatible boron delivery agents to deliver boron-10 ( 10 B) The atoms selectively gather into cancer cells; then the tumor tissue is irradiated with thermal neutrons, so that the neutrons interact with the tumor cells. 10 B undergoes a nuclear reaction, producing α-particles and lithium-7( 7 Li) atoms, and a large amount of energy is generated. The generated α-particles and 7 Li ions produce secondary irradiation, and their energy transfer is highly linear, with the particle travel distance being approximately one cell diameter. These characteristics determine that sufficient amounts of 10 B atoms can cause damage to cells. Boron neutron delivery agents can 10 B selectively accumulates in cancer cells, so this radiotherapy can selectively damage tumor cells while leaving surrounding healthy cells unaffected. BNCT greatly reduces the side effects associated with traditional cancer treatments such as chemotherapy and radiotherapy. With BNCT, patients can achieve a better quality of life during treatment. Currently, BNCT has been successfully used to treat recurrent brain gliomas, recurrent head and neck tumors, and malignant melanoma. Clinical trials have also been conducted on other common tumors such as liver cancer, lung cancer, and prostate cancer, and relatively good treatment results have been achieved. BNCT therapy brings hope to glioma patients.

[0004] Proton therapy uses the energy released after a high-energy proton beam enters the tumor tissue to kill tumor cells. After the proton enters the human body, a sharp dose peak, the Bragg peak, is formed at the end of the range. The position of the Bragg peak can be accurately controlled by modulating the energy broadening of the proton beam. In addition, the proton beam has less energy loss on the incident path, less side scattering, and a smaller beam boundary penumbra, so the normal tissue around the tumor receives less dose. However, the anatomical structure inside the tumor tissue will affect the proton beam dose distribution, and the presence of the beam boundary penumbra will also cause damage to normal tissue. Proton boron capture therapy (PBCT) is the use of 11 The nuclear transformation reaction of B with protons produces short-range high-energy alpha particles to kill tumor cells, which can improve the accuracy of proton therapy, reduce the energy and dose of proton beam, thereby reducing treatment costs and minimizing damage to normal tissues.

[0005] Whether it is BNCT or PBCT tumor treatment, the key to successful implementation is to selectively deliver sufficient boron atoms into tumor cells. Boron delivery agents used for BNCT / PBCT should have the following characteristics: (1) high tumor selectivity and a large ratio of boron concentrations in tumor tissue to normal tissue; (2) high boron uptake per gram of tumor tissue and long retention time; (3) fast blood clearance and low in vivo toxicity.

[0006] Based on the tumor targeting of heptamethine cyanine, patents CN115806567A and CN115806565A generate a conjugate of a boron compound and cyanine as a BNCT boron delivery agent by nucleophilically replacing meso-Cl in the IR-780 structure. However, removing meso-Cl from heptamethine cyanine will reduce the efficiency of boron delivery, because studies have shown that meso-Cl is essential for the tumor targeting of heptamethine cyanine and its retention time in tumor cells (Chem. Commun., 2023, 59, 13125). In addition, the two side chains of IR-780 are propyl groups with high fat solubility and high toxicity (ChemMedChem, 2023, 18, e202200561). Therefore, it is crucial to develop new tumor-targeted drugs. Summary of the invention

[0007] The purpose of the present invention is to provide a class of compounds that can be used as glioma PBCT / BNCT diagnostic and therapeutic agents, and their preparation methods and applications, in order to solve the problems existing in the above-mentioned prior art. The compound contains boron elements, which can be targeted and accumulated inside the glioma, becoming a boron carrier for PBCT and BNCT treatment of gliomas, and the compound can emit fluorescence under near-infrared light irradiation, and can be used as a glioma fluorescent probe to achieve the integration of imaging detection and PBCT or BNCT treatment.

