Borodipyrromethene compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicine, and particularly relates to a BODIPY compound and a preparation method and application thereof.The BODIPY compound is disclosed, the structure of the BODIPY compound is shown as a formula I or a formula II, and tests prove that the BODIPY compound can generate BDP-tempo and alkyl free radicals through hydrolysis, and the BDP-tempo and the alkyl free radicals can be used for preparing the BODIPY compound. The polypeptide has a good anti-tumor effect and preliminary diagnosis capability, and lays a foundation for development of anti-tumor drugs and treatment and diagnosis agents. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to a boron dipyrromethene compound, a preparation method thereof, and an application thereof. Background Art

[0002] The concept of "Theranostic" was proposed by experts and CEO Funkhouser of ParmaNetics in 1998. It is defined as a single agent that combines diagnostic and therapeutic capabilities. That is, theranostics provides both a therapeutic drug and a diagnostic imaging agent at the same dose, thereby enabling simultaneous or sequential diagnosis and treatment. Compared with using different materials for these two purposes, theranostics combines the therapeutic and diagnostic functions in the same "package", with the prospect of overcoming the adverse differences in biodistribution and selectivity existing in current different imaging agents and therapeutic agents. The primary goal in the field of theranostics is to obtain the ability to image and monitor diseased tissues and pharmacokinetics, and the long-term goal is to adjust the drug dosage and treatment method at any time according to the monitoring results, so as to provide the correct treatment for the correct patient at the correct time and correct dose in this form, thereby providing more targeted and effective drug treatment for patients and achieving personalized precision treatment.

[0003] Alkoxyamines are a class of multifunctional molecules, usually with the structure R1ONR2R3. They have the unique property of spontaneously hydrolyzing into stable nitroxide radicals and highly reactive alkyl radicals. Alkyl radicals are excellent cytotoxic agents. They change the characteristics of cell membranes by extracting labile hydrogen atoms from proteins and lipids and adding to unsaturated lipids, and also cause random changes to nucleic acids, thereby inducing oxidative stress and ultimately leading to apoptosis or necrosis of cells. This randomness greatly reduces the possibility of tumor cells acquiring drug resistance. Alkyl radical-releasing drugs such as platinum derivatives and cyclophosphamide have achieved some success in chemotherapy currently. At the same time, the generated stable nitroxide radicals can be detected by electron paramagnetic resonance imaging (EPRI) or enhanced magnetic resonance imaging (OMRI), which makes it possible to monitor the concentration and pathway of drug delivery. However, the inherently low sensitivity of magnetic resonance technology makes the dosage of the imaging agent required for imaging huge, usually much larger than the appropriate therapeutic agent dosage. This results in the uptake concentration of alkoxyamines being unable to meet the requirements of both diagnosis and treatment simultaneously. Therefore, based on the structure of such molecules, developing a more effective imaging method to overcome the deficiencies of magnetic resonance technology and meet the requirements of clinical trials is a technical problem urgently to be solved in this field.

[0004] Fluoroboron dipyrrole (BODIPY) fluorescent dyes have many advantages compared to fluorescein, rhodamine, and cyanine dyes, such as strong light absorption ability, excellent photostability, biocompatibility, and easy modification. In recent years, they have played an important role in ion recognition, biological labeling, and phototherapy. In this invention, the fluorescent dye BDPIPY is used instead of nitroxide radicals, and fluorescence detection method is used instead of enhanced magnetic resonance imaging method to complete the diagnosis work, which has the significance of innovation and exploration.

[0005] Based on the above background, this application provides a fluoroboron dipyrrole compound with diagnostic and therapeutic effects. Summary of the Invention

[0006] The technical problem to be solved by this invention is to provide a fluoroboron dipyrrole compound with diagnostic and therapeutic effects, its preparation method, and application.

[0007] To solve the above problems, the technical solutions adopted by this invention are as follows: Technical Theme 1 A fluoroboron dipyrrole compound or its pharmaceutically acceptable salt, and the structure of the compound is shown in Formula I or Formula II:

[0008] Wherein, R is a straight-chain alkyl group with 1 to 5 carbon atoms.

[0009] As a further improvement of this invention, the straight-chain alkyl group with 1 to 5 carbon atoms is —CH3, —CH2CH3, —(CH2)2CH3, —(CH2)3CH3, —(CH2)4CH3.

[0010] The "pharmaceutically acceptable salts" described in the present invention refer to the salts of the compounds of the present invention, which are prepared from the compounds of the present invention and relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, the base addition salts can be obtained by contacting the neutral form of the compounds of the present invention with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable bases include salts prepared from inorganic bases and organic bases. The salts of the inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. The salts of the organic non-toxic bases include salts of primary amines, secondary amines and tertiary amines, including substituted amines and cyclic amines. For example: N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, aminoglucose, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, etc. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, and the inorganic acids include, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, and the organic acids include, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid and other similar acids.

