A binuclear ruthenium pyridine coordination supramolecular self-assembly compound and its preparation method and application

Abstract

A binuclear ruthenium pyridine coordination supramolecular self-assembly compound and its preparation method and application, the structural formula of the compound is one of the following formulas 1, 2, and 3; its preparation method is: using a pyridine acceptor and a binuclear ruthenium acceptor as raw materials, the two raw materials are placed in a reaction vessel, an appropriate amount of an equal volume ratio of methanol / dichloromethane mixed solvent is added, the reaction is stirred for a period of time, and after the reaction is completed, the reaction solution is concentrated, and then ether is slowly added dropwise to the reaction until precipitation is complete, and the solid powder is precipitated as a self-assembly compound. This method can simply construct a large assembly and reduce production costs. The obtained binuclear ruthenium pyridine coordination supramolecular self-assembly compound is applied in the preparation of anti-tumor drugs, and can have a good inhibitory effect on lung cancer cells (A549), liver cancer cells (HepG-2), breast cancer cells (MDA-MB-231) and cervical cancer cells (HeLa), and has low toxicity while inhibiting tumor cell proliferation.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a binuclear ruthenium pyridine coordination supramolecular self-assembly compound and a preparation method and application thereof. Background Art

[0002] According to the World Health Organization, cancer is the second leading cause of death in developing countries, and the incidence of malignant tumors is very high. Chemotherapy is currently a primary treatment for malignant tumors. Commonly used anticancer drugs include cisplatin, doxorubicin, and the natural drug paclitaxel. Currently, platinum-based anticancer drugs account for approximately 50% of cancer treatments. The successful application of cisplatin and other platinum compounds in anticancer treatment has promoted the design and development of transition metal anticancer drugs as potential alternatives to platinum-based anticancer drugs. Ruthenium compounds have emerged as promising anticancer candidates. Self-assembly facilitates the construction of large-scale and high-molecular-weight supramolecules, which, due to their unique biochemical properties, may exhibit antitumor efficacy and selectivity. Summary of the Invention

[0003] The purpose of the present invention is to provide a binuclear ruthenium pyridine coordination supramolecular self-assembly compound and its preparation method and application. The method can simply construct a large assembly and reduce production costs. The prepared binuclear ruthenium pyridine coordination supramolecular self-assembly compound is used in the preparation of anti-tumor drugs. It can have a good inhibitory effect on lung cancer cells (A549), liver cancer cells (HepG-2), breast cancer cells (MDA-MB-231) and cervical cancer cells (HeLa), and has low toxicity while inhibiting tumor cell proliferation.

[0004] To achieve the above object, the technical solution adopted by the present invention is:

[0005] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound, the structural formula of which is one of the following formulas 1, 2, and 3;

[0006]

[0007] In Formulas 1 to 3, R is one of —CH3, —(CH2)2O(CH2)2OCH3, and —(CH2)2O(CH2)2O(CH2)2OCH3.

[0008] To achieve the above-mentioned object of the invention, the present invention also provides a method for preparing the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound, which specifically comprises the following steps:

[0009] A pyridine acceptor and a binuclear ruthenium acceptor are used as raw materials. The two raw materials are placed in a reaction vessel, and an appropriate amount of a methanol / dichloromethane mixed solvent in an equal volume ratio is added. The reaction is stirred for a period of time. After the reaction is completed, the reaction solution is concentrated, and then ether is slowly added dropwise to the reaction until precipitation is complete. The precipitated solid powder is a binuclear ruthenium pyridine coordination supramolecular self-assembled compound.

[0010] The pyridine acceptor is one of compound L1, compound L2, and compound L3, and the structural formulas of compound L1, compound L2, and compound L3 are respectively One of the following;

[0011] The binuclear ruthenium receptor is one of compound A1, compound A2, and compound A3, and the structural formula of compound A1, compound A2, and compound A3 is One of them.

[0012] Preferably, the molar ratio between the pyridine acceptor and the binuclear ruthenium acceptor is 1:1.

[0013] Preferably, stirring is carried out at room temperature for 24 hours.

[0014] Preferably, the ratio of the volume of the mixed solvent to the molar amount of the pyridine acceptor is 1.5 mL:1 mmol.

[0015] To achieve the above-mentioned object of the invention, the present invention also provides the use of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound in the preparation of an anti-tumor drug, which is used to inhibit the growth of cancer cells.