[0008] To achieve the above object, the present invention provides the following solutions:

[0009] One of the technical solutions of the present invention is a heptamethine cyanine dye molecule with a boric acid derivative unit at the end of the alkyl side chain, and its structural formula is as follows:

[0010]

[0011] Wherein, BG is selected from any one of alkyl boronic acid, alkyl borate and alkyl trifluoroborate; B atom includes 10 B and / or 11 B; n=0, 1, 2 or 3.

[0012] The second technical solution of the present invention is a method for preparing the heptamethine cyanine dye molecule, comprising the following steps:

[0013] (1) ω-halogenated alkyl BG reacts with 2,3,3-trimethyl-3H-indole to form an intermediate;

[0014] (2) The intermediate is reacted with 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, and after the reaction is completed, the mixture is purified and dried to obtain the heptamethine cyanine dye molecule.

[0015] The third technical solution of the present invention is the use of the heptamethine cyanine dye molecule in the preparation of drugs for treating glioma PBCT / BNCT.

[0016] The fourth technical solution of the present invention is a drug for treating glioma PBCT / BNCT, comprising the heptamethine cyanine dye molecule.

[0017] Based on the above technical solution, the present invention has the following technical effects:

[0018] The heptamethine cyanine compound with a boric acid derivative unit at the end of the alkyl side chain of the present invention can pass through the blood-brain barrier and selectively accumulate in gliomas. Due to the presence of meso-Cl in the structure, the compound has higher glioma targeting and longer retention time in the tumor; at the same time, due to the presence of a sulfonate group with greater polarity in the structure, the compound has low toxicity and high safety. In addition, the compound has a simple structure, is easy to prepare, and has low production cost.

[0019] The heptamethine cyanine compound of the present invention, whose alkyl side chain ends are boronic acid derivative units, contains boron element and can be used as a boron delivery agent for PBCT or BNCT treatment of glioma. The compound emits fluorescence under near-infrared light irradiation and can be used as a near-infrared fluorescent probe for glioma. Therefore, the compound can be used as an integrated reagent for PBCT or BNCT diagnosis and treatment of glioma. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 Schematic diagram of the molecular structure of heptamethine cyanine, one side chain of which is an alkyl boronic acid derivative.

[0022] Figure 2 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is hexylboronic acid pinacol ester.

[0023] Figure 3 This is the NMR spectrum of the heptamethine cyanine molecule whose side chain is hexylboronic acid pinacol ester.

[0024] Figure 4 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is propylboronic acid pinacol ester.

[0025] Figure 5 This is the NMR spectrum of the heptamethine cyanine molecule whose side chain is propyl boric acid pinacol ester.

[0026] Figure 6 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is potassium hexyl trifluoroborate.

[0027] Figure 7 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is hexyl (N-methyliminodiacetic acid) borate.

[0028] Figure 8 The results of the cytotoxicity experiment of the heptamethine cyanine molecule, one of whose side chains is hexylboronic acid pinacol ester.

[0029] Fig. 9 The results of the cytotoxicity experiment of the heptamethine cyanine molecule, one of whose side chains is propyl boric acid pinacol ester.

[0030] Fig.10 The results of the tumor cell uptake experiment of the heptamethine cyanine molecule, one of whose side chains is hexylboronic acid pinacol ester.

[0031] Fig.11 The results of the tumor cell uptake experiment of the heptamethine cyanine molecule, one of whose side chains is propyl boric acid pinacol ester.

[0032] Fig.12 The results of the collection and imaging experiments of heptamethine cyanine molecules, one of whose side chains is hexylboronic acid pinacol ester, in tumor-bearing mice. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0038] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art, and the reagents or raw materials used, unless otherwise specified, are purchased from commercial channels or have been disclosed.

[0039] The embodiment of the present invention provides a heptamethine cyanine dye molecule whose alkyl side chain terminal is a boric acid derivative unit, and its structural formula is as follows:

[0040]

[0041] Wherein, BG is selected from any one of alkyl boronic acid, alkyl borate and alkyl trifluoroborate; B atom includes 10 B and / or 11 B; n=0, 1, 2 or 3.

