Carbazole pyridine cuprous complex and its preparation method and application in the preparation of antitumor drugs

Abstract

The present invention relates to a copper carbazole pyridine complex, a preparation method thereof, and use thereof in the preparation of anti-tumor drugs. The copper carbazole pyridine complex is selected from one of the structures represented by formula (I): #imgabs0# In the structure represented by formula (I), at least one of R1, R2, R3, R4, or R5 is selected from the structure represented by formula (II): #imgabs1# In formula (II), R1, R2, R3, R4, R5, R6, or R7 can also be independently selected from -H, -OH, -COOH, -SH, -NH2, -N(CH3)2, -NO2, a C1-C5 alkoxy group, or a C1-C5 alkyl group, and X is selected from any one of -F, -Cl, -Br, -I, -OTf, -CN, -SCN, -AcO, and -S. The copper carbazole pyridine complex of the present invention can achieve high efficiency in inhibiting tumor cells at a relatively low concentration and has low toxicity to normal cells.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmacy, and in particular relates to a cuprous carbazole pyridine complex, a preparation method thereof, and application thereof in the preparation of anti-tumor drugs. Background Art

[0002] Metal complexes can produce a wide variety of structural changes due to their variable ligand structures, multiple optional counter anions, and diverse stereoconfigurations; and they have multiple anti-tumor mechanisms of action, making them a potential anti-tumor drug.

[0003] Since the clinical application of platinum-based metal drugs such as cisplatin, the anti-tumor advantages of metal complexes have gradually become apparent. However, their dose-limiting toxicity and drug resistance continue to hamper their clinical efficacy. Metal complexes are functional molecular systems formed by the coordination bond between a central metal ion (atom) and an organic ligand. These compounds demonstrate unique value in drug development due to their high structural designability, unique electronic properties, and diverse coordination modes. In recent years, with the advancement of supramolecular chemistry and nanotechnology, research on metal complexes has expanded into emerging fields such as metal-organic frameworks (MOFs) and coordinated supramolecular self-assembly systems. In particular, breakthroughs have been achieved in applied research in areas such as targeted tumor therapy (e.g., the construction of nano-drug delivery systems), bioimaging and tracing, and photodynamic therapy, providing new avenues for anti-tumor drug development.

[0004] Copper, as an endogenous metallic element in the body, generally has low toxic side effects on normal cells. Furthermore, research has found that cuprous complexes exhibit potent inhibitory activity against tumor cells. For example, patent CN114702441B discloses a cuprous complex synthesized from a salicylaldehyde Schiff base and quinoline as a double coordination site fragment, which inhibits tumor cells by inducing apoptosis. However, these cuprous complexes exhibit relatively low activity, necessitating the development of more novel cuprous complexes. Summary of the Invention

[0005] Based on the current situation that cuprous complexes in the prior art have relatively low tumor inhibition activity, the present invention provides a carbazole pyridine ligand, a carbazole pyridine cuprous complex, a preparation method thereof, and an application thereof in the preparation of anti-tumor drugs.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] The present invention provides a copper(II) carbazole pyridine complex selected from one of the structures represented by formula (I):

[0008] Formula (I)

[0009] In formula (I), R1, R2, R3, R4 or R5 are independently selected from -H, -OH, -COOH, -SH, -NH2, -N(CH3)2, -NO2, C1-C5 alkoxy, C1-C5 alkyl or carbazole or differently substituted carbazole of the structure shown in formula (II):

[0010] Formula (II)

[0011] In formula (II), R6 or R7 is independently selected from -H, -OH, -COOH, -SH, -NH2, -N(CH3)2, -NO2, C1-C5 alkoxy or C1-C5 alkyl; R6 or R7 is further preferably -H, -OH, -COOH, -SH, -NH2, -NO2, -OCH3 or -CH3;

[0012] In the structure represented by formula (I), at least one of R1, R2, R3, R4 or R5 is selected from the structure represented by formula (II);

[0013] X is selected from any one of -F, -Cl, -Br, -I, -OTf, -CN, -SCN, -AcO, and -S.

[0014] Preferably, in the structure represented by formula (I), R1 and R5 are both selected from the structure represented by formula (II), or R2 and R4 are both selected from the structure represented by formula (II).

[0015] More preferably, in the structure shown in formula (I), R1 and R5 are both selected from the structure shown in formula (II), and the structures of R1 and R5 are the same; R2 and R4 are both selected from the structure shown in formula (II), and the structures of R2 and R4 are the same.

[0016] In one embodiment of the present invention, the copper(II) carbazole pyridine complex is selected from one of the following compounds:

[0017] 、 、 、 、 、 、 、 、 .

