A boron-containing compound and its preparation method

Abstract

This invention discloses a boron-containing compound and its preparation method. The invention provides a compound represented by Formula 5. This compound can be used to synthesize the compound represented by Formula I, which exhibits low toxicity to tumor cells and can be combined with boron neutron capture therapy for targeted cancer treatment.

Description

[0001] This application is a divisional application of Chinese invention patent filed on August 2, 2022, with application number 2022109235473 and entitled "An alkenyl compound and its application". Technical Field

[0002] This invention relates to a boron-containing compound and its preparation method. Background Technology

[0003] The latest statistics show that malignant tumors account for 23.91% of all deaths, making cancer the leading cause of death for both urban and rural residents in my country. In 2015, approximately 3.928 million new cases of malignant tumors were diagnosed, and approximately 2.338 million people died from them, meaning that more than 10,000 people were diagnosed with cancer every day. Impoverishment due to cancer and relapse into poverty due to cancer have become serious economic and social problems in my country.

[0004] Boron neutron capture therapy (BNCT) is a novel binary radiotherapy technique for targeted cancer treatment. Its principle involves delivering boron neutrons, which have a specific affinity for tumor cells, to the tumor. 10 Drug B is injected into the patient, and this boron-containing drug specifically concentrates within the tumor. Then, the tumor site is irradiated with a superheated neutron beam. 10 The boron nucleus captures a thermal neutron to form an unstable composite nucleus. 11 B, 11 B then spontaneously splits into an alpha particle with a kinetic energy of 1.78 MeV and a particle with a kinetic energy of 1.01 MeV. 7 Li recoil nuclei (reaction cross-section of 6.3%); or an alpha particle with a kinetic energy of 1.47 MeV and a particle with a kinetic energy of 0.84 MeV. 7 The Li recoil nucleus emits a photon with an energy of 0.48 MeV (reaction cross-section of 93.7%). Due to the high energy of the released particles, they exhibit high linear energy conversion (LET) and low oxygen enhancement ratio, thus achieving highly selective and intense killing of tumor cells.

[0005] 7 The ranges of Li recoil nuclei and alpha particles in biological tissues are approximately 5 μm and 9 μm, respectively, which are smaller than the 10 μm diameter of tumor cells. 7 The lethal effects of Li and alpha particles are limited to uptake. 10 B's cells and their immediate neighbors. On the other hand, 10 B(n, α) 7 The Li reaction cross-section reaches 3840 keV, far exceeding the reaction cross-section of neutrons with nuclides that make up normal human tissues and blood. Therefore, neutrons have a greater potential to react with untaken nuclides. 10The radiation damage to normal tissues in β-radiotherapy can be controlled within a safe dose level. Therefore, this treatment is hailed as cellular-level radiotherapy, combining the advantages of both external and internal radiation therapy, while effectively avoiding the side effects of conventional external and internal radiation therapy.

[0006] Compared to other cancer treatment technologies, BNCT has unique advantages:

[0007] 1) Targeting of BNCT therapy

[0008] Boron-containing drugs possess targeting properties. They have a high affinity for tumor cells and are primarily absorbed by cancerous tissue, entering the tumor cells. 10 The content of B is higher than that in normal cells. 10 The content of B is high, so the therapeutic dose for tumor cells is much greater than that for normal cells.

[0009] 2) High Linear Energy Transfer Density (LET) Characteristics

[0010] Traditional radiotherapy methods, such as X-rays and gamma rays, are of low LET (relative biological effect), meaning they have a low RBE (relative biological effect on the body). They require oxygen to enhance the biological radiation effect. However, due to the rapid and invasive growth of malignant tumors, tumor tissue often suffers from insufficient blood supply, leading to local hypoxia and relatively poor treatment outcomes. Alpha particles and... 7 Li particles are highly LET-charged particles that can kill tumor cells in both oxygen-rich and hypoxic environments. Even for heavy ions such as protons and carbon ions, the RBE of alpha particles, which are products of the BNCT nuclear reaction, remains the highest.

[0011] 3) The therapeutic effect does not depend on the state of cancer cells.

[0012] Chemotherapy, X-knife, gamma knife, and conventional radiotherapy generally work on tumor cells in the proliferative phase (G1, S, G2, and M phases), but are insensitive to tumor cells in the quiescent phase (G0 phase). The continued growth of G0 phase tumor cells is the root cause of tumor recurrence. High-LET-based BNCT, alpha particle therapy, and... 7 Li particles' killing effect on tumor cells is independent of the cell cycle and can kill quiescent tumor cells as well. Hypoxic tumors become more resistant to conventional radiotherapy such as low-LET gamma rays and electron beams, while BNCT-based treatments can kill hypoxic cancer cells as long as sufficient boron is introduced.

[0013] 4) Precise treatment standards

[0014] To date, among cancer therapies, only neutron capture therapy has achieved a treatment scale precise to the micrometer level. Even the most precise minimally invasive surgical procedures are limited to the millimeter level, and the Bragg peak focused dose depth in heavy ion therapy is approximately 2.5–3.0 cm. The finer the treatment area, the fewer the potential side effects.