[0016] Preferably, the cancer cells are lung cancer cells, liver cancer cells, breast cancer cells and cervical cancer cells.

[0017]

[0018] The binuclear ruthenium pyridine coordination supramolecular self-assembly compound of the present invention is composed of two pyridine acceptors L (upper and lower) and two binuclear ruthenium acceptors A (left and right), such as Figure 1 shown.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] This invention successfully developed a novel ruthenium-containing self-assembly compound using a pyridine acceptor and a binuclear ruthenium receptor as ligands. This binuclear ruthenium pyridine-coordinated supramolecular self-assembly compound exhibits excellent inhibitory effects against lung cancer (A549) cells, liver cancer cells (HepG-2), breast cancer cells (MDA-MB-231), and cervical cancer cells (HeLa), while also suppressing tumor cell proliferation and exhibiting low toxicity. The method employed in this invention allows for the simple construction of large assemblies, reducing production costs, achieving relatively high reaction yields, and exhibiting strong substrate universality. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Schematic diagram of the structure of the binuclear ruthenium pyridine coordinated supramolecular self-assembly compound of the present invention;

[0022] Figure 2 Schematic diagram of the change in relative fluorescence intensity of zebrafish tumors with the injection time of self-assembling compound M3;

[0023] Figure 3 Schematic diagram of the change in relative fluorescence intensity of zebrafish tumors with the time of injection of the self-assembling compound M6. DETAILED DESCRIPTION

[0024] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0025] Unless otherwise specified, the materials and reagents used in the examples can be obtained from commercial sources.

[0026] Compound A1, Compound A2, and Compound A3 used in the following examples were prepared by the method reported in the literature (Proceedings of the National Academy of Sciences, 2019, 116(10), 4090-4098).

[0027] The synthesis and characterization of Compound L1, Compound L2, and Compound L3 used in the following examples are as follows:

[0028] (1) Synthesis and characterization of compound L1:

[0029] m-Dibromophenol (4.8 mmol, 1.2 g) and iodomethane (5.7 mmol, 0.811 g) were weighed and added to a 100 mL eggplant-shaped flask. 20 mL of acetone was added, and the nitrogen atmosphere was purged three times before stirring and refluxed overnight. After the reaction, the acetone was dried, dissolved in ethyl acetate, and washed three times with water and saturated sodium chloride solution. The product was dried over magnesium sulfate, the solvent was dried, and column chromatography using a PE / EA = 25 / 1 solvent was performed to obtain 1.02 g of the pure solid product, 3,5-dibromoanisole, with a yield of 80%. 3,5-Dibromoanisole (3.6 mmol, 0.95 g), 4-pyridineboronic acid (7.9 mmol, 1.1076 g), tetrakistriphenylphosphine palladium (0.36 mmol, 0.4128 g), and potassium carbonate (36 mmol, 5 g) were weighed and added to a 100 mL eggplant-shaped flask. DMF:H2O = 50:1 mL was then added, the nitrogen atmosphere was replaced three times, and the mixture was heated to 100°C and refluxed for 48 h. After the reaction, the mixed solution was dissolved in chloroform and filtered. The filtrate was washed three times with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent was dried by spin drying. Column chromatography with DCM / MeOH = 15 / 1 solvent was performed to obtain 1.10.5 g of a white solid powder with a yield of 52%.

[0030] Characterization: 1 HNMR(400MHz, CDCl3): δ8.71(s,4H,H α-Py ),7.57(d,J=4.6Hz,4H,H β-Py ),7.45(t,J=1.4Hz,1H,Ph),7.22(d,J=1.4Hz,2H,Ph),3.95(s,3H,CH3). 13 C NMR (100MHz, CDCl3): δ161.0,150.1,148.4,140.6,122.1,118.5,113.5,55.8.Elemental analysis:calcd(%)for C 17 H 14 N2O: C 77.84, H 5.37, N 10.68; found: C 77.80, H 5.54, N 9.96.

[0031] (2) Synthesis and characterization of compound L2:

[0032] S1. Diethylene glycol methyl ether (10 mmol, 1.201 g), p-toluene cyclopentyl chloride (11 mmol, 2.097 g) and triethylamine (10.8 mmol, 1.093 g) were weighed in 10 mL of dry DCM and stirred at room temperature for 2 h. After completion of the reaction, the product was washed with 1 mol / L hydrochloric acid, washed three times with saturated sodium bicarbonate aqueous solution and saturated sodium chloride solution, and then dried over anhydrous sodium sulfate. The product a was spin-dried to give 2.33 g of a yellow liquid in 85% yield.