[0018] Preferably, the copper carbazole pyridine complex is selected from Cu-1, Cu-2, Cu-4, Cu-6, Cu-7, and Cu-8. More preferably, the copper carbazole pyridine complex is selected from Cu-7.

[0019] The copper carbazole pyridine complex provided by the present invention can achieve the purpose of inhibiting various tumor cells under relatively low concentration conditions.

[0020] The present invention further provides a method for preparing the copper carbazole pyridine complex, comprising the following steps:

[0021] (1) The bromine-substituted pyridine reacts with carbazole to first obtain a carbazole-pyridine ligand;

[0022] (2) The carbazole pyridine ligand reacts with a monovalent Cu salt in a solvent to obtain the carbazole pyridine cuprous complex.

[0023] The carbazole pyridine ligand is selected from one of the structures shown in formula (III):

[0024] Formula (III)

[0025] In formula (III), R1, R2, R3, R4 or R5 is selected from -H, -OH, -COOH, -SH, -NH2, -N(CH3)2, -NO2, C1-C5 alkoxy, C1-C5 alkyl or carbazole or differently substituted carbazole of the structure shown in formula (II):

[0026] Formula (II)

[0027] In formula (II), R6 or R7 is independently selected from -H, -OH, -COOH, -SH, -NH2, -N(CH3)2, -NO2, C1-C5 alkoxy or C1-C5 alkyl; R6 or R7 is further preferably -H, -OH, -COOH, -SH, -NH2, -NO2, -OCH3 or -CH3;

[0028] In the structure shown in formula (III), at least one of R1, R2, R3, R4 or R5 is selected from the structure shown in formula (II).

[0029] In one embodiment of the present invention, the solvent is selected from an organic solvent and / or water; the organic solvent is selected from one or more solvents such as acetonitrile, methanol, ethanol, dichloromethane, tetrahydrofuran or dioxane.

[0030] In one embodiment of the present invention, the Cu salt is selected from the anhydrous form or crystalline water form of CuF, CuCl, CuBr, CuI, CuOTf, CuCN, CuSCN, CuAcO, and CuS.

[0031] In one embodiment of the present invention, a method for preparing the copper(II) carbazole pyridine complex is provided, comprising the following steps:

[0032] (1) Preparation of carbazole pyridine ligand: Under the protection of inert gas (such as nitrogen), pyridine and carbazole substituted with different positions and amounts of bromine are reacted with CuI catalyst in DMPU as solvent for a certain time. After cooling, dilute HCl is added and the mixture is extracted with DCM. The organic layers are combined and washed with dilute ammonia water, dried over anhydrous Na2SO4, and the solvent is removed in vacuo. The carbazole pyridine ligand is obtained by column chromatography.

[0033] (2) Preparation of cuprous carbazole pyridine ligand complex: a monovalent copper salt reacts with a carbazole pyridine ligand in a reaction solvent to obtain the cuprous carbazole pyridine complex.

[0034] In one embodiment of the present invention, in step (1), the reaction conditions are: heating at 230° C. overnight for 48 hours.

[0035] In one embodiment of the present invention, in step (1), the concentration of the added dilute HCl is 2 M.

[0036] In one embodiment of the present invention, in step (2), the molar ratio of the carbazole pyridine ligand to the copper salt is (1.0-5.0):1, which can be 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, etc.

[0037] In one embodiment of the present invention, in step (2), the reaction temperature is 0-100°C, preferably 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, etc., more preferably 25-60°C. The reaction time is 10 min-24 h, which can be 2 h, 4 h, 6 h, 8 h, 10 h, 13 h, 15 h, 17 h, 19 h, 21 h, 23 h, etc., more preferably 4-8 h.

[0038] In one embodiment of the present invention, in step (2), the reaction solvent is selected from an organic solvent and / or water; the organic solvent is selected from one or more solvents such as acetonitrile, methanol, ethanol, dichloromethane, tetrahydrofuran or dioxane.

[0039] The present invention further provides application of the carbazole-pyridine cuprous complex in the preparation of anti-tumor drugs.