[0015] Ideal boron-containing targeted therapies for BNCT treatment should meet the following requirements:

[0016] It is non-toxic to humans at clinical doses;

[0017] 10 Drug B has a high affinity for tumor tissue, and the concentration ratio between tumor and normal tissue can reach 4:1 to 3:1.

[0018] per gram of tumor tissue 10 Drug B concentration reaches 20~35μg;

[0019] It can maintain a certain therapeutic concentration in tumor tissue during treatment.

[0020] Currently, the development of BNCT is relatively slow, which is closely related to the relatively lagging research and development of boron-containing drugs. To date, only disodium mercaptododecanoate (BSH) and p-dicarboxyboronylphenylalanine (BPA) are available for clinical use. BPA was approved for marketing in Japan in 2020, becoming the world's first approved boron-containing drug for BNCT. As a key breakthrough in BNCT technology, the efficacy of BSH and BPA falls far short of requirements, exhibiting insufficient tumor-targeting specificity and inadequate affinity for tumor cells. In-depth research has revealed that the ratio of boron concentration in tumors to normal tissues measured by BSH in human glioma samples is generally <1 (average 0.6), indicating it is considered a non-specific boron carrier. BPA, on the other hand, generally produces a boron concentration ratio no greater than 2.4, and it cannot penetrate the internal components of tumor cells, thus only remaining transiently within the cells. Furthermore, some tumor cells do not take up BPA. Clinically, boron-targeted therapies (BNCTs) are mainly used to treat cancers such as brain tumors, gliomas, and melanomas. This is primarily limited by the limited variety and indications of currently available boron-containing drugs. Therefore, developing boron-targeted drugs with broader indications, lower toxicity, and better targeting is imperative.

[0021] Fibroblast activating protein (FAP) is a specific biomarker for tumor-associated fibroblasts (TAF), possessing unique biological characteristics, stable genome, and abundant, specific expression in the tumor stroma. Existing FAP inhibitors are primarily used for cancer treatment. However, there are currently no FAP-targeting drugs applicable to boron neutron capture therapy. Summary of the Invention

[0022] The technical problem to be solved by the present invention is that the existing compounds suitable for BNCT have a single structure. To this end, the present invention provides a boron-containing compound and its preparation method, which can be used to synthesize the compound shown in Formula I. The compound shown in Formula I has low toxicity to tumor cells and can be combined with boron neutron capture therapy for targeted cancer treatment.

[0023] This invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof:

[0024]

[0025] Among them, R 1 Single bond, C1~C 10 Straight-chain alkylene, C2~C 10 Straight-chain subalkenyl groups, C2~C 10 Branched alkylene, C5-C8 cycloalkylene, C6-C 10 arylene-C1~C4 alkylene, C1~C4 alkylene-C6~C 10 arylene-C1~C4 alkylene Or C6~C 10 Alpha-aryl;

[0026] R 2 It is a hydroxyl group, a C1-C5 straight-chain alkyl group, a C3-C6 branched alkyl group, a C5-C8 cycloalkyl group, or a C6-C8 cycloalkyl group. 10 aryl-C1~C4 alkyl, C1~C4 alkyl-C6~C 10 Aryl-C1~C4 alkyl Or C6~C 10 Aryl;

[0027] R 3 It is a hydroxyl group, a C1-C5 straight-chain alkyl group, a C3-C6 branched alkyl group, a C5-C8 cycloalkyl group, or a C6-C8 cycloalkyl group. 10 aryl-C1~C4 alkyl, C1~C4 alkyl-C6~C 10 Aryl-C1~C4 alkyl Or C6~C 10 Aryl;

[0028] R 4 It can be H, F, or Cl;

[0029] R 5 It can be H, F, or Cl;

[0030] The above n1, n2 and n3 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0031] In one embodiment, in the compound represented by Formula I or a pharmaceutically acceptable salt thereof, some groups are defined as follows, and the remaining groups are defined as in any other embodiment (hereinafter referred to as "in one embodiment"): The R 1 It can be a single key.

[0032] In one scheme, the R... 2 It can be a hydroxyl group.

[0033] In one scheme, the R... 3 It can be a hydroxyl group.

[0034] In one scheme, the R... 4 It can be F or Cl.

[0035] In one scheme, the R... 4 It can be F.

[0036] In one scheme, the R... 5 It can be F or Cl.

[0037] In one scheme, the R... 5 It can be F.

[0038] In one embodiment, the compound represented by Formula I may be compound I-1:

[0039] .

[0040] The present invention also provides a compound as shown in Formula 2, Formula 3, Formula 4, Formula 5 or Formula 6:

[0041] , , , or

[0042] Among them, R 51 It is a C1~C4 alkyl group;

[0043] In the compound shown in Formula 3, R 32 It is a C1~C3 alkyl group; R 33 It is a C1~C3 alkyl group; R 34 It is a C1~C3 alkyl group;

[0044] In the compound shown in Formula 5, R52 It is a C1~C3 alkyl group; R 53 It is a C1~C3 alkyl group; R 54 It is a C1~C3 alkyl group; R 55 It is a C1~C3 alkyl group;

[0045] R 1 R 2 R 3 Defined as described above.