[0011] Technical Theme 2 A method for preparing a fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof as described in Technical Theme 1, comprising the following steps: S1:

[0012] In solvent A, 2,4-dimethylpyrrole reacts with chloroacetyl chloride, and then triethylamine and boron trifluoride diethyl ether are added successively and continue to react to obtain BDP-1; S2:

[0013] In solvent B, BDP-1 reacts with sodium azide to obtain BDP-2; S3:

[0014] In solvent C, BDP-2 and compound X are reacted through a Click reaction to obtain BDP-Py; The R1 is .

[0015] As a further improvement of the present invention, the solvent A is selected from one of dichloromethane or toluene; the solvent B is ethanol; the solvent C is selected from one of ethanol, chloroform or water.

[0016] As a further improvement of the present invention, the molar ratio of chloroacetyl chloride, 2,4-dimethylpyrrole and boron trifluoride diethyl ether is 1~1.2:2:6~8; the molar ratio of BDP-1 and sodium azide is 1:4~5; the molar ratio of BDP-2 and compound X is 1:1.5~2.

[0017] As a further improvement of the present invention, when the solvent in S1 is dichloromethane, 2,4-dimethylpyrrole and chloroacetyl chloride are heated under reflux in a nitrogen atmosphere, the dropping temperatures of triethylamine and boron trifluoride diethyl ether are -10~0°C, and after addition, they are heated under reflux; when the solvent in S1 is toluene, 2,4-dimethylpyrrole and chloroacetyl chloride react at 40~50°C in a nitrogen atmosphere, the dropping temperatures of triethylamine and boron trifluoride diethyl ether are -10~0°C, and after addition, they react at 40~50°C; Sodium azide in S2 is added dropwise at room temperature, and the reaction temperature after dropping is 40~45°C; S3 reacts under nitrogen at room temperature, and the reaction time is 10~14h.

[0018] As a further improvement of the present invention, compound X is prepared by the following method:

[0019] a. Raw material 1 and 3-bromopropyne are reacted through a Williamson etherification reaction to obtain compound 2; b. Raw material 3 and NBS are reacted through a bromination reaction to obtain compound 4; c. Compound 2 and compound 4 are reacted through a Williamson etherification reaction to obtain compound 5; d. Compound 5 and R-I are reacted to obtain compound 6; The obtained compound 5 and compound 6 are compound X.

[0020] Technical theme three A pharmaceutical composition comprising a fluoroboron dipyrrole compound as described in Technical Theme One or a pharmaceutically acceptable salt thereof, which further comprises at least one pharmaceutically acceptable carrier or excipient.

[0021] As used herein, a "pharmaceutical composition" contains a therapeutically effective amount of the compound in Technical Subject One, its pharmaceutically acceptable salts or esters, solvates, isomers, polymorphs, isotopically labeled compounds, metabolites or prodrugs, and one or more pharmaceutically acceptable carriers, and is prepared in the form of tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, medicinal wines, decoction extracts, lozenges, mixtures, suppositories, injections, inhalants or sprays, etc. The pharmaceutical composition preferably contains 0.1% to 99.5% by weight of the poly-substituted benzheterocyclic compound of the present invention or its pharmaceutically acceptable salt as an active ingredient, and more preferably contains 0.5% to 99.5% by weight of the active ingredient.

[0022] As used herein, "pharmaceutically acceptable carriers or excipients" include: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifying agents, sweetening agents, flavoring agents, taste masking agents, coloring agents, anti-caking agents, humectants, chelating agents, plasticizers, thickening agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents. Those skilled in the art will understand that certain pharmaceutically acceptable excipients can be used with more than one function and with alternative functions, depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation. For example: When used orally, oral formulations can be prepared, such as tablets, capsules, granules, pills, etc., containing fillers (such as sugar derivatives like lactose, sucrose, glucose, mannitol and sorbitol; starch derivatives like corn starch, potato starch, dextrin and carboxymethyl starch; cellulose derivatives like crystalline cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose; gum arabic; dextran; silicate derivatives like magnesium aluminum metasilicate; phosphate derivatives like calcium phosphate; carbonate derivatives like calcium carbonate; sulfate derivatives like calcium sulfate, etc.), binders (such as gelatin, polyvinylpyrrolidone and polyethylene glycol), disintegrants (such as cellulose derivatives like sodium carboxymethyl cellulose, polyvinylpyrrolidone), lubricants (such as talc, calcium stearate, magnesium stearate, cetyl, boric acid, sodium benzoate, leucine), stabilizers (methyl paraben, propyl paraben, etc.), flavoring agents (such as common sweeteners, sour agents and fragrances, etc.). When used parenterally, injectables can be prepared, including sterile powders for injection and solvents for injection, and the carriers or excipients used include sterile water, Ringer's solution and isotonic sodium chloride solution. Appropriate additional agents such as antioxidants, buffering agents and bacteriostatic agents can also be added according to the nature of the drug. When used for rectal administration, the drug can be made into suppositories, etc. When used for pulmonary administration, the drug can be made into inhalants or sprays, etc. There are many resources available to those skilled in the art that describe pharmaceutically acceptable excipients and can be used to select suitable pharmaceutically acceptable excipients, such as books like "Remington: The Science and Practice of Pharmacy", "Chinese Pharmaceutical Yearbook", "Pharmaceutics", etc.

