Quinoline-functionalized n-heterocyclic carbene ruthenium complexes with multiple anticancer activities and methods of making the same

Abstract

The present application relates to a kind of quinoline functional group nitrogen heterocyclic carbene ruthenium complex with multiple anticancer activity and its preparation method, the complex of the present application is prepared by 2-chloromethyl quinoline hydrochloride as starting compound, by with N-substituted imidazole or benzimidazole salt and metal exchange reaction, structural general formula is [LRu (p-cymene) Cl] (PF6). The present application nitrogen heterocyclic carbene ruthenium complex in lung cancer cell, cervical cancer cell, liver cancer cell, intestinal cancer cell, ovarian cancer cell and breast cancer cell cytotoxicity experiment all show strong cytotoxicity;In intestinal cancer cell, the present application nitrogen heterocyclic carbene ruthenium complex can effectively inhibit cell proliferation and migration;Mechanism research shows that the nitrogen heterocyclic carbene ruthenium complex mainly by interfering with mitochondrial membrane potential, induces cancer cell apoptosis;The present application nitrogen heterocyclic carbene ruthenium complex structure is variable, synthesis is convenient, stable, and has good application prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of organic synthesis and relates to a nitrogen heterocyclic carbene ruthenium complex, in particular to a quinoline functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anti-cancer activities and a preparation method thereof. BACKGROUND

[0002] Metal drugs, such as platinum drugs, are a kind of main anticancer drugs, which are widely used in the treatment of various cancers and prolong the life expectancy of numerous patients. However, the clinical application of platinum drugs is affected by serious toxicity, non-selectivity, intrinsic or acquired drug resistance and other reasons.

[0003] Compared with platinum drugs, ruthenium complexes have the characteristics of low systemic toxicity, high anti-proliferation and anti-metastasis activities, etc., so in recent years they have attracted continuous exploration and made great progress. In addition, the cytotoxic effect of ruthenium complexes involves multiple action modes, has the therapeutic potential of inducing cancer cell death through multiple action modes, and is expected to overcome the problem of drug resistance of cancer cells. As an excellent δ ligand, functionalized nitrogen heterocyclic carbene can be easily derived into various different molecules, making it an ideal ligand for metal drug screening. Quinoline is a group existing in many natural compounds, and its derivatives have a wide range of biological activities, such as anti-tumor and anti-bacterial effects. However, metal drugs based on quinoline groups are rarely used in anti-cancer research. In order to fully combine the characteristics of carbene ligand, ruthenium metal compound and quinoline group, the present application synthesizes a quinoline functionalized nitrogen heterocyclic carbene ruthenium complex, and applies such metal complex to anti-cancer research. SUMMARY

[0004] The present application provides a quinoline functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anti-cancer activities and a preparation method thereof. The complex of the present application can effectively inhibit the proliferation, migration of cancer cells and induce apoptosis of cancer cells. Under the action of the double active centers of quinoline active groups and ruthenium metal ions, the present application can effectively overcome the problem that single drug is easy to produce cancer cell drug resistance, and the multiple active centers can produce multiple anti-cancer mechanisms, which has important significance in the research and development of anti-cancer drugs.

[0005] A quinoline functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anti-cancer activities, the structure general formula of the complex is [LRu(p-cymene)Cl](PF6), wherein L is a quinoline functionalized nitrogen heterocyclic carbene ligand, and p-cymene is p-methyl isopropyl benzene; the molecular structure formula of the complex is:

[0006]

[0007] R is any one of phenyl, p-methylphenyl, p-trifluoromethylphenyl, 2,4,6-trimethylphenyl, 2,3,4,5,6-pentamethylphenyl.

[0008] The application discloses a novel quinoline functionalized azolide ruthenium complex, wherein the substituent on the imidazole ring or the benzimidazole ring is easy to change, easy to synthesize, stable in performance and environment-friendly, and can exist stably in air for a long time.

[0009] As preferred, the molecular structural formula of the quinoline functionalized azolide ligand is as follows:

[0010]

[0011] The application further provides a preparation method of the quinoline functionalized azolide ruthenium complex with multiple anti-cancer activities, which comprises the following steps: taking 2-chloromethyl quinoline hydrochloride as raw material, removing hydrochloride by alkaline hydrolysis, and then reacting with nitrogen-substituted imidazole or nitrogen-substituted benzimidazole to obtain a quinoline functionalized imidazole salt ligand or a quinoline functionalized benzimidazole salt ligand; then taking acetonitrile or acetone as a solvent, adding the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand and silver oxide, and carrying out a light-avoiding reaction, adding [Ru (p-cymene) Cl2]2, and carrying out a room-temperature reaction, and finally carrying out post-treatment to obtain the quinoline functionalized azolide ruthenium complex.

[0012] As preferred, the molar ratio of the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand to silver oxide is 2:1.

[0013] As preferred, the light-avoiding reaction is carried out at a temperature of 50-60 DEG C for 4-6 hours.

[0014] As preferred, the molar ratio of [Ru (p-cymene) Cl2]2 to the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand is 1:2, and the room-temperature reaction is carried out for 2-4 hours.

[0015] As preferred, the post-treatment comprises the following steps: filtering, concentrating the filtrate, separating by chromatography or adding diethyl ether, and recrystallizing to obtain the quinoline functionalized azolide ruthenium complex.

[0016] As preferred, the quinoline functionalized imidazole salt ligand or the benzimidazole salt ligand is prepared by the following steps: stirring 2-chloromethylquinoline hydrochloride 10 mmol, sodium hydroxide 12.5 mmol in 20 mL of acetonitrile solvent at room temperature for 30 minutes, filtering to remove insoluble impurities; adding nitrogen-substituted imidazole or nitrogen-substituted benzimidazole 10 mmol to the filtrate, heating to 90 degrees, continuing to react for 6 hours, salifying; after cooling to room temperature, removing the acetonitrile solvent under reduced pressure, dissolving in 50 mL of deionized water, filtering to remove insoluble impurities; adding ammonium hexafluorophosphate 10 mmol to the filtrate, performing anion exchange, and precipitating white solids; after drying, the quinoline functionalized imidazole salt ligand or the benzimidazole salt ligand is obtained.

[0017] As preferred, the nitrogen-substituted imidazole is one of N-benzyl imidazole, N-(4-methylbenzyl)imidazole, N-(4-trifluoromethylbenzyl)imidazole, N-(2,4,6-trimethylbenzyl)imidazole, and N-(2,3,4,5,6-pentamethylbenzyl)imidazole; the nitrogen-substituted benzimidazole is one of N-benzyl benzimidazole, N-(2,4,6-trimethylbenzyl)benzimidazole, and N-(2,3,4,5,6-pentamethylbenzyl)benzimidazole.