[0042] In some specific embodiments, the heptamethine cyanine dye molecule having a boronic acid derivative unit at the end of the alkyl side chain includes:

[0043] The embodiment of the present invention also provides a method for preparing the heptamethine cyanine dye molecule, comprising the following steps:

[0044] (1) ω-halogenated alkyl BG reacts with 2,3,3-trimethyl-3H-indole to form an intermediate;

[0045] (2) The intermediate is reacted with 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, and after the reaction is completed, the mixture is purified and dried to obtain the heptamethine cyanine dye molecule.

[0046] In some specific embodiments, in step (1), the ω-halogenated alkyl BG includes 2-(6-bromohexyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 4-(2-bromoethoxy)2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 6-bromohexyl potassium trifluoroborate and 2,3,3-trimethyl-1-(6-(6-methyl-4,8-dioxo-1,3,6,2-dioxaborol-2-yl)hexyl)-3H-indol-1-ium bromide.

[0047] In some specific embodiments, in step (1), the molar ratio of the ω-halogenated alkyl BG to 2,3,3-trimethyl-3H-indole is (1-10):1.

[0048] In some specific embodiments, in step (1), the reaction conditions include: reaction temperature of 80-120° C., reaction time of 12-36 hours, and reaction solvent including at least one of water, benzene, toluene, ethylbenzene, xylene, dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, acetonitrile, ether, tetrahydrofuran, ethylene glycol dimethyl ether, 1,4-dioxane, acetone, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, tert-butanol, N-methylpyrrolidone, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide.

[0049] In some specific embodiments, in step (2), the molar ratio of the intermediate to 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-en-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate is 1:(1 to 6).

[0050] In some specific embodiments, in step (2), the reaction conditions include: reacting in the presence of a base, the reaction temperature is 80-120° C., the reaction time is 12-36 hours, and the reaction solvent includes at least one of water, benzene, toluene, ethylbenzene, xylene, dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, acetonitrile, ether, tetrahydrofuran, ethylene glycol dimethyl ether, 1,4-dioxane, acetone, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, tert-butanol, N-methylpyrrolidone, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide;

[0051] The molar ratio of the base to the intermediate is 1:1;

[0052] The base includes at least one of sodium hydroxide, potassium hydroxide, sodium acetate, potassium acetate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, triethylamine, triethylenediamine, 1,8-diazobispiro[5.4.0]undec-7-ene (DBU), 1,5-diazabicyclo[4.3.0]-5-nonene (DBN), pyridine, 4-dimethylaminopyridine (DMAP), N-methylmorpholine, tetramethylethylenediamine and tetramethylguanidine (TMG).

[0053] The embodiment of the present invention also provides the use of the heptamethine cyanine dye molecule in the preparation of a drug for treating glioma PBCT / BNCT.

[0054] The embodiment of the present invention also provides a drug for treating glioma PBCT / BNCT, comprising the heptamethine cyanine dye molecule.

[0055] In some specific embodiments, the ω-halogenated alkyl BG includes 3-halopropyl BG (n=0), 4-halobutyl BG (n=1), 5-halopentyl BG (n=2), 5-halohexyl BG (n=3), wherein the halogen includes bromine (Br) and iodine (I).

[0056] The test materials of the embodiments of the present invention are as follows:

[0057] The CAS number of 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-en-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate is 1802321-08-9.

[0058] Example 1 Synthesis of Heptamethine Cyanine Dye Molecule with Hexylboronic Acid Pinacol Ester as One Side Chain

[0059] Combined with Figure 1 , Figure 1 The figure is a schematic diagram of the molecular structure of heptamethine cyanine, one of whose side chains is an alkyl boronic acid derivative. Figure 2 conduct, Figure 2 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is hexylboronic acid pinacol ester.

[0060] In the first step, 2,3,3-trimethyl-3H-indole (318 mg, 2 mmol), 2-(6-bromohexyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1164 mg, 4 mmol) were placed in a 50 ml round bottom flask, 15 ml acetonitrile was added, and the mixture was stirred and refluxed for 18 hours under a nitrogen atmosphere at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 10-2% methanol / dichloromethane to obtain a solid intermediate product 1-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane)hexyl)-2,3,3-trimethyl-3H-indol-1-ium bromide.