[0023] Technical Subject Four Use of a boron dipyrromethene compound as described in Technical Subject One in the preparation of an anti-tumor drug.

[0024] Technical Subject Five Use of a boron dipyrromethene compound as described in Technical Subject One in the preparation of a therapeutic diagnostic agent.

[0025] The beneficial effects produced by adopting the above technical solution are as follows: The present application discloses a boron dipyrromethene compound for treatment and imaging. It has been experimentally verified that the boron dipyrromethene compound disclosed in the present application can generate BDP-tempo and alkyl free radicals through hydrolysis, and has good anti-tumor effects and preliminary diagnostic capabilities, laying a foundation for the development of anti-tumor drugs and theranostics. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is the ultraviolet absorption spectrum of BDP-Py-5, BDP-Py-4, and BDP-tempo of the present invention in methanol; Figure 2 is the fluorescence emission diagram of BDP-Py-5, BDP-Py-4, and BDP-tempo of the present invention in methanol; Figure 3 is the test result diagram of the singlet oxygen generation ability of BDP-Py-5 and BDP-Py-4 of the present invention in methanol, where the result change diagrams per minute from top to bottom are for 0 - 10 minutes; Figure 4 is the test result diagram of the photostability of BDP-Py-5 and BDP-Py-4 of the present invention in DMSO, where the measurement results every 10 minutes from top to bottom are for 0 - 60 minutes; Figure 5 is the hydrolysis rate diagram of BDP-Py-4 of the present invention at pH = 6.5. From top to bottom, the fluorescence intensities detected at 0 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h are shown in sequence; Figure 6 is the hydrolysis rate diagram of BDP-Py-4 of the present invention at pH = 7.4. From top to bottom, the fluorescence intensities detected at 0 min, 30 min, 1 h, 2 h, 4 h, 8 h, 24 h, 36 h, 48 h, and 72 h are shown in sequence; Figure 7 is the hydrolysis rate diagram of BDP-Py-5 of the present invention at pH = 6.5. From top to bottom, the fluorescence intensities detected at 0 min, 30 min, 1 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h are shown in sequence; Figure 8 is the hydrolysis rate diagram of BDP-Py-5 of the present invention at pH = 7.4. From top to bottom, the fluorescence intensities detected at 0 min, 30 min, 1 h, 4 h, 8 h, 12 h, 24 h, 36 h, and 48 h are shown in sequence; Figure 9These are the A375 cell inhibitory activity graphs of different concentrations of BDP-Py-5 and BDP-Py-4 in the present invention under illuminated and non-illuminated states. The illumination condition is a 525 nm green light lamp with a radiation intensity of 20 W / m 2 ; Figure 10 These are the bar graphs of the survival rates of human vascular endothelial cells of different concentrations of BDP-Py-5 and BDP-Py-4 in the present invention under illuminated (525 nm green light, radiation intensity of 20 W / m 2 ) and non-illuminated states; Figure 11 These are the test graphs of the ability of different concentrations of BDP-Py-5 and BDP-Py-4 in the present invention to generate intracellular ROS under illuminated (525 nm green light, radiation intensity of 20 W / m 2 ). Rousp is the positive control group in the kit; Figure 12 These are the mouse tumor graphs in the anti-tumor experiment in mice of the present invention. In the figure, A - E correspond to groups A - E in Effect Example 8; Figure 13 These are the graphs of the changes in the body weights of mice in the anti-tumor experiment in mice of BDP-Py-5 and BDP-Py-4 in the present invention; Figure 14 These are the graphs of the tumor volumes of mice in the anti-tumor experiment in mice of BDP-Py-5 and BDP-Py-4 in the present invention. Detailed Embodiments

[0027] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be clearly and completely described below in conjunction with specific embodiments.