[0018] The present application has the following beneficial effects:

[0019] The carbene ruthenium complexes of the present application exhibit strong cytotoxicity in cytotoxicity experiments of lung cancer cells (A549), cervical cancer cells (HeLa), liver cancer cells (7402), intestinal cancer cells (HCT-116), ovarian cancer cells (ES-2), and breast cancer cells (4T-1); in intestinal cancer cells HCT-116, the carbene ruthenium complexes of the present application can effectively inhibit cell proliferation and migration; mechanism studies show that the carbene ruthenium complexes mainly interfere with the mitochondrial membrane potential to induce cancer cell apoptosis; the carbene ruthenium complexes of the present application have variable structures, are easy to synthesize, stable in performance, environmentally friendly, and have good application prospects; under the action of the dual active centers of the quinoline active group and the ruthenium metal ion, the present application can effectively overcome the problem of cancer cell drug resistance of single drugs, and multiple active centers can produce multiple anticancer mechanisms, which is of great significance in the research and development of anticancer drugs. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a structure diagram of Ru1 cation (hydrogen atoms are omitted);

[0021] Figure 2 is a structure diagram of Ru5 single crystal cation (hydrogen atoms are omitted);

[0022] Figure 3 is a structure diagram of Ru7 single crystal cation (hydrogen atoms are omitted);

[0023] Figure 4 Figure 2 is a graph showing the effect of quinoline functionalized N-heterocyclic carbene ruthenium complexes Ru4, Ru8 and cis-Pt (2 and 4 μΜ) on the proliferation of colon cancer cells HCT-116;

[0024] Figure 5 Figure 3 is a graph showing the effect of quinoline functionalized N-heterocyclic carbene ruthenium complexes Ru4, Ru8 and cis-Pt (4 μΜ) on the migration of colon cancer cells HCT-116;

[0025] Figure 6 Figure 4 is a graph showing the effect of quinoline functionalized N-heterocyclic carbene ruthenium complexes Ru4, Ru8 and cis-Pt (4 μΜ) on the mitochondrial membrane potential of colon cancer cells HCT-116;

[0026] Figure 7 Figure 5 is a graph showing the effect of quinoline functionalized N-heterocyclic carbene ruthenium complexes Ru4, Ru8 and cis-Pt (4 μΜ) on the apoptosis of colon cancer cells HCT-116. DETAILED DESCRIPTION

[0027] The application will be further described in conjunction with the specific examples below, but the scope of the application is not limited thereto. It is obvious to those skilled in the art that any simple change or replacement based on the essential spirit of the application shall fall within the scope of the protection claimed by the application.

[0028] The experimental methods described in the examples are all conventional methods unless otherwise specified; the reagents and materials described are all commercially available unless otherwise specified.

[0029] Example 1

[0030]

[0031] Synthesis of quinoline functionalized imidazole salt ligand HL1PF6. 2- chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. N-benzylimidazole (1.58 g, 10 mmol) was added to the filtrate, which was heated to 90 degrees and reacted for 6 hours to form a salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the solution was dissolved in 50 mL of deionized water, filtered, and the insoluble impurities were removed. Ammonium hexafluorophosphate (1.63 g, 10 mmol) was added to the filtrate for anion exchange, and white solids were precipitated. After drying, HL1PF6 white powder was obtained. 1H NMR (400MHz, DMSO-d6): δ9.48 (s, imidazole C2-H, 1H), 8.48 (d, J=8.0Hz, imidazole CH, 1H), 8.03 (d, J=8.0Hz, imidazole CH, 1H), 7.92-7.76 (m, quinoline CH,4H),7.70-7.58(m,quinoline CH,2H),7.54-7.36(m,phenyl CH,5H),5.83(s,CH2,2H),5.54(s,CH2,2H). 13 CNMR (100MHz, DMSO): δ154.70,147.29,138.11,138.05,135.53,130.72,129.52,129 .24,128.95,128.57,128.52,127.76,127.51,124.47,122.96,120.38,53.95,52.50.

[0032]

[0033] At 50-60°C, quinoline-functionalized imidazolium salt ligand HL1PF6 (445 mg, 1.0 mmol), 10 mL of acetonitrile, and silver oxide (116 mg, 0.5 mmol) were added and reacted for 4 hours. [Ru(p-cymene)Cl2]2 (306 mg, 0.5 mmol) was added and the reaction was continued at room temperature for 2 hours. The reaction was centrifuged and the filtrate was concentrated to 2 mL. Ether was added to precipitate a yellow solid, which was washed twice with ether and dissolved in acetonitrile. Ether was slowly added and recrystallized to obtain 475 mg of a ruthenium complex Ru1 coordinated by a nitrogen heterocyclic carbene ligand, with a yield of 65%. 1H NMR (400 MHz, DMSO-d6): δ 9.52 (d, J = 9.2 Hz, CH, 1H), 8.71 (d, J = 8.4 Hz, CH, 1H), 8.12 (d, J = 8.0 Hz, CH, 1H), 8.00 (t, J = 7.2 Hz, CH, 1H), 7.86 (d, J = 8.4 Hz, CH, 1H), 7.84-7.75 (m, CH, 2H), 7.43-7.31 (m, CH, 5H), 7.27 (d, J = 2.0 Hz, CH, 1H), 6.02 (d, J = 4.8 Hz, CH, 1H), 6.00-5.91 (m, CH+CH2, 2H), 5.73 (d, J = 6.0 Hz, CH, 1H), 5.65-5.56 (m, CH+CH2, 2H), 5.45 (d, J = 15.2 Hz, CH2, 1H), 5.14 (d, J = 15.2 Hz, CH2, 1H), 2.69-2.62 (m, CH(CH3)2, 1H), 2.13 (s, CH3, 3H), 1.03 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.92 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 CNMR (100 MHz, DMSO): δ 174.65 (Ru-C), 160.13, 149.73, 141.42, 137.33, 132.50, 130.90, 129.20, 129.09, 128.49, 128.46, 128.28, 123.78, 123.58, 123.41, 108.67, 103.03, 91.48, 85.43, 85.07, 84.88, 55.69, 53.28, 31.46, 23.00, 22.38, 18.26 ppm.

[0034] Crystallization gave Ru1 crystals, the structure of Ru1 cation is shown as Figure 1 32 H 30 ClF6N4PRu; Fw: 752.09; crystal system: Triclinic space group: P-1; 13.4495(5); 14.5842(7); 19.3263(9); β / deg: 95.224(4); 3309.7(3); Z: 4; D calcd , Mg / m 3 ​:1.509; Refls collected: 43459; Independent reflections, R int: 11650, 0.0755; Goodness-of-fit: 1.071; R1, wR2 [I > 2σ(I)]: 0.0613, 0.1331; R1, wR2 (all data): 0.1004, 0.1584; Largest diff. peak and hole 1.013 and -0.777. Selected bond lengths bond angles are: Ru(l)-C(l) 2.031(6), Ru(l)-C(23) 2.188(6), Ru(l)-N(3) 2.196(4), Ru(l)-C(24) 2.201(6), Ru(l)-C(26) 2.210(6), Ru(l)-C(25) 2.237(6), Ru(l)-C(27) 2.280(6), Ru(l)-C(22) 2.301(6), Ru(l)-Cl(l) 2.4088(15), C(l)-Ru(l)-C(23) 119.1(2), C(l)-Ru(l)-N(3) 85.41(19), C(23)-Ru(l)-N(3) 155.3(2), C(l)-Ru(l)-C(24) 92.9(2), C(23)-Ru(l)-C(24) 37.4(2), N(3)-Ru(l)-C(24) 154.79(19), C(l)-Ru(l)-C(26) 119.9(2).

[0035] Example 2

[0036]

[0037] Synthesis of quinoline functionalized imidazole salt ligand HL2PF6. 2- chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) solvent at room temperature for 30 minutes, filtered to remove insoluble impurities. To the filtrate was added N-(4- methylbenzyl)imidazole (1.72 g, 10 mmol) and heated to 90 degrees and the reaction continued for 6 hours to form the salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure and the residue dissolved in 50 mL of deionized water, filtered to remove insoluble impurities. To the filtrate was added ammonium hexafluorophosphate (1.63 g, 10 mmol) and anion exchange was performed to precipitate a white solid. After drying, HL2PF6was obtained as a white powder. 1HNMR (400 MHz, DMSO-d6): δ 9.46 (s, imidazole C2-H, 1H), 8.48 (d, J = 8.0 Hz, imidazole CH, 1H), 8.03 (d, J = 8.0 Hz, imidazole CH, 1H), 7.89-7.76 (m, quinoline CH, 4H), 7.70-7.57 (m, quinoline CH, 2H), 7.42-7.21 (m, phenyl CH, 4H), 5.82 (s, CH2, 2H), 5.47 (s, CH2, 2H), 3.38 (s, CH3, 3H). 13 C NMR (100 MHz, DMSO): δ 154.71, 147.28, 138.75, 138.05, 137.94, 132.48, 130.71, 130.04, 128.96, 128.71, 128.51, 127.76, 127.51, 124.40, 122.86, 120.38, 53.93, 52.32, 21.19.