[0033] S2. The obtained product a (1.2 mmol, 0.3292 g), m-dibromophenol (1 mmol, 0.2519 g), and potassium carbonate (2 mmol, 0.2764 g) were weighed and dissolved in DMF, stirred, and heated to 100° C. for overnight reaction. After the reaction, water and EA were added, the mixture was extracted, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was spin-dried to obtain a transparent liquid product b, weighing 0.26 g, with a yield of 75%.

[0034] S3, weighed the synthesized product b (3mmol, 1.062g), 4-pyridineboronic acid (6mmol, 0.846g), tetrakistriphenylphosphine palladium (0.15mmol, 0.173g) and potassium carbonate (6mmol, 0.829g) were added to a 100mL eggplant flask, nitrogen was replaced three times, the temperature was raised to 80 ° C, and stirred at reflux for 4h. After the reaction, the solvent was evaporated, extracted with DCM, washed three times with saturated sodium chloride solution, the solvent was spin-dried, and DCM / MeOH = 100 / 1 to 50 / 1 was used as the developing solvent for column chromatography to give a white solid product L20.588 g with a yield of 56%.

[0035] Characterization: 1 H NMR (400MHz, CD3OD): δ8.62 (d, J=6.2Hz, 4H, H α-Py ),7.79(d,J=6.4Hz,4H,H β-Py ),7.69(t,J=1.4Hz,1H,Ph),7.43(d,J=1.4Hz,2H,Ph),4.32(t,J=4.6Hz,2H,OCH2),3.90 (t,J=4.6Hz,2H,CH2),3.76-3.68(m,2H,CH2),3.61–3.53(m,2H,CH2),3.36(s,3H,CH3). 13 C NMR (100MHz, CDCl3): δ161.0,150.1,148.2,140.5,121.9,118.6,114.2,72.0,70.9,69.8,68.0,59.2.ESI-MS: m / z calcd for[L2+H] +:351.1703; found:351.1618.Elemental analysis:calcd(%)for C 21 H 22 N2O3: C 71.98, H 6.33, N 7.99; found: C 71.32, H 6.12, N 7.47.

[0036] (3) Synthesis and characterization of compound L3:

[0037] The synthesis steps of compound L3 were obtained by replacing "diethylene glycol methyl ether" with "triethylene glycol" in step S1 of the synthesis of compound L2, while keeping the other steps unchanged. 0.28 g of white solid powder L3 was obtained with a yield of 53%.

[0038] Characterization: 1 H NMR (400MHz, CD3CN): δ8.65 (d, J=5.9Hz, 4H, H α-Py ),7.68(d,J=5.0Hz,4H,H β-Py ),7.63(s,1H,Ph),7.35(s,2H,Ph),4.28(t,J=4.1Hz,2H,OCH2),3.83(t,J=4.5Hz,2H,CH2) ,3.69-3.62(m,2H,CH2),3.60-3.52(m,4H,CH2),3.47-3.41(m,2H,CH2),3.27(s,3H,CH3). 13 C NMR (100MHz, CDCl3): δ161.0,151.2,148.1,141.3,122.7,119.2,114.7,72.5,71.3,71.1,71.0,70.2,68.9,58.8.ESI-MS: m / z calcd for[L3+H] + :395.1965; found:395.1953.Elemental analysis:calcd(%)for C 23 H 26 N2O4: C 70.03, H 6.64, N 7.10; found: C 70.14, H 6.45, N 6.83.

[0039] Example 1

[0040] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M1 has a structural formula of Formula 1, wherein R is -CH3.