[0040] In one embodiment of the present invention, the tumor cells are selected from A549 (human non-small cell lung cancer cells), A-498 (human renal cancer cells), A-431 (human epidermal cancer cells), MCF-7 (human breast cancer cells), HeLa (human cervical cancer cells), HCT 116 (human colon cancer cells), PC-3 (human prostate cancer cells), SiHa (human cervical cancer cells), SK-OV-3 (human ovarian adenocarcinoma cells), SW480 (human colon cancer cells), T24 (human bladder cancer cells), HL-60 (human promyelocytic leukemia cells), HT1080 (human fibrosarcoma cells), Hep G2 (human liver cancer cells), LNCaP (human prostate cancer cells), etc., as well as drug-resistant tumor cells A549 / DDP, MCF7 / DDP, HeLa / DDP, HCT 116 / DDP, PC-3 / DDP, SiHa / DDP, SK-OV-3 / DDP, SW480 / DDP, T24 / DDP, HL-60 / DDP, HT1080 / DDP, Hep G2 / DDP, LNCaP / DDP, etc. Preferably, the tumor cells are selected from A549 (human non-small cell lung cancer cells), MCF7 (human breast cancer cells), HeLa (human cervical cancer cells), HCT116 (human colon cancer cells), PC-3 (human prostate cancer cells), SiHa (human cervical cancer cells), SK-OV-3 (human ovarian adenocarcinoma cells), L-60 (human promyelocytic leukemia cells), Hep G2 (human liver cancer cells).

[0041] Furthermore, the copper carbazole pyridine complex obtained by the present invention is studied for its biological activity in different tumor diseases and has good inhibitory activity against cisplatin-resistant tumor cells.

[0042] The invention prepares a cuprous complex by chelating a carbazole pyridine ligand with cuprous ions, and utilizes the π electrons of the carbazole in the ligand and the metal center cuprous to carry out electron transfer and dispersion, thereby obtaining a cuprous complex with better pharmacological activity and greater stability.

[0043] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0044] (1) The copper complex provided by the present invention is prepared from a carbazole pyridine ligand and a corresponding cuprous salt. Since the large π-conjugated system of the carbazole ligand can cause electron transfer with the metal center cuprous salt, this is very different from the structural type of ligands in traditional cuprous complexes, enriching the diversity of ligand structures in the anti-tumor application of cuprous complexes.

[0045] (2) The copper carbazole pyridine complex prepared by the present invention can achieve high efficiency in inhibiting tumor cells at relatively low concentrations (1.7-51.2 μM), and has low toxicity to normal cells, very low inhibitory activity (greater than 55 μM), and has an inhibitory selectivity of more than 32 times for normal cells and tumor cells, which greatly increases the selective inhibition of the complex on tumor cells, reduces the toxic side effects of chemotherapy drugs on patients, and increases the effect of practical clinical applications.

[0046] (3) The copper-containing carbazole pyridine complex provided by the present invention also has good inhibitory ability against cisplatin-resistant tumor cells. Therefore, this type of copper-containing carbazole pyridine complex achieves inhibition of tumor cells based on catalyzing the generation of reactive oxygen species in tumor cells, which is different from the mechanism by which cisplatin exerts its anti-tumor activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 is the hydrogen spectrum of carbazole pyridine ligand L-1;

[0048] Figure 2 is the carbon spectrum of carbazole pyridine ligand L-1;

[0049] Figure 3 is the hydrogen spectrum of carbazole pyridine ligand L-2;

[0050] Figure 4 is the carbon spectrum of carbazole pyridine ligand L-2;

[0051] Figure 5 is the hydrogen spectrum of carbazole pyridine ligand L-3;

[0052] Figure 6 is the carbon spectrum of carbazole pyridine ligand L-3;

[0053] Figure 7 is the hydrogen spectrum of carbazole pyridine ligand L-4;

[0054] Figure 8 This is the carbon spectrum of the carbazole pyridine ligand L-4. DETAILED DESCRIPTION

[0055] The technical solutions of the present invention are further described below in conjunction with specific embodiments. However, the embodiments described below are intended to explain the present invention and are not to be construed as limiting the present invention. If no specific techniques or conditions are specified in the embodiments, the techniques or conditions described in the literature in the art or the product instructions shall be followed.

[0056] Unless otherwise defined, the technical terms used in the following examples have the same meanings as commonly understood by those skilled in the art to which the present invention belongs; the test reagents used are conventional biochemical reagents unless otherwise specified; the experimental methods described are conventional methods unless otherwise specified.

[0057] Example 1

[0058] A carbazole pyridine copper complex Cu-1 has the following structure:

[0059]

[0060] The preparation method is as follows:

[0061] (1) Preparation of carbazole pyridine ligand L-1: 2,6-Dibromopyridine (377 mg, 1.59 mmol), carbazole (586 mg, 3.5 mmol), CuI (61 mg, 0.32 mmol), 18-crown-6 (84 mg, 0.32 mmol), and K2CO3 (549 mg, 3.98 mmol) were added to DMPU (2 mL) at room temperature and stirred at 230 °C for 72 h. After the reaction, the temperature was lowered and 2M HCL (100 mL) was added. The mixture was extracted with DCM (3×125 mL), and the combined organic phases were washed with NH3·H2O (25%, 60 mL) and water (100 mL), dried over anhydrous Na2SO4, and the solvent was removed in vacuo. 503 mg of white carbazole pyridine ligand L-1 was obtained by column chromatography.

