Synthesis method and application of a benzisoindolinone compound

Abstract

This invention belongs to the field of organic chemistry, specifically relating to a method for synthesizing benzisoindolinone compounds. The method is characterized by the use of acetic acid as a solvent and phenyl selenide as a catalyst to react 2-phenylynylbenzaldehyde compounds with N-(benzyloxy)acrylamide compounds in a reaction flask. This synthetic scheme efficiently achieves the synthesis of benzisoindolinone compounds through [4+2] cycloaddition and intramolecular condensation cyclization of ketones under the action of the non-metallic catalyst phenyl selenide. This scheme eliminates the need for halogenation or pre-activation of substrates, and is environmentally friendly, with mild and controllable reaction conditions and excellent reaction efficiency. Bioactivity evaluation results show that some of the obtained compounds exhibit good bioactivity, which can contribute to the research of novel benzisoindolinone molecular skeleton drugs in my country and has significant application value.

Description

Technical Field

[0001] This invention relates to the field of organic chemistry, specifically to a method for synthesizing and applying a benzisoindolone compound. Background Technology

[0002] Isoindolinones, a class of nitrogen-containing heterocyclic skeletal molecules, have become a hot research topic at the intersection of natural product chemistry and drug development due to their stable six-membered and five-membered ring structures, tunable substitution sites, and good biocompatibility. Their research value has been fully demonstrated through decades of exploration. In the field of anti-tumor therapy, these compounds have shown clear clinical translational potential: Midostaurin, developed by Novartis, is a typical example. As a multi-target kinase inhibitor (acting on targets such as FLT3 and KIT), it works by inhibiting tumor cell proliferation signaling pathways, becoming a milestone drug for targeted therapy of FLT3-mutant acute myeloid leukemia (AML). In the field of anti-inflammatory and immunomodulatory therapy, indobufen's core structure is an isoindolinone derivative. It exerts its antiplatelet aggregation effect by inhibiting the active site of platelet cyclooxygenase-1 (COX-1), offering advantages over traditional aspirin, including reversible action and fewer gastrointestinal side effects. In the fields of anti-inflammatory and antibacterial properties, pomalidomide, an analogue of thalidomide, uses phthalimide as its core skeleton. However, its isoindolinone structural analogues exhibit stronger activity in the treatment of multiple myeloma by regulating the release of inflammatory factors such as TNF-α, and also reduce the risk of neurotoxicity. Numerous studies have confirmed that these compounds can exhibit multiple activities, including antitumor, antiepileptic, anti-inflammatory, and antibacterial effects, by acting on different pathological pathways. The progress in the development of these specific drugs and candidate molecules fully demonstrates the drug development potential of the isoindolinone skeleton. The core skeleton formed by the fusion of the isoindolinone ring and the benzene ring endows benzene-isoindolinone compounds with a unique fused-ring structure. This molecular architecture, constructed by the direct fusion of two types of aromatic ring systems, not only endows these compounds with excellent chemical stability and flexible structural modifiability but also enables them to exhibit rich diversity of biological activities. It is this structural advantage derived from ring fusion that has driven these compounds to become a research focus in many fields, such as the development of organic synthesis methodologies, total synthesis of natural products, and drug molecule design and screening, making them one of the most promising research hotspots in the field of heterocyclic chemistry in recent years.

[0003] In the prior art, there are some reported methods for synthesizing benzisoindololinone compounds: Jiang Bo et al. (Chemical Communications, 2017, 53, 3369-3372) utilized iodine-mediated oxygen migration and benzene cyclization to generate 1-aramido-2-naphthaldehyde derivatives containing conjugated 1,4-dicarbonyl structural units. This compound can be cyclized with aromatic amines via a [4+1] cyclization reaction to obtain benzisoindololinone compounds; Xu Liwen et al. (ACS Catalysis, 2019, 9, 2, 1431-1436) found that under a carbon monoxide / oxygen reaction atmosphere, a palladium / copper bimetallic / amino acid ligand catalytic system can catalyze the asymmetric C(sp) cyclization of chiral arylsulfonamides. 2 The Li Yanzhong research group at Central China Normal University (Organic Letters, 2019, 21, 16, 6264-6269) reported a Co(OAc)2 / AgOAc synergistic catalytic system that can achieve β-functionalization of α-diazoketones, followed by intramolecular cyclization to obtain benzisoindolone compounds in excellent yields.

