Binuclear ruthenium pyridine coordination supramolecular self-assembly compound as well as preparation method and application thereof

Abstract

The invention discloses a binuclear ruthenium pyridine coordination supramolecular self-assembly compound as well as a preparation method and application thereof. The structural formula of the compound is one of a formula 1, a formula 2 and a formula 3. The preparation method comprises the following steps: by taking a pyridine acceptor and a binuclear ruthenium acceptor as raw materials, putting the two raw materials into a reaction container, adding a proper amount of methanol / dichloromethane mixed solvent with the same volume ratio, stirring and reacting for a period of time, concentrating reaction liquid after the reaction is finished, then slowly dropwise adding diethyl ether into the reaction until the precipitation is complete, and drying to obtain the target product. And separating out solid powder which is the self-assembled compound. According to the method, a large assembly can be simply constructed, the production cost is reduced, and the obtained binuclear ruthenium pyridine coordination supramolecular self-assembly compound has a good inhibition effect on lung cancer cells (A549), liver cancer cells (HepG-2), breast cancer cells (MDA-MB-231) and cervical cancer cells (HeLa) when being applied to preparation of antitumor drugs, and can be used for preparing antitumor drugs. The toxicity is relatively low while the tumor cell proliferation is inhibited. # imgabs0 #

Description

Technical Field

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

[0002] According to statistics from 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 major measure for the treatment of malignant tumors. Commonly used anticancer drugs include cisplatin, doxorubicin, and the natural drug paclitaxel. At present, the use of platinum anticancer drugs accounts for about 50% of cancer treatment. The successful application of cisplatin and other platinum compounds in anticancer treatment has promoted the design and development of transition metal anticancer drugs. As a potential alternative to platinum-based anticancer drugs, ruthenium compounds have become promising anticancer candidates. Self-assembly helps to construct supramolecules of large size and high molecular weight. Due to their unique biochemical properties, these supramolecules may show 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 a preparation method and application thereof. 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, and has 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 Formulae 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 purpose 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 container, an appropriate amount of a methanol / dichloromethane mixed solvent of equal volume ratio is added, the reaction is stirred for a period of time, the reaction solution is concentrated after the reaction is completed, and then ether is slowly added dropwise to the reaction until precipitation is complete, and the precipitated solid powder is a binuclear ruthenium pyridine coordination supramolecular self-assembly 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;

[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 purpose 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 anti-tumor drugs, and the anti-tumor drugs are 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] The present invention uses a pyridine acceptor and a binuclear ruthenium receptor as ligands, and successfully develops a new type of ruthenium-containing self-assembly compound. The binuclear ruthenium pyridine coordination supramolecular self-assembly compound has 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. The method used in the present invention can simply construct a large assembly, reduce production costs, the reaction yield is relatively high, and the substrate universality is strong. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a 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 of relative fluorescence intensity of zebrafish tumors with the injection time of self-assembly compound M3;

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

[0024] The present invention is 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] Weigh m-dibromophenol (4.8mmol, 1.2g) and iodomethane (5.7mmol, 0.811g) into a 100mL eggplant-shaped bottle, add 20mL acetone, replace nitrogen three times, and stir and reflux overnight. After the reaction is completed, spin-dry acetone, dissolve in ethyl acetate, and wash three times with water and saturated sodium chloride solution, dry with magnesium sulfate, spin-dry the solvent, and use PE / EA=25 / 1 developing agent column chromatography to obtain 1.02g of pure solid product 3,5-dibromoanisole, with a yield of 80%. Continue to weigh 3,5-dibromoanisole (3.6mmol, 0.95g), 4-pyridine boronic acid (7.9mmol, 1.1076g), tetrakistriphenylphosphine palladium (0.36mmol, 0.4128g) and potassium carbonate (36mmol, 5g) and add them to a 100mL eggplant-shaped bottle, then add DMF:H2O=50:1mL, replace nitrogen three times, heat to 100℃ and reflux for 48h. After the reaction is completed, the mixed solution is dissolved in chloroform, filtered, and the filtrate is washed three times with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and the solvent is spin-dried and chromatographed with DCM / MeOH=15 / 1 as a developing agent to obtain a white solid powder L10.5 g, 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, weigh diethylene glycol methyl ether (10mmol, 1.201g), p-toluene cycloacyl chloride (11mmol, 2.097g) and triethylamine (10.8mmol, 1.093g) in 10mL dry DCM, stir at room temperature for 2h. After the reaction is completed, wash with 1mol / L hydrochloric acid, wash three times with saturated sodium bicarbonate aqueous solution and saturated sodium chloride solution, then dry with anhydrous sodium sulfate, spin dry to obtain 2.33g of yellow liquid product a, with a yield of 85%.