[0062] 1 H NMR (400 MHz, CDCl3): δ8.16 (d, J=7.3Hz, 4H), 8.12 (d, J=7.8Hz,1H), 8.05 (d, J=8.2Hz, 4H), 7.65 (d, J=7.6Hz, 2H), 7.44 (t, J=8.6Hz, 6.8Hz, 4H), 7.36 (t, J=7.8Hz, 7.3Hz, 4H), such as Figure 1 shown. 13 C NMR (100 MHz, CDCl3) δ 151.57, 140.48, 139.54, 126.45, 124.61, 120.95, 120.25, 114.99, 112.04, as Figure 2 shown.

[0063] (2) Preparation of carbazole pyridine copper complex Cu-1: 99.0 mg (1.0 mmol) of CuCl was dissolved in 8 mL of dichloromethane under nitrogen protection, and 409.5 mg (1.0 mmol) of carbazole pyridine ligand L-1 was added. The mixture was stirred in a constant temperature oil bath at 20 °C for 12 h. After the reaction, the solvent was dried by spin drying and washed three times with dichloromethane and methanol respectively. The mixture was then dried under vacuum to obtain 397.4 mg of reddish-brown carbazole pyridine copper complex Cu-1.

[0064] The element content tested is: C, 68.50%; H, 3.77%; Cl, ​​6.97%; Cu, 12.50%; N, 8.26%.

[0065] Example 2

[0066] A copper(II) carbazole pyridine complex Cu-2 has the following structure:

[0067]

[0068] The preparation method is as follows:

[0069] (1) using the carbazole pyridine ligand L-1 prepared in Example 1;

[0070] (2) Preparation of carbazole pyridine copper complex Cu-2: 190.5 mg (1.0 mmol) of CuI was dissolved in 8 mL of methanol under nitrogen protection, and 409.5 mg (1.0 mmol) of carbazole pyridine ligand L-1 was added. The mixture was stirred in a constant temperature oil bath at 25 °C for 12 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 349.3 mg of reddish-brown carbazole pyridine copper complex Cu-2.

[0071] The element content tested is: C, 58.06%; H, 3.19%; Cu, 10.59%; I, 21.15%; N, 7.00%.

[0072] Example 3

[0073] A copper(II) carbazole pyridine complex Cu-3 has the following structure:

[0074]

[0075] The preparation method is as follows:

[0076] (1) Preparation of carbazole pyridine ligand L-2: 3,5-Dibromopyridine (377 mg, 1.59 mmol), carbazole (586 mg, 3.5 mmol), CuI (61 mg, 0.32 mmol), 18-crown-6 (84 mg, 0.32 mmol), and K2CO3 (549 mg, 3.98 mmol) were added to DMPU (2 mL) at room temperature, and the mixture was stirred at 230 °C for 48 h. After the reaction, the temperature was lowered and 2M HCL (100 mL) was added. The mixture was extracted with DCM, and the combined organic phases were washed with dilute aqueous ammonia, dried over anhydrous Na2SO4, and the solvent was removed in vacuo. The white carbazole pyridine ligand L-2 was obtained by column chromatography.

[0077] 1 H NMR (400 MHz, CDCl3): δ9.02 (d, J=2.3Hz, 2H), 8.18 (s, 1H), 8.17 (d, J=7.9Hz, 4H), 7.53 (d, J=8.1Hz, 4H), 7.50 - 7.46 (m, 4H), 7.36 (t, J=7.9Hz, 6.9Hz, 4H), such as Figure 3 shown. 13 C NMR (100 MHz, CDCl3) δ 146.60, 140.34, 135.61, 131.96, 126.56, 123.98, 121.06, 120.73, 109.32, Figure 4 shown.

[0078] (2) Preparation of carbazole pyridine copper complex Cu-3: 99.0 mg (1 mmol) of CuCl was dissolved in 6 mL of methanol under nitrogen protection, and 450.5 mg (1.1 mmol) of carbazole pyridine ligand L-2 was added. The mixture was stirred in a 30°C oil bath for 9 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 369.7 mg of reddish-brown pyridine carbazole copper complex Cu-3.

[0079] The element content tested is: C, 68.50%; H, 3.77%; Cl, ​​6.97%; Cu, 12.50%; N, 8.26%.