[0038]

[0039] The quinoline functionalized imidazole salt ligand HL2PF6(459 mg, 1.0 mmol) was added at 50-60 °C in acetonitrile 10 mL, silver oxide (116 mg, 0.5 mmol) was added and the reaction was continued for 4 hours, then [Ru(p-cymene)Cl2]2(306 mg, 0.5 mmol) was added and the reaction was continued for 2 hours at room temperature, the reaction mixture was centrifuged and filtered, the filtrate was concentrated to 2 mL, ether was added to precipitate a yellow solid, the yellow solid was washed twice with ether, then dissolved in acetonitrile and slowly added to ether to recrystallize, obtaining 454 mg of the ruthenium complex Ru2 coordinated with the carbene ligand, with a yield of 61%.

[0040] 1H NMR (400 MHz, DMSO-d6): δ 9.52 (d, J = 9.2 Hz, CH, 1H), 8.70 (d, J = 8.4 Hz, CH, 1H), 8.12 (d, J = 8.4 Hz, CH, 1H), 8.05 - 7.96 (m, CH, 1H), 7.86 (d, J = 8.4 Hz, CH, 1H), 7.80 (t, J = 7.6 Hz, CH, 1H), 7.76 (d, J = 2.0 Hz, CH, 1H), 7.27 - 7.16 (m, CH + phenyl CH, 5H), 6.03 (d, J = 6.0 Hz, CH, 1H), 5.96 (d, J = 9.2 Hz, CH, 1H), 5.93 (s, CH2, 1H), 5.74 (d, J = 6.0 Hz, CH, 1H), 5.59 (d, J = 4.8 Hz, CH, 1H), 5.55 (s, CH2, 1H), 5.38 (d, J = 15.2 Hz, CH2, 1H), 5.13 (d, J = 15.2 Hz, CH2, 1H), 2.69 - 2.62 (m, CH(CH3)2, 1H), 2.30 (s, CH3, 3H), 2.13 (s, CH3, 3H), 1.02 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.93 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 C NMR (100 MHz, DMSO): δ 174.46 (Ru-C), 160.14, 149.73, 141.41, 137.76, 134.21, 132.51, 130.89, 129.74, 129.21, 129.09, 128.53, 128.28, 123.63, 123.50, 123.40, 108.69, 103.00, 91.54, 85.37, 85.02, 84.90, 55.67, 53.04, 31.47, 22.98, 22.39, 21.18, 18.26 ppm.

[0041] Example 3

[0042]

[0043] Synthesis of the quinoline-functionalized imidazolium salt ligand HL3PF6. 2-Chloromethylquinoline hydrochloride (1.88 g, 10 mmol) and sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and insoluble impurities were removed by filtration. N-(4-trifluoromethylbenzyl)imidazole (2.26 g, 10 mmol) was added to the filtrate, and the mixture was heated to 90°C and allowed to react for 6 hours to form the salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the mixture was dissolved in 50 mL of deionized water and filtered to remove insoluble impurities. Ammonium hexafluorophosphate (1.63 g, 10 mmol) was added to the filtrate for anion exchange, resulting in the precipitation of a white solid. After drying, HL3PF6 was obtained as a white powder. 1 HNMR (400MHz, DMSO-d6): δ9.53 (s, imidazole C2-H, 1H), 8.50 (d, J=8.0Hz, imidazole CH, 1H), 8.04 (d, J=8.0Hz, imidazole CH, 1H), 7.94 (d, J=9.6Hz, quinoline CH,2H),7.88(d,J=8.0Hz,quinoline CH,2H),7.85-7.77(m,quinoline CH,2H),7.71(d,J=8.0Hz,phenyl CH,2H),7.66(t,J=7.2Hz,phenyl CH,2H),5.86(s,CH2,2H),5.68(s,CH2,2H). 13 CNMR (100MHz, DMSO): δ154.65,147.26,140.14,138.36,138.06,130.67,129.46,128.96,128 .51,127.76,127.52,126.47,126.44,126.40,126.36,124.57,123.06,120.40,54.00,51.86.

[0044]

[0045] At 50-60°C, quinoline-functionalized imidazolium salt ligand HL3PF6 (513 mg, 1.0 mmol), 10 mL of acetonitrile, and silver oxide (116 mg, 0.5 mmol) were added and reacted for 4 hours. [Ru(p-cymene)Cl2]2 (306 mg, 0.5 mmol) was added and the reaction was continued at room temperature for 2 hours. The reaction was centrifuged and filtered. The filtrate was concentrated to 2 mL. Ether was added to precipitate a yellow solid. The yellow solid was washed twice with ether and dissolved in acetonitrile. Ether was slowly added and recrystallized to obtain 463 mg of a ruthenium complex Ru3 coordinated by a nitrogen heterocyclic carbene ligand, with a yield of 58%.

[0046] 1 H NMR (400 MHz, DMSO-d6): δ 9.51 (d, J = 9.2 Hz, imidazole CH, 1H), 8.71 (d, J = 8.4 Hz, imidazole CH, 1H), 8.13 (d, J = 8.4 Hz, quinoline CH, 1H), 8.00 (t, J = 8.0 Hz, quinoline CH, 1H), 7.87 (d, J = 8.4 Hz, quinoline CH, 1H), 7.83-7.78 (m, quinoline CH, 2H), 7.74 (d, J = 8.0 Hz, phenyl CH, 2H), 7.50 (d, J = 8.0 Hz, phenyl CH, 2H), 7.33 (d, J = 2.0 Hz, quinoline CH, 1H), 6.04 (d, J = 6.4 Hz, p-cymene CH, 1H), 5.99-5.96 (m, p-cymene CH + CH2, 2H), 5.79-5.72 (m, p-cymene CH + CH2, 2H), 5.64 (d, J = 6.0 Hz, p-cymene CH, 1H), 5.57 (d, J = 15.6 Hz, CH2, 1H), 5.16 (d, J = 15.6 Hz, CH2, 1H), 2.71-2.64 (m, CH(CH3)2, 1H), 2.14 (s, CH3, 3H), 1.04 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.95 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 CNMR (100 MHz, DMSO): δ 175.31 (Ru-C), 160.12, 149.70, 142.20, 141.45, 132.46, 130.90, 129.22, 129.10, 128.30, 125.96, 125.92, 123.88, 123.82, 123.43, 109.06, 103.16, 91.31, 85.51, 85.22, 84.97, 55.74, 52.94, 31.46, 22.98, 22.37, 18.30 ppm.