[0041] The preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M1 is as follows: Compound A1 (0.004mmol, 3.4269mg) and Compound L1 (0.004mmol, 1.0492mg) are accurately weighed using an electronic balance with a precision of 1 / 10 million and placed in an 8mL catalytic vial. 6mL of methanol solvent is then added and stirred at room temperature for 24h. After the reaction is completed, the solution is dried to 1mL and then ether is slowly added. Solid powder gradually precipitates from the solution, which is washed twice with ether and dried to obtain an orange powder with a yield of 75%. The data are characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.04 (d, J=5.0Hz, 8H, H α-Py ),7.71(d,J=5.0Hz,8H,H β-Py ),7.47(s,2H,Ph),7.18(s,4H,Ph),5.93(t,J=4.6Hz,8H,H p-cymene ),5.75(d,J=5.6Hz,8H,H p-cymene ),3.99(s,6H,CH3),2.92-2.77(m,4H,CH),2.23(s,12H,CH3),1.38(dd,J=6.9,2.0Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ172.4,153.8,151.1,138.6,124.8,121.9(q,J=316.9Hz, CF3),118.9,115.7,103.7,98.8,83.5,83.0,56.2,32.5,22.5,18.1.ESI-MS:m / z calcd for[M1-2OTf] 2+ :970.57;found:970.55.

[0042] Example 2

[0043] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M2 has a structural formula of formula 2, wherein R is -CH3.

[0044] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M2, the raw materials A2 (0.004 mmol, 3.6272 mg) and compound L1 (0.004 mmol, 1.0492 mg) were selected. The other steps were kept consistent with Example 1 to obtain an orange solid powder with a yield of 68%. The data were characterized as follows: 1 HNMR (400MHz, CD3OD): δ8.22 (d, J=6.2Hz, 8H, H α-Py ),7.73(d,J=6.0Hz,8H,Hβ-Py ),7.51(s,2H,Ph),7.19(s,4H,Ph),6.04(d,J=6.2Hz,8H,H p-cymene ),5.81(d,J=6.4Hz,12H,H p-cymene ,H dobq ),3.90(s,6H,CH3),2.97-2.82(m,4H,CH),2.21(s,12H,CH3),1.37(d,J=6.9Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ185.2,162.4,154.2,151.2,138.8,124.7,118.9,115.5,105.2,102.7,100.0,84.8,83.3,56.4,32.6,22.6,18.2.ESI-MS: m / z calcd for[M2-2OTf] 2+ :1020.60; found:1020.53.Elementalanalysis:calcd(%)for C 90 H 88 O 22 N4S4F 12 Ru4:C 46.23,H 3.79,N 2.40;found:C 44.78,H4.10,N 1.93.

[0045] Example 3

[0046] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M3 has a structural formula of formula 3, wherein R is -CH3.

[0047] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M3, the raw materials A3 (0.004 mmol, 3.8274 mg) and compound L1 (0.004 mmol, 1.0492 mg) were selected. The other steps were kept consistent with Example 1 to obtain an orange solid powder with a yield of 78%. The data were characterized as follows: 1 HNMR (400MHz, CD3OD): δ8.40 (d, J=6.4Hz, 8H, H α-Py ),7.75(d,J=6.4Hz,8H,H β-Py ),7.64(s,2H,Ph),7.24(s,12H,Ph,H donq ),5.84(d,J=6.3Hz,8H,H p-cymene ),5.61(d,J=6.3Hz,8H,H p-cymene),3.81(s,6H,CH3),2.91-2.75(m,4H,CH),2.11(s,12H,CH3),1.34(d,J=6.9Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ172.2,162.5,153.3,150.8,138.6,138.5,124.3,115. 4,112.7,104.9,103.4,101.1,85.9,84.0,56.3,32.0,22.5,17.4.ESI-MS:m / z calcd for[M3-2OTf] 2+ :1070.60; found:1070.52.Elemental analysis:calcd(%)for C 98 H 92 O 22 N4S4F 12 Ru4:C 48.27,H 3.80,N2.30;found:C 26.80,H 2.12,N 1.25.

[0048] Example 4

[0049] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M4 has a structural formula of Formula 1, wherein R is -(CH2)2O(CH2)2OCH3.