[0046] In one embodiment, the compound represented by Formula 2 is compound 2-1:

[0047] .

[0048] In one embodiment, the compound represented by Formula 3 is compound 3-1:

[0049] .

[0050] In one embodiment, the compound represented by Formula 4 is compound 4-1:

[0051] .

[0052] In one embodiment, the compound represented by Formula 5 is compound 5-1:

[0053] .

[0054] In one embodiment, the compound represented by Formula 6 is compound 6-1:

[0055] .

[0056] The present invention also provides a method for preparing the compound represented by Formula I, comprising the following steps: in a solvent, performing an amidation reaction as shown below with the compound represented by Formula 6 and the compound represented by Formula a in the presence of an amidating agent to obtain the compound represented by Formula I.

[0057] .

[0058] In one embodiment, the amidation reaction can be carried out under normal pressure.

[0059] In one embodiment, the amidation reaction can be carried out in the presence of oxygen.

[0060] In one embodiment, the amidating agent may be a combination of N,N,N',N'-tetramethylchloroformamidin hexafluorophosphate (TCFH) and 1-methylimidazole.

[0061] The solvent is a commonly used solvent for this type of reaction in the art.

[0062] In one embodiment, the solvent may be acetonitrile.

[0063] In one embodiment, the compound represented by Formula 6 may be compound 6-1 as described above.

[0064] In one embodiment, the molar volume ratio of the compound represented by Formula 6 to the solvent can be 0.121 mol / L.

[0065] In one embodiment, the molar ratio of the compound represented by Formula 6 to N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (TCFH) in the amidating agent may be 0.845.

[0066] In one embodiment, the molar volume ratio of the compound represented by Formula 6 to 1-methylimidazole in the amidating agent can be 2.417 mol / L.

[0067] In one embodiment, the reaction temperature can be 25°C.

[0068] In one embodiment, the reaction conditions may be a stirred reaction for 2 hours.

[0069] In one embodiment, the preparation method may further include the separation of the reaction products, wherein the separation may be performed sequentially by acidification, extraction, washing, drying, and column chromatography.

[0070] In one embodiment, the acidification may be performed using trifluoroacetic acid. Preferably, 1% trifluoroacetic acid is used.

[0071] In one embodiment, the extraction may be ethyl acetate extraction. Preferably, an equal volume of ethyl acetate is used for extraction.

[0072] In one embodiment, the washing may be performed with a sodium bicarbonate solution. Preferably, washing is done with an equal volume of sodium bicarbonate solution.

[0073] In one embodiment, the drying process may be anhydrous sodium sulfate drying.

[0074] In one embodiment, the developing solvent used in the column chromatography may be petroleum ether or ethyl acetate.

[0075] In one embodiment, the acidification may be achieved using 1% trifluoroacetic acid; the extraction may be performed three times with an equal volume of ethyl acetate; the washing may be performed twice with an equal volume of sodium bicarbonate solution; the drying may be performed using anhydrous sodium sulfate; and the developing solvent used in the column chromatography may be petroleum ether or ethyl acetate.

[0076] In one embodiment, the compound represented by Formula I may be the compound represented by Formula I-2: .

[0077] The present invention also provides a method for preparing the compound shown in Formula I-2, which includes the following steps: (1) in a solvent, the compound shown in Formula I and the compound shown in Formula D are reacted as shown below to obtain the compound shown in Formula 2;

[0078] (2) In a solvent, the compound shown in Formula 2 and the compound shown in Formula B are reacted as shown below to prepare the compound shown in Formula 3;

[0079] (3) The compound shown in Formula 3 was reacted in a solvent as shown below to prepare the compound shown in Formula 4;

[0080] (4) The compound shown in Formula 4 and the compound shown in Formula C are reacted in a solvent as shown below to prepare the compound shown in Formula 5-2;

[0081] (5) The compound shown in Formula 5-2 was reacted in a solvent as shown below to prepare the compound shown in Formula 6-2;

[0082] (6) In a solvent, the compound shown in Formula 6-1 and the compound shown in Formula A are subjected to an amidation reaction as shown below in the presence of an amidating agent to obtain the compound shown in Formula I-2. .

[0083] The solvent is a commonly used solvent for this type of reaction in the art.

[0084] In one embodiment, in step (1), the reaction can be carried out at atmospheric pressure; the reaction can be carried out in the presence of oxygen; the solvent can be methanol; the compound represented by formula D can be methanol; the reaction can be carried out in the presence of thionyl chloride; the reaction temperature can be 80°C; the reaction time can be 4 hours; the reaction can be quenched by adding water; the reaction may also include the separation of the reaction products, which may be carried out by adjusting the pH of the reaction solution and filtering in sequence.