[0004] Although there have been numerous reports on the synthesis of benzisoindolones, existing synthetic methods generally suffer from drawbacks such as the need for expensive transition metal catalysts and complex raw material preparation. Therefore, guided by the concept of green chemistry, developing a direct, efficient, and environmentally friendly synthetic strategy for benzisoindolone compounds is of great significance for the research and development of drugs with independent intellectual property rights. Summary of the Invention

[0005] The purpose of this invention is to provide a novel synthetic method for benzisoindolone compounds and its applications. This synthetic scheme efficiently synthesizes benzisoindolone compounds via [4+2] cycloaddition and intramolecular condensation cyclization of ketones using a non-metallic catalyst, phenyl selenide bromide. This method eliminates the need for halogenation or pre-activated substrates, and is characterized by its environmental friendliness, mild and controllable reaction conditions, and excellent reaction efficiency.

[0006] We also obtained the molecular skeletons of some novel benzisoindolone compounds with good bioactivity through bioactivity evaluation tests.

[0007] The objective of this invention is achieved through the following technical solution:

[0008] A novel synthetic method for benzisoindolone compounds, which have the following structures:

[0009]

[0010] Where R 1R represents hydrogen or halogen. 2 R represents hydrogen or halogen. 3 It indicates hydrogen, trifluoromethyl, or halogen.

[0011] The novel preparation method (synthetic reaction formula) of this benzisoindolone compound is as follows:

[0012]

[0013] Step 1: First, prepare 2-phenylynylbenzaldehyde compounds: Under nitrogen protection, o-bromobenzaldehyde, phenylacetylene compounds, cuprous iodide, and palladium dichloride of bis(triphenylphosphine) were added to triethylamine solvent and reacted at 50 °C for 24 h. After cooling to room temperature, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylynylbenzaldehyde compounds.

[0014] Step 2: Preparation of N-(benzyloxy)acrylamide compounds: O-benzylhydroxylamine hydrochloride compounds were added to a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ and stirring for 10 minutes. Acrylyl chloride was then slowly added dropwise to the reaction system, and stirring continued at room temperature for several hours. The organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 5:1-2:1) to obtain N-(benzyloxy)acrylamide compounds.

[0015] Step 3 (cyclization reaction): The 2-phenylynylbenzaldehyde compounds synthesized in Step 1, the N-(benzyloxy)acrylamide compounds synthesized in Step 2, phenyl selenide bromide, and acetic acid are added to a reaction flask to react. After the reaction is complete, the reaction solution is successively cooled to room temperature, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated column chromatography to obtain the benzisoindolone compound. The molar ratio of the o-2-phenylynylbenzaldehyde compounds to the N-(benzyloxy)acrylamide compounds is between 1:1 and 1:3, preferably 1:2; the reaction ratio of the 2-phenylynylbenzaldehyde compounds to the acetic acid is 0.1 mmol / mL to 0.2 mmol / mL, preferably 0.13 mmol / mL.

[0016] The 2-phenylynylbenzaldehyde compounds are 2-phenylynylbenzaldehyde, 2-phenylynyl-5-bromobenzaldehyde, 2-phenylynyl-5-chlorobenzaldehyde, 2-phenylynyl-4-fluorobenzaldehyde, 2-(3-fluorophenylynyl)benzaldehyde, and 2-(4-chlorophenylynyl)benzaldehyde.

[0017] The N-(benzyloxy)acrylamide compounds are N-(benzyloxy)acrylamide, N-(2-chlorobenzyloxy)acrylamide, and N-(4-(trifluoromethyl)benzyloxy)acrylamide.

