Selenized isoquinoline compounds, their synthesis methods and applications

Abstract

This invention discloses a selenized isoquinoline compound, its synthesis method, and its applications. The selenized isoquinoline compound has the structure shown in Formula I and exhibits excellent antitumor activity. The synthesis method includes the following steps: Compounds of Formula I-1 and Formula I-2 are electrolyzed in an electrolytic cell containing an electrolyte to obtain the selenized isoquinoline compound with the structure shown in Formula I.

Description

Technical Field

[0001] This invention belongs to the fields of organic synthesis and pharmaceutical technology, and in particular relates to a selenized isoquinoline compound, its synthesis method, and its application. Background Technology

[0002] Isoquinolines are widely found in natural products and bioactive molecules, particularly in antibacterial and antitumor active molecules. However, the rapid and efficient synthesis of functionalized isoquinoline skeletons remains a significant challenge. Therefore, there is an urgent need to develop new reactions to synthesize functionalized isoquinolines to meet the needs of drug development.

[0003] Gelson Perin et al (Araujo DR, Goulart HA, Barcellos AM, Cargnelutti R., EJ, and Perin G. J. J. R. G. Chem., 2021, 86, 1721-1729. reported the intramolecular electrophilic cyclization of 3,4-disubstituted isoquinoline N-oxides from alkynylbenzaldehyde oxime and diselenyl ether under ultrasonic irradiation, followed by deoxygenation at high temperature to prepare isoquinolines. However, this method cannot directly prepare isoquinolines, and the preparation of isoquinolines requires reaction with dichloroethane, a solvent with extremely high ozone depletion potential, at 85°C, which is environmentally unfriendly.

[0004] Currently, there are no reports in domestic or international literature on the synthesis of selenized isoquinoline compounds through electrochemical methods. Summary of the Invention

[0005] Based on this, the present invention provides a class of selenized isoquinoline compounds, their synthesis methods, and their applications in antitumor activity.

[0006] This invention includes the following technical solutions:

[0007] The use of selenized isoquinoline compounds having the structure shown in Formula I, or their pharmaceutically acceptable salts, or their stereoisomers, in the preparation of drugs for the treatment or prevention of tumors;

[0008]

[0009] Wherein, ring A is selected from: one or more R1-substituted or unsubstituted benzene rings, or one or more R1-substituted or unsubstituted 5-6-membered aromatic heterocycles;

[0010] R1 is selected from: hydrogen, C1~C1 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1–C6 alkoxy groups;

[0011] R2 is selected from: C1~C 10 Alkyl, C3-C8 cycloalkyl, one or more R4-substituted or unsubstituted C6-C 18 Aryl, one or more R4-substituted or unsubstituted 5- to 10-membered heteroaryl groups;

[0012] R3 is selected from: C1~C 10 Alkyl, one or more R5-substituted or unsubstituted C6-C 10 Aryl;

[0013] R4 is selected from: hydrogen, C1~C1 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1–C6 alkoxy, amino, and phenyl groups;

[0014] R5 is selected from: hydrogen, C1~C 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1-C6 alkoxy groups.

[0015] In some embodiments, the selenized isoquinoline compound has the structure shown in Formula II or Formula III:

[0016]

[0017] Among them, X1, X2, X3 and X4 are independently selected from: N, CR1; X5 is selected from: O, S, NR6;

[0018] R6 is selected from: hydrogen, C1 to C6 alkyl groups.

[0019] In some embodiments, X1, X3, and X4 are all CH; X2 is CR1.

[0020] In some embodiments, X4 is N; X1, X2 and X3 are all CH.

[0021] In some of these embodiments, R1 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy.

[0022] In some of these embodiments, R1 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and trifluoromethyl.

[0023] In some of these embodiments, R2 is selected from: C1-C3 alkyl, C3-C6 cycloalkyl, one or more R4-substituted or unsubstituted phenyl, one or more R4-substituted or unsubstituted naphthyl, one or more R4-substituted or unsubstituted phenanthryl, one or more R4-substituted or unsubstituted anthracel, one or more R4-substituted or unsubstituted 5-membered heteroaryl.

[0024] In some of these embodiments, R2 is selected from: cyclopropyl, one or more R4-substituted or unsubstituted phenyl, naphthyl, phenanthryl, anthraceneyl, thiophene.

[0025] In some of these embodiments, R4 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, and phenyl.

[0026] In some of these embodiments, R4 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and phenyl.

[0027] In some of these embodiments, R3 is selected from: C1-C4 alkyl groups, one or more R5-substituted or unsubstituted phenyl groups.

[0028] In some of these embodiments, R5 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy.

[0029] In some of these embodiments, R5 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and trifluoromethyl.

[0030] In some embodiments, the selenized isoquinoline compound has the structure shown in Formula IV or Formula V:

[0031]

[0032] In some embodiments, the selenized isoquinoline compound has the structure shown in Formula VI:

[0033]

[0034] In some embodiments, the selenized isoquinoline compound has the structure shown in formula VII or VIII:

[0035]

[0036] In some embodiments, the selenized isoquinoline compound has the structure shown in Formula IX or Formula X:

[0037]

[0038] In some embodiments, the tumor is: lung cancer, breast cancer, or cervical cancer.

[0039] The present invention also provides an antitumor pharmaceutical composition prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is the selenized isoquinoline compound or its pharmaceutically acceptable salt or its stereoisomer.

[0040] The present invention also provides a method for synthesizing the aforementioned selenized isoquinoline compounds, comprising the following steps:

[0041] Compounds of Formula I-1 and Formula I-2 are electrolyzed in an electrolytic cell containing an electrolyte to obtain selenized isoquinoline compounds with the structure shown in Formula I.

[0042]

[0043] Wherein, rings A, R2, and R3 are as described in any one of claims 1-18;

[0044] R6 is selected from: H, C1-C6 alkyl acyl groups.

[0045] In some embodiments, the solvent in the electrolyte is selected from one or more of acetonitrile, trifluoroacetic acid, hexafluoroisopropanol, and methanol.

[0046] In some embodiments, the solvent is a mixture of acetonitrile and trifluoroacetic acid.

[0047] In some embodiments, the solvent is a mixture of acetonitrile and trifluoroacetic acid in a volume ratio of 4 to 20:1.

[0048] In some embodiments, the solvent is a mixture of acetonitrile and trifluoroacetic acid in a volume ratio of 6 to 12:1.

[0049] In some embodiments, the solvent is a mixture of acetonitrile and trifluoroacetic acid in a volume ratio of 8 to 10:1.

[0050] In some embodiments, the electrolyte in the electrolyte solution is one or more tetrabutylammonium salts.

[0051] In some embodiments, the tetrabutylammonium salt is selected from at least one of tetrabutylammonium hexafluorophosphate, tetrabutylammonium hydrogen sulfate, tetrabutylammonium perchlorate, and tetrabutylammonium iodide.

[0052] In some embodiments, the molar ratio of the compound of formula I-1 to the compound of formula I-2 is 1:0.8 to 1.5.

[0053] In some embodiments, the molar ratio of the compound of formula I-1 to the compound of formula I-2 is 1:1.1 to 1.3.

[0054] In some embodiments, the concentration of the electrolyte in the electrolyte solution is 0.03 mmol / mL to 0.15 mmol / mL.

[0055] In some embodiments, the concentration of the electrolyte in the electrolyte solution is 0.08 mmol / mL to 0.12 mmol / mL.

[0056] In some embodiments, the molar ratio of the electrolyte to the compound of formula I-1 in the electrolyte is 1.0 to 2.0:1.