[0047] Example 4

[0048]

[0049] Synthesis of quinoline functionalized imidazole salt ligand HL4PF6. 2-Chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. To the filtrate, N-(2,4,6-trimethylbenzyl)imidazole (2.00 g, 10 mmol) was added, and the mixture was heated to 90 °C and stirred for 6 hours to form the salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the residue was dissolved in 50 mL of deionized water, filtered, and the insoluble impurities were removed. To the filtrate, ammonium hexafluorophosphate (1.63 g, 10 mmol) was added, and anion exchange was performed to precipitate a white solid. After drying, HL4PF6white powder was obtained. 1 H NMR (400 MHz, DMSO-d6): δ 9.08 (s, imidazole C2-H, 1H), 8.46 (d, J = 8.0 Hz, imidazole CH, 1H), 8.02 (d, J = 8.0 Hz, imidazole CH, 1H), 7.83 (t, J = 1.6 Hz, quinoline CH, 1H), 7.81-7.75 (m, quinoline CH, 2H), 7.69 (s, quinoline CH, 1H), 7.64 (t, J = 7.2 Hz, quinoline CH, 1H), 7.58 (d, J = 8.4 Hz, quinoline CH, 1H), 7.01 (s, phenyl CH, 2H), 5.77 (d, J = 2.0 Hz, CH2, 2H), 5.49 (s, CH2, 2H), 2.33 (s, CH3, 6H), 2.26 (s, CH3, 3H). 13 C NMR (101 MHz, DMSO): δ 154.88, 147.22, 139.12, 138.57, 137.95, 137.53, 130.73, 129.91, 128.75, 128.53, 127.71, 127.44, 127.40, 124.31, 122.75, 120.18, 53.65, 47.55, 21.07, 19.75.

[0050]

[0051] The quinoline functionalized imidazole salt ligand HL4PF6(487 mg, 1 mmol) was added at 50-60 °C in acetonitrile 10 mL, silver oxide (116 mg, 0.5 mmol) and the reaction was continued for 4 h, then [Ru(p-cymene)Cl2]2(306 mg, 0.5 mmol) was added and the reaction was continued at room temperature for 2 h, the reaction mixture was centrifuged and filtered, the filtrate was concentrated to 2 mL, diethyl ether was added to precipitate a yellow solid, the yellow solid was washed twice with diethyl ether, then dissolved in acetonitrile and slowly added to diethyl ether to recrystallize the product, obtaining 463 mg of the ruthenium complex coordinated with the carbene ligand Ru4, with a yield of 60%. 1 H NMR (400 MHz, DMSO-d6): δ 9.46 (d, J = 9.2 Hz, CH, 1H), 8.70 (d, J = 8.0 Hz, CH, 1H), 8.12 (d, J = 8.0 Hz, CH, 1H), 8.04-7.98 (m, CH, 1H), 7.84 (d, J = 8.4 Hz, CH, 1H), 7.81 (d, J = 8.0 Hz, CH, 1H), 7.63 (d, J = 2.0 Hz, CH, 1H), 6.98 (s, CH, 2H), 6.47 (d, J = 2.0 Hz, CH, 1H), 6.13 (d, J = 6.4 Hz, CH, 1H), 6.07 (d, J = 6.4 Hz, CH, 1H), 5.95 (d, J = 15.6 Hz, CH2, 1H), 5.83 (d, J = 15.6 Hz, CH2, 1H), 5.75 (d, J = 6.4 Hz, CH, 2H), 5.19 (d, J = 15.6 Hz, CH2, 1H), 5.12 (d, J = 15.6 Hz, CH2, 1H), 2.75-2.68 (m, CH(CH3)2, 1H), 2.34 (s, CH3, 3H), 2.28-2.23 (m, CH3, 9H), 1.05 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.99 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 C NMR (100 MHz, DMSO): δ 173.26 (Ru-C), 160.00, 149.69, 141.36, 139.03, 138.63, 132.37, 130.76, 129.73, 129.19, 129.13, 128.25, 127.98, 123.37, 122.70, 121.12, 108.12, 105.25, 93.34, 84.69, 83.71, 83.52, 55.78, 48.21, 31.63, 24.41, 21.12, 21.05, 20.01, 18.39 ppm.

[0052] Example 5

[0053]

[0054] Synthesis of quinoline functionalized imidazole salt ligand HL5PF6. 2-Chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. N-(2,3,4,5,6-Pentamethylbenzyl)imidazole (2.28 g, 10 mmol) was added to the filtrate, and the reaction was heated to 90 degrees and continued for 6 hours to form a salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the solution was dissolved in 50 mL of deionized water, filtered, and the insoluble impurities were removed. Ammonium hexafluorophosphate (1.63 g, 10 mmol) was added to the filtrate to exchange anions, and white solids were precipitated. After drying, HL5PF6 white powder was obtained. 1 H NMR (400 MHz, DMSO-d6): δ 8.99 (s, imidazole C2-H, 1H), 8.45 (d, J = 8.0 Hz, imidazole CH, 1H), 8.01 (d, J = 8.0 Hz, imidazole CH, 1H), 7.84 (s, quinoline CH, 1H), 7.82-7.74 (m, quinoline CH, 2H), 7.71 (s, quinoline CH, 1H), 7.65 (d, J = 8.0 Hz, quinoline CH, 1H), 7.57 (d, J = 8.4 Hz, quinoline CH, 1H), 5.77 (s, CH2, 2H), 5.58 (s, CH2, 2H), 2.28 (s, CH3, 6H), 2.22-2.18 (m, CH3, 9H). 13 C NMR (101 MHz, DMSO): δ 154.38, 147.09, 143.06, 138.01, 136.89, 134.37, 133.51, 132.28, 131.91, 130.70, 128.58, 128.53, 127.72, 127.45, 127.09, 126.29, 120.37, 114.45, 114.19, 51.14, 46.81, 17.46, 17.20, 16.88.

[0055]

[0056] At 50-60°C, quinoline-functionalized imidazolium salt ligand HL5PF6 (515 mg, 1.0 mmol), 10 mL of acetonitrile, and silver oxide (116 mg, 0.5 mmol) were added and reacted for 4 hours. Then, [Ru(p-cymene)Cl2]2 (306 mg, 0.5 mmol) was added and the reaction was continued at room temperature for 2 hours. The reaction was centrifuged and the filtrate was concentrated to 2 mL. Ether was added to precipitate a yellow solid, which was washed twice with ether and dissolved in acetonitrile. Ether was slowly added and recrystallized to obtain 520 mg of a ruthenium complex Ru5 coordinated by a nitrogen heterocyclic carbene ligand, with a yield of 65%. 1 H NMR (400MHz, DMSO-d6): δ9.46(d,J=9.2Hz,CH,1H),8.70(d,J=8.4Hz,CH,1H),8.13(d,J=8.4Hz,CH,1H),8.02(t,J=8.0Hz,CH,1H),7.84(d,J=8.4H z,CH,1H),7.81(t,J=7.6Hz,CH,1H),7.61(d,J=2.0Hz,CH,1H),6.48(d,J=2.0Hz,CH,1H),6.15(d,J=7.2Hz,CH,1H),6.09(d,J=7.2Hz,CH,1H),5.95 (d,J=15.6Hz,CH2,1H),5.88(d,J=15.6Hz,CH2,1H),5.75(d,J=6.4Hz,CH,2H),5.23(d,J=15.6Hz,CH2,1H),5.12(d,J=15.6Hz,CH2,1H),2.74-2.67 (m,CH(CH3)2,1H),2.33(s,CH3,3H),2.24(s,CH3,3H),2.22-2.04(m,CH3,12H),1.04(d,J=6.8Hz,CH(CH3)2,3H),1.00(d,J=6.8Hz,CH(CH3)2,3H). 13 CNMR(100MHz,DMSO): δ172.93(Ru-C),159.99,149.71,141.36,135.98,134.50,133.19,132.37,130.78,129.20,129.14,128.24,127.99, 123.37,122.50,121.60,107.79,105.18,93.29,84.76,83.89,83.61,55.80,49.51,31.67,24.39,21.14,18.37,17.36,17.10,17.07ppm.