[0018] The cyclization reaction (i.e., step three) is carried out by stirring at a temperature of 100-120 °C for 1-12 h.

[0019] Furthermore, the cyclization reaction was carried out with stirring at 110 °C for 8 h.

[0020] In step three, the eluent for column chromatography purification of the reaction solution is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 7:1.

[0021] The advantages of this invention lie in providing a novel method for synthesizing benzisoindolone compounds. This method uses acetic acid as a solvent and phenyl selenide as a catalyst to react 2-phenylynylbenzaldehyde compounds with N-(benzyloxy)acrylamide compounds in a reaction flask. This synthetic scheme efficiently achieves the synthesis of benzisoindolone compounds through [4+2] cycloaddition and intramolecular condensation cyclization of ketones under the action of the non-metallic catalyst phenyl selenide. The raw materials used in this synthetic method are all commercially available chemical products, making them inexpensive and readily available. Furthermore, the novel synthetic scheme involved in this invention is environmentally friendly, with mild and controllable reaction conditions and excellent reaction efficiency. In addition, the in vitro growth inhibitory activity of some compounds against two cancer cell lines, HGC-27 (human gastric cancer) and PC-3 (human prostate cancer), was evaluated by MTT assay. Detailed Implementation

[0022] The present invention will be further described below with reference to examples:

[0023] The structural formula of the benzo[i]indolineone compound of the present invention is as follows:

[0024]

[0025] Where R 1 R represents hydrogen or halogen. 2 R represents hydrogen or halogen. 3 It indicates hydrogen, trifluoromethyl, or halogen.

[0026] The preparation method is as follows: 2-Phenylacetylbenzaldehyde, N-(benzyloxy)acrylamide, phenyl selenide bromide, and acetic acid are added to a reaction flask and reacted. The molar ratio of 2-phenylylacetylbenzaldehyde to N-(benzyloxy)acrylamide is 1:2, and the reaction ratio of 2-phenylylacetylbenzaldehyde to acetic acid is 0.13 mmol / mL. The reaction is carried out by stirring at 110 °C for 8 h. After the reaction is complete, the reaction solution is successively cooled, extracted with ethyl acetate, dried with anhydrous sodium sulfate, and purified by column chromatography to obtain the benzisoindolone compound. The eluent for the column chromatography purification in the above reaction is a mixture of petroleum ether and ethyl acetate, with a volume ratio of petroleum ether to ethyl acetate of 7:1.

[0027] In addition, the in vitro growth inhibitory activity of some compounds against two cancer cell lines, HGC-27 (human gastric cancer) and PC-3 (human prostate cancer), was evaluated by MTT assay.

[0028] Example 1

[0029] Step 1: Under nitrogen protection, 2,5-dibromobenzaldehyde (20 mmol), phenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylynyl-5-bromobenzaldehyde.

[0030] Step 2: O-benzylhydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 5:1-2:1) to obtain N-(benzyloxy)acrylamide.

[0031] Step 3: The 2-phenylynyl-5-bromobenzaldehyde (0.2 mmol), N-(benzyloxy)acrylamide (0.4 mmol), phenylselenide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain 2-(benzyloxy)-7-bromo-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 64%, and its structural formula is as follows:

[0032]

[0033] The spectral data for 2-(benzyloxy)-7-bromo-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0034] Column chromatography eluent (petroleum ether: ethyl acetate = 20:1), yellow solid, melting point 149-151°C. o C.