[0080] Example 4

[0081] A copper-4 carbazole pyridine complex has the following structure:

[0082]

[0083] The preparation method is as follows:

[0084] (1) Preparation of carbazole pyridine ligand L-3: 2,6-dibromo-N,N-dimethylpyridin-4-amine (733 mg, 2.54 mmol), carbazole (833 mg, 4.88 mmol), CuI (91.5 mg, 0.48 mmol), 18-crown-6 (126 mg, 0.48 mmol), and K2CO3 (823.5 mg, 5.97 mmol) were added to DMPU (3 mL) at room temperature, and the mixture was stirred at 240 °C for 72 h. After the reaction, the temperature was lowered and 2M HCl was added. The mixture was extracted with DCM, and the combined organic phases were added with diluted ammonia water, dried over anhydrous Na2SO4, and the solvent was removed in vacuo. The white carbazole pyridine ligand L-3 was obtained by column chromatography.

[0085] 1 H NMR (400 MHz, CDCl3): δ8.11 (d, J=7.5Hz, 4H), 7.96 (d, J=8.1Hz,4H), 7.43 -7.39 (m, 4H), 7.29 (t, J=7.6Hz, 7.3Hz, 4H), 6.83 (s, 2H), 3.12 (s,6H), such as Figure 5 shown. 13 C NMR (100 MHz, CDCl3) δ 157.99, 152.10, 140.03, 126.12,124.19, 120.58, 120.14, 111.84, 99.98, 39.8, e.g. Figure 6 shown.

[0086] (2) Preparation of carbazole pyridine copper complex Cu-4: 99.0 mg (1 mmol) of CuCl was dissolved in 8 mL of dioxane under nitrogen protection, and 543.1 mg (1.2 mmol) of carbazole pyridine ligand L-3 was added. The mixture was stirred in a 40°C oil bath for 7 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 490.7 mg of reddish-brown carbazole pyridine copper complex Cu-4.

[0087] The element content tested is: C, 67.51%; H, 4.39%; Cl, ​​6.43%; Cu, 11.52%; N, 10.16%.

[0088] Example 5

[0089] A copper(II) carbazole pyridine complex Cu-5 has the following structure:

[0090]

[0091] The preparation method is as follows:

[0092] (1) Preparation of carbazole pyridine ligand L-4: 4-Bromopyridine (401.3 mg, 2.54 mmol), carbazole (398.8 mg, 2.385 mmol), CuI (91.5 mg, 0.48 mmol), 18-crown-6 (126 mg, 0.48 mmol), and K2CO3 (823.5 mg, 5.97 mmol) were added to DMPU (3 mL) at room temperature, and the mixture was stirred at 230 °C for 72 h. After the reaction, 2 M HCL was added, and the mixture was extracted with DCM. The organic phases were combined and diluted ammonia was added. The organic phases were combined and dried over anhydrous Na2SO4. The solvent was removed in vacuo, and the mixture was purified by column chromatography to obtain white carbazole pyridine ligand L-4.

[0093] 1 H NMR (400 MHz, CDCl3): δ8.86 (s, 2H), 8.14 (d, J=8.2Hz, 2H), 7.60 (d, J=6.3Hz, 2H), 7.57 (d, J=8.2Hz, 2H), 7.45 (t, J=8.4Hz, 7.0Hz, 2H), 7.34(t, J=7.8Hz, 7.2Hz, 2H), such as Figure 7 shown. 13 C NMR (100 MHz, CDCl3) δ 151.72, 145.56,139.48, 126.44, 124.24, 121.16, 120.61, 109.82, as Figure 8 shown.

[0094] (2) Preparation of carbazole pyridine copper complex Cu-5: 121.6 mg (1 mmol) of CuSCN was dissolved in 8 mL of dichloromethane under nitrogen protection, and 342.0 mg (1.4 mmol) of carbazole pyridine ligand L-4 was added. The mixture was stirred in a 50°C oil bath for 5 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 205.9 mg of a reddish-brown carbazole pyridine copper complex Cu-5.

[0095] The element content tested is: C, 59.08%; H, 3.31%; Cu, 17.37%; N, 11.48%; S, 8.76%.