[0057] In some embodiments, the molar ratio of the electrolyte to the compound of formula I-1 in the electrolyte is 1.5 to 1.8:1.

[0058] In some embodiments, the anode for electrolysis is a graphite sheet, a mesh of glassy carbon, a carbon felt, or a platinum sheet, and the cathode is a platinum sheet, an iron sheet, a lead sheet, a nickel sheet, or a zinc sheet.

[0059] In some embodiments, electrolysis is performed using a constant current with an intensity of 4 mA to 16 mA, and the amount of charge is 2.0 F / mol to 9.0 F / mol based on the amount of substance of the compound of formula I-1.

[0060] In some embodiments, the current intensity is 6 mA to 10 mA, and the charge is 2.0 F / mol to 5.0 F / mol in terms of the amount of substance of the compound of formula I-1.

[0061] In some embodiments, the electrolysis temperature is 10°C to 70°C.

[0062] In some embodiments, the electrolysis temperature is 20°C to 50°C.

[0063] This invention provides an electrochemical synthesis method for selenized isoquinoline compounds. This method is simple, low-cost, and enables the green and efficient synthesis of selenized isoquinoline compounds. This invention synthesizes a series of novel selenized isoquinoline compounds using this method. These compounds exhibit good antitumor effects and can be used to prepare therapeutic drugs against various tumors (such as lung cancer, breast cancer, and cervical cancer).

[0064] This invention employs an electrochemical method to directly electrolyze alkynylbenzaldehyde oxime compounds and diselenate ether compounds. Electrolysis generates selenium ether ions, which then attack the alkyne, followed by nitrogen cyclization, and finally cleavage of the nitrogen-oxygen bond to yield selenized isoquinoline compounds. This novel reaction mechanism significantly improves upon the shortcomings of the two-step synthesis of isoquinolines previously used by Gelson Perin et al., avoiding high temperatures and the use of dichloroethane, thus reducing the number of reaction steps. The reaction uses electrons as the oxidant, eliminating the need for an oxidant, resulting in a clean and environmentally friendly reaction system that avoids chemical waste generation and contributes to atom economy. The reaction conditions are mild, requiring neither high temperatures nor inert gas protection, and a high-purity product can be obtained through simple purification steps, making it more suitable for industrial production.

[0065] The yield of the obtained selenide isoquinoline compounds can be further improved by further optimizing the specific reaction conditions (solvent, electrolyte, electrolysis time and temperature, current intensity, concentration of reactants, etc.). Attached Figure Description

[0066] Figure 1 The results of the scratch assay were used to determine the anti-tumor cell migration effect of the compound.

[0067] Figure 2 The results are based on the quantitative analysis of the scratch test.

[0068] Figure 3 The results of the Transwell assay were used to determine the anti-tumor cell migration effect of the compound.

[0069] Figure 4 The results are for quantitative analysis of Transwell experiments. Detailed Implementation

[0070] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments are merely illustrative of the present invention and should not be construed as limiting the invention.

[0071] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0072] The terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, apparatus, product, or device that includes a series of steps is not limited to the steps or modules listed, but may optionally include steps not listed, or may optionally include other steps inherent to such process, method, product, or device.

[0073] In this invention, "multiple" refers to two or more. "And / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone. The character " / " generally indicates that the preceding and following related objects have an "or" relationship.

[0074] In the compounds described in this invention, when any variable (e.g., R) 4 If a component (e.g., a substituent) appears more than once in any component, the definition of each occurrence is independent of the definition of each subsequent occurrence. Similarly, combinations of substituents and variables are permitted, provided such combinations stabilize the compound. A line drawn from a substituent into the ring system indicates that the bond referred to can be attached to any substituted ring atom. If the ring system is polycyclic, it means that such a bond is attached only to any suitable carbon atom of a neighboring ring. It should be understood that those skilled in the art can select the substituents and substitution patterns of the compounds of this invention to provide chemically stable compounds that can be readily synthesized from readily available starting materials using techniques in the art and the methods described below. If a substituent is itself substituted by more than one group, it should be understood that these groups can be on the same carbon atom or on different carbon atoms, as long as the structure is stable.

[0075] As used in this invention, the term "alkyl" refers to a saturated aliphatic hydrocarbon group, including branched and straight-chain groups having a specific number of carbon atoms. For example, the definition of "C1-C6 alkyl" includes groups having 1, 2, 3, 4, 5, or 6 carbon atoms arranged in a straight or branched chain. Specifically, "C1-C6 alkyl" includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, and hexyl.

[0076] As used in this invention, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic, fused-ring, or bridged-ring hydrocarbon group having a specific number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, etc.

[0077] The term "alkoxy" as used in this invention refers to a group having an -O-alkyl structure, such as -OCH3, -OCH2CH3, -OCH2CH2CH3, -O-CH2CH(CH3)2, -OCH2CH2CH2CH3, -O-CH(CH3)2, etc.

[0078] The term "heteroaryl" as used in this invention refers to an aromatic ring containing one, two, or three heteroatoms selected from O, N, or S. Heteroaryl groups within the scope of this invention include, but are not limited to: quinazoline, quinolinyl, pyrazolyl, pyrroleyl, thiopheneyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl, triazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridazinyl, etc.

[0079] As will be understood by those skilled in the art, the term “halo” or “halogen” as used herein refers to chlorine, fluorine, bromine, and iodine.

[0080] This invention includes the free form of compounds of Formula I, as well as their pharmaceutically acceptable salts and stereoisomers. The stereoisomers described in this invention are (depending on their structure) enantiomers, diastereomers, syn- / anti-isomers, cis- / trans-isomers, epimers, and (E)- / (Z)-isomers. Compounds of Formula I may be used in the context of this invention as pure stereoisomers or as any mixture of stereoisomers, preferably racemic in the latter case.

[0081] The starting materials and reagents used in the following examples are all commercially available conventional materials and reagents, or the substrates required for electrolysis can be synthesized by known methods reported in the literature.

[0082] In the following examples, room temperature refers to 23–25°C.

[0083] Example 1: Electrochemical synthesis of compound 3a

[0084]

[0085] In a single-chamber electrolytic cell with graphite sheet as anode and platinum sheet as cathode, raw material 1a (0.3 mmol), raw material 2a (0.36 mmol), and electrolyte tetrabutylammonium hexafluorophosphate (n-Bu4NPF6 0.5 mmol) were added to a mixed solvent system of 4.5 mL acetonitrile and 0.5 mL trifluoroacetic acid. Electrolysis was started with a constant current of 8 mA and stirred at room temperature for 3 hours (the charge was 3.0 F / mol based on the amount of raw material 1a). Electrolysis was then stopped, the reaction solution was transferred and concentrated, and 3a was obtained by column chromatography as a pale yellow solid with a yield of 72%.

[0086] The characterization data of compound 3a are as follows: 1 H NMR (400MHz, CDCl3) δ9.25 (s, 1H), 8.56 (dd, J = 9.4, 5.2Hz, 1H), 7.59-7.57 (m, 3H ),7.50-7.42(m,4H),6.99(d,J=9.0Hz,2H),6.63(d,J=8.7Hz,2H),3.69(s,3H);

[0087] 13 C NMR (100MHz, CDCl3) δ 161.1 (d, 1 J C-F=249Hz), 158.8, 157.7 (d, 4 J C-F =3Hz), 152.1(d, 4 J C-F =5Hz),142.0,135.6,132.6,132.0(d, 3 J C-F =8Hz), 130.1, 128.9, 128.3 (d, 3 J C-F =8Hz),127.8,123.0,122.7,121.9(d, 2 J C-F =25Hz), 115.0, 111.1(d, 2 J C-F =20Hz), 55.3;

[0088] HR-MS(ESI)m / z calcd for C 22 H 17 FNOSe[M+H] + 410.0454, found 410.0451.