[0035] 1 H NMR (500 MHz, CDCl3) δ 8.00 (d, J = 1.5 Hz, 1H), 7.90 (d, J = 8.0Hz, 1H), 7.90 (d, J = 8.5 Hz, 1H), 7.33 (dd, J = 9.0, 2.0 Hz, 1H), 7.30 –7.18 (m, 9H), 7.07 (dd, J = 7.5, 1.5 Hz, 2H), 5.51 (s, 1H), 5.08 (d, J = 10.5Hz, 1H), 4.62 (d, J = 10.5 Hz, 1H);

[0036] 13 C NMR (126 MHz, CDCl3) δ 165.6, 140.9, 136.9, 136.0, 135.2, 131.4,130.8, 129.8, 129.4, 129.3, 129.1, 129.0, 128.6, 128.6, 128.2, 126.0, 125.4,122.2, 121.0, 79.1, 65.6;

[0037] IR (KBr): 2918, 2848, 1691, 1582, 1450, 1386, 1267, 1071, 991, 906,875, 729, 694 cm -1 ;

[0038] HRMS for C 25 H 19 BrNO2 + (M+H) + Calculated value: 444.05937, measured value: 444.05945.

[0039] Example 2

[0040] Step 1: Under nitrogen protection, 2-bromo-5-chlorobenzaldehyde (20 mmol), phenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylynyl-5-chlorobenzaldehyde.

[0041] Step 2: O-benzylhydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 5:1-2:1) to obtain N-(benzyloxy)acrylamide.

[0042] Step 3: The 2-phenylynyl-5-chlorobenzaldehyde (0.2 mmol), N-(benzyloxy)acrylamide (0.4 mmol), phenyl selenide bromide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain 2-(benzyloxy)-7-chloro-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 57%, and its structural formula is as follows:

[0043]

[0044] The spectral data for 2-(benzyloxy)-7-chloro-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0045] Column chromatography eluent (petroleum ether: ethyl acetate = 30:1-20:1), yellow solid, melting point 116-118°C. o C.

[0046] 1 H NMR (500 MHz, CDCl3) δ 7.98 (d, J = 7.5 Hz, 1H), 7.89 – 7.86 (m,2H), 7.37 – 7.30 (m, 9H), 7.26 (dd, J = 9.0, 2.0 Hz, 1H), 7.16 – 7.13 (m,2H), 5.58 (s, 1H), 5.15 (d, J = 10.5 Hz, 1H), 4.70 (d, J = 11.0 Hz, 1H);

[0047] 13 C NMR (126 MHz, CDCl3) δ 165.6, 140.8, 136.5, 136.0, 135.2, 133.9,129.8, 129.3, 129.3, 129.2, 129.0, 128.6, 128.6, 128.3, 128.1, 128.0, 125.7,125.3, 121.0, 79.0, 65.6;

[0048] IR (KBr): 2982, 2922, 1718, 1580, 1452, 1415, 1374, 1260, 1229, 1073,902, 877, 731 cm -1 ;

[0049] HRMS for C 25 H 19 ClNO2 + (M+H) + Calculated value: 400.10988, measured value: 400.10959.

[0050] Example 3

[0051] Step 1: Under nitrogen protection, 2-bromo-4-fluorobenzaldehyde (20 mmol), phenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylynyl-4-fluorobenzaldehyde.

[0052] Step 2: O-benzylhydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 5:1-2:1) to obtain N-(benzyloxy)acrylamide.

[0053] Step 3: The 2-phenylynyl-4-fluorobenzaldehyde (0.2 mmol), N-(benzyloxy)acrylamide (0.4 mmol), phenyl selenide bromide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain 2-(benzyloxy)-8-fluoro-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 55%, and its structural formula is as follows:

[0054]

[0055] The spectral data for 2-(benzyloxy)-8-fluoro-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0056] Column chromatography eluent (petroleum ether: ethyl acetate = 30:1-20:1), yellow solid, melting point 146-148°C. o C.