[0033] S2, weigh the obtained product a (1.2mmol, 0.3292g), m-dibromophenol (1mmol, 0.2519g) and potassium carbonate (2mmol, 0.2764g), dissolve in DMF, stir and heat to 100°C overnight. After the reaction, add water and EA for extraction, wash with saturated sodium chloride solution, dry with anhydrous sodium sulfate, spin dry the solvent to obtain a transparent liquid product b, weighing 0.26g, with a yield of 75%.

[0034] S3, weigh the synthesized product b (3mmol, 1.062g), 4-pyridine boronic acid (6mmol, 0.846g), tetrakistriphenylphosphine palladium (0.15mmol, 0.173g) and potassium carbonate (6mmol, 0.829g) and add them to a 100mL eggplant-shaped bottle, replace nitrogen three times, heat to 80°C, and stir and reflux for 4h. After the reaction is completed, evaporate the solvent, extract with DCM, wash three times with saturated sodium chloride solution, spin dry the solvent, and use DCM / MeOH=100 / 1 to 50 / 1 as the developing solvent column chromatography to obtain 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 are as follows: "diethylene glycol methyl ether" in step S1 of the synthesis of compound L2 is replaced by "triethylene glycol", and the other steps remain unchanged. 0.28 g of white solid powder L3 is obtained, and the yield is 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 binuclear ruthenium pyridine coordination supramolecular self-assembly compound M1 is as follows: accurately weigh compound A1 (0.004mmol, 3.4269mg) and compound L1 (0.004mmol, 1.0492mg) using an electronic balance of one ten-millionth, and place them in an 8mL catalytic vial, then add 6mL of methanol solvent, and stir 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. The yield is 75%, and the data is 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 binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M2, the raw materials A2 (0.004mmol, 3.6272mg) and compound L1 (0.004mmol, 1.0492mg) were selected, and the other steps were kept consistent with Example 1 to obtain an orange solid powder with a yield of 68%. The data are 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 binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M3, the raw materials A3 (0.004mmol, 3.8274mg) and compound L1 (0.004mmol, 1.0492mg) were selected, and the other steps were kept consistent with Example 1 to obtain an orange solid powder with a yield of 78%. The data are 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 binuclear ruthenium pyridine coordination supramolecular self-assembly compound M4, the raw materials are compound A1 (0.004mmol, 3.4269mg) and compound L2 (0.004mmol, 1.4016mg), and the other steps are kept consistent with Example 1 to obtain a dark red solid powder with a yield of 71%. 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=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 binuclear ruthenium pyridine coordination supramolecular self-assembly compound M5, the raw materials are compound A2 (0.004mmol, 3.6272mg) and compound L2 (0.004mmol, 1.4016mg), and the other steps are kept consistent with Example 1 to obtain a dark red solid powder with a yield of 69%. The data are 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 binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M6, the raw materials A3 (0.004mmol, 3.8274mg) and compound L2 (0.004mmol, 1.4016mg) were selected, and the other steps were kept consistent with Example 1 to obtain a dark red solid powder with a yield of 69%. The data are 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 binuclear ruthenium pyridine coordinated supramolecular self-assembly compound M7, the raw materials are compound A1 (0.004mmol, 3.4269mg) and compound L3 (0.004mmol, 1.5778mg), and the other steps are kept consistent with Example 1 to obtain a green solid powder with a yield of 73%. The data are 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] In the preparation method of the above binuclear ruthenium pyridine coordination supramolecular self-assembly compound M8, the raw materials are compound A2 (0.004mmol, 3.6272mg) and compound L3 (0.004mmol, 1.5778mg), and the other steps are kept consistent with Example 1 to obtain a green solid powder with a yield of 72%. The data are 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 binuclear ruthenium pyridine coordination supramolecular self-assembly compound M9, the raw materials are compound A3 (0.004mmol, 3.8274mg) and compound L3 (0.004mmol, 1.5778mg), and the other steps are kept consistent with Example 1 to obtain a green solid powder with a yield of 69%. The data are 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 prepared. The cells were sourced from Nanjing-Shulihua Biotechnology Co., Ltd.