[0050] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M4, the raw materials were selected from compound A1 (0.004 mmol, 3.4269 mg) and compound L2 (0.004 mmol, 1.4016 mg). The other steps were kept consistent with Example 1, and a dark red solid powder was obtained with a yield of 71%. The data were characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.04 (d, J=5.0Hz, 8H, H α-Py ),7.71(d,J=4.9Hz,8H,H β-Py ),7.48(s,2H,Ph),7.20(s,4H,Ph),5.93(t,J=5.3Hz,8H,H p-cymene ),5.75(d,J=5.9Hz,8H,H p-cymene),4.31(t,J=4.3Hz,4H,CH2),3.96(t,J=4.3Hz,4H,CH2),3.80-3.71(m,4H,CH2),3.65-3.58(m,4H,CH2 ),3.39(s,6H,CH3),2.93-2.79(m,4H,CH),2.23(s,12H,CH3),1.38(dd,J=6.9,2.6Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ172.5,161.6,153.9,151.1,138.6,124.8,116.5,103.7,9 8.8,83.6,83.0,73.1,71.6,70.9,69.3,59.2,32.5,22.5,22.4,18.1.ESI-MS:m / z calcd for[M4-2OTf] 2+ :1058.57; found:1058.62.Elemental analysis:calcd(%)forC 90 H 100 O 26 N4S4F 12 Ru4:C 44.77,H 4.17,N 2.32;found:C 44.00,H 4.07,N 2.19.

[0051] Example 5

[0052] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M5 has a structural formula of Formula 2, wherein R is -(CH2)2O(CH2)2OCH3.

[0053] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M5, the raw materials were selected from compound A2 (0.004 mmol, 3.6272 mg) and compound L2 (0.004 mmol, 1.4016 mg), and the other steps were kept consistent with Example 1 to obtain a dark red solid powder with a yield of 69%. The data were characterized as follows: 1 H NMR (400MHz, Acetonitrile-d3): δ8.20 (d, J = 6.6Hz, 8H, H α-Py ),7.78(d,J=6.7Hz,8H,H β-Py ),7.66(s,2H,Ph),7.31(s,4H,Ph),5.89(d,J=5.3Hz,8H,H p-cymene ),5.73(s,4H,H dobq ),5.67(d,J=6.4Hz,8H,Hp-cymene ),4.17(t,J=4.3Hz,4H,CH2),3.76(t,J=4.2Hz,4H,CH2),3.63-3.57(m,4H,CH2),3.49-3.43(m,4H ,CH2),3.22(s,6H,CH3),2.87-2.74(m,4H,CH),2.15(s,12H,CH3),1.31(d,J=6.9,24H,CH(CH3)2). 13 C NMR (100MHz, Acetonitrile-d3): δ185.2,161.3,153.9,149.8,138.0,124.3,120.5,116.3, 104.4,102.4,99.9,84.6,82.7,72.6,71.2,70.2,69.2,58.9,32.1,22.4,18.3.ESI-MS:m / z calcd for[M5-2OTf] 2+ :1108.64; found:1108.57.Elemental analysis:calcd(%)for C 98 H 104 O 26 N4S4F 12 Ru4:C 46.81,H 4.17,N 2.23;found:C 44.36,H 4.75,N 1.15.

[0054] Example 6

[0055] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M6 has a structural formula of Formula 3, wherein R is -(CH2)2O(CH2)2OCH3.

[0056] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M6, the raw materials A3 (0.004 mmol, 3.8274 mg) and compound L2 (0.004 mmol, 1.4016 mg) were selected. The other steps were kept consistent with Example 1 to obtain a dark red solid powder with a yield of 69%. The data were characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.41 (d, J=6.6Hz, 8H, H α-Py ),7.76(d,J=6.5Hz,8H,H β-Py ),7.65(s,2H,Ph),7.32(s,4H,Ph),7.24(s,8H,H donq ),5.85(d,J=6.3Hz,8H,H p-cymene),5.61(d,J=6.3Hz,8H,H p-cymene ),4.18(t,J=4.4Hz,4H,CH2),3.81(t,J=4.3Hz,4H,CH2),3.68-3.61(m,4H,CH2),3.53-3.46(m,4H, CH2),3.21(s,6H,CH3),2.87-2.78(m,4H,CH),2.11(s,12H,CH3),1.34(d,J=6.9Hz,24H,CH(CH3)2). 13 CNMR (100MHz, CD3OD): δ172.3,162.8,153.3,150.8,138.6,138.6,124.3,116.3,112.8,10 4.9,102.6,101.1,85.9,83.9,72.9,71.5,70.8,69.2,59.0,32.0,22.5,17.4.ESI-MS:m / z calcd for[M6-2OTf] 2+ :1158.66; found:1158.57.Elemental analysis:calcd(%)forC 106 H 108 O 26 N4S4F 12 Ru4:C 48.69,H 4.16,N 2.14;found:C 45.37,H 4.47,N 1.32.