[0089] Example 2: Electrochemical synthesis of compound 3b

[0090]

[0091] 3b was obtained by the same procedure as in Example 1, and was a pale yellow solid with a yield of 69%.

[0092] The characterization data of compound 3b are as follows: 1 H NMR (400MHz, CDCl3) δ9.20 (s, 1H), 8.46 (d, J = 9.1Hz, 1H), 7.94 (d, J = 2.1Hz, 1H), 7.62-7. 56(m,3H),7.45-7.41(m,3H),6.97(d,J=8.7Hz,2H),6.62(d,J=8.7Hz,2H),3.68(s,3H);

[0093] 13 C NMR (100MHz, CDCl3) δ158.9,158.4,151.9,141.9,137.0,133.3,132.6,13 2.4,130.8,130.1,128.8,128.4,127.9,126.7,123.0,122.6,115.1,55.3;

[0094] HR-MS(ESI)m / z calcd for C 22 H 17 ClNOSe[M+H] + 426.0158, found 426.0155.

[0095] Example 3: Electrochemical Synthesis of Compounds

[0096]

[0097] 3c was obtained by the same operation as in Example 1. It was a pale yellow solid with a yield of 44%.

[0098] The characterization data of compound 3c are as follows: 1 H NMR(400MHz, CDCl3) δ9.20(s,1H),8.38(d,J=9.1Hz,1H),8.12(s,1H),7.74(d,J=9.1Hz,1H),7 .59-7.55(m,2H),7.45-7.40(m,3H),6.97(d,J=8.2Hz,2H),6.62(d,J=8.2Hz,2H),3.69(s,3H);

[0099] 13 C NMR (100MHz, CDCl3) δ158.9,158.4,151.8,141.9,137.2,134.9,132.6,130.8,130.1,129.2,128.4,127.9,123.0,122.6,121.4,115.0,55.3;

[0100] HR-MS(ESI)m / z calcd for C 22 H 17 BrNOSe[M+H] + 469.9653, found 469.9644.

[0101] Example 4: Electrochemical synthesis of compound 3d

[0102]

[0103] The same procedure as in Example 1 was followed to obtain 3d, which was a pale yellow solid with a yield of 68%.

[0104] The characterization data of compound 3d are as follows: 1H NMR (400MHz, CDCl3) δ9.38(s,1H),8.67(d,J=8.8Hz,1H),8.31(s,1H),7.87(dd,J=8.9,1.9Hz,1H) ,7.62-7.58(m,2H),7.46-7.45(m,3H),7.00(d,J=8.7Hz,2H),6.64(d,J=8.9Hz,2H),3.70(s,3H)..

[0105] 13 C NMR (100MHz, CDCl3) δ160.0,159.0,153.4,141.7,140.1,132.8,130.2,130.1,129.3(d, 2 J C-F =33Hz),128.6,127.9,127.1,127.0(d, 3 J C-F =3Hz), 126.1(d, 3 J C-F =5Hz), 123.9(d, 1 J C-F =271Hz),123.2,122.3,115.1,55.3.;

[0106] HR-MS(ESI)m / z calcd for C 23 H 17 F3NOSe[M+H] + 460.0422, found 460.0415.

[0107] Example 5: Electrochemical synthesis of compound 3e

[0108]

[0109] 3e was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 88%.

[0110] The characterization data of compound 3e are as follows: 1 H NMR (400MHz, CDCl3) δ9.23 (s, 1H), 8.41 (d, J = 8.6Hz, 1H), 7.76 (s, 1H), 7.59-7.54 (m, 3H), 7.45-7.40(m,3H),7.00(d,J=8.7Hz,2H),6.63(d,J=8.7Hz,2H),3.70(s,3H),2.55(s,3H);

[0111] 13C NMR (100MHz, CDCl3) δ158.6,157.4,152.4,142.4,137.5,136.9,133.9,132.3 ,130.1,128.6,128.5,128.0,127.7,127.0,123.1,122.5,114.9,55.3,21.6;

[0112] HR-MS(ESI)m / z calcd for C 23 H 20 NOSe[M+H] + 406.0705, found 406.0744.

[0113] Example 6: Electrochemical synthesis of compound 3f

[0114]

[0115] 3f was obtained by the same operation as in Example 1, which was a pale yellow solid with a yield of 79%.

[0116] The characterization data of compound 3f are as follows: 1 H NMR (400MHz, CDCl3) δ9.14 (s, 1H), 7.88 (d, J = 9.1Hz, 1H), 7.72 (s, 1H), 7.58-77.55 (m, 2H), 7.47-7.35 (m,3H),7.21(dd,J=9.0,2.1Hz,1H),7.07-7.02(m,2H),6.67-6.61(m,2H),3.81(s,3H),3.71(s,3H);

[0117] 13 C NMR (100MHz, CDCl3) δ162.0,158.7,158.6,151.8,142.5,140.5,132.5,130.0 ,130.0,128.1,127.8,124.0,122.9,121.8,120.5,115.0,106.9,55.6,55.3.

[0118] HR-MS(ESI)m / z calcd for C 23 H 20 NO2Se[M+H] + 422.0654, found 422.0645.

[0119] Example 7: Electrochemical synthesis of 3g of compound

[0120]

[0121] 3g of a pale yellow solid was obtained by the same procedure as in Example 1, with a yield of 78%.

[0122] The characterization data for compound 3g are as follows: 1 H NMR(400MHz, CDCl3) δ9.30(s,1H),8.48(dd,J=9.4,5.0Hz,1H),7.63(d,J=8.1Hz,1H),7.58 (d,J=4.2Hz,2H),7.44(dd,J=20.6,7.6Hz,4H),7.09(d,J=4.4Hz,3H),7.05-6.99(m,2H).;

[0123] 13 C NMR (100MHz, CDCl3) δ 161.1 (d, 1 J C-F =250Hz), 158.2(d, 4 J C-F =2Hz), 152.5(d, 4 J C-F =5Hz),141.9,135.7,133.0,131.9(d, 3 J C-F =8Hz),129.9,129.8,129.4,128.9(d, 3 J C-F =9Hz),128.3,127.8,126.4,122.1(d, 2 J C-F =25Hz), 121.5(d, 4 J C-F =2Hz),111.1(d, 2 J C-F =20Hz);

[0124] 19 F NMR (376MHz, CDCl3) δ = -110.5

[0125] HR-MS(ESI)m / z calcd for C 21 H 15 FNSe[M+H] + 380.0348, found 380.0341.

[0126] Example 8: Electrochemical synthesis of compounds for 3 hours

[0127]

[0128] The same procedure as in Example 1 was followed to obtain a 3h solid, which was a pale yellow solid with a yield of 78%.

[0129] The characterization data of compound 3h are as follows: 1 H NMR (400MHz, CDCl3) δ9.27 (s, 1H), 8.40 (d, J = 9.0Hz, 1H), 7.98 (s, 1H), 7.62-7 .58(m,3H),7.43-7.42(d,J=5.3Hz,3H),7.10-7.09(m,3H),7.03-7.01(m,2H);

[0130] 13 C NMR (100MHz, CDCl3) δ158.9,152.3,141.8,137.1,133.4,132.9,132.6,130.8,130.0,129.8,129.4,128.7,128.4,127.9,126.8,126.5,121;

[0131] HR-MS(ESI)m / z calcd for C 21 H 15 ClNSe[M+H] + 396.0053, found 396.0048.