[0061] In the second step, the intermediate obtained in the first step (370 mg, 1 mmoL) and 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (525 mg, 1 mmoL) were placed in a 50 ml round-bottom flask, sodium acetate (82 mg, 1 mmoL) and 15 ml of anhydrous ethanol were added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 12-6% methanol / dichloromethane to obtain a solid product.

[0062] The compound drug obtained in this example was subjected to characterization tests, as shown in the attached Figure 3 As shown, Figure 3 The 1H-NMR spectrum of the boron-containing cyanine compound is shown in FIG. Figure 3 The difference in peaks between substances can be seen in the hydrogen nuclear magnetic resonance (1H-NMR) spectrum, and the expected new structure molecule was formally formed through comparative analysis.

[0063] Example 2 Synthesis of Heptamethine Cyanine Dye Molecule with Propyl Boronic Acid Pinacol Ester as One Side Chain

[0064] Combined with Figure 1 , Figure 1 The figure is a schematic diagram of the molecular structure of heptamethine cyanine, one of whose side chains is an alkyl boronic acid derivative. Figure 4 conduct, Figure 4 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is hexylboronic acid pinacol ester.

[0065] In the first step, 2,3,3-trimethyl-3H-indole (318 mg, 2 mmol), 4-(2-bromoethoxy)2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (992 mg, 4 mmol) were placed in a 50 ml round bottom flask, 15 ml acetonitrile was added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 10-2% methanol / dichloromethane to obtain a solid intermediate product 1-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane)propyl)-2,3,3-trimethyl-3H-indol-1-ium bromide.

[0066] In the second step, the intermediate obtained in the first step (328 mg, 1 mmoL) and 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (525 mg, 1 mmoL) were placed in a 50 ml round-bottom flask, sodium acetate (82 mg, 1 mmoL) and 15 ml of anhydrous ethanol were added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 12-6% methanol / dichloromethane to obtain a solid product.

[0067] The compound drug obtained in this example was subjected to characterization tests, as shown in the attached Figure 5 As shown, Figure 5 The 1H-NMR spectrum of the boron-containing cyanine compound is shown in FIG. Figure 5The difference in peaks between substances can be seen in the hydrogen nuclear magnetic resonance (1H-NMR) spectrum, and the expected new structure molecule was formally formed through comparative analysis.

[0068] Example 3 Synthesis of Heptamethine Cyanine Dye Molecule with Potassium Hexyl Trifluoroborate as One Side Chain

[0069] Combined with Figure 1 , Figure 1 The figure is a schematic diagram of the molecular structure of heptamethine cyanine, one of whose side chains is an alkyl boronic acid derivative. Figure 6 conduct, Figure 6 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is potassium hexyl trifluoroborate.

[0070] In the first step, 2,3,3-trimethyl-3H-indole (318 mg, 2 mmol), 6-bromohexyl trifluoroborate potassium (1084 mg, 4 mmol) were placed in a 50 ml round bottom flask, 15 ml acetonitrile was added, and the mixture was stirred and refluxed for 18 hours under a nitrogen atmosphere at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 10-2% methanol / dichloromethane to obtain a solid intermediate product 6-(2,3,3-trimethyl-3H-indolium-1-)hexyl trifluoroborate.

[0071] In the second step, the intermediate obtained in the first step (311 mg, 1 mmoL) and 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (525 mg, 1 mmoL) were placed in a 50 ml round-bottom flask, sodium acetate (82 mg, 1 mmoL) and 15 ml of anhydrous ethanol were added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 18-6% methanol / dichloromethane to obtain a solid product.