[0085] In one embodiment, in step (1), the molar volume ratio of the compound represented by Formula 1 to the solvent can be 0.8 mol / L; the molar volume ratio of the compound represented by Formula 1 to the compound represented by Formula D can be 0.8 mol / L; the molar volume ratio of the compound represented by Formula 1 to sulfoxide can be 6.6 mol / L; and the pH of the reaction solution can be adjusted using sodium bicarbonate.

[0086] In one embodiment, in step (2), the reaction can be carried out under normal pressure; the reaction can be carried out under an inert atmosphere; the solvent can be tetrahydrofuran; the compound represented by formula 2 can be compound 2-1 above; the compound represented by formula B can be trimethylethynylsilane; the reaction can also be carried out in the presence of bis(triphenylphosphine)palladium(II) chloride, cuprous iodide and triethylamine; the reaction temperature can be 50°C; the reaction time can be 3 hours; the reaction can also include the separation and purification of the reaction product, which can be carried out sequentially as follows: filtering the reaction solution, dilution, extraction, washing, drying, filtration, concentration and purification.

[0087] In one embodiment, in step (2), the molar volume ratio of the compound represented by Formula 2 to the solvent can be 0.278 mol / L; the molar ratio of the compound represented by Formula 2 to the compound represented by Formula B can be 0.5; the molar ratio of the compound represented by Formula 2 to trimethylethynylsilane can be 20; the molar ratio of the compound represented by Formula 2 to cuprous iodide can be 20; the molar ratio of the compound represented by Formula 2 to triethylamine can be 0.67; the extraction in the separation and purification is ethyl acetate extraction; and the purification in the separation and purification is column chromatography purification.

[0088] In one embodiment, in step (2), the column chromatography purification process during separation and purification uses petroleum ether and ethyl acetate as the developing solvent.

[0089] In one embodiment, in step (3), the reaction can be carried out under normal pressure; the reaction can be carried out under an inert atmosphere; the compound represented by formula 3 can be compound 3-1; the solvent can be a mixture of methanol and dichloromethane; the reaction can be carried out in the presence of potassium fluoride; the reaction temperature can be room temperature; the reaction time can be 12 hours; the reaction can also include the separation and purification of the reaction product, which can be carried out sequentially as follows: filtering the reaction solution, dilution, extraction, washing, drying, filtration, concentration, pulping, filtration and drying.

[0090] In one embodiment, in step (3), the molar volume ratio of the compound shown in formula 3 to the solvent can be 0.45 mol / L; the molar ratio of the compound shown in formula 3 to potassium fluoride can be 0.33; the extraction in the separation and purification is ethyl acetate extraction; and the pulping in the separation and purification is pulping with petroleum ether and ethyl acetate.

[0091] In one embodiment, in step (4), the reaction can be carried out under normal pressure; the reaction can be carried out under an inert atmosphere; the solvent can be toluene; the compound represented by formula 4 can be compound 4-1 above; the compound represented by formula C can be pinacolborane; the reaction can be carried out in the presence of carbonyl chloride tris(triphenylphosphine)ruthenium(II); the reaction temperature can be 50°C; the reaction time can be 14 hours; the reaction can also include the separation and purification of the reaction product, which can be carried out sequentially as follows: filtering the reaction solution, dilution, extraction, washing, drying, filtration, concentration and purification.

[0092] In one embodiment, in step (4), the molar volume ratio of the compound represented by Formula 4 to the solvent can be 0.47 mol / L; the molar ratio of the compound represented by Formula 4 to the compound represented by Formula C can be 0.26; the molar ratio of the compound represented by Formula 4 to carbonyl chloride tris(triphenylphosphine)ruthenium(II) can be 71; the extraction in the separation and purification is ethyl acetate extraction; and the purification in the separation and purification is column chromatography purification.

[0093] In one embodiment, in step (4), the column chromatography purification process during separation and purification uses petroleum ether and ethyl acetate as the developing solvent.

[0094] In one embodiment, in step (5), the reaction can be carried out under normal pressure; the reaction can be carried out in the presence of oxygen; the solvent can be 1,4-dioxane; the compound represented by formula 5-2 can be the above-mentioned compound 5-1; the reaction can be carried out in the presence of hydrochloric acid; the reaction temperature can be 85°C; the reaction time can be 12 hours; the reaction can also include the separation and purification of the reaction product, which can be carried out sequentially as extraction of the reaction solution, separation, drying, pulping, filtration and drying.

[0095] In one embodiment, in step (5), the molar volume ratio of the compound represented by formula 5-2 to the solvent can be 0.59 mol / L; the concentration of hydrochloric acid is 6 mol / L; the molar volume ratio of the compound represented by formula 5-2 to hydrochloric acid can be 0.39 mol / L; the extraction in the separation and purification is ethyl acetate extraction; and the pulping in the separation and purification is pulping with methyl tert-butyl ether.

[0096] In one embodiment, the reaction conditions in step (6) can refer to the reaction conditions in the preparation method of the compound shown in Formula I above.

[0097] The present invention also provides a pharmaceutical composition comprising substance X and pharmaceutical excipients; wherein substance X is a compound as shown in Formula I above or a pharmaceutically acceptable salt thereof.