[0132] Example 9: Electrochemical synthesis of compound 3i

[0133]

[0134] 3i was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 78%.

[0135] The characterization data of compound 3i are as follows: 1 H NMR (400MHz, CDCl3) δ9.26 (s, 1H), 8.32 (d, J = 9.0Hz, 1H), 8.16 (s, 1H), 7.73 (d, J = 9.1 Hz,1H),7.62-7.56(m,2H),7.43-7.42(m,3H),7.12-7.05(m,3H),7.05-6.99(m,2H);

[0136] 13 C NMR (100MHz, CDCl3) δ158.9,152.2,141.8,137.3,135.1,132.9,130.8,130.2,130.0,129.9,129.4,129.1,128.4,127.9,126.5,121.6,121.6;

[0137] HR-MS(ESI)m / z calcd for C 21 H 15 BrNSe[M+H] + 439.9548 found 439.9541.

[0138] Example 10: Electrochemical synthesis of compound 3j

[0139]

[0140] 3j was obtained by the same operation as in Example 1. It was a pale yellow solid with a yield of 80%.

[0141] The characterization data of compound 3j are as follows: 1 H NMR (400MHz, CDCl3) δ9.44(s,1H),8.59(d,J=8.8Hz,1H),8.33(s,1H),7.86(d,J=9.0H z,1H),7.64-7.57(m,2H),7.47-7.39(m,3H),7.10(d,J=5.2Hz,3H),7.07-7.00(m,2H);

[0142] 13 C NMR (100MHz, CDCl3) δ160.5,153.8,141.6,140.2,132.6,130.2,130.0,129.5,129.4(d, 2 J C-F =33Hz), 128.7, 127.9, 127.2 (d, 3 J C-F =2Hz), 127.1, 126.6 (d, 3 J C-F =4Hz), 126.1, 123.8 (d, 1 J C-F =270Hz), 121.8;

[0143] 19 F NMR (376MHz, CDCl3) δ = -62.3.

[0144] HR-MS(ESI)m / z calcd for C 22 H 15 F3NSe[M+H] + 430.0316, found 430.0315.

[0145] Example 11: Electrochemical synthesis of compound 3k

[0146]

[0147] 3k was obtained by the same procedure as in Example 1, and was a pale yellow solid with a yield of 81%.

[0148] The characterization data of compound 3k are as follows: 1 H NMR (400MHz, CDCl3) δ9.37(s,1H),8.47(d,J=8.3Hz,1H),8.04(d,J=8.0Hz,1H),7.72(dd,J=8.7,Hz,1H ),7.64(dd,J=7.5Hz,1H),7.61-7.58(m,2H),7.45-7.38(m,3H),7.10-7.08(m,3H),7.06-7.04(m,2H);

[0149] 13 C NMR (100MHz, CDCl3) δ158.6,153.4,142.2,138.7,133.2,131.8,129.9,129.7,129.3,128.7,128.3,128.2,127.8,127.6,126.2,121.4;

[0150] HR-MS(ESI)m / z calcd for C 21 H 16 NSe[M+H] + 362.0442, found 362.0438.

[0151] Example 12: Electrochemical synthesis of compound 3l

[0152]

[0153] 3l of a pale yellow solid was obtained by the same procedure as in Example 1, with a yield of 51%.

[0154] The characterization data of compound 3l are as follows: 1 H NMR (400MHz, CDCl3) δ9.19 (s, 1H), 7.89 (d, J = 8.8Hz, 1H), 7.64 (s, 1H), 7.59-7. 58(m,2H),7.43-7.41(m,3H),7.21(d,J=8.8Hz,1H),7.10(s,5H),3.73(s,3H);

[0155] 13C NMR (100MHz, CDCl3) δ162.1,159.0,152.1,142.4,140.6,133.0,130.0,130.0,129.9,129.3,128.2,127.8,126.3,123.9,120.6,106.8,55.6;

[0156] HR-MS(ESI)m / z calcd for C 22 H 18 NOSe[M+H] + 392.0548, found 392.0545.

[0157] Example 13: Electrochemical synthesis of compound 3m

[0158]

[0159] The same procedure as in Example 1 was followed to obtain 3m, a pale yellow solid, with a yield of 64%.

[0160] The 3m characterization data of the compound are as follows: 1 H NMR(400MHz, CDCl3) δ9.36(s,1H),8.39(d,J=8.5Hz,1H),8.03(d,J=8.2Hz,1H),7.73(dd,J=7.9Hz,1H),7 .64(dd,J=7.6Hz,1H),7.59-7.54(m,2H),7.42-7.40(m,3H),7.18(d,J=8.1Hz,2H),6.87(d,J=8.3Hz,2H);

[0161] 13 C NMR (100MHz, CDCl3) δ158.7,153.6,142.0,138.4,132.3,132.1,132.0,131.3,129.9,128.4,128.4,128.3,128.3,127.9,127.8,121.1,120.2;

[0162] HR-MS(ESI)m / z calcd for C 21 H 15 BrNSe[M+H] + 396.0053 found 396.0052.

[0163] Example 14: Electrochemical synthesis of compound 3n

[0164]

[0165] 3n was obtained by the same operation as in Example 1, which was a yellow solid with a yield of 66%.

[0166] The characterization data of compound 3n are as follows: 1 H NMR(400MHz, CDCl3) δ9.36(s,1H),8.39(d,J=8.5Hz,1H),8.03(d,J=8.2Hz,1H),7.73(dd,J=7.9Hz,1H),7 .64(dd,J=7.6Hz,1H),7.59-7.54(m,2H),7.42-7.40(m,3H),7.18(d,J=8.1Hz,2H),6.87(d,J=8.3Hz,2H);

[0167] 13 C NMR (100MHz, CDCl3) δ158.7,153.6,142.0,138.4,132.3,132.1,132.0,131.3,129.9,128.4,128.4,128.3,128.3,127.9,127.8,121.1,120.2;

[0168] HR-MS(ESI)m / z calcd for C 21 H 15 BrNSe[M+H] + 439.9548 found 439.9546.

[0169] Example 15: Electrochemical synthesis of compound 3o

[0170]

[0171] 3o was obtained by the same operation as in Example 1, which was a pale yellow solid with a yield of 67%.

[0172] The characterization data of compound 3o are as follows: 1 H NMR (400MHz, CDCl3) δ9.39(d,J=2.7Hz,1H),8.36(d,J=8.4Hz,1H),8.06(d,J=7.9Hz,1H),7.74(dd,J=7.6Hz ,1H),7.67(dd,J=7.6Hz,1H),7.55(m,2H),7.43-7.38(m,3H),7.31(d,J=7.9Hz,2H),7.09(d,J=7.8Hz,2H);

[0173] 13C NMR (100MHz, CDCl3) δ159.0,153.9,141.9,138.5,132.2,129.8,129.1,128.5,128.4,128.3,128.2(d, 2 J C-F =33Hz), 127.9, 126.0 (d, 4 J C-F =4Hz), 124.1(d, 1 J C-F =270Hz), 120.2;

[0174] 19 F NMR (376MHz, CDCl3) δ = -62.4.

[0175] HR-MS(ESI)m / z calcd for C 22 H 15 F3NSe[M+H] + 430.0316 found 430.0303.

[0176] Example 16: Electrochemical synthesis of compound 3p

[0177]

[0178] 3p was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 66%.