[0057] 1H NMR (500 MHz, CDCl3) δ 7.97 – 7.89 (m, 3H), 7.38 – 7.30 (m, 8H), 7.25 (td, J = 8.5, 2.5 Hz, 1H), 7.16 (dd, J = 8.0, 2.0 Hz, 2H), 7.00 (dd, J =9.5, 2.0 Hz, 1H), 5.53 (s, 1H), 5.17 (d, J = 10.5 Hz, 1H), 4.71 (d, J = 10.5Hz, 1H);

[0058] 13 C NMR (126 MHz, CDCl3) δ 165.6, 160.9 (d, JC-F = 249.5 Hz), 140.1(d, JC-F = 5.0 Hz), 135.7, 135.2, 132.8, 131.9 (d, JC-F = 8.8 Hz), 129.9,129.8, 129.4, 129.3, 129.0, 128.8, 128.6, 128.4 (d, JC-F = 10.0 Hz), 119.1 (d, JC-F = 2.5 Hz), 118.1 (d, JC-F = 25.2 Hz), 107.5 (d, JC-F = 21.4 (Hz), 79.0, 65.6;

[0059] 19 F NMR (470 MHz, CDCl3): δ -110.3.

[0060] IR (KBr): 2959, 2918, 1697, 1625, 1596, 1460, 1367, 1260, 1180, 1009,937, 857, 754, 692 cm -1 ;

[0061] HRMS for C 25 H 19 FNO2 + (M+H) + Calculated value: 384.13943, measured value: 384.13968.

[0062] Example 4

[0063] Step 1: Under nitrogen protection, 2-bromobenzaldehyde (20 mmol), 1-ethynyl-3-fluorobenzene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-(3-fluorophenylynyl)benzaldehyde.

[0064] Step 2: O-benzylhydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 5:1-2:1) to obtain N-(benzyloxy)acrylamide.

[0065] Step 3: The 2-(3-fluorophenylynyl)benzaldehyde (0.2 mmol), N-(benzyloxy)acrylamide (0.4 mmol), phenyl selenide bromide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain the product 2-(benzyloxy)-1-(3-fluorophenyl)-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 61%, and its structural formula is as follows:

[0066]

[0067] The spectral data for 2-(benzyloxy)-1-(3-fluorophenyl)-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0068] Column chromatography eluent (petroleum ether: ethyl acetate = 20:1) is a yellow solid with a melting point of 91-93°C. o C.

[0069] 1H NMR (500 MHz, CDCl3) δ 8.00 – 7.94 (m, 2H), 7.92 (d, J = 8.0 Hz,1H), 7.54 – 7.50 (m, 1H), 7.40 – 7.29 (m, 8H), 7.04 (td, J = 8.5, 2.0 Hz,1H), 6.97 (d, J = 8.0 Hz, 1H), 6.79 (d, J = 9.0 Hz, 1H), 5.55 (s, 1H), 5.17(d, J = 11.0 Hz, 1H), 4.81 (d, J = 10.5 Hz, 1H);

[0070] 13 C NMR (126 MHz, CDCl3) δ 166.3, 163.07 (d, JC-F = 248.2 Hz), 140.3,138.9 (d, JC-F = 6.3 Hz), 135.9, 135.3, 130.7 (d, JC-F = 8.8 Hz), 130.3,129.8, 129.5, 129.1, 128.7, 128.0, 127.7, 127.5, 127.4, 124.4 (d, JC-F = 3.8Hz), 123.6, 119.8, 116.2 (d, JC-F = 21.4 Hz), 115.6 (d, JC-F = 22.7 Hz), 79.0, 65.1;

[0071] IR (KBr): 2922, 2887, 1701, 1593, 1505, 1378, 1223, 1151, 984, 846,762, 743, 698, 527, 498 cm -1 ;

[0072] HRMS for C 25 H 19 FNO2 + (M+H) + Calculated value: 384.13943, measured value: 384.13962.

[0073] Example 5

[0074] Step 1: Under nitrogen protection, o-bromobenzaldehyde (20 mmol), 4-chlorophenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-(4-chlorophenylacetyl)benzaldehyde.

[0075] Step 2: O-benzylhydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 5:1-2:1) to obtain N-(benzyloxy)acrylamide.