[0028] Example 1 Preparation of Intermediate BDP-1

[0029] 2,4-Dimethylpyrrole (30.8 g) was added to 2000 ml of dry dichloromethane and stirred evenly. After replacing with nitrogen, chloroacetyl chloride (12.8 ml) was added under nitrogen protection and stirred evenly. The mixture was heated to reflux and stirred for 1.5 h. After monitoring the reaction by TLC until 2,4-dimethylpyrrole was completely reacted, triethylamine (110 ml) was added at 0 °C. After 30 min, boron trifluoride diethyl etherate (140 ml) was added by syringe at 0 °C. After the addition was completed, the mixture was heated to reflux and reacted for 1.5 h. After monitoring the reaction by TLC until it was complete, it was concentrated to obtain a solid residue. The obtained residue was dissolved in dichloromethane, washed three times with 1000 ml of saturated sodium chloride, the organic phases were combined, dried over an appropriate amount of anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure from the filtrate. The sample was separated and purified by silica gel column chromatography with gradient elution. The ratio of the eluent petroleum ether:ethyl acetate (PE:EA) = 100:1 - 20:1 (v:v), and the solvent was evaporated under reduced pressure. 9 g of a brick-red solid powder (BDP-1) was obtained, with a yield of 18.7%. HRMS (ESI): Calcd for: 297.1136 Found: 297.1129. Preparation of Intermediate BDP-2 in Example 2

[0030] Compound BDP-1 (6.66 g) was added to 4000 ml of ethanol. Sodium azide (6.86 g) was added at room temperature under nitrogen protection, and the mixture was heated to 40 °C and reacted for 12 h. After monitoring the reaction of BDP-1 by LC-MS until it was complete, the solvent was concentrated under reduced pressure, diluted with water, extracted three times with 400 ml of dichloromethane, the organic phases were combined, washed with 400 ml of saturated brine, dried over an appropriate amount of anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The sample was separated and purified by silica gel column chromatography with gradient elution. The eluent was petroleum ether:dichloromethane (PE:DCM) = 100:1 - 1:1 (v:v), and 4 g of a red solid powder (BDP-2) was obtained, with a yield of 58.8%. HRMS (ESI): Calcdfor: 304.1540 Found: 304.1537. Preparation of Intermediate 6 in Example 3

[0031] Add raw material 1 (40.0 g) to a 1000 ml single-necked flask equipped with magnetic stirring, add 600 ml of CCl4. After dissolution, add NBS (73.0 g) and AIBN (6.2 g), heat under reflux for 1 h, cool to room temperature, add 500 ml of water for dilution, separate the layers, extract the aqueous phase with 500 ml of dichloromethane, combine the organic phases, wash three times with saturated brine, collect the organic phase, dry over anhydrous sodium sulfate, and further purify the product by silica gel column chromatography. The eluent ratio is petroleum ether: ethyl acetate (PE:EA) = 100:1 - 20:1. Obtain 38.0 g of a pale yellow oil (Compound 2) with a yield of 54.7%. HRMS (ESI): Calcd for: 210.1494 Found: 210.1497. Dissolve raw material 3 (30.0 g) in 300 ml of anhydrous DMF. Add NaH (9.0 g) portionwise at 0 °C, stir for 30 min, keep at 0 °C and dropwise add 3-bromopropyne (30.9 g). Stir at room temperature for 3 h, cool to 0 °C, quench the reaction with water, extract three times with 300 ml of EtOAc, combine the organic phases, wash three times with 300 ml of saturated brine, dry with an appropriate amount of anhydrous sodium sulfate, and concentrate under reduced pressure to evaporate the solvent. Further separate and purify the sample by silica gel column chromatography with gradient elution. The eluent ratio is petroleum ether: ethyl acetate (PE:EA) = 100:1 - 10:1 (v:v). Obtain 25 g of a white solid (Compound 4) with a yield of 68%. Calcd for: 185.9913 Found: 185.9911. Dissolve Compound 2 (16.3 g) and Compound 4 (14.4 g) in 300 ml of dry toluene, add PMDEAT (16.1 g) and copper iodide (13.3 g), protect with nitrogen, heat to 50 °C and react for 12 h. Cool to room temperature, filter by suction, wash the filter cake with 50 ml of ethyl acetate, wash the filtrate with saturated brine, dry over anhydrous sodium sulfate, concentrate, and separate and purify the sample by silica gel column chromatography. The eluent ratio is petroleum ether: ethyl acetate (PE:EA) = 100:1 - 8:1. Obtain 13.0 g of a white solid (Intermediate 5) with a yield of 53.1%. HRMS (ESI): Calcd for: 317.2224 Found: 317.2221. In a 100 ml single-necked flask, dissolve Intermediate 5 (5.5 g) in 55 ml of acetonitrile, add methyl iodide (3.87 g) at room temperature, stir at room temperature for 12 h, and concentrate to obtain 3.0 g of a white solid (crude Intermediate 6).