[0072] Example 4 Synthesis of Heptamethine Cyanine Dye Molecule with Hexyl (N-methyliminodiacetic Acid) Borate as One Side Chain

[0073] Combined with Figure 1 , Figure 1 The figure is a schematic diagram of the molecular structure of heptamethine cyanine, one of whose side chains is an alkyl boronic acid derivative. Figure 7 conduct, Figure 7 The invention provides a synthetic route for a heptamethine cyanine molecule in which one side chain is hexyl (N-methyliminodiacetic acid) borate.

[0074] In the first step, 2,3,3-trimethyl-3H-indole (318 mg, 2 mmol), 2,3,3-trimethyl-1-(6-(6-methyl-4,8-dioxo-1,3,6,2-dioxaborol-2-yl)hexyl)-3H-indol-1-ium bromide (1280 mg, 4 mmol) were placed in a 50 ml round bottom flask, 15 ml of acetonitrile was added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 10-2% methanol / dichloromethane to obtain a solid intermediate product 2,3,3-trimethyl-1-(6-(6-methyl-4,8-dioxo-1,3,6,2-dioxaborol-2-yl)hexyl)-3H-indol-1-ium bromide.

[0075] In the second step, the intermediate obtained in the first step (399 mg, 1 mmoL) and 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (525 mg, 1 mmoL) were placed in a 50 ml round-bottom flask, sodium acetate (82 mg, 1 mmoL) and 15 ml of anhydrous ethanol were added, and the mixture was stirred and refluxed under nitrogen atmosphere for 18 hours at a reaction temperature of 100°C. After the reaction was completed, the mixture was cooled to room temperature. The reaction solution was concentrated and then eluted with 20-10% methanol / dichloromethane to obtain a solid product.

[0076] Example 5

[0077] 1 This example demonstrates the toxicity of a heptamethine cyanine dye molecule whose side chain is hexylboronic acid pinacol ester to human microglial cells HMC3, human glioma blast cells U251, and human glioma blast cells U87MG through experiments.

[0078] The three types of cells were seeded in a 96-well plate at a density of 5000 cells per well and incubated for 24 hours at 37±0.5°C and 5% CO2. The drug was then added to the wells at a concentration gradient of (10, 20, 30, 40, 50) μM and incubated for another 24 hours. The proportion of live cells was determined by the CCK-8 method to evaluate the toxicity of the drug.

[0079] Through experiments and Figure 8 The results show that through the cytotoxicity test graphic analysis, the side chain boron-containing cyanine compound was incubated with human microglial cells HMC3, human glioma blast cells U251, and human glioma blast cells U87MG at a concentration of 50μM for 24 hours, and the cell viability reached 63%, 74%, and 59%, respectively. This shows that the side chain boron-containing cyanine compound has little toxicity to normal cell glioma cells and is highly safe.

[0080] 2 This example demonstrates the toxicity of a heptamethine cyanine dye molecule whose side chain is propyl boric acid pinacol ester to human microglial cells HMC3, human glioma blast cells U251, and human glioma blast cells U87MG through experiments.

[0081] The three types of cells were seeded in a 96-well plate at a density of 5000 cells per well and incubated for 24 hours at 37±0.5°C and 5% CO2. The drug was then added to the wells at a concentration gradient of (10, 20, 30, 40, 50) μM and incubated for another 24 hours. The proportion of live cells was determined by the CCK-8 method to evaluate the toxicity of the drug.

[0082] Through experiments and Fig. 9 The results show that through the cytotoxicity test graphic analysis, the side chain boron-containing cyanine compound was incubated with human microglial cells HMC3, human glioma blast cells U251, and human glioma blast cells U87MG at a concentration of 50 μM for 24 hours, and the cell viability reached 79%, 75%, and 90%, respectively. This shows that the side chain boron-containing cyanine compound has little toxicity to normal cell glioma cells and is highly safe.

[0083] The present invention also carried out cytotoxicity tests using a heptamethine cyanine dye molecule whose side chain is potassium hexyl trifluoroborate and a heptamethine cyanine dye molecule whose side chain is hexyl (N-methyliminodiacetic acid) borate. The results show that these two compounds have low toxicity to normal cell glioma cells and are highly safe.