[0057] Crystallization gave Ru5 crystals with the cationic structure shown below and crystallographic data of Formula: C Figure 2 H 37 H 40 ClF6N4PRu; Fw: 822.22; crystal system: Triclinic; space group: P-1; 11.4285(6); 12.9636(5); 13.9466(7); β / deg: 99.786(4); 1866.52(15); Z: 2; D calcd ,Mg / m 3 : 1.463; Refls collected: 14146; Independent reflections, R int: 6579, 0.0235; Goodness-of-fit: 1.046; R1, wR2 [I > 2σ(I)]: 0.0481, 0.1295; R1, wR2 (all data): 0.0572, 0.1377; Largest diff. peak and hole 1.553, -0.907. Selected bond lengths bond angles are: Ru(l)-C(l) 2.047(4), Ru(l)-N(l) 2.186(3), Ru(l)-C(32) 2.189(4), Ru(l)-C(31) 2.209(4), Ru(l)-C(27) 2.228(4), Ru(l)-C(28) 2.248(4), Ru(l)-C(30) 2.252(4), Ru(l)-C(29) 2.285(4), Ru(l)-Cl(l) 2.4028(11), C(l)-Ru(l)-N(l) 85.31(14), C(l)-Ru(l)-C(32) 91.04(15), N(l)-Ru(l)-C(32) 147.44(15), C(l)-Ru(l)-C(31) 97.42(16), N(l)-Ru(l)-C(31) 110.41(14), C(32)-Ru(l)-C(31) 38.01(16), C(l)-Ru(l)-C(27) 111.95(16), N(l)-Ru(l)-C(27) 162.72(15).

[0058] Example 6

[0059]

[0060] Synthesis of quinoline functionalized imidazole salt ligand HL6PF6. 2-Chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. To the filtrate was added N-benzylbenzoimidazole (2.08 g, 10 mmol), and the mixture was heated to 90 °C and stirred for 6 hours to form the salt. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the residue was dissolved in 50 mL of deionized water, filtered, and the insoluble impurities were removed. To the filtrate was added ammonium hexafluorophosphate (1.63 g, 10 mmol) to perform anion exchange, and a white solid was precipitated. After drying, HL6PF6 was obtained as a white powder. 1 HNMR (400 MHz, DMSO-d6): δ 10.19 (s, benzoimidazole C2-H, 1H), 8.51 (d, J = 8.4 Hz, benzoimidazole CH, 1H), 8.06-7.99 (m, benzoimidazole CH + quinoline CH, 3H), 7.81-7.72 (m, benzoimidazole CH + quinoline CH, 3H), 7.68-7.60 (m, benzoimidazole CH + quinoline CH, 3H), 7.58 (d, J = 7.2 Hz, phenyl CH, 2H), 7.50-7.47 (m, phenyl CH, 2H), 7.44-7.42 (m, phenyl CH, 1H), 6.21 (s, CH2, 2H), 5.92 (s, CH2, 2H). 13 CNMR (100 MHz, DMSO): δ 154.22, 147.23, 144.45, 138.14, 134.67, 132.19, 131.27, 130.74, 129.54, 129.27, 128.86, 128.58, 128.53, 127.81, 127.53, 127.35, 127.20, 120.63, 114.43, 114.36, 51.60, 50.36.

[0061]

[0062] Quinoline functionalized benzimidazole salt ligand HL6PF6(495 mg, 1.0 mmol), acetonitrile 10 mL, silver oxide (116 mg, 0.5 mmol) at 50-60 °C, reaction for 4 h, then add [Ru(p-cymene)Cl2]2(306 mg, 0.5 mmol) continue to react at room temperature for 2 h, centrifugal filtration, the filtrate was concentrated to 2 mL, add ether to precipitate yellow solid, the yellow solid was washed with ether for 2 times, then dissolved with acetonitrile, slowly add ether, recrystallization to get 406 mg of the ruthenium complex coordinated with the N-heterocyclic carbene ligand Ru6, yield 52%. 1 H NMR (400 MHz, DMSO-d6): δ 9.55 (d, J = 9.2 Hz, CH, 1H), 8.78 (d, J = 8.4 Hz, CH, 1H), 8.23 (d, J = 8.4 Hz, CH, 1H), 8.19 (d, J = 8.4 Hz, CH, 1H), 8.13 (d, J = 8.0 Hz, CH, 1H), 8.02 (t, J = 8.0 Hz, CH, 1H), 7.82 (t, J = 8.0 Hz, CH, 1H), 7.46 (t, J = 8.0 Hz, CH, 1H), 7.39-7.34 (m, CH, 3H), 7.30 7.22 (m, CH, 3H), 7.15 (d, J = 8.0 Hz, CH, 1H), 6.55 (d, J = 16.0 Hz, CH2, 1H), 6.20 (d, J = 6.4 Hz, CH, 1H), 6.11 (d, J = 6.4 Hz, CH, 1H), 5.92 (s, CH2, 2H), 5.82 (d, J = 6.4 Hz, CH, 1H), 5.62 (d, J = 6.4 Hz, pCH, 1H), 5.25 (d, J = 16.0 Hz, CH2, 1H), 2.68-2.61 (m, CH(CH3)2, 1H), 2.05 (s, CH3, 3H), 1.07 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.81 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 C NMR (100 MHz, DMSO): δ 191.04 (Ru-C), 160.00, 149.88, 141.68, 136.96, 134.96, 134.52, 132.40, 131.17, 129.29, 129.21, 128.38, 128.15, 127.06, 123.98, 123.93, 123.52, 112.35, 111.37, 109.44, 102.08, 91.57, 87.08, 86.46, 86.33, 65.40, 52.35, 51.91, 31.48, 23.24, 21.94, 18.28, 15.64 ppm.

[0063] Example 7

[0064]

[0065] Synthesis of quinoline functionalized imidazole salt ligand HL7PF6. 2- chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. To the filtrate was added N-(2,4,6- trimethylbenzyl)benzoimidazole (2.50 g, 10 mmol), and the mixture was heated to 90 °C and stirred for 6 hours. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the residue was dissolved in 50 mL of deionized water and filtered to remove insoluble impurities. To the filtrate was added ammonium hexafluorophosphate (1.63 g, 10 mmol), and the mixture was stirred for 30 minutes. The white solid was collected by filtration and dried to give HL7PF6 as a white powder. 1 H NMR (400 MHz, DMSO-d6): δ 9.43 (s, benzoimidazole C2-H, 1H), 8.47 (d, J = 8.4 Hz, benzoimidazole CH, 1H), 8.18 (d, J = 8.4 Hz, benzoimidazole CH, 1H), 8.02-7.96 (m, quinoline CH, 2H), 7.76-7.71 (m, benzoimidazole CH + quinoline CH, 3H), 7.68-7.59 (m, benzoimidazole CH + quinoline CH, 3H), 7.08 (s, phenyl CH, 2H), 6.11 (s, CH2, 2H), 5.79 (s, CH2, 2H), 2.36 (s, CH3, 6H), 2.30 (s, CH3, 3H). 13 CNMR (101 MHz, DMSO): δ 154.32, 147.10, 143.18, 139.39, 138.89, 138.01, 132.21, 131.93, 130.73, 130.08, 128.59, 128.51, 127.73, 127.44, 127.12, 126.31, 120.38, 114.28, 114.21, 51.15, 45.59, 21.12, 19.78.