[0098] The present invention also provides the use of substance X in the preparation of a drug; wherein substance X is a compound as shown in Formula I above or a pharmaceutically acceptable salt thereof; and wherein the drug is a drug for treating tumors.

[0099] In one particular approach, the drug used to treat tumors is a radiotherapy-targeted drug.

[0100] In one particular approach, the radiotherapy-targeting drug is a boron neutron capture therapy drug.

[0101] Unless otherwise specified, the terms used in this invention have the following meanings:

[0102] The term "alkyl" refers to a straight-chain or branched alkyl group having a specified number of carbon atoms (e.g., C1 to C4). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, etc.

[0103] The term "alkylene" refers to a straight-chain or branched divalent alkyl group having a specified number of carbon atoms (e.g., C1-C4) and connected to other groups through two sites. Alkylenes include, but are not limited to, those with a specific number of carbon atoms (e.g., C1-C4). , , , wait.

[0104] The term "alkenyl" refers to a straight-chain or branched divalent alkenyl group having a specified number of carbon atoms (e.g., C2-C4) and connected to other groups through two sites. Alkenyl groups include, but are not limited to, those with a specific number of carbon atoms (e.g., C2-C4). , , wait.

[0105] The term "cycloalkyl" refers to a saturated monocyclic cyclic group consisting only of carbon atoms and having a specified number of carbon atoms (e.g., C5 to C8). Cycloalkyl groups include, but are not limited to, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0106] The term "aryl" refers to an aryl group having a specified number of carbon atoms (e.g., C6~C5). 10 Aryl groups are cyclic groups consisting solely of carbon atoms, which can be monocyclic or polycyclic, and each ring is aromatic (conforming to Hückel's rule). Aryl groups include, but are not limited to, phenyl and naphthyl groups.

[0107] The term "aryl" refers to a compound with a specified number of carbon atoms (e.g., C6~C5). 10 A aryl group is a divalent cyclic group consisting solely of carbon atoms, connected to other groups through two sites. It can be monocyclic or polycyclic, and each ring is aromatic (following Hückel's rule). Aryl groups include, but are not limited to, […]. , wait.

[0108] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for patient use) acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, aluminum, magnesium, bismuth, and ammonium salts. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, sulfate, and mesylate salts. See Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002) for details.

[0109] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0110] The reagents and raw materials used in this invention are all commercially available.

[0111] The positive and progressive effects of this invention are that the compound has low toxicity to tumor cells and can be combined with boron neutron capture therapy for targeted cancer treatment. Detailed Implementation

[0112] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0113] The A549 cells, HepG2 cells, U87MG cells, SCC9 cells, and A375 cells used in the following examples were all obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0114] The following examples

[0115] The manufacturer of the CCK-8 cell viability assay kit is Shanghai Donghuan Biotechnology Co., Ltd.

[0116] The manufacturer of PBS is Shanghai Shuangru Biotechnology Co., Ltd.

[0117] The manufacturer of DMEM high-glucose culture medium is Shanghai Donghuan Biotechnology Co., Ltd.

[0118] Example 1: Preparation of Compound I-1 of the Present Invention

[0119]

[0120] 1. Preparation of compound I-1

[0121] The preparation route of I-1 is shown below:

[0122]

[0123] Step 1: Compound 1 (50 g, 198 mmol) was dissolved in methanol (250 ml), and thionyl chloride (30 ml) was slowly added at room temperature. The reaction solution was reacted at 80 °C for 4 hours. The reaction solution was quenched with water (50 ml), the pH was adjusted to 7 with sodium bicarbonate solution, filtered, and the solid was collected. White compound 2-1 was obtained, with a yield of 93%.

[0124] The identification data for compound 2-1 are as follows: LC-MS: M (C 11 H8BrNO2) = 266.09 (m / z), [M+H] + 267.2

[0125] Step 2: Compound 2-1 (37.0 g, 139 mmol) and trimethylethynylsilane(b) (27.3 g, 278 mmol) were dissolved in tetrahydrofuran (500 mL). Then, under nitrogen protection, Pd(PPh3)2Cl2 (4.88 g, 6.95 mmol), CuI (1.32 g, 6.95 mmol), and Et3N (208 mmol, 29.0 mL) were slowly added. The reaction mixture was reacted at 50 °C for 3 hours under nitrogen protection. The reaction mixture was filtered, collected, diluted with water (100 mL), and extracted twice with ethyl acetate (250 mL each time). The organic phases were combined, washed twice with saturated brine (100 mL each time), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (eluent: petroleum ether, ethyl acetate). Compound 3-1 was obtained as a yellow solid, with a yield of 96%.