[0084] Example 6

[0085] This example demonstrates the ability of human microglial cells, human glioma blast cells U251 and human glioma blast cells U87MG to absorb heptamethine cyanine dye molecules whose side chain is hexylboronic acid pinacol ester through experiments.

[0086] The three types of cells were seeded in a 6-well plate at a density of 100,000 cells per well and incubated for 24 hours at 37±0.5°C and 5% CO2. The drug was then added to the 6-well plate at a concentration of 50 μM and incubated for 6 hours and 12 hours, respectively.

[0087] The cells were treated by first removing the original culture medium, washing with PBS, digesting the cells from the 6-well plate, centrifuging, and counting. Finally, the boron content was detected by ICP-MS.

[0088] Through experiments and Fig.10The results showed that at a concentration of 50 μM, the compound had a high uptake in human glioma cells U251 and U87, especially in U87 cells, where the boron content could reach up to 0.23 μg / 10 6 This content is consistent with the minimum concentration of 20ng / 10 required to achieve BNCT. 6 This further demonstrates that the compound is very suitable for the treatment of brain glioma BNCT.

[0089] 2 This example demonstrates the ability of human glioma blast cells U251 and human glioma blast cells U87MG to take up heptamethine cyanine dye molecules whose side chain is propyl boric acid pinacol ester through experiments.

[0090] The three types of cells were seeded in a 6-well plate at a density of 100,000 cells per well and incubated for 24 hours at 37±0.5°C and 5% CO2. The drug was then added to the 6-well plate at a concentration of 50 μM and incubated for 6 hours and 12 hours, respectively.

[0091] The cells were treated by first removing the original culture medium, washing with PBS, digesting the cells from the 6-well plate, centrifuging, and counting. Finally, the boron content was detected by ICP-MS.

[0092] Through experiments and Fig.11 The results showed that at a concentration of 50 μM, the compound had a high uptake in human glioma cells U251 and U87, especially in U87 cells, where the highest boron content could reach 0.19 μg / 10 6 This content is consistent with the minimum concentration of 20ng / 10 required to achieve BNCT. 6 This further demonstrates that the compound is very suitable for the treatment of brain glioma BNCT.

[0093] The present invention also carried out an uptake capacity test using a heptamethine cyanine dye molecule whose side chain is potassium hexyl trifluoroborate and a heptamethine cyanine dye molecule whose side chain is hexyl (N-methyliminodiacetic acid) borate, and achieved technical effects comparable to those of a heptamethine cyanine dye molecule whose side chain is hexyl pinacol borate and a heptamethine cyanine dye molecule whose side chain is propyl pinacol borate, indicating that the compound is highly suitable for the treatment of brain glioma BNCT.

[0094] Example 7

[0095] This example demonstrates through experiments that a heptamethine cyanine dye molecule whose side chain is hexylboronic acid pinacol ester can accumulate at the glioma site in the brain of tumor-bearing mice and can be imaged by near-infrared fluorescence.

[0096] Weigh 20 mg of a heptamethine cyanine dye molecule compound whose side chain is hexylboronic acid pinacol ester, add 26 mg of D-fructose, then add 1 ml of animal injection water and 1 mol / l NaOH solution, add drop by drop, and blow the pipette fully to dissolve the compound at the bottom as completely as possible. Use pH test paper to measure the pH value of the solution, add 0.1 mol / l hydrochloric acid solution drop by drop, and adjust the pH value to about 7-8. Inject 300-400 ul of the prepared solution into mice with brain gliomas through tail vein injection. Use animal in vivo fluorescence imaging to measure the imaging results of mouse brain tumors at 0.5, 1, 2, 3, 4, 6, 9, 12, and 24 hours.

[0097] Through experiments and Fig.12 The results show that the heptamethine cyanine dye molecule with hexylboronic acid pinacol ester as one side chain can accumulate in the glioma site in the brain of tumor-bearing mice and can be imaged by near-infrared fluorescence. The amount of the compound accumulated in the glioma reached a peak in 3 hours, and then the fluorescence value gradually decayed and basically disappeared after 24 hours. This shows that the compound can be cleared through metabolism.