[0057] Example 7

[0058] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M7 has a structural formula of Formula 1, wherein R is -(CH2)2O(CH2)2O(CH2)2OCH3.

[0059] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M7, the raw materials were selected from compound A1 (0.004 mmol, 3.4269 mg) and compound L3 (0.004 mmol, 1.5778 mg). The other steps were kept consistent with Example 1, and a green solid powder was obtained with a yield of 73%. The data were characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.04 (d, J=5.2Hz, 8H, H α-Py ),7.71(d,J=5.1Hz,8H,H β-Py ),7.48(s,2H,Ph),7.21(s,4H,Ph),5.93(t,J=5.4Hz,8H,H p-cymene),5.76(d,J=5.9Hz,8H,H p-cymene ),4.32(t,J=4.3Hz,4H,CH2),3.97(t,J=4.3Hz,4H,CH2),3.80-3.73(m,4H,CH2),3.72-3.62(m,8H,CH2),3.57-3.5 2(m,4H,CH2),3.35(s,6H,CH3),2.92-2.80(m,4H,CH),2.23(s,12H,CH3),1.38(dd,J=6.9,2.7Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ172.5,161.6,153.9,151.1,138.6,124.8,119.1,116.5,103.7,9 8.8,83.5,83.0,73.00,71.8,71.6,71.4,70.9,69.3,59.1,32.5,22.5,18.1.ESI-MS:m / z calcd for[M7-2OTf] 2+ :1102.65; found:1102.59.Elemental analysis:calcd(%)for C 94 H 108 O 28 N4S4F 12 Ru4:C 45.12,H 4.35,N 2.24;found:C 44.83,H 4.17,N 2.04.

[0060] Example 8

[0061] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M8 has a structural formula of Formula 2, wherein R is -(CH2)2O(CH2)2O(CH2)2OCH3.

[0062] The above-mentioned preparation method of the binuclear ruthenium pyridine coordination supramolecular self-assembly compound M8 was prepared using compound A2 (0.004 mmol, 3.6272 mg) and compound L3 (0.004 mmol, 1.5778 mg) as raw materials. The other steps were the same as those in Example 1. A green solid powder was obtained with a yield of 72%. The data were characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.23 (d, J=5.9Hz, 8H, H α-Py ),7.70(d,J=5.6Hz,8H,H β-Py ),7.48(s,2H,Ph),7.21(s,4H,Ph),6.04(d,J=6.3Hz,8H,Hp-cymene ),5.81(d,J=6.2Hz,12H,H p-cymene ,H dobq ),4.25(s,4H,CH2),3.90(s,4H,CH2),3.78-3.63(m,8H,CH2),3.61-3.54(m,4H,CH2),3.50-3.43(m,4 H,CH2),3.23(s,6H,CH3),2.95-2.84(m,4H,CH),2.21(s,12H,CH3),1.38(d,J=6.9Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ185.3,161.6,154.3,151.3,138.9,124.8,119.1,116.4,105.2,102.7,1 00.0,84.8,83.2,72.9,71.7,71.6,71.3,70.8,69.3,59.0,32.6,22.6,18.2.ESI-MS:m / zcalcd for[M8-2OTf] 2+ :1152.67; found:1152.54.Elemental analysis:calcd(%)forC 102 H 112 O 28 N4S4F 12 Ru4:C 47.07,H 4.34,N 2.15;found:C 45.19,H 4.83,N 1.36.

[0063] Example 9

[0064] A binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M9 has a structural formula of Formula 3, wherein R is -(CH2)2O(CH2)2O(CH2)2OCH3.