[0068] Take out the above cells from liquid nitrogen, then dissolve the cells in a 37°C water bath, add new culture fluid after centrifugation, shake well and drop the cell suspension into DMEM medium composed of 10% FBS fetal bovine serum and 1% penicillin-streptomycin, then place the inoculated cell culture medium in a 37°C 5% CO2 incubator and culture for a period of time (all sterile operation). Suck the passaged cells into a 1mL centrifuge tube, discard the culture fluid, wash with PBS, then add trypsin to the centrifuge tube for wall removal, and evenly divide the cells into two centrifuge tubes after wall removal.

[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 The 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 continuously cultured in the incubator for 72 h.

[0070] Dissolve MTT in phosphate buffered saline (PBS, pH = 7.2), add 20 μL of MTT solution to each well, and culture in an incubator for 4 hours. After the culture is completed, discard the excess liquid in the well plate, and then use dimethyl sulfoxide to dissolve the cell metabolites. Use an enzyme marker to test the absorbance of the solution. The IC can be calculated by the absorbance test. 50 value

[0071] The detection principle of the MTT assay is that succinate dehydrogenase in living cells reduces MTT to water-insoluble blue-purple crystalline formazan, which is then dissolved in DMSO and the absorbance of the solution is measured to determine the number of living 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, up to 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 assembled 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 use a disposable vacuum filter to filter and prepare Holtbuffer solution, which is stored at room temperature for later use. Use purchased wild-type AB strain zebrafish, select half of the male and female fish and let them mate freely, collect the embryos after 1 hour, add Holt buffer solution, and place them in a 28.5℃ constant temperature incubator for 48 hours for later 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) to stain for 30 minutes, and place them in a 25℃ carbon dioxide incubator for later use. The hatched zebrafish were anesthetized with 3-aminobenzoic acid ethyl ester methanesulfonate, then sucked into a plate and spread out. The fish belly was injected with 6nL / fish of stained tumor cells using a MicroSyringe Pump Controller. The fish were then placed in a culture medium in an incubator, and PTU was added to the culture medium to inhibit the growth of zebrafish pigments. After 24 hours, the same method was used to inject 6nL / fish of compounds at concentrations of 10ng / nL and 5ng / nL (30 zebrafish were injected at each concentration), and each zebrafish was placed in a 96-well plate, and PTU and culture medium were continued to be added. After the end, the fish were placed under an inverted fluorescence microscope to take fluorescent photos of stained tumor cells in the fish at 0h, 24h and 48h.

[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, with the same number of tumor cells injected into each fish on average. 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 approximately 30 fish at each concentration. Using the blank group as a control, a fluorescence microscope was used to observe changes in the size of zebrafish tumor cells over time. Figure 2 and Figure 3 As shown in the figure, compared with the blank group, the A549 and MDA-MB-231 tumor tissues were reduced under the action of self-assembly compounds M3 and M6. Among them, the effect of self-assembly compound M6 was the most obvious, and the compound did not cause the death of fish 48 hours after injection into the fish, indicating that it has ultra-low toxicity while inhibiting the proliferation of tumor cells.

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 Formulae 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 as claimed in 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 container, an appropriate amount of a methanol / dichloromethane mixed solvent of equal volume ratio is added, the reaction is stirred for a period of time, the reaction solution is concentrated after the reaction is completed, and then ether is slowly added dropwise to the reaction until precipitation is complete, and the precipitated solid powder is a binuclear ruthenium pyridine coordination supramolecular self-assembly 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 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. A 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 a binuclear ruthenium pyridine coordination supramolecular self-assembly compound as claimed in 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 anti-tumor drugs, wherein the cancer cells are lung cancer cells, liver cancer cells, breast cancer cells and cervical cancer cells.

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