[0126] The identification data for compound 3-1 are as follows: LC-MS: M (C 16 H 17 NO2Si) = 283.40 (m / z), [M+H] + 284.2

[0127] Step 3: Compound 3-1 (38 g, 134 mmol) was dissolved in methanol (150 ml) and dichloromethane (150 ml). Under nitrogen protection, potassium fluoride (23.4 g, 402 mmol) was slowly added, and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was filtered and collected, diluted with water (100 ml), and extracted twice with ethyl acetate (250 ml each time). The organic phases were combined and washed twice with saturated brine (100 ml each time). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was stirred with petroleum ether and ethyl acetate (1:3, 30 ml) at room temperature for 30 minutes. After filtration and drying, compound 4-1 was obtained as a yellow solid, with a yield of 53%.

[0128] The identification data for compound 4-1 are as follows: LC-MS: M (C 13 H9NO2) = 211.22 (m / z), [M+H] + 212.0

[0129] Step 4: Compound 4-1 (15 g, 71 mmol) was dissolved in toluene (150 mL). Under nitrogen protection, ruthenium(II) carbonyl chloride (triphenylphosphine) (16971-33-8) (1 g, 1 mmol) and pinacol borane (c) (35.3 g, 275 mmol) were slowly added. The reaction mixture was stirred at 50 °C for 14 hours under nitrogen protection. The reaction mixture was filtered, collected, diluted with water (50 mL), and extracted twice with ethyl acetate (100 mL each time). The organic phases were combined, washed twice with saturated brine (50 mL each time), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (evolving solvent: petroleum ether, ethyl acetate). Compound 5-1 was obtained as a yellow solid, in 85% yield.

[0130] The identification data for compound 5-1 are as follows: LC-MS: M (C 19 H 22 BNO4) = 339.2 (m / z), [M+H] + 340.0

[0131] Step 5: Compound 5-1 (20 g, 59 mmol) was dissolved in 1,4-dioxane (100 ml), and hydrochloric acid (6 mol / L, 150 ml) was slowly added. The reaction mixture was stirred at 85 °C for 12 hours. The reaction mixture was extracted three times with ethyl acetate (80 ml each time). The aqueous phase was collected, freeze-dried, and the crude product was slurried with methyl tert-butyl ether at room temperature for 30 minutes. After filtration and drying, compound 6-1 was obtained as a yellow solid, with a yield of 91%.

[0132] The structural identification data of compound 6-1 are as follows: LC-MS: M (C 12 H10 BNO4) = 243.03 (m / z), [M+H] + 244.1

[0133] Step 6: Compound 6-1 (8 g, 29 mmol) and compound a (18.9 g, 15.7 mmol) were dissolved in acetonitrile (240 ml). N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (TCFH) (9.6 g, 34.3 mmol) and 1-methylimidazole (12 ml, 12.4 g) were slowly added. The reaction mixture was stirred at 25 °C for 2 hours. The pH of the reaction mixture was adjusted to 5 with 1% trifluoroacetic acid, and the reaction mixture was extracted three times with ethyl acetate (500 ml each time). The organic phases were combined and washed twice with sodium bicarbonate solution (200 ml each time). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (developing solvent: petroleum ether, ethyl acetate). Product I-1 was obtained as a yellow solid, with a yield of 46%.

[0134] 2. Structural identification

[0135] The LC-MS and NMR identification data of compound I-1 are as follows:

[0136] LC-MS: M (C 19 H 17 BF2N4O4) =414.18 (m / z), [M+H]+ 415.2

[0137] 1 H NMR: (400 MHz, DMSO-d6) δ 9.18-9.15 (m, 1H), 8.98 (d, J = 4.4 Hz,1H), 8.29 (s, 1H), 8.09-7.60 (m, 5H), 7.45-7.40 (m, 1H), 6.36-6.32 (m, 1H), 5.22 (d, J = 9.6 Hz, 1H), 4.34-4.16 (m, 4H), 2.97-2.83 (m, 2H).

[0138] F NMR: (400 MHz, DMSO-d6) δ -95.054, -103.651.

[0139] Example 2: Cytotoxicity of Compound I-1 on A549 Cells

[0140] A549 cells in the logarithmic growth phase were seeded into 96-well plates at a density of 1 x 10⁶ cells per well. 4Cells were cultured overnight in DMEM high-glucose medium. The medium was then aspirated, and the cells were washed twice with PBS (0.01 mol / L). Compound I-1 was added to fresh medium to prepare a series of solutions containing compound I-1 at concentrations of 0, 100, 250, 500, and 750 µg / ml. 100 µl of each solution was added to the wells of the cell plate to be tested. After 48 hours of drug treatment, the cell culture medium was aspirated, and the cells were washed twice with PBS (0.01 mol / L). 100 µL of CCK8 working solution was added, and the plate was incubated for 2 hours. The absorbance at 450 nm was measured using a microplate reader, and the effect of the drug on cell viability was calculated.

[0141] The experimental results are shown in Table 1. The low cytotoxicity of compound I-1 is an important indicator of BNCT, ensuring maximum intracellular boron concentration in tumor cells. Compound I-1 described in this invention exhibits low cytotoxicity to tumor cells, and its effect on the IC50 concentration of A549 cells is relatively low. 50 The concentration was 458.1 µg / ml, which meets the requirement for low toxicity of boron-containing drugs in boron neutron capture therapy.