[0032] Preparation of Example 4 BDP-Py-4

[0033] In a 1000 ml three-necked flask equipped with magnetic stirring, add BDP-2 (1.0 g) and compound 6 (2.26 g) dissolved in 92 ml of chloroform. At room temperature, add 5 ml of triethylamine and copper(I) iodide (0.63 g). Replace nitrogen three times. After reacting at room temperature for 12 h, dilute with water, extract with 200 ml of dichloromethane. Combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and perform silica gel column chromatography with an eluent ratio of petroleum ether:methanol (PE:MeOH) = 100:1 - 1:1. Obtain 1.5 g of a brick-red solid powder. Further purify by thin-layer chromatography to obtain 0.37 g of a brick-red powder. The yield is 14.7%. 1 H NMR (600 MHz, DMSO-d6) δ 8.92 (d, J = 6.2 Hz, 2H), 8.10 - 8.02 (m, 3H), 6.28 (s, 2H), 5.80 (s, 2H), 5.08 (q, J = 6.7 Hz, 1H), 4.51 (s, 2H), 4.31 (s, 3H), 3.64 (tt, J = 11.2, 4.0 Hz, 1H), 2.45 (s, 6H), 2.18 (s, 6H), 1.85 (d, J = 12.6 Hz, 1H), 1.74 (d, J = 12.1 Hz, 1H), 1.45 (d, J = 6.7 Hz, 3H), 1.25 (s, 4H), 1.22 - 1.15 (m, 2H), 1.12 (s, 3H), 1.01 (s, 3H), 0.66 (s, 3H). 13 C NMR (151 MHz, DMSO-d6) δ 163.28, 156.57, 145.99, 142.33, 132.58, 125.18, 123.78, 123.03, 81.39, 69.01, 60.87, 60.34, 60.19, 47.94, 45.84, 45.51, 40.55, 34.54, 23.24, 21.35, 15.54, 14.82. HRMS (ESI): Calcd for: 634.3847 Found: 634.3845. Example 5 Preparation of BDP-Py-5

[0034] In a 1000 ml three-necked flask equipped with magnetic stirring, add BDP-2 (1.0 g) and compound 5 (1.56 g) dissolved in ethanol / chloroform / water. In this application, 292 ml of ethanol is used. At room temperature, add 5 ml of triethylamine and copper(I) iodide (0.63 g), displace nitrogen three times. After reacting at room temperature for 12 h, dilute with water, extract with 200 ml of dichloromethane, combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, evaporate the solvent under reduced pressure, and perform silica gel column chromatography. The eluent ratio is petroleum ether:methanol (PE:MeOH) = 100:1 - 25:1. Obtain 0.67 g of a brick-red solid powder. Further purify by thin-layer chromatography to obtain 0.47 g of a brick-red powder. The yield is 23.1%. 1 1H-NMR (600 MHz, DMSO-d6) δ 8.55 - 8.45 (m, 2H), 8.05 (s, 1H), 7.35 - 7.26 (m, 2H), 6.28 (s, 2H), 5.80 (s, 2H), 4.76 (q, J = 6.7 Hz, 1H), 4.50 (s, 2H), 3.61 (tt, J = 11.2, 4.1 Hz, 1H), 2.45 (s, 6H), 2.17 (s, 6H), 1.82 (d, J = 12.7 Hz, 1H), 1.75 - 1.62 (m, 1H), 1.40 (d, J = 6.7 Hz, 3H), 1.25 (d, J = 12.0 Hz, 4H), 1.16 (t, J = 12.1 Hz, 1H), 1.09 (s, 3H), 0.97 (s, 3H), 0.61 (s, 3H). 13 13C-NMR (151 MHz, DMSO-d6) δ 156.55, 150.04, 142.34, 132.59, 123.79, 123.03, 121.93, 81.95, 69.09, 60.85, 60.14, 60.05, 45.83, 45.55, 40.55, 34.60, 34.28, 23.40, 21.33, 21.29, 15.53, 14.81. HRMS (ESI): Calcd for: 619.3618 Found: 619.3621. Example 6 Preparation of BDP-tempo

[0035] In a 500 mL single-necked flask, dissolve BDP-2 (1.9 g) and Compound 2 (1.32 g) in a mixed solvent of ethanol, chloroform, and water (38 mL / 478 mL / 38 mL). At room temperature, add 16 mL of triethylamine and copper(I) iodide (1.19 g). Stir overnight at room temperature under N2 protection. Dilute with water, extract with dichloromethane (150 mL × 3), combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, concentrate, and purify by column chromatography (DCM:MeOH = 500:1 - 25:1) to obtain 1.47 g of a brick-red solid. Purify by thin-layer chromatography (PE:EA = 1:1) to obtain 0.23 g of a brick-red solid. HRMS (ESI): Calcd for: 513.2961 Found: 513.2957. Effect Example 1 Using MeOH as the solvent, prepare stock solutions of compounds BDP-Py-4, BDP-Py-5, and BDP-tempo at 1 mmol / L respectively. Take 300 μL of the stock solution and dilute it with 2700 μL of MeOH to prepare a 100 μmol / L test solution. Measure its ultraviolet absorption spectrum (100 μM) using a UV-visible spectrophotometer. The results are as Figure 1 shown.