[0098] The inventors also used the compounds prepared in Examples 2 to 4 to conduct a collection test at the glioma site in the brain of tumor-bearing mice, and achieved a technical effect equivalent to that of the heptamethine cyanine dye molecule with hexylboronic acid pinacol ester as a side chain.

[0099] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation methods of the present invention. For ordinary technical users in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A heptamethine cyanine dye molecule having a boric acid derivative unit at the end of the alkyl side chain, characterized in that: Its structural formula is as follows: Wherein, BG is selected from any one of alkyl boronic acid, alkyl borate and alkyl trifluoroborate; B atom includes 10 B and / or 11 B; n=0, 1, 2 or 3.

2. The heptamethine cyanine dye molecule according to claim 1, characterized in that The heptamethine cyanine dye molecule whose alkyl side chain ends are boric acid derivative units includes:

3. The method for preparing the heptamethine cyanine dye molecule according to claim 1 or 2, characterized in that: The following steps are involved: (1) ω-halogenated alkyl BG reacts with 2,3,3-trimethyl-3H-indole to form an intermediate; (2) The intermediate is reacted with 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-ene-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, and after the reaction is completed, the mixture is purified and dried to obtain the heptamethine cyanine dye molecule.

4. The preparation method according to claim 3, characterized in that: In step (1), the ω-halogenated alkyl BG includes 2-(6-bromohexyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 4-(2-bromoethoxy)2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane, 6-bromohexyl potassium trifluoroborate and 2,3,3-trimethyl-1-(6-(6-methyl-4,8-dioxo-1,3,6,2-dioxaborol-2-yl)hexyl)-3H-indol-1-ium bromide.

5. The preparation method according to claim 3, characterized in that: In step (1), the molar ratio of the ω-halogenated alkyl BG to 2,3,3-trimethyl-3H-indole is (1-10):

1.

6. The preparation method according to claim 3, characterized in that: In step (1), the reaction conditions include: a reaction temperature of 80-120° C., a reaction time of 12-36 hours, and a reaction solvent including at least one of water, benzene, toluene, ethylbenzene, xylene, dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, acetonitrile, ether, tetrahydrofuran, ethylene glycol dimethyl ether, 1,4-dioxane, acetone, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, tert-butanol, N-methylpyrrolidone, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide.

7. The preparation method according to claim 3, characterized in that: In step (2), the molar ratio of the intermediate to 4-(2-((E)-2-((E)-2-chloro-3-((phenylamino)methylene)cyclohex-1-en-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate is 1:(1 to 6).

8. The preparation method according to claim 3, characterized in that: In step (2), the reaction conditions include: reacting in the presence of a base, the reaction temperature is 80-120° C., the reaction time is 12-36 hours, and the reaction solvent includes at least one of water, benzene, toluene, ethylbenzene, xylene, dichloromethane, chloroform, carbon tetrachloride, 1,2-dichloroethane, acetonitrile, ether, tetrahydrofuran, ethylene glycol dimethyl ether, 1,4-dioxane, acetone, methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, tert-butanol, N-methylpyrrolidone, N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide; The molar ratio of the base to the intermediate is 1:1; The base includes at least one of sodium hydroxide, potassium hydroxide, sodium acetate, potassium acetate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, triethylamine, triethylenediamine, 1,8-diazobispiro[5.4.0]undec-7-ene (DBU), 1,5-diazabicyclo[4.3.0]-5-nonene (DBN), pyridine, 4-dimethylaminopyridine (DMAP), N-methylmorpholine, tetramethylethylenediamine and tetramethylguanidine (TMG).

9. Use of the heptamethine cyanine dye molecule according to claim 1 or 2 in the preparation of a drug for treating glioma PBCT / BNCT.

10. A drug for treating glioma PBCT / BNCT, characterized in that: Including the heptamethine cyanine dye molecule according to claim 1 or 2.

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