[0142] Table 1. Inhibition rate of different concentrations of the compound on A549 cells

[0143]

[0144] Example 3: Cytotoxicity of Compound I-1 on HepG2 Cells

[0145] HepG2 cells in the logarithmic growth phase were seeded into 96-well plates at a density of 1 x 10⁶ cells per well. 4 Cells were cultured overnight in DMEM high-glucose medium. The medium was then aspirated, and the cells were washed twice with PBS (0.01 mol / L). A series of solutions containing compound I-1 at concentration gradients of 0, 100, 250, 500, and 750 µg / ml were prepared by adding compound I-1 to fresh medium. 100 µl of each solution was added to the wells of the culture plate. After 48 hours of treatment, the medium was aspirated, and the cells were washed twice with PBS (0.01 mol / L). 100 µL of CCK8 working solution was added, and the plate was incubated for 2 hours. The absorbance at 450 nm was measured using a microplate reader, and the effect of the drug on cell viability was calculated.

[0146] The experimental results are shown in Table 2. The low cytotoxicity of compound I-1 is an important indicator of BNCT, ensuring maximum intracellular boron concentration in tumor cells. Compound I-1 described in this invention exhibits low cytotoxicity to tumor cells, and its effect on the IC50 concentration of HepG2 cells is relatively low. 50The concentration was 340.1 µg / ml, which meets the requirement for low toxicity of boron-containing drugs in boron neutron capture therapy.

[0147] Table 2. Inhibition rate of different concentrations of compounds on HepG2 cells

[0148]

[0149] Example 4: Cytotoxicity of Compound I-1 on U87MG Cells

[0150] U87MG cells in the logarithmic growth phase were seeded into 96-well plates at a density of 1 x 10⁶ cells per well. 4 Cells were cultured overnight in DMEM high-glucose medium. The medium was then aspirated, and the cells were washed twice with PBS (0.01 mol / L). Compound I-1 was added to fresh medium to prepare a series of solutions containing compound I-1 at concentrations of 0, 100, 250, 500, and 750 µg / ml. 100 µl of each solution was added to the wells of the cell plate to be tested. After 48 hours of drug treatment, the cell culture medium was aspirated, and the cells were washed twice with PBS (0.01 mol / L). 100 µL of CCK8 working solution was added, and the plate was incubated for 2 hours. The absorbance at 450 nm was measured using a microplate reader, and the effect of the drug on cell viability was calculated.

[0151] The experimental results are shown in Table 3. The low cytotoxicity of compound I-1 is an important indicator of BNCT, ensuring maximum intracellular boron concentration in tumor cells. Compound I-1 described in this invention exhibits low cytotoxicity to tumor cells, and its effect on the IC50 concentration of U87MG cells is significant. 50 The concentration was 411.8 µg / ml, which meets the requirement for low toxicity of boron-containing drugs in boron neutron capture therapy.

[0152] Table 3. Inhibition rate of different concentrations of compounds on U87MG cells

[0153]

[0154] Example 5: Cytotoxicity of Compound I-1 on SCC9 Cells

[0155] SCC9 cells in the logarithmic growth phase were seeded into 96-well plates at a density of 1 x 10⁶ cells per well. 4Cells were cultured overnight in DMEM high-glucose medium. The medium was then aspirated, and the cells were washed twice with PBS (0.01 mol / L). A series of solutions containing compound I-1 at concentration gradients of 0, 100, 250, 500, and 750 µg / ml were prepared by adding compound I-1 to fresh medium. 100 µl of each solution was added to the wells of the culture plate. After 48 hours of treatment, the medium was aspirated, and the cells were washed twice with PBS (0.01 mol / L). 100 µL of CCK8 working solution was added, and the plate was incubated for 2 hours. The absorbance at 450 nm was measured using a microplate reader, and the effect of the drug on cell viability was calculated.

[0156] The experimental results are shown in Table 4. The low cytotoxicity of compound I-1 is an important indicator of BNCT, ensuring maximum intracellular boron concentration in tumor cells. Compound I-1 described in this invention exhibits low cytotoxicity to tumor cells, and its effect on the IC50 concentration of SCC9 cells is relatively low. 50 The concentration was 289.1 µg / ml, which meets the requirement for low toxicity of boron-containing drugs in boron neutron capture therapy.

[0157] Table 4. Inhibition rate of different concentrations of compounds on SCC9 cells

[0158]

[0159] Example 6: Cytotoxicity of Compound I-1 on A375 Cells

[0160] A375 cells in the logarithmic growth phase were seeded into 96-well plates at a density of 1 x 10⁶ cells per well. 4 Cells were cultured overnight in DMEM high-glucose medium. The medium was then aspirated, and the cells were washed twice with PBS (0.01 mol / L). A series of solutions containing compound I-1 at concentration gradients of 0, 100, 250, 500, and 750 µg / ml were prepared by adding compound I-1 to fresh medium. 100 µl of each solution was added to the wells of the culture plate. After 48 hours of treatment, the medium was aspirated, and the cells were washed twice with PBS (0.01 mol / L). 100 µL of CCK8 working solution was added, and the plate was incubated for 2 hours. The absorbance at 450 nm was measured using a microplate reader, and the effect of the drug on cell viability was calculated.