[0096] Example 6

[0097] A copper(II) carbazole pyridine complex Cu-6 has the following structure:

[0098]

[0099] The preparation method is as follows:

[0100] (1) Preparation of carbazole pyridine ligand L-5: 2,6-dibromo-N,N-dimethylpyridin-4-amine (733 mg, 2.54 mmol), 3,8-di-tert-butylcarbazole (1363.5 mg, 4.88 mmol), CuI (91.5 mg, 0.48 mmol), 18-crown-6 (126 mg, 0.48 mmol), and K2CO3 (823.5 mg, 5.97 mmol) were added to DMPU (3 mL) at room temperature, and the mixture was stirred at 230 °C for 72 h. After the reaction, 2M HCL was added, and the mixture was extracted with DCM. The organic phases were combined and diluted ammonia was added. The organic phases were combined and dried over anhydrous Na2SO4. The solvent was removed in vacuo, and the mixture was purified by column chromatography to obtain white carbazole pyridine ligand L-5.

[0101] 1 H NMR (400 MHz, CDCl3): δ8.12 -7.95 (m, 6H), 7.47-7.25 (m, 6H), 6.87(s, 6H), 3.05(s, 6H), 1.42(s, 12H). 13 C NMR (100 MHz, CDCl3) δ 157.80, 152.01,139.55, 126.15, 124.26, 120.47, 120.01, 111.85, 99.70, 39.87, 34.87, 31.85.

[0102] (2) Preparation of carbazole pyridine copper complex Cu-6: 82.5 mg (1 mmol) of CuF was dissolved in 8 mL of dichloromethane under nitrogen protection, and 947.8 mg (1.4 mmol) of carbazole pyridine ligand L-5 was added. The mixture was stirred in a 50°C oil bath for 5 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 479.1 mg of reddish-brown carbazole pyridine copper complex Cu-6.

[0103] The element content tested is: C, 74.32%; H, 7.43%; Cu, 8.37%; F, 2.50%; N, 7.38%.

[0104] Example 7

[0105] A copper(II) carbazole pyridine complex Cu-7 has the following structure:

[0106]

[0107] The preparation method is as follows:

[0108] (1) Using the carbazole pyridine ligand L-5 prepared in Example 6;

[0109] (2) Preparation of carbazole pyridine copper complex Cu-7: 99.0 mg (1 mmol) of CuCl was dissolved in 8 mL of dichloromethane under nitrogen protection, and 812.4 mg (1.2 mmol) of carbazole pyridine ligand L-5 was added. The mixture was stirred in a 50°C oil bath for 5 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 515.2 mg of reddish-brown carbazole pyridine copper complex Cu-7.

[0110] The element content tested is: C, 72.75%; H, 7.27%; Cl, ​​4.57%; Cu, 8.19%; N, 7.22%.

[0111] Example 8

[0112] A copper(II) carbazole pyridine complex Cu-8 has the following structure:

[0113]

[0114] The preparation method is as follows:

[0115] (1) Using the carbazole pyridine ligand L-5 prepared in Example 6;

[0116] (2) Preparation of carbazole pyridine copper complex Cu-8: 143.5 mg (1 mmol) of CuBr was dissolved in 8 mL of dichloromethane under nitrogen protection, and 744.7 mg (1.1 mmol) of carbazole pyridine ligand L-5 was added. The mixture was stirred in a 50°C oil bath for 5 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 524.8 mg of reddish-brown carbazole pyridine copper complex Cu-8.

[0117] The element content tested is: C, 68.81%; H, 6.88%; Br, 9.74%; Cu, 7.75%; N, 6.83%.

[0118] Example 9

[0119] A copper(II) carbazole pyridine complex Cu-9 has the following structure:

[0120]

[0121] The preparation method is as follows:

[0122] (1) Using the carbazole pyridine ligand L-5 prepared in Example 6;

[0123] (2) Preparation of carbazole pyridine copper complex Cu-8: 190.5 mg (1 mmol) of CuI was dissolved in 8 mL of dichloromethane under nitrogen protection, and 677.0 mg (1.0 mmol) of carbazole pyridine ligand L-5 was added. The mixture was stirred in a 50°C oil bath for 5 h. After the reaction, the solvent was dried and washed three times with dichloromethane and methanol, respectively. The mixture was then dried under vacuum to obtain 801.7 mg of a reddish-brown carbazole pyridine copper complex Cu-9.

[0124] The element content tested is: C, 65.08%; H, 6.51%; Cu, 7.33%; I, 14.63%; N, 6.46%.

[0125] From the different copper carbazole pyridine complexes prepared in the above examples and their structural characterization, it can be seen that the present invention successfully prepared copper carbazole pyridine complexes. The copper complexes of the present invention have a wide reaction temperature range for their preparation process, more optional solvents, and a relatively simple reaction step method.