[0065] In the preparation method of the above-mentioned binuclear ruthenium pyridine coordination supramolecular self-assembly compound M9, the raw materials were selected from compound A3 (0.004 mmol, 3.8274 mg) and compound L3 (0.004 mmol, 1.5778 mg). The other steps were kept consistent with Example 1 to obtain a green solid powder with a yield of 69%. The data were characterized as follows: 1 H NMR (400MHz, CD3OD): δ8.41 (d, J=6.5Hz, 8H, H α-Py ),7.76(d,J=6.5Hz,8H,H β-Py ),7.65(s,2H,Ph),7.31(s,4H,Ph),7.24(s,8H,Hdonq ),5.85(d,J=6.3Hz,8H,H p-cymene ),5.61(d,J=6.3Hz,8H,H p-cymene ),4.18(t,J=4.3Hz,4H,CH2),3.81(t,J=4.3Hz,4H,CH2),3.68-3.52(m,8H,CH2),3.50-3.43(m,4H,CH2),3.37- 3.33(m,4H,CH2),3.11(s,6H,CH3),2.90-2.79(m,4H,CH),2.11(s,12H,CH3),1.34(d,J=6.9Hz,24H,CH(CH3)2). 13 C NMR (100MHz, CD3OD): δ171.0,160.4,152.0,149.4,137.3,137.2,123.0,117.4,115.0,111.4,1 03.5,99.7,84.6,82.6,71.5,70.3,70.2,69.9,69.5,67.8,57.6,30.7,21.2,16.1.ESI-MS:m / z calcd for[M9-2OTf] 2+ :1202.68; found:1202.60.Elementalanalysis:calcd(%)for C 110 H 116 O 28 N4S4F 12 Ru4:C 48.88, H 4.33, N 2.07; found: C 45.56, H5.47, N 0.70.

[0066] Example 10 Cytotoxicity Test

[0067] Four human cancer cells, including lung cancer cells (A549), liver cancer cells (HepG-2), breast cancer cells (MDA-MB-231), and cervical cancer cells (HeLa), were used as standby cells. The cells were sourced from Nanjing Shulihua Biotechnology Co., Ltd.

[0068] The cells were removed from the liquid nitrogen and lysed in a 37°C waterbath. After centrifugation, fresh culture medium was added and the cell suspension was shaken until well mixed. The inoculated cell culture medium was then added dropwise to DMEM medium consisting of 10% FBS fetal bovine serum and 1% penicillin-streptomycin. The inoculated cell culture medium was then placed in a 37°C 5% CO2 incubator and cultured for a period of time (aseptic operation throughout). The passaged cells were aspirated into a 1 mL centrifuge tube, the culture medium was discarded, and the tubes were washed with PBS. The tubes were then delaminated with trypsin. After delaminization, the cells were evenly divided into two centrifuge tubes.

[0069] Compounds L1-L3, compounds A1-A3, self-assembled compounds M1-M9 and the control group were dissolved in dimethyl sulfoxide at a concentration of 5 mg mL -1 Cells were added to a 96-well plate, and the number of cells in each well was controlled at approximately 0.5×10 4 –1.0×10 4 The wells with separated cells were placed in an incubator for 24 h, and then compounds L1-L3, compounds A1-A3 and self-assembly compounds M1-M9 were added in a gradient manner. The cells after administration were further cultured in the incubator for 72 h.

[0070] MTT was dissolved in phosphate buffered saline (PBS, pH = 7.2), 20 μL of MTT solution was added to each well, and cultured in an incubator for 4 hours. After the culture was completed, the excess liquid in the well plate was discarded, and then the cell metabolites were dissolved with dimethyl sulfoxide. The absorbance of the solution was tested with a microplate reader. The IC value can be calculated by the absorbance test. 50 value

[0071] The MTT assay is based on the principle that succinate dehydrogenase in living cells reduces MTT to water-insoluble blue-purple crystalline formazan. The formazan is then dissolved in DMSO and the absorbance of the solution is measured to determine the number of viable cells. The experimental results are shown in Table 1 below:

[0072] Table 1 Cancer cell inhibition test results

[0073]

[0074] As can be seen from Table 1, M3, M6 and M9 of the assembly A3 containing ruthenium receptors have a high inhibitory effect on cell proliferation, that is, good anti-cancer activity. Among them, for A549 cells, the anti-cancer effect of assembly M6 is better than cisplatin and doxorubicin. For HepG-2 cells, assemblies M3 and M9 are more cytotoxic than cisplatin and paclitaxel, but are still slightly worse than doxorubicin. For MDA-MB-231 cells, assembly M3 has the strongest cytotoxicity, reaching 1.72μM, which is 2-5 times lower than paclitaxel, indicating that the assembly has high anti-cancer activity. By comparing the pyridine acceptor and the binuclear ruthenium receptor, it can be seen that the anti-cancer effect of the assembly after assembly is significantly improved, which may be due to the synergistic effect of the pyridine acceptor and the assembly.