[0161] The experimental results are shown in Table 5. The low cytotoxicity of compound I-1 is an important indicator of BNCT, ensuring maximum intracellular boron concentration in tumor cells. Compound I-1 described in this invention exhibits low cytotoxicity to tumor cells, and its effect on the IC50 concentration of A375 cells is relatively low. 50The concentration was 586.5 µg / ml, which meets the requirement for low toxicity of boron-containing drugs in boron neutron capture therapy.

[0162] Table 5. Inhibition rate of different concentrations of the compound on A375 cells

[0163]

Claims

1. A compound as shown in formula 5: ###0001### Formula 5 wherein n1 is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. ， wherein R 1 Single bond, C1~C 10 Straight-chain alkylene, C2~C 10 Straight-chain subalkenyl groups, C2~C 10 Branched alkylene, C5-C8 cycloalkylene, C6-C 10 arylene-C1~C4 alkylene, C1~C4 alkylene-C6~C 10 arylene-C1~C4 alkylene Or C6~C 10 Alpha-aryl; R 51 R is C1-C4alkyl; R 52 R is C1-C3alkyl; R 53 is C1-C3alkyl; R 54 is C1-C3alkyl; R 55 is C1-C3alkyl; The compound shown in formula 4 is reacted with a compound shown in formula C in a solvent to produce a compound shown in formula 5-2, ###0002### Formula 5-2 wherein the reaction satisfies one or more of the following conditions:

2. The compound of claim 1, having the formula 5, ###0002### 5 R 1 is a single bond.

3. The compound of claim 1, having the formula 5, ###0002### 5 The compound represented by the formula 5 is compound 5-1: .

4. A method of preparing a compound as shown in formula 5-2, characterized in that, (1) the reaction is carried out under normal pressure; ， wherein R 51 is C1-C4 alkyl; R 52 is C1-C3 alkyl; R 53 is C1-C3 alkyl; R 54 is C1-C3 alkyl; R 55 is C1-C3 alkyl.

5. The preparation method according to claim 4, characterized in that, (2) the reaction is carried out under inert atmosphere; (3) the solvent is toluene; (5) the compound shown in formula C is pinacolborane; (6) the reaction is carried out in the presence of carbonylchlorohydridotriphenylphosphine ruthenium (II); (4) the compound represented by formula 4 is compound 4-1: ; (7) the reaction temperature is 50°C; (8) the reaction further comprises isolation and purification of the reaction product, which comprises filtration of the reaction solution, dilution, extraction, washing, drying, filtration, concentration and purification in sequence. The compound shown in formula 4 is reacted with a compound shown in formula C in a solvent to produce a compound shown in formula 5-2, ###0002### Formula 5-2 wherein the reaction satisfies one or more of the following conditions: (1) the molar volume ratio of the compound shown in formula 4 to the solvent is 0.47 mol / L; 6. The preparation method according to claim 5, characterized in that, (2) the molar ratio of the compound shown in formula 4 to the compound shown in formula C is 0.26; (3) the molar ratio of the compound shown in formula 4 to carbonylchlorohydridotriphenylphosphine ruthenium (II) is 71; (4) the extraction in the isolation and purification is ethyl acetate extraction; and the purification in the isolation and purification is column chromatography purification, wherein in the column chromatography purification in the isolation and purification, the developing agent is petroleum ether and ethyl acetate. The preparation method of the compound shown in formula 4 comprises the following steps: (1) the compound shown in formula 1 is reacted with a compound shown in formula D in a solvent to produce a compound shown in formula 2, ###0003### Formula 2 wherein the reaction satisfies one or more of the following conditions:

7. The preparation method according to claim 4, characterized in that, (1) the reaction is carried out under normal pressure; (2) the reaction is carried out under inert atmosphere; (3) the solvent is toluene; (5) the compound shown in formula D is pinacolborane; ， wherein R is C1-C4alkyl; and 51 C1-C4alkyl; and In the compound shown in Formula 3, R 32 It is a C1~C3 alkyl group; R 33 It is a C1~C3 alkyl group; R 34 It is a C1~C3 alkyl group.

8. A process for the preparation of a compound of formula I-2, characterized in that, (6) the reaction is carried out in the presence of carbonylchlorohydridotriphenylphosphine ruthenium (II); (7) the reaction temperature is 50°C; (8) the reaction further comprises isolation and purification of the reaction product, which comprises filtration of the reaction solution, dilution, extraction, washing, drying, filtration, concentration and purification in sequence. The preparation method of the compound shown in formula 4 comprises the following steps: The preparation method of the compound shown in formula 5-2 according to any one of claims 4-7; and, the preparation method of the compound shown in formula I-2 from the compound shown in formula 5-2, ###0004### Formula I-2 ， Among them, R 32 R 33 R 34 and R 51 As described in claim 7, R 4 It can be H, F, or Cl; R 5 is H, F or Cl.

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