[0076] Step 3: The 2-(4-chlorophenylynyl)benzaldehyde (0.2 mmol), N-(benzyloxy)acrylamide (0.4 mmol), phenyl selenide bromide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain the product 2-(benzyloxy)-1-(4-chlorophenyl)-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 61%, and its structural formula is as follows:

[0077]

[0078] The spectral data for 2-(benzyloxy)-1-(4-chlorophenyl)-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0079] Column chromatography eluent (petroleum ether: ethyl acetate = 20:1-10:1), yellow solid, melting point 142-144°C. o C.

[0080] 1H NMR (500 MHz, CDCl3) δ 8.02 – 7.97 (m, 2H), 7.95 (d, J = 8.5 Hz,1H), 7.56 – 7.53 (m, 1H), 7.40 – 7.34 (m, 7H), 7.33 – 7.30 (m, 2H), 7.09 –7.07 (m, 2H), 5.57 (s, 1H), 5.19 (d, J = 11.0 Hz, 1H), 4.85 (d, J = 10.5 Hz, 1H);

[0081] 13 C NMR (126 MHz, CDCl3) δ 166.3, 140.4, 135.9, 135.3, 134.9, 134.9,130.3, 130.0, 129.8, 129.4, 129.4, 129.0, 128.6, 128.0, 127.6, 127.4, 127.3,123.6, 119.7, 79.0, 65.0;

[0082] IR (KBr): 2954, 2923, 1701, 1587, 1490, 1382, 1258, 1084, 988, 832,742, 693 cm -1 ;

[0083] HRMS for C 25 H 19 ClNO2 + (M+H) + Calculated value: 400.10988, measured value: 400.11008.

[0084] Example 6

[0085] Step 1: Under nitrogen protection, o-bromobenzaldehyde (20 mmol), phenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylacetylbenzaldehyde.

[0086] Step 2: O-(2-chlorobenzyl)hydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 5:1-2:1) to obtain N-(2-chlorobenzyloxy)acrylamide.

[0087] Step 3: The 2-phenylynylbenzaldehyde (0.2 mmol), N-(2-chlorobenzyloxy)acrylamide (0.4 mmol), phenylselenide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 12 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain 2-(2-chlorobenzyloxy)-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 57%, and its structural formula is as follows:

[0088]

[0089] The spectral data for 2-(2-chlorobenzyloxy)-1-phenyl-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0090] Column chromatography eluent (petroleum ether: ethyl acetate = 30:1-20:1), yellow solid, melting point 194-196°C. o C.

[0091] 1 H NMR (500 MHz, CDCl3) δ 7.89 – 7.85 (m, 2H), 7.82 (d, J = 8.0 Hz,1H), 7.43 – 7.40 (m, 1H), 7.36 – 7.34 (m, 2H), 7.27 – 7.20 (m, 5H), 7.18 –7.08 (m, 4H), 5.63 (s, 1H), 5.17 (d, J = 11.0 Hz, 1H), 4.90 (d, J = 11.0 Hz, 1H);

[0092] 13C NMR (126 MHz, CDCl3) δ 166.4, 141.0, 136.2, 135.9, 134.7, 133.1,131.9, 130.2, 130.1, 129.6, 129.4, 129.1, 129.1, 128.7, 127.9, 127.6, 127.5,127.3, 127.0, 123.9, 119.7, 75.5, 65.6;

[0093] IR (KBr): 2988, 2900, 1697, 1452, 1378, 1254, 1232, 1059, 982, 894,867, 754 cm -1 ;

[0094] HRMS for C 25 H 19 ClNO2 + (M+H) + Calculated value: 400.10988, measured value: 400.10999.

[0095] Example 7

[0096] Step 1: Under nitrogen protection, o-bromobenzaldehyde (20 mmol), phenylacetylene (24 mmol), cuprous iodide (0.2 mmol), and bis(triphenylphosphine)palladium dichloride (0.4 mmol) were added to 50 mL of triethylamine solvent and reacted at 50 °C for 24 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with ethyl acetate (60 mL), dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio of 100:1-50:1) to obtain 2-phenylacetylbenzaldehyde.