[0075] Example 11 Zebrafish in vivo toxicity test

[0076] Weigh 7g of sodium chloride, 0.4g of sodium bicarbonate, 0.1g of potassium chloride and 0.235g of calcium chloride and dissolve them in 2000mL of redistilled water, stir to dissolve and filter using a disposable vacuum filter to prepare Holt buffer solution, which is stored at room temperature for later use. Use purchased wild-type AB strain zebrafish, select half of the male and half of the female fish and allow them to mate freely. After 1 hour, collect the embryos, add Holt buffer solution, and place them in a 28.5℃ constant temperature incubator for 48 hours before use. Resuscitate tumor cells A549 and MDAMB-231. When the cells fill the culture dish, digest and collect the cells, add DiI (cell membrane red fluorescent probe) for staining for 30 minutes, and place them in a 25℃ carbon dioxide incubator for later use. Hatched zebrafish were anesthetized with ethyl 3-aminobenzoate methanesulfonate, then aspirated to a plate and spread out. Using a MicroSyringe Pump Controller, 6 nL of stained tumor cells were injected into the fish's abdomen. The fish were then placed in culture medium in an incubator, and PTU was added to the medium to inhibit pigmentation. Twenty hours later, the same method was repeated, with 6 nL of compound injected into each fish at concentrations of 10 ng / nL and 5 ng / nL (30 zebrafish were injected at each concentration). Each zebrafish was then placed in a 96-well plate, and PTU and culture medium were added again. Fluorescence images of the stained tumor cells were captured under an inverted fluorescence microscope at 0, 24, and 48 hours.

[0077] 48 hours after the zebrafish laid eggs, the hatched zebrafish were collected. The cultured tumor cells A549 and MDA-MB-231 were stained with fluorescence and then injected into the zebrafish with a needle. On average, the same number of tumor cells was injected into each fish. After 24 hours, 6nL of high concentration 10mg / mL and low concentration 5mg / mL solutions of the self-assembly compounds M3 and M6 were injected into the treated zebrafish, with about 30 fish at each concentration. Using the blank group as a control, the changes in the size of zebrafish tumor cells over time were observed using a fluorescence microscope. Figure 2 and Figure 3 As shown, compared to the blank control group, the self-assembling compounds M3 and M6 reduced the size of A549 and MDA-MB-231 tumor tissues. The self-assembling compound M6 had the most pronounced effect, and did not cause death in fish 48 hours after injection, demonstrating its ultra-low toxicity while inhibiting tumor cell proliferation.

Claims

1. A binuclear ruthenium pyridine coordination supramolecular self-assembly compound, characterized in that: Its structural formula is one of the following formulas 1, 2, and 3; In Formulas 1 to 3, R is one of —CH3, —(CH2)2O(CH2)2OCH3, and —(CH2)2O(CH2)2O(CH2)2OCH3.

2. A method for preparing the binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 1, characterized in that: The specific steps are: A pyridine acceptor and a binuclear ruthenium acceptor are used as raw materials. The two raw materials are placed in a reaction vessel, and an appropriate amount of a methanol / dichloromethane mixed solvent in an equal volume ratio is added. The reaction is stirred for a period of time. After the reaction is completed, the reaction solution is concentrated, and then ether is slowly added dropwise to the reaction until precipitation is complete. The precipitated solid powder is a binuclear ruthenium pyridine coordination supramolecular self-assembled compound. The pyridine acceptor is one of compound L1, compound L2, and compound L3, and the structural formulas of compound L1, compound L2, and compound L3 are respectively One of the following; The binuclear ruthenium receptor is one of compound A1, compound A2, and compound A3, and the structural formulas of compound A1, compound A2, and compound A3 are respectively One of them.

3. The method for preparing a binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 2, characterized in that: The molar ratio between the pyridine acceptor and the binuclear ruthenium acceptor is 1:

1.

4. The method for preparing a binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 2 or 3, characterized in that: Stir at room temperature for 24 h.

5. The method for preparing a binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 2 or 3, characterized in that: The ratio of the volume of the mixed solvent to the molar amount of the pyridine acceptor is 1.5 mL:1 mmol.

6. Use of the binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 1 in the preparation of an anti-tumor drug, wherein the anti-tumor drug is used to inhibit the growth of cancer cells.

7. Use of a binuclear ruthenium pyridine coordination supramolecular self-assembly compound according to claim 6 in the preparation of an anti-tumor drug, wherein the cancer cells are lung cancer cells, liver cancer cells, breast cancer cells and cervical cancer cells.

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