[0179] The characterization data of compound 3p are as follows: 1 H NMR (400MHz, CDCl3) δ9.35(s,1H),8.48(d,J=8.6Hz,1H),8.01(d,J=7.9Hz,1H),7.71(dd,J=7.6Hz,1 H),7.63-7.60(m,3H),7.45-7.41(m,3H),6.96(d,J=8.1Hz,2H),6.90(d,J=8.0Hz,2H),2.22(s,3H);

[0180] 13 C NMR (100MHz, CDCl3) δ158.5,153.3,142.3,138.7,136.1,131.8,130.1,13 0.0,130.0,129.9,129.4,128.8,128.3,128.2,127.8,127.6,121.8,21.1;

[0181] HR-MS(ESI)m / z calcd for C22H18NSe[M+H]+ 376.0599 found 376.0596.

[0182] Example 17: Electrochemical synthesis of compound 3q

[0183]

[0184] 3q was obtained by the same operation as in Example 1, which was a yellow solid with a yield of 78%.

[0185] The 3q characterization data of the compound are as follows: 1 H NMR (400MHz, CDCl3) δ9.31 (s, 1H), 8.56-8.49 (m, 1H), 7.99 (d, J = 8.1Hz, 1H), 7.72 (dd, J = 7.8Hz, 1H ),7.63-7.57(m,3H),7.44-7.42(m,3H),7.00(d,J=8.8Hz,2H),6.63(d,J=8.7Hz,2H),3.68(s,3H);

[0186] 13 C NMR (100MHz, CDCl3) δ158.6,158.1,153.0,142.3,138.6,132.4,131.7,13 0.1,128.7,128.3,128.2,128.2,127.8,127.5,122.9,122.8,114.9,55.3;

[0187] HR-MS(ESI)m / z calcd for C 22 H 18 NOSe[M+H] + 392.0548 found 392.0545...

[0188] Example 18: Electrochemical synthesis of compound 3r

[0189]

[0190] 3r was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 72%.

[0191] The characterization data of compound 3r are as follows: 1H NMR (400MHz, CDCl3) δ9.34(s,1H),8.47(d,J=8.2Hz1H),8.02(d,J=7.8Hz,1H),7.73(dd,J= 7.8Hz,1H),7.65(dd,J=7.8Hz,1H),7.58-7.54(m2H),7.11-7.07(m,5H),7.03-7.01(m,2H);

[0192] 13 C NMR (100MHz, CDCl3) δ 162.8 (d, 1 J C-F =246Hz),157.5,153.4,138.8(d, 4 J C-F =2Hz)138.2,133.0,132.0(d, 3 J C-F =8Hz),131.8,129.7,129.3,128.3,128.2,128.2,126.3,121.6,114.7(d, 2 J C-F =21Hz);

[0193] 19 F NMR (376MHz, CDCl3) δ = -110.7.

[0194] HR-MS(ESI)m / z calcd for C 21 H 15 FNSe[M+H] + 380.0348, found 380.0348.

[0195] Example 19: Electrochemical synthesis of compound 3s

[0196]

[0197] The same procedure as in Example 1 was followed to obtain 3s, which is a pale yellow solid with a yield of 75%.

[0198] The characterization data of compound 3s are as follows: 1H NMR (400MHz, CDCl3) δ9.34(s,1H),8.47(d,J=8.5Hz,1H),8.03(d,J=8.0Hz,1H),7.73(dd,J=7.9Hz,1H),7. 65(dd,J=7.5Hz,1H),7.51(d,J=8.3Hz,2H),7.36(d,J=8.4Hz,2H),7.10-7.08(m,3H),7.03-7.00(m,2H).;

[0199] 13 C NMR (100MHz, CDCl3) δ157.3,153.5,140.6,138.7,134.2,132.9,132.0,131.4,129.7,129.3,128.7,128.3,128.3,127.9,127.8,126.4,121.7;

[0200] 19 F NMR (376MHz, CDCl3) δ = -110.7.

[0201] HR-MS(ESI)m / z calcd for C 21 H 15 ClNSe[M+H] + 396.0053, found 396.0041.

[0202] Example 20: Electrochemical synthesis of 3t of compound

[0203]

[0204] 3 tons of a pale yellow solid were obtained through the same procedure as in Example 1, with a yield of 85%.

[0205] The characterization data of compound 3t are as follows: 1 H NMR (400MHz, CDCl3) δ9.38 (s, 1H), 8.47 (d, J = 8.4Hz, 1H), 8.04 (d, J = 8.0Hz, 1H), 7. 76-7.62(m,8H),7.47(dd,J=7.6Hz,2H),7.37(dd,J=7.4Hz,1H),7.11-7.0(m,5H);

[0206] 13C NMR (100MHz, CDCl3) δ158.2,153.5,141.1,141.0,140.9,138.7,133.3,131.9,130.5, 129.7,129.3,128.9,128.7,128.3,128.3,127.7,127.5,127.3,126.6,126.3,121.4;

[0207] HR-MS(ESI)m / z calcd for C 27 H 20 NSe[M+H] + 438.0755, found 438.0755.

[0208] Example 21: Electrochemical synthesis of compound 3u

[0209]

[0210] 3u was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 85%.

[0211] The characterization data of compound 3u are as follows: 1 HNMR (400MHz, CDCl3) δ9.34(s,1H),8.42(d,J=8.5Hz,1H),7.99(d,J=8.1Hz,1H),7.67(dd,J=7.8Hz,1H ),7.59(dd,J=7.5Hz,1H),7.50(d,J=7.7Hz,2H),7.22(d,J=7.9Hz,2H),7.09-7.02(m,5H),2.40(s,3H);

[0212] 13 C NMR (100MHz, CDCl3) δ158.7,153.4,139.3,138.7,138.1,133.4,131.7,12 9.9,129.6,129.3,128.7,128.5,128.2,128.2,127.5,126.1,121.1,21.5;

[0213] HR-MS(ESI)m / z calcd for C 22 H 18 NSe[M+H] + 376.0599, found 376.0591.

[0214] Example 22: Electrochemical synthesis of compound 3v

[0215]

[0216] 3V was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 61%.

[0217] The characterization data of compound 3v are as follows: 1 HNMR (400MHz, CDCl3) δ9.34(s,1H),8.41(d,J=8.4Hz,1H),8.01(d,J=8.0Hz,1H),7.68(d ,J=8.1Hz,1H),7.62-7.56(m,3H),7.14-7.01(m,5H),6.95(d,J=8.1Hz,2H),3.85(s,3H);

[0218] 13 C NMR (100MHz, CDCl3) δ159.7,158.2,153.4,138.8,134.7,133.4,131.8,13 1.4,129.5,129.3,128.7,128.3,128.1,127.4,126.1,120.8,113.8,55.4;

[0219] HR-MS(ESI)m / z calcd for C 22 H 18 NOSe[M+H] + 392.0548, found 392.0536.

[0220] Example 23: Electrochemical synthesis of compound 3w

[0221]

[0222] 3w was obtained by the same operation as in Example 1, which was a pale yellow solid with a yield of 73%.

[0223] The characterization data of compound 3w are as follows: 1 H NMR (400MHz, CDCl3) δ9.29(s,1H),8.55(d,J=8.6Hz,1H),8.00(d,J=8.0Hz,1H),7.75(dd,J=7.7Hz,1H),7.64(dd,J= 7.5Hz,1H),7.51(d,J=8.4Hz,2H),7.39(d,J=8.4Hz,2H),6.98(d,J=8.8Hz,2H),6.63(d,J=8.9Hz,2H),3.71(s,3H).;

[0224] 13C NMR (100MHz, CDCl3) δ158.8,156.8,153.1,141.2,138.6,132.5,131.8,130.9,128.7,128.3,128.2,127.7,123.1,122.6,122.5,115.0,55.3;

[0225] HR-MS(ESI)m / z calcd for C 22 H 17 ClNOSe[M+H] + 426.0158, found 426.0148.