[0126] Pharmacological activity test

[0127] The method for determining the antitumor activity of different copper carbazole pyridine complexes on A549 cells is as follows:

[0128] A549 cells in the logarithmic growth phase were inoculated into a 96-well plate, and an equal volume of PBS was added to the edge of the plate to prevent edge effects. The cells were placed in an incubator and taken out after culturing for 24 hours. A549 cells were treated with different cuprous complexes at concentrations of 0.125 μM, 0.25 μM, 0.5 μM, 1.0 μM, 2.0 μM, 4.0 μM, 8.0 μM, 16.0 μM, 32.0 μM, and 100 μM and incubated for 48 hours. The control wells were added with DMSO at the same concentration as the drug and incubated for 48 hours. 20 μL of CCK8 reagent at a concentration of 5 mg / mL was added to each well, and the wells were replicated for 3 times before incubation in the incubator for 1 hour. The absorbance at 450 nm was monitored using a microplate reader to calculate the IC values ​​of the cuprous complexes Cu-1 to Cu-9 to be tested. 50 For details, see Table 1.

[0129] Table 1: Half-inhibitory activity of each compound in the examples on A549 cells, i.e. IC50 Value (μM)

[0130] Copper complexes <![CDATA[IC 50 ]]> Copper complexes <![CDATA[IC 50 ]]> Cu-1 9.6 Cu-6 8.9 Cu-2 16.1 Cu-7 1.7 Cu-3 42.3 Cu-8 13.4 Cu-4 7.2 Cu-9 44.5 Cu-5 51.2

[0131] From the examples and test results, it can be seen that when the cuprous carbazole pyridine complexes provided by the present invention are used as anti-tumor drugs, most cuprous complexes have relatively high inhibitory activity against A549 cells, among which Cu-7 has the highest inhibitory activity against A549 cells, IC 50 is 1.7 μM.

[0132] In addition, the inhibitory activity of the carbazole pyridine copper complex Cu-7 on different tumor cells was further tested using the carbazole pyridine copper complex Cu-7 as the object.

[0133] The method for determining the inhibitory activity of the carbazole pyridine copper complex Cu-7 on different tumor cells is as follows: different cells in the logarithmic growth phase are inoculated into a 96-well plate, and an equal volume of PBS is added to the edge of the plate to prevent edge effects. Place the plate in an incubator and take it out after culturing the cells for 24 hours. The cells are treated with copper complex Cu-7 at concentrations of 0.125 μM, 0.25 μM, 0.5 μM, 1.0 μM, 2.0 μM, 4.0 μM, 8.0 μM, 16.0 μM, 32.0 μM, and 100 μM and incubated for 48 hours. The control wells are added with DMSO at the same concentration as the drug and incubated for 48 hours. 20 μL of CCK8 reagent at a concentration of 5 mg / mL is added to each well, and the wells are replicated three times, and then incubated in the incubator for 1 hour. The absorbance at 450 nm is monitored using a microplate reader to calculate the IC value of the cuprous complex Cu-7. 50 .

[0134] In this experiment, for the copper complex Cu-7, the tumor cells are MCF-7, IC 50 is 2.3 μM; HCT116, IC 50 is 1.7 μM; SiHa, IC 50 is 50.4 μM; SK-OV-3, IC 50 is 36.8 μM; SW480, IC 50 is 82.2 μM; A549 / DDP, IC 50 The test results are shown in Table 2:

[0135] Table 2 Half inhibitory activity of compound Cu-7 on other tumor and normal cells, i.e. IC 50 Value (μM)

[0136] Copper complexes cell <![CDATA[IC 50 ]]> Copper complexes cell <![CDATA[IC 50 ]]> Cu-7 MCF-7 5.1 Cu-7 SK-OV-3 43.2 Cu-7 HCT116 3.8 Cu-7 SW480 50.7 Cu-7 SiHa 41.3 Cu-7 A549 / DDP 17.4 Cu-7 EBAS-2B 55.0

[0137] From the examples and performance tests, it can be seen that the carbazole pyridine copper complex Cu-7 has good inhibitory activity against different tumor cells, and also has good inhibitory activity against cisplatin-resistant tumor cells. The IC values ​​of several tumor cell lines tested are 50 The IC values ​​for EBAS-2B in normal cells range from 1.2 μM to 51.2 μM. 50 The selectivity for tumor and normal cells was greater than 32-fold.