[0097] Step 2: O-(4-trifluoromethylbenzyl)hydroxylamine hydrochloride (8 mmol) was added to 30 mL of a mixed solvent of ethyl acetate / water (volume ratio 2:1), followed by the addition of K₂CO₃ (8.8 mmol) and stirring for 10 minutes. Acryloyl chloride (8.4 mmol) was then slowly added dropwise to the reaction system, and stirring was continued at room temperature for 12 h. After dilution with ethyl acetate (20 mL), the organic layer was collected, washed with saturated brine, dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 5:1-2:1) to obtain N-(4-(trifluoromethyl)benzyloxy)acrylamide.

[0098] Step 3: The 2-phenylynylbenzaldehyde (0.2 mmol), N-(4-(trifluoromethyl)benzyloxy)acrylamide (0.4 mmol), phenylselenide (0.3 eq), and acetic acid (1.5 mL) synthesized in Steps 1 and 2 were added to a reaction flask and reacted. The mixture was stirred at 110 °C for 8 h in the presence of air. After the reaction was completed, the reaction system was cooled to room temperature. The product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by concentrated silica gel column chromatography (petroleum ether to ethyl acetate volume ratio 7:1) to obtain 1-phenyl-2-((4-(trifluoromethyl)benzyl)oxy)-1,2-dihydro-3H-benzo[e]isoindole-3-one. The yield was 53%, and its structural formula is as follows:

[0099]

[0100] The spectral data for 1-phenyl-2-((4-(trifluoromethyl)benzyl)oxy)-1,2-dihydro-3H-benzo[e]isoindole-3-one are as follows:

[0101] Column chromatography eluent (petroleum ether: ethyl acetate = 20:1), yellow solid, melting point 183-185°C. o C.

[0102] 1 H NMR (500 MHz, CDCl3) δ 7.91 – 7.82 (m, 3H), 7.47 – 7.41 (m, 3H), 7.34 (d, J = 8.5 Hz, 1H), 7.30 – 7.23 (m, 6H), 7.11- 7.09 (m, 2H), 5.66 (s,1H), 5.08 (d, J = 11.5 Hz, 1H), 4.69 (d, J = 11.5 Hz, 1H);

[0103] 13 C NMR (126 MHz, CDCl3) 166.9, 140.9, 139.2, 136.3, 136.0, 130.9 (q,JC-F = 32.4 Hz), 130.3, 129.6, 129.4, 129.3, 128.6, 128.0, 127.5, 127.5,127.4, 125.4 (q, JC-F = 3.7 Hz), 124.1 (q, JC-F = 272.8 Hz), 123.9, 119.7,78.0, 65.9;

[0104] 19F NMR (470 MHz, CDCl3): δ -62.6

[0105] IR (KBr): 2986, 2902, 1691, 1615, 1409, 1318, 1246, 1122, 1067, 898,822, 756 cm -1 ;

[0106] HRMS for C 26 H 19 F3NO2 + (M+H) + Calculated value: 434.13624, measured value: 434.13666.

[0107] Test example:

[0108] The compounds obtained above were selected for activity assay.

[0109] Experimental materials:

[0110] 1. Human cancer cell lines: all purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences.

[0111] 2. Test drug: Dissolved in DMSO and prepared to an initial concentration of 10000 μg / mL for later use.

[0112] 3. 0.9% physiological saline: 250 mL; 2.25 g, Zhengzhou Yonghe Pharmaceutical Co., Ltd.