[0226] Example 24: Electrochemical synthesis of compound 3x

[0227]

[0228] 3x was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 88%.

[0229] The characterization data of compound 3x are as follows: 1 H NMR (400MHz, CDCl3) δ9.28(s,1H),8.55(d,J=8.3Hz,1H),8.00(dd,J=8.2,1.3Hz,1H),7.75(dd,J=7.1Hz,1H),7.64(dd ,J=7.5Hz,1H),7.54(d,J=8.4Hz,2H),7.44(d,J=8.5Hz,2H),6.97(d,J=8.9Hz,2H),6.63(d,J=8.8Hz,2H),3.71(s,3H);

[0230] 13 C NMR (100MHz, CDCl3) δ158.8,156.8,153.1,140.7,138.6,134.2,132.5,131 .8,131.5,128.7,128.3,128.2,127.9,127.7,123.1,122.7,115.0,55.3.;

[0231] HR-MS(ESI)m / z calcd for C 22 H 17 BrNOSe[M+H] + 469.9654, found 469.9646.

[0232] Example 25: Electrochemical synthesis of compound 3y

[0233]

[0234] 3y was obtained by the same procedure as in Example 1, which was a pale yellow solid with a yield of 70%.

[0235] The characterization data of compound 3y are as follows: 1 H NMR (400MHz, CDCl3) δ9.31 (s, 1H), 9.21-9.12 (m, 1H), 8.33 (d, J = 8.1Hz, 1H), 7.56-7.55 (m, 3H), 7.35-7.33 (m, 3H), 7.09-6.95 (m, 5H);

[0236] 13 C NMR (100MHz, CDCl3) δ160.8,155.3,152.6,151.7,141.4,136.3,132.6,131.7,129.8,128.8,128.3,127.8,126.4,125.8,122.9,122.8;

[0237] HR-MS(ESI)m / z calcd for C 20 H 15 N2Se[M+H] + 363.0395 found 363.0391.

[0238] Example 26: Electrochemical synthesis of compound 3z

[0239]

[0240] Following the same procedure as in Example 1, 3z was obtained as a pale yellow solid with a yield of 56%.

[0241] The characterization data of compound 3z are as follows: 1 H NMR (400MHz, CDCl3) δ99.25 (s, 1H), 9.19 (dd, J=4.2, 1.8Hz, 1H), 8.30 (dd, J=8.2, 1.8Hz, 1H) ,7.57-7.52(m,3H),7.37-7.34(m,,3H),7.08-7.04(m,2H),6.55-6.51(m,2H),3.69(s,3H);

[0242] 13C NMR (100MHz, CDCl3) δ160.2,158.9,155.0,152.1,151.6,141.6,136.2,134.8,129.8,128.2,127.9,127.2,122.9,122.7,122.3,114.4,55.3.;

[0243] HR-MS(ESI)m / z calcd for C 21 H 17 N2OSe[M+H] + 393.0504, found 393.0494.

[0244] Example 27: Electrochemical synthesis of compound 3aa

[0245]

[0246] 3aa was obtained by the same procedure as in Example 1, which was a yellow solid with a yield of 57%.

[0247] The characterization data of compound 3aa are as follows: 1 H NMR (400MHz, CDCl3) δ9.21 (s, 1H), 7.68 (d, J = 5.3Hz, 1H), 7.57 (d, J = 5.7Hz, 2H), 7.49 (d, J = 5.3Hz, 1H), 7.41 (d, J = 6.1Hz, 3H), 7.11 (s, 5H);

[0248] 13 C NMR (100MHz, CDCl3) δ157.5,149.9,144.4,141.2,135.1,133.0,132.7,130.3,129.9,129.4,128.3,127.9,126.6,126.2,118.9;

[0249] HR-MS(ESI)m / z calcd for C 19 H 14 NSSe[M+H] + 368.0007, found 368.0010.

[0250] Example 28: Electrochemical synthesis of compound 3ab

[0251]

[0252] 3ab was obtained by the same procedure as in Example 1, and it was a yellow solid with a yield of 90%.

[0253] The characterization data of compound 3ab are as follows: 1 H NMR (400MHz, CDCl3) δ9.16 (s, 1H), 7.67 (d, J = 5.5Hz, 1H), 7.57 (d, J = 2.1Hz, 1H), 7.56 (d, J = 1.5Hz, 1H) ,7.54(d,J=5.5Hz,1H),7.45-7.40(m,3H),7.07(d,J=8.8Hz,2H),6.66(d,J=8.7Hz,2H),3.72(s,3H).;

[0254] 13 C NMR (100MHz, CDCl3) δ159.0,157.0,149.5,144.0,141.4,135.1,133.2,132.7,130.0,128.2,127.9,126.1,122.4,120.4,115.0,55.3;

[0255] HR-MS(ESI)m / z calcd for C 20 H 16 NOSSe[M+H] + 398.0112, found 398.0107.

[0256] Example 29: Electrochemical synthesis of compound 3ac

[0257]

[0258] 3ac was obtained by the same procedure as in Example 1, which was a yellow solid with a yield of 57%.

[0259] The characterization data of compound 3ac are as follows: 1 H NMR (400MHz, CDCl3) δ9.21 (s, 1H), 7.68 (d, J = 5.3Hz, 1H), 7.57 (d, J = 5.7Hz, 2H), 7.49 (d, J = 5.3Hz, 1H), 7.41 (d, J = 6.1Hz, 3H), 7.11 (s, 5H);

[0260] 13 C NMR (100MHz, CDCl3) δ153.7,153.6,142.8,139.0,133.2,131.9,129.7,129.4,129.4,128.8,128.2,128.1,127.5,126.6,126.2,124.4,120.7;

[0261] HR-MS(ESI)m / z calcd for C 19 H 14 NSSe[M+H] + 368.0007, found 368.0010.

[0262] Example 30: Electrochemical synthesis of compound 3ad

[0263]

[0264] 3ad was obtained by the same procedure as in Example 1, which was a yellow solid with a yield of 78%.

[0265] The characterization data of compound 3ad are as follows: 1 H NMR (400MHz, CDCl3) δ9.40 (s, 1H), 8.50 (d, J = 8.4Hz, 1H), 8.06-8.03 (m, 2H), 7.88 (d, J = 8.0Hz, 2H), 7. 82(d,J=7.4Hz,1H),7.76-7.72(m,2H),7.65(dd,J=7.6Hz,1H),7.53-7.49(m,2H),7.12-7.00(m,5H);

[0266] 13 C NMR (100MHz, CDCl3) δ158.5,153.5,139.6,138.8,133.3,133.1,132.9,131.9,129.8,129 .5,129.3,128.7,128.6,128.3,127.8,127.8,127.7,127.2,126.4,126.3,126.1,121.7;

[0267] HR-MS(ESI)m / z calcd for C 25 H 18 NSe[M+H] + 412.0599, found 412.0593.

[0268] Example 31: Electrochemical synthesis of compound 3ae

[0269]

[0270] 3ae, a yellow solid, was obtained by the same procedure as in Example 1, with a yield of 89%.

[0271] The characterization data of compound 3ae are as follows: 1H NMR (400MHz, CDCl3) δ.44 (s, 1H), 8.76 (dd, J = 12.6, 8.4Hz, 2H), 8.53 (d, J = 8.4Hz, 1H ),8.11(d,J=8.0Hz,1H),7.81-7.57(m,7H),7.51-7.40(m,2H),7.06-6.95(m,5H).;

[0272] 13 C NMR (100MHz, CDCl3) δ157.9,153.4,138.5,132.9,131.9,131.3,131.3,130.6,130.5,129.1,129 .1,128.6,128.5,128.4,128.3,127.9,126.9,126.7,126.6,126.5,126.4,124.5,123.0,122.7;

[0273] HR-MS(ESI)m / z calcd for C 29 H 20 NSe[M+H] + 462.0755, found 462.0745.