[0066]

[0067] Quinoline functionalized benzimidazole salt ligand HL7PF6(537 mg, 1.0 mmol) was added at 50-60 °C, acetonitrile 10 mL, silver oxide 116 mg (0.5 mmol), reaction for 4 hours, then add [Ru(p-cymene)Cl2] 2306 mg (0.5 mmol) continue to react at room temperature for 2 hours, centrifugal filtration, the filtrate was concentrated to 2 mL, add ether to precipitate yellow solid, the yellow solid was washed with ether twice, then dissolved with acetonitrile, slowly add ether, recrystallization to obtain 559 mg of nitrogen heterocyclic carbene ligand coordinated ruthenium complex Ru7, yield 68%. 1 H NMR (400 MHz, DMSO-d6): δ 9.51 (d, J = 9.2 Hz, CH, 1H), 8.77 (d, J = 8.4 Hz, CH, 1H), 8.19 (d, J = 8.4 Hz, CH, 2H), 8.14 (d, J = 8.0 Hz, CH, 1H), 8.02 (t, J = 8.0 Hz, CH, 1H), 7.82 (t, J = 8.0 Hz, CH, 1H), 7.38 (t, J = 8.0 Hz, CH, 1H), 7.11-7.03 (m, CH, 2H), 6.77 (s, CH, 1H), 6.52 (d, J = 16.0 Hz, CH2, 1H), 6.29-6.24 (m, CH+CH2, 3H), 6.20 (d, J = 6.0 Hz, CH, 1H), 6.04 (d, J = 6.4 Hz, CH, 1H), 5.94 (d, J = 6.4 Hz, CH, 1H), 5.74 (d, J = 14.8 Hz, CH2, 1H), 5.20 (d, J = 16.0 Hz, CH2, 1H), 2.81-2.74 (m, CH(CH3)2, 1H), 2.27 (s, CH3, 3H), 2.16 (s, CH3, 3H), 1.67 (s, CH3, 3H), 1.10 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.99 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 C NMR (100 MHz, DMSO): δ 189.04 (Ru-C), 159.72, 149.77, 141.69, 138.77, 138.07, 138.03, 135.35, 134.34, 132.43, 130.96, 130.19, 129.86, 129.27, 128.38, 128.28, 123.82, 123.61, 123.47, 111.38, 111.31, 110.48, 102.66, 92.06, 86.82, 86.42, 85.50, 52.37, 49.58, 31.50, 22.79, 22.43, 21.03, 19.93, 18.35 ppm.

[0068] Crystallization gave Ru7 crystals with the cationic structure as shown below, and crystallographic data as follows: Figure 3 37 H 35 ClF6N3PRu; Fw: 803.17; crystal system: Triclinic; space group: P-l; 9.8512(5); 11.5915(5); 16.7355(8); β / deg: 74.237(4); 1764.51(14); Z: 2; D calcd ,Mg / m 3 : 1.512; Refls collected: 23377; Independent reflections, R int: 6208, 0.0349; Goodness-of-fit: 1.047; R1, wR2 [I > 2σ(I)]: 0.0434, 0.1113; R1, wR2 (all data): 0.0518, 0.1198; Largest diff. peak and hole ​1.272, -0.728. The partial bond lengths bond angles are: Ru(l)-C(l) 2.031(4), Ru(l)-N(3) 2.179(3), Ru(l)-C(30) 2.183(4), Ru(l)-C(33) 2.188(4), Ru(l)-C(31) 2.194(4), Ru(l)-C(32) 2.230(4), Ru(l)-C(34) 2.278(4), Ru(l)-C(29) 2.294(4), Ru(l)-Cl(l) 2.3907(11), C(l)-Ru(l)-N(3) 86.66(13), C(l)-Ru(l)-C(30) 118.55(15), N(3)-Ru(l)-C(30) 154.61(14), C(l)-Ru(l)-C(33) 118.84(15), N(3)-Ru(l)-C(33) 91.49(14), C(30)-Ru(l)-C(33) 79.24(16), C(l)-Ru(l)-C(31) 92.34(14), N(3)-Ru(l)-C(31) 154.84(14), C(30)-Ru(l)-C(31) 37.61(15), C(33)-Ru(l)-C(31) 67.04(15), C(l)-Ru(l)-C(32) 92.16(14), N(3)-Ru(l)-C(32) 117.44(13).

[0069] Example 8

[0070]

[0071] Synthesis of quinoline functionalized imidazole salt ligand HL8PF6. 2- chloromethylquinoline hydrochloride (1.88 g, 10 mmol), sodium hydroxide (500 mg, 12.5 mmol) were stirred in acetonitrile (20 mL) at room temperature for 30 minutes, and the insoluble impurities were removed by filtration. To the filtrate was added N-(2,3,4,5,6-pentamethylbenzyl)benzimidazole (2.78 g, 10 mmol), and the mixture was heated to 90 °C and stirred for 6 hours. After cooling to room temperature, the acetonitrile solvent was removed under reduced pressure, and the residue was dissolved in 50 mL of deionized water and filtered to remove insoluble impurities. To the filtrate was added ammonium hexafluorophosphate (1.63 g, 10 mmol), and the mixture was stirred for 30 minutes. The white solid was collected by filtration and dried to give HL8PF6 as a white powder. 1H NMR (400 MHz, DMSO-d6): δ 9.30 (s, benzoimidazole C2-H, 1H), 8.45 (d, J = 8.4 Hz, benzoimidazole CH, 1H), 8.28 (d, J = 8.4 Hz, benzoimidazole CH, 1H), 7.98 (d, J = 8.0 Hz, quinoline CH 2H), 7.74 (d, J = 8.0 Hz, quinoline CH 2H), 7.68 (d, J = 8.4, quinoline CH, 2H), 7.63-7.58 (m, benzoimidazole CH, 2H), 6.07 (s, CH2, 2H), 5.83 (s, CH2, 2H), 2.29 (s, CH3, 6H), 2.25 (d, J = 2.7 Hz, CH3, 9H). 13 CNMR (101 MHz, DMSO): δ 154.88, 147.21, 137.95, 137.31, 136.60, 134.01, 133.38, 130.69, 128.75, 128.53, 127.70, 127.44, 127.26, 124.21, 122.72, 120.17, 53.66, 48.78, 17.39, 17.13, 16.80.

[0072]

[0073] The quinoline functionalized benzoimidazole salt ligand HL8PF6(565 mg, 1.0 mmol), acetonitrile 10 mL, silver oxide 116 mg (0.5 mmol) was added at 50-60 °C temperature, reaction for 4 hours, then [Ru(p-cymene)Cl2] 2306 mg (0.5 mmol) was added and continued to react at room temperature for 2 hours, centrifugal filtration, the filtrate was concentrated to 2 mL, added ether to precipitate yellow solid, the yellow solid was washed with ether for 2 times, then dissolved with acetonitrile, slowly added ether, recrystallized to obtain 561 mg of ruthenium complex coordinated with azacyclic carbene ligand Ru8, yield 66%. 11H NMR (400 MHz, DMSO-d6): δ 9.51 (d, J = 9.2 Hz, CH, 1H), 8.77 (d, J = 8.4 Hz, CH, 1H), 8.19 (d, J = 8.4 Hz, CH, 2H), 8.14 (d, J = 8.0 Hz, CH, 1H), 8.07 - 8.01 (m, CH, 1H), 7.82 (t, J = 8.0 Hz, CH, 1H), 7.33 (t, J = 8.0 Hz, CH, 1H), 7.00 (t, J = 8.0 Hz, CH, 1H), 6.51 (d, J = 16.0 Hz, CH2, 1H), 6.35 (d, J = 15.2 Hz, CH2, 1H), 6.27 - 6.25 (m, CH, 2H), 6.08 (d, J = 8.4 Hz, CH, 1H), 6.02 (d, J = 6.4 Hz, CH, 1H), 5.95 (d, J = 7.6 Hz, CH, 1H), 5.70 (d, J = 15.2 Hz, CH2, 1H), 5.20 (d, J = 16.0 Hz, CH2, 1H), 2.80 - 2.73 (m, CH(CH3)2, 1H), 2.43 (s, CH3, 3H), 2.30 (s, CH3, 3H), 2.28 (s, CH3, 3H), 2.21 (s, CH3, 3H), 2.08 (s, CH3, 3H), 1.81 (s, CH3, 3H), 1.09 (d, J = 6.8 Hz, CH(CH3)2, 3H), 0.99 (d, J = 6.8 Hz, CH(CH3)2, 3H). 13 13C NMR (100 MHz, DMSO): δ 189.10 (Ru-C), 159.70, 149.78, 141.67, 135.99, 135.44, 135.05, 134.33, 133.18, 133.07, 132.36, 130.95, 129.28, 128.34, 128.25, 123.84, 123.46, 123.41, 111.55, 111.23, 109.56, 103.23, 92.52, 86.56, 86.27, 85.25, 52.38, 51.14, 31.65, 23.18, 22.16, 18.37, 17.66, 17.49, 17.33, 17.15, 17.10, 16.86, 16.79 ppm.