[0036] Take 300 μL of the stock solutions of BDP-Py-4, BDP-Py-5, and BDP-tempo at 1 mmol / L respectively and 2700 μL of MeOH to prepare a 100 μmol / L test solution. Measure its fluorescence emission spectrum (100 μM) using a fluorescence spectrophotometer. The results are as Figure 2 shown.

[0037] The ultraviolet spectrum and fluorescence emission spectrum further prove the successful preparation of BDP-Py-4 and BDP-Py-5. The fluorescence intensities of BDP-Py-4 and BDP-Py-5 are much greater than those of their decomposition product BDP-tempo.

[0038] Effect Example 2 Test on the singlet oxygen generation ability of BDP-Py-4 and BDP-Py-5 Use DPBF (1,3-diphenylisobenzofuran) as a singlet oxygen scavenger and verify the singlet oxygen generation ability of the target compound by ultraviolet monitoring of its characteristic absorption peak at 415 nm.

[0039] The specific procedure is as follows. Add DPBF (final concentration 1.0×10 -4 M) and methylene blue (MB, final concentration 0.2×10 -4 M) to a 25 mL two-necked round-bottom flask, and then add 10 ml of a solution with a concentration of 1.0×10 -5A DMSO solution of BDP-Py-4 or BDP-Py-5 at a concentration of [[mol / L]] and a magnetic stir bar were used. A xenon lamp with a power of 35 W was used as the light source. An aqueous solution of 0.72 M NaNO2 was placed between the light source and the two-necked round-bottom flask as an absorber for light below 385 nm to filter out ultraviolet light and heat. The light intensity irradiating on the round-bottom flask was 20 W / m 2 . The magnetic stirring was started. After 60 s of light irradiation, a sample was taken from the two-necked round-bottom flask to measure the ultraviolet absorption spectrum. After the measurement was completed, the sample was poured back into the round-bottom flask and stirring was continued. After another 60 s of light irradiation, the above steps were repeated until there was no obvious change in the ultraviolet absorption spectrum. The results are shown in Figure 3 , which proved that BDP-Py-4 and BDP-Py-5 have good singlet oxygen generation ability.

[0040] Effect Example 3: Photostability Test of BDP-Py-4 and BDP-Py-5 The MeOH solutions (10 μM) of compounds BDP-Py-4 and BDP-Py-5 were exposed to green light irradiation at 525 nm (20 W / m 2 ) for 60 min, and the ultraviolet absorption intensity was measured every 10 min. The results are as shown in Figure 4 . The ultraviolet absorption intensity of the compounds hardly decreased significantly within 60 min, proving that this series of compounds has good photostability and is suitable for photodynamic therapy.

[0041] Effect Example 4: Test of the Hydrolysis Rate of BDP-Py-4 and BDP-Py-5 in Aqueous Solution PBS solutions with pH = 6.5 or pH = 7.4 containing 10 μM of compound BDP-Py-4 or BDP-Py-5 were incubated in the dark in a 37 °C water bath, and the changes in fluorescence intensity at different times were detected.

[0042] The results are as shown in Figures 5 - 8As shown, BDP-Py-4 and BDP-Py-5 hydrolyze in PBS to generate BDP-tempo and alkyl radicals, and the fluorescence intensity will decrease. By monitoring the rate of decrease in fluorescence intensity, the hydrolysis rates of BDP-Py-4 and BDP-Py-5 can be reflected. It can be seen that the hydrolysis rate of BDP-Py-5 is greater than that of BDP-Py-4, proving that the hydrolysis rate can be regulated by adjusting the carbon chain length, and it can play a better role at the tumor site. For BDP-Py-4, under the condition of pH = 6.5, in the first 2 h, the fluorescence intensity did not decrease significantly. Starting from 4 h, the fluorescence intensity decreased. Under the condition of pH = 7.4, in the first 1 h, the fluorescence intensity did not decrease significantly. The decomposition rate of BDP-Py-5 at pH = 6.5 was also significantly greater than that at pH = 7.4. This shows that the hydrolysis rate of the BDP-Py series compounds disclosed in this application at pH = 6.5 is greater than that at pH = 7.4. Because the interior of solid tumors is a weak acid environment, the hydrolysis rate at the tumor site will be faster than that at other sites, and it can play a better anti-tumor effect. At the same time, the change in fluorescence intensity generated by the decomposition of the BDP-Py series compounds also provides a certain degree of reference for the diagnosis of tumors.