[0274] Example 32: Electrochemical synthesis of compound 3af

[0275]

[0276] 3af was obtained by the same operation as in Example 1, which was a pale yellow solid with a yield of 43%.

[0277] The characterization data of compound 3af are as follows: 1 H NMR (400MHz, CDCl3) δ9.15(s,1H),8.44(d,J=8.3Hz,1H),7.88(d,J=8.0Hz,1H),7.65(dd,J=7.8Hz,1H),7.50(dd,J= 7.5Hz,1H),7.22-7.07(m,5H),3.15(tt,J=8.7,4.7Hz,1H),1.20(dt,J=5.8,3.1Hz,2H),1.00(dq,J=6.7,3.4Hz,2H);

[0278] 13C NMR (100MHz, CDCl3) δ160.4,153.9,138.9,132.6,131.5,129.4,129.3,128.1,127.7,127.6,126.3,126.1,120.3,17.5,10.7;

[0279] HR-MS(ESI)m / z calcd for C 18 H 16 NSe[M+H] + 326.0442, found 326.0437.

[0280] Example 33: Electrochemical synthesis of compound 3k

[0281]

[0282] The difference between this embodiment and embodiment 11 is that:

[0283] [1] Tetrabutylammonium hydrogen sulfate was used instead of tetrabutylammonium hexafluorophosphate, with a yield of 80%;

[0284] [2] Tetrabutylammonium tetrafluoroborate was used instead of tetrabutylammonium hexafluorophosphate, with a yield of 17%;

[0285] [3] Tetrabutylammonium perchlorate was used instead of tetrabutylammonium hexafluorophosphate, with a yield of 65%;

[0286] [4] Using pure acetonitrile instead of the mixed solvent of acetonitrile and trifluoroacetic acid, the yield was 13%;

[0287] [5] Using a mixed solvent of acetonitrile and methanol with a volume ratio of 9:1 instead of a mixed solvent of acetonitrile and trifluoroacetic acid, the yield was 20%.

[0288] [6] Replacing 8mA with 20mA resulted in a 0% yield;

[0289] [7] Heating to 50°C instead of room temperature yields 80% of the product.

[0290] Example 34: Determination of the antitumor activity of selenoisoquinoline compounds by MTT assay

[0291] First, human lung cancer H460 cells, breast cancer MCF-7 cells, human cervical cancer HeLa cells, and human lung cancer cisplatin-resistant cell line A549 / DDP were cultured to the logarithmic growth phase. The cells were then digested with trypsin, centrifuged to obtain cell pellets, resuspended in culture medium, and seeded into 96-well plates at 5000 cells per well (100 μL / well). After cell attachment, the original culture medium was discarded, and 100 μL of culture medium containing different concentrations (100, 50, 25, 12.5, 6.25, 3.125 μM) of the test compound was added to each well (experimental wells). Gefitinib was used as a positive control, and the blank control wells contained only an equal volume of culture medium without the compound. The cells were cultured for another 48 hours. After 48 hours of culture, 10 μL of 5 mg / mL MTT (thiazolyl bromide blue tetrazolium) was added to each well, and the plate was incubated in a cell culture incubator for 4 hours. The liquid in the wells was then aspirated, and 100 μL of DMSO was added to each well. The plate was then incubated for 15 minutes. After the formazan was completely dissolved, the OD value of each well at 570 nm was measured using a microplate reader, and the inhibition rate was calculated based on the OD value. Inhibition rate = (OD...) 空白对照孔 -OD 实验孔 ) / OD 空白对照孔 ×100%. Plot the concentration-cell survival rate curves for each compound and perform non-linear fitting to obtain the half-maximal inhibitory concentration (IC50) of the compound on tumor cells. 50 ).

[0292] The test results are shown in Tables 1-9: The selenoisoquinoline compounds synthesized in this invention have good antitumor activity.

[0293] Table 1. Half-maximal inhibitory concentration (IC50) of compounds against tumor cells. 50 (μM)

[0294]

[0295]

[0296] Table 2. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0297]

[0298]

[0299] Table 3. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0300]

[0301]

[0302] Table 4. Half-maximal inhibitory concentrations (IC50, μM) of the compounds against tumor cells.

[0303]

[0304]

[0305]

[0306] Table 5. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0307]

[0308]

[0309] Table 6. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0310]

[0311]

[0312] Table 7. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0313]

[0314]

[0315] Table 8. Half-maximal inhibitory concentrations (IC50) of compounds against tumor cells. 50 (μM)

[0316]

[0317]

[0318] Table 9. Half-maximal inhibitory concentrations (IC50) of compounds against drug-resistant cell lines. 50 (μM)

[0319]

[0320] As shown in Tables 1-9, the selenoisoquinoline compounds synthesized in this invention have good antitumor activity. Among them, compounds 3q, 3f, and 3z have more significant inhibitory effects on three types of human tumor cells, and also have significant inhibitory effects on the human lung cancer cisplatin-resistant cell line A549 / DDP.

[0321] Example 35 Scratch Test

[0322] 1. Human lung cancer H460 cells were seeded into 6-well plates. After the cells adhered, they were treated with the test compound or the positive control drug gefitinib. The concentrations of compounds 3q and 3f were 2.5 μM, the concentration of compound 3z was 0.5 μM, and the concentration of gefitinib was 10 μM. The blank control wells were not treated.

[0323] 2. When the cells reach 100% confluence, remove the original culture medium, wash twice with PBS, and starve the cells for 12 hours with serum-free culture medium containing the corresponding compound or positive control drug.

[0324] 3. After starvation treatment, perform cell streaking. Using a ruler as a guide, gently streak the cell streaks with a 10μL pipette tip, ensuring the streaks are perpendicular to the two horizontal lines previously drawn on the bottom of the well plate with a marker, resulting in four intersection points between the streaks and the marker lines. Take microscopic photographs at 0h and 24h after streaking. To ensure consistent photographic coverage, take one photograph above and one below each intersection point. Calculate the migration rate based on the streak width at 0h and 24h. Cell migration distance S = (L... 0h -L 24h Cell migration rate = S experimental group / S blank control group × 100% (L is the scar width).

[0325] from Figure 1 Cell scratch images and Figure 2 The results of scratch analysis showed that, compared with the blank control group, compounds 3f and 3q at a concentration of 2.5 μM and compound 3z at a concentration of 0.5 μM could significantly inhibit the migration of H460 cells (P<0.05), and their inhibitory effect was comparable to that of 10 μM positive control gefitinib.

[0326] Example 36 Transwell Cell Migration Assay

[0327] 1. H460 lung cancer cells treated with the test compound or positive control drug gefitinib (blank control cells were untreated) were digested, dispersed into single cells, centrifuged, and resuspended in PBS. After centrifugation again, the PBS was discarded to remove serum from the original culture medium. The cells were resuspended in serum-free culture medium containing the corresponding test compound or positive control drug gefitinib and counted. 600 μL of serum-containing culture medium was added to each well of a 24-well plate (lower chamber), and 40,000 cells / well were added to each upper chamber (upper chamber), for a total cell suspension volume of 200 μL. The plates were incubated for 24 h.

[0328] 2. After culturing for 24 hours, remove the culture medium from the Transwell chamber, wash twice with PBS, and then fix the Transwell chamber in 4% paraformaldehyde for 15 minutes.