[0074] Example 9

[0075] The quinoline-functionalized azolyl carbene ruthenium complexes (Ru1-Ru8) obtained in the examples were used to illustrate the anticancer bioactivity of the azolyl carbene ruthenium complexes of the present application by in vitro cytotoxicity experiments: cancer cells (A549, HeLa, 7402, HCT-116, ES-2, and 4T-1) in the growth phase were trypsinized, and then seeded (96-well plate, 5x10 3 cells / well) and incubated at 37°C for 24 h to allow the cells to adhere. Different concentrations of the quinoline-functionalized azolyl carbene ruthenium complexes prepared in the examples were added (100 μL / well), and the cells were incubated for 48 h. Then, 30 μL of thiazolyl blue (MTT, 5 mg / mL) was added to each well, and the cells were incubated for another 4 h. The supernatant was removed, 100 μL of dimethyl sulfoxide (DMSO) was added to each well, and the cells were shaken to dissolve the crystal product. The absorbance of each well was measured at 490 nm using a microplate reader. The cell survival curves were plotted, and the half-inhibitory concentration (IC 50 ) values of the nanoparticles for the cells were calculated. The in vitro toxicity of the quinoline-functionalized azolyl carbene ruthenium complexes to various tumor cells is shown in Table 1.

[0076] Table 1: IC 50 (μM)

[0077] A549 7402 HCT-116 ES-2 4T-1 Ru1 74.2±5.2 28.0±1.4 29.8±1.3 48.5±3.0 46.9±1.8 Ru2 18.2±1.1 10.4±0.5 14.3±0.6 14.7±0.4 11.3±11.3 Ru3 9.9±0.4 4.1±0.2 7.4±0.2 9.1±0.32 14.4±0.5 Ru4 7.6±0.4 3.0±0.2 4.3±0.2 4.5±0.2 6.9±0.2 Ru5 3.96±0.2 2.2±0.1 2.3±0.1 .2±0.1 2.5±0.1 Ru6 10.8±0.7 5.7±0.3 9.5±0.3 13.7±0.3 11.7±0.5 Ru7 2.4±0.1 2.6±0.1 2.2±0.1 2.9±0.1 2.8±0.1 Ru8 1.4±0.1 1.7±0.1 1.0±0.1 1.3±0.1 1.2±0.1

[0078] As can be seen from Table 1, Ru2-Ru8 all showed good activity in different cancer cell lines. Among the Ru1-Ru5 compounds synthesized based on imidazole ligands, the Ru5 compound modified with a pentamethylbenzyl group showed the best activity, with an IC 50 value of 3.96 ± 0.2 μM in the lung cancer cell line A549. The Ru4 compound modified with a trimethylbenzyl group and the Ru3 compound modified with a trifluoromethylbenzyl group were second, with IC 50 values of 7.6 ± 0.4 μM and 9.9 ± 0.4 μM, respectively. The Ru2 compound modified with a 4-methylbenzyl group showed moderate cytotoxicity in the A549 line, with an IC 50 value of 18.2 ± 1.1 μM, while the Ru1 compound modified with a benzyl group showed lower cytotoxicity in the A549 line, with an IC 50 value of 74.2 ± 5.2. The activity order of the Ru1-Ru5 compounds was Ru5 > Ru4 > Ru3 > Ru2 > Ru1. The Ru6-Ru8 compounds synthesized based on benzimidazole showed better anticancer activity, and the Ru8 compound showed the highest activity, with an IC 50 value of only 1.4 ± 0.1 μM in the A549 line and superior anticancer activity in other cell lines.

[0079] Example 10

[0080] Colon cancer cell line HCT-116 was placed in a 48-well plate (2 × 10 5 Then, cisplatin, Ru4, and Ru8 at concentrations of 2 μM and 4 μM were added to the cells, and a blank (untreated) control was added and incubated at 37°C for 24 hours. At the end of the drug treatment, DNA synthesis was quantified using the Click iT EdU Alexa Fluor488 detection kit (Invitrogen). Finally, the total number of cells was counted in 5 areas with a total number of 1500-2000 using a fluorescence microscope (Olympus, IX72, Japan) to evaluate the presence of cell proliferation. The results are shown in Figure 2. Figure 4 As shown, the average proliferation rate of untreated HCT-116 cells (light-colored cells) was 37%. After treatment with 2 μM cisplatin, Ru4, and Ru8, the average proliferation rates of HCT-116 cells were 34%, 25%, and 10%, respectively. After treatment with 4 μM cisplatin, Ru4, and Ru8, the average proliferation rates of HCT-116 cells decreased to 30%, 15%, and 5%, respectively. After treatment with the quinoline-functionalized nitrogen heterocyclic carbene ruthenium complexes Ru4 or Ru8, the average proliferation rate of HCT-116 cells was significantly lower than that of cells treated with the same concentration of cisplatin (cis-Pt).

[0081] Example 11

[0082] Colon cancer cell HCT-116 cells were seeded into 6-well plates and incubated at 37°C for 24 hours to reach 90% coverage. A line was formed on the cell monolayer using a pipette tip (200 μL). After washing with PBS, the cells were cultured in serum-free medium containing Ru4, Ru8 and cisplatin (4 μM). Scratch images were acquired by optical microscopy (Olympus, IX72, Japan) at 0 and 24 hours of incubation. Migration rate (%) = change in width value of each group / average width of initial wound × 100%. Figure 5 As shown, only slight wound closure was observed in Ru3- and Ru4-treated samples compared to untreated and cisplatin-treated cells. Analysis revealed that the average cell migration rates in untreated and cisplatin-treated (cis-Pt) cells were 33% and 27%, respectively, while wound closure rates in Ru4- and Ru8-treated cells were significantly reduced to 17% and 11%, respectively. This result clearly demonstrates that the quinoline-functionalized nitrogen heterocyclic carbene ruthenium complexes Ru4 and Ru8 can effectively inhibit HCT-116 cell migration.