[0043] Effect Example 5 Determination of in vitro anti-tumor activity The phototoxicity and dark toxicity of BDP-Py-4 and BDP-Py-5 were investigated by the CCK-8 method.

[0044] In this application, A375 cells were selected and cultured in DMEM medium containing 10% fetal bovine serum (FBS) at 37 °C. At 5000 cells / well, they were added to a 96-well plate, incubated with 100 μL of DMEM medium at 37 °C for 24 h, and divided into 1. BDP-Py-4 group and 2. BDP-Py-5 group. The original medium was discarded, and the medium in each group was replaced with 100 μL of fresh DMEM medium containing 10% FBS with the corresponding drug. The drug concentrations in each medium were 0.01 μM, 0.05 μM, 0.1 μM, 0.5 μM, 1 μM, 5 μM, 10 μM, 50 μM, and 100 μM respectively. The phototoxicity and dark toxicity were measured for each group. After adding the drug for 40 h, the light was irradiated for 30 min (525 nm green light lamp, radiation intensity 20 W / m 2 ), and then tested after incubating for another 7.5 h. The dark toxicity was tested after incubating for 48 h after drug administration. After incubation, 10 μL of CCK-8 detection solution was added to each well, incubated for another 1 h, placed in an enzyme-linked immunosorbent assay (ELISA) reader, and the OD value of each well was read at a wavelength of 450 nm to calculate the cell survival rate. The survival rate = (OD 给药 - OD 空白 ) / (OD 对照 - OD 空白 ).

[0045] The results are as follows Figure 9 shown. It can be seen that under normoxic conditions, the compound described in the present application has good anti-tumor effects, and its phototoxicity is significantly stronger than its dark toxicity.

[0046] Effect Example 6 In vitro cytotoxicity test on normal cells In the present application, the CCK-8 method was used to investigate the phototoxicity and dark toxicity of BDP-Py-4 and BDP-Py-5 in human vascular endothelial cells.

[0047] In the present application, HUVECS cells were selected and cultured in ECM medium containing 10% FBS at 37 °C. At a density of 5000 cells / well, they were added to a 96-well plate and incubated with 100 μL of ECM medium containing 10% FBS at 37 °C for 24 h. The original medium was discarded, and the medium in each group was replaced with 100 μL of fresh ECM medium containing the corresponding drug and 10% FBS. The drug concentrations in each medium were 0.01 μM, 0.05 μM, 0.1 μM, 0.5 μM, 1 μM, 5 μM, 10 μM, 50 μM, and 100 μM, respectively. The phototoxicity and dark toxicity were measured for each group. Phototoxicity was measured after irradiating with light for 30 min (the radiation intensity of 525 nm green light was 20 W / m 2 ²) 40 h after adding the drug and then incubating for another 7.5 h. Dark toxicity was measured 48 h after administering the drug. After incubation, 10 μL of CCK-8 detection solution was added to each well, and incubation was continued for 1 h. Then, it was placed in an enzyme-linked immunosorbent assay (ELISA) reader, and the OD value of each well was read at a wavelength of 450 nm to calculate the cell survival rate. The survival rate = (OD 给药 - OD 空白 of the experimental group) / (OD 对照 - OD 空白 of the blank control group).

[0048] The results are as follows Figure 10 shown. It can be seen that the IC50 values of both the phototoxicity and dark toxicity of BDP-Py-4 and BDP-Py-5 are greater than 100 μmol / L, indicating that they cause less damage to normal tissues.

[0049] Effect Example 7 Test for the ability of tumor cells to generate ROS The probe kit used in this effect example was purchased from APExBiO Company, with the product number Catalog No.K2065, and the positive control Rousp was assembled in this kit.

[0050] In this application, A375 cells were selected and cultured in DMEM medium containing 10% FBS at 37°C. At a density of 5000 cells / well, they were added to a black-walled and bottom-transparent 96-well plate and incubated with 100 μL of DMEM medium containing 10% FBS at 37°C for 24 h. The original medium was discarded and replaced with 100 μL of fresh medium containing the corresponding drugs BDP-Py-4, BDP-Py-5, and Rousp (drug concentrations were 1 μM, 10 μM, and 20 μM respectively). After 48 h of drug incubation, they were irradiated with light for 30 min (green light at 525 nm, radiation intensity was 20 W / m 2 ), 100 μL of DCFH probe was added, and after incubation at 37°C for 30 min, the original medium was discarded, and the cells were washed three times with PBS, and then photographed using a confocal microscope.