[0329] 3. After fixation, stain the cells that have migrated to the submural region with 0.5% crystal violet dye for 15 minutes. After staining, wash away any residual crystal violet dye with PBS, and gently remove any remaining crystal violet and droplets from the chambers with a cotton swab. Photograph the cells that have migrated to the submural region using a microscope, count them, and calculate the cell migration rate. Cell migration rate = (Number of cells in experimental wells / Number of cells in control wells) × 100%.

[0330] from Figure 3 Cell migration images and Figure 4 Quantitative analysis showed that, compared with the blank control group, compounds 3f and 3q at a concentration of 2.5 μM and compound 3z at a concentration of 0.5 μM could significantly inhibit the migration of H460 cells (P<0.01), and their inhibitory effect was comparable to that of 10 μM positive control gefitinib.

[0331] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0332] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A method for synthesizing a selenized isoquinoline compound having the structure shown in Formula I, characterized in that, Includes the following steps: Compounds of Formula I-1 and Formula I-2 are electrolyzed in an electrolytic cell containing an electrolyte to obtain selenized isoquinoline compounds with the structure shown in Formula I. ； Wherein, ring A is selected from: one or more R1-substituted or unsubstituted benzene rings, or one or more R1-substituted or unsubstituted 5-6-membered aromatic heterocycles; R1 is selected from: hydrogen, C1~C1 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1–C6 alkoxy groups; R2 is selected from: C1~C 10 Alkyl, C3-C8 cycloalkyl, one or more R4-substituted or unsubstituted C6-C 18 Aryl, one or more R4-substituted or unsubstituted 5- to 10-membered heteroaryl groups; R3 is selected from: C1~C 10 Alkyl, one or more R5-substituted or unsubstituted C6-C 10 Aryl; R4 is selected from: hydrogen, C1~C1 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1–C6 alkoxy, amino, and phenyl groups; R5 is selected from: hydrogen, C1~C 10 Alkyl, C1-C6 alkoxy, halogen, halogen-substituted C1-C6 10 Alkyl or halogen-substituted C1–C6 alkoxy groups; R6 is selected from: H, C1-C6 alkyl acyl group; The molar ratio of the compound of formula I-1 to the compound of formula I-2 is 1:0.8 to 1.5; The electrolyte in the electrolyte solution is one or more tetrabutylammonium salts, wherein the tetrabutylammonium salt is selected from at least one of tetrabutylammonium hexafluorophosphate, tetrabutylammonium hydrogen sulfate and tetrabutylammonium perchlorate; The solvent in the electrolyte is a mixture of acetonitrile and trifluoroacetic acid in a volume ratio of 6 to 12:

1. The concentration of the electrolyte in the electrolyte solution is 0.03 mmol / mL to 0.15 mmol / mL; The molar ratio of the electrolyte to the compound of formula I-1 in the electrolyte solution is 1.0 to 2.0:1; Electrolysis is performed using a constant current with an intensity of 6 mA to 10 mA, and the amount of charge, expressed as the amount of substance of the compound of formula I-1, is 2.0 F / mol to 5.0 F / mol; the electrolysis temperature is 10 °C to 70 °C.

2. The method for synthesizing selenide isoquinoline compounds according to claim 1, characterized in that, The selenized isoquinoline compounds have the structures shown in Formula II or Formula III as follows: Among them, X1, X2, X3 and X4 are independently selected from: N, CR1; X5 is selected from: O, S, NR7; R7 is selected from: hydrogen, C1 to C6 alkyl groups.

3. The method for synthesizing selenized isoquinoline compounds according to claim 2, characterized in that, X1, X3, and X4 are all CH; X2 is CR1.

4. The method for synthesizing selenide isoquinoline compounds according to claim 2, characterized in that, X4 is N; X1, X2 and X3 are all CH.

5. The method for synthesizing selenized isoquinoline compounds according to claim 1, characterized in that, R1 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy.

6. The method for synthesizing selenized isoquinoline compounds according to claim 5, characterized in that, R1 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and trifluoromethyl.

7. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, R2 is selected from: C1-C3 alkyl, C3-C6 cycloalkyl, one or more R4-substituted or unsubstituted phenyl, one or more R4-substituted or unsubstituted naphthyl, one or more R4-substituted or unsubstituted phenanthryl, one or more R4-substituted or unsubstituted anthracel, one or more R4-substituted or unsubstituted 5-membered heteroaryl.

8. The method for synthesizing selenized isoquinoline compounds according to claim 7, characterized in that, R2 is selected from: cyclopropyl, one or more R4-substituted or unsubstituted phenyl, naphthyl, phenanthryl, anthracene, thiophene.

9. The method for synthesizing selenized isoquinoline compounds according to claim 1, characterized in that, R4 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, and phenyl.

10. The method for synthesizing selenized isoquinoline compounds according to claim 9, characterized in that, R4 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and phenyl.

11. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, R3 is selected from: C1 to C4 alkyl groups, one or more R5-substituted or unsubstituted phenyl groups.

12. The method for synthesizing selenized isoquinoline compounds according to claim 1, characterized in that, R5 is selected from: hydrogen, C1-C3 alkyl, C1-C3 alkoxy, halogen, halogen-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy.

13. The method for synthesizing selenized isoquinoline compounds according to claim 12, characterized in that, R5 is selected from: hydrogen, methyl, methoxy, fluorine, chlorine, bromine, and trifluoromethyl.

14. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The selenized isoquinoline compounds have the structure shown in Formula IV or Formula V: 。 15. The method for synthesizing selenized isoquinoline compounds according to claim 1, characterized in that, The selenized isoquinoline compound has the structure shown in Formula VI: 。 16. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1, 9-10, characterized in that, The selenized isoquinoline compounds have the structure shown in formula VII or VIII as follows: 。 17. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1, 12-13, characterized in that, The selenized isoquinoline compounds have the structure shown in Formula IX or Formula X: 。 18. The method for synthesizing selenized isoquinoline compounds according to claim 1, characterized in that, The selenized isoquinoline compounds are selected from the following compounds: 。 19. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The solvent is a mixture of acetonitrile and trifluoroacetic acid in a volume ratio of 8 to 10:

1.

20. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The tetrabutylammonium salt is selected from at least one of tetrabutylammonium hexafluorophosphate and tetrabutylammonium hydrogen sulfate.

21. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The anode used for electrolysis is a graphite sheet, a mesh of glassy carbon, a carbon felt, or a platinum sheet, and the cathode is a platinum sheet, an iron sheet, a lead sheet, a nickel sheet, or a zinc sheet.

22. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The molar ratio of the compound of formula I-1 to the compound of formula I-2 is 1:1.1 to 1.

3.

23. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The concentration of the electrolyte in the electrolyte solution is 0.08 mmol / mL to 0.12 mmol / mL.

24. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The molar ratio of the electrolyte to the compound of formula I-1 in the electrolyte solution is 1.5 to 1.8:

1.

25. The method for synthesizing selenide isoquinoline compounds according to any one of claims 1-6, characterized in that, The electrolysis temperature is 20℃~50℃.

26. A selenized isoquinoline compound or a pharmaceutically acceptable salt thereof, characterized in that, The selenized isoquinoline compounds are selected from the following compounds: 。 27. The use of the selenized isoquinoline compound of claim 26 or its pharmaceutically acceptable salt in the preparation of a medicament for treating or preventing tumors, wherein the tumor is: lung cancer, breast cancer, or cervical cancer.

28. An antitumor pharmaceutical composition, characterized in that, It is prepared from an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is a selenized isoquinoline compound as described in claim 26 or a pharmaceutically acceptable salt thereof.

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