[0083] Example 12

[0084] Colon cancer HCT-116 cells were seeded in 6-well plates (2×10 5cells / well) and incubated overnight. After treatment with Ru4, Ru8 or cisplatin (4 μM) for 12 hours, the cells were incubated with 5 μM JC-1 dye for 30 minutes. After incubation, the cells were washed twice with preheated PBS, harvested, and analyzed by flow cytometry. JC-1 fluorescence data were recorded and analyzed using CytExpert software. Parallel batches of treated cells were stained with JC-1 and washed with PBS before observation under a fluorescence microscope (Olympus, IX72, Japan). Figure 6 As shown, JC-1 dye accumulates in the mitochondrial matrix of untreated HCT-116 cells and exhibits orange fluorescence. After treatment with Ru4 or Ru8 (4 μM) for 12 hours, the orange fluorescence of HCT-116 cells gradually weakened, and green fluorescence emerged. This result demonstrates that the quinoline-functionalized nitrogen heterocyclic carbene ruthenium complexes Ru4 and Ru8 effectively depolarize the mitochondrial membrane potential of HCT-116 cells and reduce the accumulation of JC-1 dye in mitochondria. Compared with Ru4 and Ru8, cisplatin (cis-Pt) has a negligible effect on the mitochondrial membrane potential of cells.

[0085] Example 13

[0086] Colon cancer cells HCT-116 were seeded in 6-well plates (1×10 6 cells / well) and grown overnight at 37°C. The cells were then incubated with Ru4, Ru8, and cisplatin (4 μM) for another 24 hours. Untreated cells were used as controls. After drug treatment, the cells were centrifuged and washed repeatedly with cold PBS, and the cells (1×10 5 ) Cell apoptosis was tested using the Alexa Fluor 488 annexin V / PI kit and analyzed using CytExpert software. Quinoline-functionalized nitrogen heterocyclic carbene ruthenium complexes Ru4, Ru8 and cisplatin (cis-Pt) (4 μM) induced apoptosis in colon cancer cells HCT-116. Figure 7 As shown, flow cytometric analysis showed ( Figure 7 a and 7b), after 24 hours of treatment of HCT-116 cells with Ru4 and Ru8 (4 μM), the apoptotic cells increased to 24.0% and 34.0%, respectively. The proportion of apoptotic cells was significantly higher than that of HCT-116 cells treated with cisplatin (cis-Pt) at the same concentration. Subsequently, in western blot tests, the expression levels of pro-apoptotic proteins PARP, c-PARP, casepase-3, and casepase-9 in cells treated with Ru4 and Ru8 were significantly increased compared with cisplatin and untreated cells. The above results indicate that quinoline-functionalized nitrogen heterocyclic carbene ruthenium complexes Ru4 and Ru8 have a strong ability to induce apoptosis in colon cancer cells HCT-116.

[0087] The present application complex takes 2-chloromethyl quinoline hydrochloride as a starting compound, is prepared by salification with N-substituted imidazole or benzimidazole and metal exchange reaction, and the structural general formula is [LRu(p-cymene)Cl](PF6)(L=quinoline functionalized nitrogen heterocyclic carbene ligand, and p-cymene is aryl p-methyl isopropyl benzene). The nitrogen heterocyclic carbene ruthenium complex shows strong cytotoxicity in the cytotoxicity experiment of lung cancer cells (A549), cervical cancer cells (HeLa), liver cancer cells (7402), intestinal cancer cells (HCT-116), ovarian cancer cells (ES-2), and breast cancer cells (4T-1). In the intestinal cancer cells HCT-116, the nitrogen heterocyclic carbene ruthenium complex can effectively inhibit cell proliferation and migration. Mechanism research shows that the nitrogen heterocyclic carbene ruthenium complex mainly interferes with the mitochondrial membrane potential to induce cancer cell apoptosis. The nitrogen heterocyclic carbene ruthenium complex has good application prospect with variable structure, convenient synthesis, stable performance and environmental friendliness.

Claims

1. A quinoline-functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anticancer activities, characterized in that: The structural general formula of the complex is [LRu(p-cymene)Cl](PF6), wherein L is a quinoline functionalized nitrogen heterocyclic carbene ligand, and p-cymene is para-methyl isopropyl benzene; the molecular structural formula of the complex is: or ; R is any one of a phenyl group, a para-methyl phenyl group, a para-trifluoromethyl phenyl group, a 2,4,6-trimethyl phenyl group, and a 2,3,4,5,6-pentamethyl phenyl group.

2. The quinoline-functionalized nitrogen-heterocyclic carbene ruthenium complex with multiple anticancer activities according to claim 1, characterized in that: The molecular structural formula of the quinoline functionalized nitrogen heterocyclic carbene ligand is: or .

3. A method for preparing the quinoline-functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anticancer activities according to claim 1, characterized in that: 2-Chloromethyl quinoline hydrochloride is used as a raw material, and then the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand is obtained by alkaline hydrochloride removal and reaction with nitrogen-substituted imidazole or nitrogen-substituted benzimidazole; then, acetonitrile or acetone is used as a solvent, the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand and silver oxide are added, and the reaction is carried out in the dark; then, p-cymene chlororuthenate [Ru(p-cymene)Cl2]2 is added, and the reaction is carried out at room temperature; and finally, the quinoline functionalized nitrogen heterocyclic carbene ruthenium complex is obtained through post-treatment.

4. The method of preparing quinoline-functionalized N-heterocyclic carbene ruthenium complexes having multiple anticancer activities according to claim 3, characterized in that: The molar ratio of the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand to silver oxide is 2:

1.

5. The method of preparing quinoline-functionalized N-heterocyclic carbene ruthenium complexes having multiple anticancer activities according to claim 3, characterized in that: The temperature of the reaction in the dark is 50-60 DEG C, and the reaction time in the dark is 4-6 hours.

6. The method of preparing quinoline-functionalized N-heterocyclic carbene ruthenium complexes having multiple anticancer activities according to claim 3, characterized in that: The molar ratio of [Ru(p-cymene)Cl2]2 to the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand is 1:2, and the reaction is carried out at room temperature for 2-4 hours.

7. The method of preparing quinoline-functionalized N-heterocyclic carbene ruthenium complexes having multiple anticancer activities according to claim 3, characterized in that The post-treatment is filtration, concentration of the filtrate, chromatographic separation or addition of ether, and recrystallization to obtain the quinoline functionalized nitrogen heterocyclic carbene ruthenium complex.

8. The method of preparing quinoline-functionalized N-heterocyclic carbene ruthenium complexes having multiple anticancer activities according to claim 3, characterized in that The quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand is prepared by the following steps: 2-chloromethyl quinoline hydrochloride 10 mmol and sodium hydroxide 12.5 mmol are stirred in 20 mL of acetonitrile solvent at room temperature for 30 minutes, and then the insoluble impurities are removed by filtration; 10 mmol of nitrogen-substituted imidazole or nitrogen-substituted benzimidazole is added to the filtrate, heated to 90 DEG C, and the reaction is continued for 6 hours to form a salt; after cooling to room temperature, the acetonitrile solvent is removed under reduced pressure, dissolved in 50 mL of deionized water, filtered, and the insoluble impurities are removed; 10 mmol of ammonium hexafluorophosphate is added to the filtrate, and anion exchange is carried out to precipitate white solids; and after drying, the quinoline functionalized imidazole salt ligand or the quinoline functionalized benzimidazole salt ligand is obtained.

9. The method for preparing the quinoline-functionalized nitrogen heterocyclic carbene ruthenium complex with multiple anticancer activities according to claim 8, characterized in that: The nitrogen-substituted imidazole is one of N-benzyl imidazole, N-(4-methylbenzyl)imidazole, N-(4-trifluoromethylbenzyl)imidazole, N-(2,4,6-trimethylbenzyl)imidazole, and N-(2,3,4,5,6-pentamethylbenzyl)imidazole; and the nitrogen-substituted benzimidazole is one of N-benzyl benzimidazole, N-(2,4,6-trimethylbenzyl)benzimidazole, and N-(2,3,4,5,6-pentamethylbenzyl)benzimidazole.

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