[0051] The results are as Figure 11 shown. It can be seen that BDP-Py-4 and BDP-Py-5 have good ROS generation ability, and with the increase of concentration, the fluorescence intensity is greater, indicating that their ROS generation ability is stronger.

[0052] Effect Example 8: Antitumor ability test in mice Four-week-old C57 black mice were selected. Each mouse was inoculated with 1×10 6 cells of MC38 cells under the right shoulder and continued to be incubated. When the tumor volume of the mouse reached 100 mm 3 , they were randomly divided into 5 groups, with 3 mice in each group (Group A: normal saline + light, Group B: BDP-Py-4 2.5 mg / kg + light, Group C: BDP-Py-4 5 mg / kg + light, Group D: BDP-Py-5 2.5 mg / kg + light, Group E: BDP-Py-5 5 mg / kg + light). Administration intervention was carried out by tail vein injection, and the drug was administered every other day. At the same time every day, the tumor volume was measured and the weight was measured. They were irradiated with a green light at 525 nm with an intensity of 20 W / m 2 for 30 min every day. After five administrations, the mice were sacrificed, the tumor tissues were removed and photographed ( Figure 12 ).

[0053] The graph of the change in mouse body weight is as Figure 13 shown. The body weight of the mice remained basically unchanged, proving that the compounds disclosed in this application are almost non-toxic. The change in the tumor volume of the mice is as Figure 14 shown, proving that BDP-Py-4 and BDP-Py-5 have good antitumor effects, laying a foundation for the development of antitumor drugs.

[0054] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof, characterized in that, The structure of the said compound is shown in Formula I or Formula II: ； Wherein, R is a straight-chain alkyl group with 1 to 5 carbon atoms.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein, The straight-chain alkyl group with 1 to 5 carbon atoms is —CH3, —CH2CH3, —(CH2)2CH3, —(CH2)3CH3, —(CH2)4CH3.

3. A method for preparing a fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof as described in claim 1, characterized in that, It includes the following steps: S1： ； In solvent A, 2,4-dimethylpyrrole reacts with chloroacetyl chloride, and then triethylamine and boron trifluoride diethyl ether are successively added and continue to react to obtain BDP-1; S2： ； In solvent B, BDP-1 reacts with sodium azide to obtain BDP-2; S3： ； In solvent C, BDP-2 and compound X undergo a Click reaction to obtain BDP-Py; The R1 is .

4. The preparation method according to claim 3, characterized in that, The said solvent A is selected from one of dichloromethane or toluene; the said solvent B is ethanol; the said solvent C is selected from one of ethanol, chloroform or water.

5. The preparation method according to claim 3, characterized in that, The molar ratio of the chloroacetyl chloride, 2,4-dimethylpyrrole and boron trifluoride diethyl ether is 1~1.2:2:6~8; the molar ratio of BDP-1 and the sodium azide is 1:4~5; the molar ratio of BDP-2 and compound X is 1:1.5~2.

6. The preparation method according to claim 3, characterized in that, When the solvent in S1 is dichloromethane, 2,4-dimethylpyrrole and chloroacetyl chloride react under nitrogen protection by heating under reflux, the dropping temperature of triethylamine and boron trifluoride diethyl ether is -10~0 °C, and after addition, react by heating under reflux; when the solvent in S1 is toluene, 2,4-dimethylpyrrole and chloroacetyl chloride react at 40~50 °C under nitrogen protection, the dropping temperature of triethylamine and boron trifluoride diethyl ether is -10~0 °C, and after addition, react at 40~50 °C; In S2, sodium azide is added dropwise at room temperature, and the reaction temperature after the addition is 40~45 °C; S3 reacts under nitrogen conditions at room temperature, and the reaction time is 10~14 h.

7. The preparation method according to claim 3, characterized in that, The said compound X is prepared by the following method: ； a. Raw material 1 and 3-bromopropyne undergo a Williamson etherification reaction to obtain compound 2; b. Raw material 3 and NBS undergo a bromination reaction to obtain compound 4; c. Compound 2 and compound 4 undergo a Williamson etherification reaction to obtain compound 5; d. Compound 5 reacts with R-I to obtain compound 6; The obtained compound 5 and compound 6 are the above-mentioned compound X.

8. A pharmaceutical composition comprising the fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof as described in claim 1, characterized in that, It further includes at least one pharmaceutically acceptable carrier or excipient.

9. Use of a fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof as described in claim 1 in the preparation of an anti-tumor drug.

10. Use of a fluoroboron dipyrrole compound or a pharmaceutically acceptable salt thereof as described in claim 1 in the preparation of a tumor treatment diagnostic agent.

Images