[0138] Mechanism research

[0139] The method for verifying the anti-tumor mechanism of the copper(II) carbazole pyridine complex Cu-7 on A549 cells is as follows:

[0140] A549 cells were cultured in a 6-well plate and incubated in a culture incubator for 4 hours. The culture medium was aspirated, and DCFH-DA diluted in serum-free medium was added. The cells were incubated in the dark for 30 minutes. 1 mL of 10 μmol / L diluted DCFH-DA was added to cover the cells, and the cells were incubated in a 37°C cell culture incubator for 20 minutes. The cells were washed three times with serum-free cell culture medium to fully remove the DCFH-DA that had not entered the cells. 1 μL of the reactive oxygen species (ROS) positive control Rosup and various concentrations of the preferred carbazole pyridine ligand cuprous complex Cu-7 were added. After stimulation for 20-30 minutes, the cells were harvested and analyzed using a fluorescence microplate reader and flow cytometry. The results showed that the carbazole pyridine ligand cuprous complex Cu-7 catalyzed an approximately 20% increase in the level of ROS in A549 cells compared to a system without the cuprous complex. This preliminary validation suggests that these cuprous complexes may achieve tumor inhibition by catalyzing the generation of ROS in tumor cells.

[0141] As can be seen from the examples and test results, the cuprous carbazole pyridine complex prepared from the carbazole pyridine ligand provided by the present invention has good inhibitory activity against various tumor cells when used as an anti-tumor agent. Among them, the inhibitory activity against A549 cells is relatively high, and it also has good inhibitory activity against cisplatin-resistant tumor cells.

[0142] As can be seen from the examples and test results, in the present invention, different cuprous complexes prepared have different inhibitory activities depending on the structure of the corresponding carbazole pyridine ligand, the overall electrical properties of the ligand, and the anion. The same cuprous complex also has different inhibitory activities against different types of tumor cells.

[0143] The above description of the embodiments is intended to facilitate understanding and use of the invention by those skilled in the art. It will be apparent that those skilled in the art can readily make various modifications to these embodiments and apply the general principles described herein to other embodiments without requiring inventive effort. Therefore, the present invention is not limited to the above-described embodiments. Improvements and modifications made by those skilled in the art based on the disclosure of the present invention, without departing from the scope of the present invention, should be within the scope of protection of the present invention.

Claims

1. A copper(II) carbazole pyridine complex, characterized in that: The copper carbazole pyridine complex is selected from one of the following compounds: 、 、 、 、 、 、 、 、 。 2. The method for preparing the copper(I) carbazole-pyridine complex according to claim 1, wherein: The following steps are involved: (1) The bromine-substituted pyridine reacts with carbazole and its derivatives to first obtain the carbazole-pyridine ligand; (2) reacting a carbazole pyridine ligand with a monovalent copper salt in a solvent to obtain the carbazole pyridine cuprous complex according to claim 1; The carbazole pyridine ligand is selected from the structure shown in formula (III): Formula (III) The copper(II) carbazole pyridine complex is selected from the structure shown in formula (I): Formula (I) Wherein, R1, R2, R3, R4, R5 and X are the corresponding groups of the copper(II) carbazole pyridine complex in claim 1.

3. The method for preparing the copper(II) carbazole pyridine complex according to claim 2, wherein: The following steps are involved: (1) Preparation of carbazole pyridine ligand: Under the protection of inert gas, pyridine and carbazole substituted with different positions and amounts of bromine are reacted with CuI catalyst in DMPU as solvent for a certain time. After cooling, dilute HCl is added and the mixture is extracted with DCM. The organic layers are combined and washed with dilute ammonia water, dried over anhydrous Na2SO4, and the solvent is removed in vacuo. The carbazole pyridine ligand is obtained by column chromatography. (2) Preparation of cuprous carbazole pyridine ligand complex: a monovalent copper salt reacts with the carbazole pyridine ligand in a solvent to obtain the cuprous carbazole pyridine complex according to claim 1.

4. The method for preparing the copper(I) complex of carbazole pyridine according to claim 3, wherein: The copper salt is selected from the anhydrous form or the crystalline water form of CuCl, CuBr, CuI or CuSCN.

5. The method for preparing the copper(I) complex of carbazole pyridine according to claim 3, wherein: In step (2), the molar ratio of the carbazole pyridine ligand to the copper salt is (1.0-5.0):

1.

6. The method for preparing the copper(I) complex of carbazole pyridine according to claim 3, wherein: In step (2), the reaction temperature is 0-100°C; The reaction time is 10 min-24 h; In step (2), the solvent is selected from one or more of acetonitrile, methanol, ethanol, dichloromethane, tetrahydrofuran or dioxane.

7. Use of the copper(II) carbazole pyridine complex according to claim 1 in the preparation of anti-tumor drugs, characterized in that: The tumor is selected from one or more of A549, MCF-7, HCT 116, SiHa, SK-OV-3 or SW480.

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