[0113] Test method:

[0114] Cells were routinely seeded in complete culture medium and cultured at 37 °C with 5% CO2 and saturated humidity for expansion. After digestion with 0.25% trypsin, the cells were diluted with culture medium to a concentration of 1 × 10⁻⁶ cells / mL. 5 Cell suspension (trypan blue staining, viable cell count >95%) was used for experiments. Negative control wells, positive control wells, and wells with different concentrations of the test sample were prepared in 96-well sterile culture plates. The concentrations were set at 64, 32, 16, 8, 4, 2, 1, and 0.5 μg / mL, with three replicates for each concentration. The prepared cell suspension was seeded into 96-well sterile culture plates and cultured for 24 h. Different concentrations of the compound were then added. An equal volume of culture medium was added to the negative control wells, and the plates were incubated. After 72 h of culture, the plates were removed, and 20 μL of MTT was added to each well. The plates were incubated for another 4 h, then centrifuged, and the supernatant was discarded. 150 μL of DMSO was added to each well, and the plates were shaken to completely dissolve the purple-blue formazan crystals. The OD value of each well was measured using a microplate reader, and the IC50 was calculated using SPSS. 50 .

[0115] Test results

[0116] The anticancer activity evaluation data of the above compounds against two human cancer cells are as follows (IC50). 50 ):

[0117] 1 3.93+0.40 59.04+6.34 2 4.06±0.81 36.26±3.03 3 2.83±0.40 19.55±0.20 4 2.11±0.01 16.18±2.38 5 2.55±0.01 12.83±0.78 6 6.92±1.98 50.96±0.72 7 2.29±0.01 12.55±5.16

[0118] The results showed that ① the synthesized new compounds 1 to 7 had a significant inhibitory effect on the proliferation of human prostate cancer cells (PC-3); ② the synthesized new compounds 5 and 7 had a significant inhibitory effect on the proliferation of human gastric cancer cells (HGC-27).

Claims

1. A method for synthesizing a benzisoindolone compound, characterized in that: The compound has the following structure: Where R 1 R represents hydrogen or halogen. 2 R represents hydrogen or halogen. 3 The compound, representing hydrogen, trifluoromethyl, or halogen, is synthesized by the following method: 2-phenylynylbenzaldehyde, N-(benzyloxy)acrylamide, phenyl selenide bromide, and acetic acid are added to a reaction flask and reacted; the reaction solution is then cooled to room temperature, extracted with ethyl acetate, dried with anhydrous sodium sulfate, and purified by concentrated column chromatography to obtain the benzisoindolone compound.

2. The synthesis method according to claim 1, characterized in that, 2-Phenyneylbenzaldehyde compounds include 2-phenylyneylbenzaldehyde, 2-phenylyneyl-5-bromobenzaldehyde, 2-phenylyneyl-5-chlorobenzaldehyde, 2-phenylyneyl-4-fluorobenzaldehyde, 2-(3-fluorophenylyneyl)benzaldehyde, and 2-(4-chlorophenylyneyl)benzaldehyde.

3. The synthesis method according to claim 1, characterized in that, The N-(benzyloxy)acrylamide compounds are N-(benzyloxy)acrylamide, N-(2-chlorobenzyloxy)acrylamide, and N-(4-(trifluoromethyl)benzyloxy)acrylamide.

4. The synthesis method according to claim 1, 2, or 3, characterized in that, The molar ratio of 2-phenylynylbenzaldehyde, N-(benzyloxy)acrylamide, and phenylselenide added is between 1:1:0.2 and 1:3:0.6, preferably 1:2:0.3; the reaction ratio of 2-phenylynylbenzaldehyde with the acetic acid is between 0.1 mmol / mL and 0.2 mmol / mL, preferably 0.13 mmol / mL.

5. The synthesis method according to claim 1, characterized in that, The reaction occurs at a temperature of 100-120°C. o Stirring reaction at C for 1-12 h, preferably 110 h. o C reaction for 8 hours.

6. The synthesis method according to claim 1, characterized in that, The eluent used for column chromatography purification was a mixture of petroleum ether and ethyl acetate, with a volume ratio of 7:

1.

7. An application of a benzisoindolone compound synthesized by the method described in claim 1, characterized in that, Biological evaluation of its in vitro antiproliferative activity showed that this benzisoindolone compound effectively inhibited the proliferation of human gastric cancer cells (HGC-27) and human prostate cancer cells (PC-3), and had a significant inhibitory effect.

Images