Ion transport molecule, pharmaceutical composition and application

Abstract

The invention relates to ion transport molecules, pharmaceutical compositions and uses. The ion transport molecule has a structure as shown in the following formula I, and x is equal to 1, 2, 3, 4 or 5; r1 and R2 are respectively and independently a crown ether group, an aza-crown ether group, a cyclodextrin group, a thiourea group, an amide ring group, a benzamide ring group, an aryl triazolyl group, a cholic acid group, a cholesteryl group, CH3CH2 (OCH2CH2) k-O-, hydrogen or an alkyl group, and k is equal to 0-10; and R3 and R4 are each independently a linking group, an alkylene group, an alkylene oxy group, an alkylene acyl group, an alkylene amino group, an alkylene triazole group, an alkynylene group, an imidoalkyl acyl group, a sulfinyl alkyl acyl group, a phosphonite alkyl acyl group, an arylene group, or a heteroarylene group. The ion transport molecule provided by the invention can be inserted into a phospholipid bilayer to inhibit the growth of cancer cells or inhibit the growth of bacteria, so that the effect of treating cancer or bacterial infection is achieved. # imgabs0 #

Description

Technical Field

[0001] The invention relates to the technical field of artificial ion transmission, in particular to an ion transmission molecule with the activity of treating bacterial infection and cancer. Background Art

[0002] As the basic unit of life, cells need to exchange nutrients and transmit biological signals with the surrounding environment to ensure the orderly progress of various life activities of organisms. The ion balance inside and outside the cell is completed by ion transporters embedded in the cell surface, which is of great significance for realizing various cell functions. Abnormal function of ion transporters can cause many diseases. For example, many ion channel diseases are related to the transmembrane transport of chloride ions, such as cystic fibrosis (CF), congenital ankylosing spondylosis (Thomsen) and hereditary kidney stones.

[0003] However, most natural ion transporters are composed of proteins. Due to the complex molecular structure and extremely low content of channel proteins, they are difficult to synthesize artificially and extract naturally, which seriously limits the research and development of transport proteins in physiology and biophysics. In order to achieve the treatment of related diseases, scientists began to use chemical total synthesis methods to construct ion transporters with similar structures and functions to natural transport proteins.

[0004] To date, many artificial ion transporters have shown anticancer and antibacterial activities (Saha T., et al. (2016) J. Am. Chem. Soc., 138(24): 7558-7567; Peters, AD, et al. (2020) Chem. Sci., 11(27): 7023-7030). However, the transport activity and cell specificity of most reported artificial ion transporters are still far from meeting medical requirements.

[0005] Therefore, it is urgent to develop an artificial ion transport system that can stably exist on the cell membrane, achieve efficient ion transport, and selectively kill cancer cells and / or bacteria. Summary of the invention

[0006] After in-depth research on the above problems, the present invention found that the ion transport molecule with a structure of pyrimidine-hydrazone-pyrimidine (pym-hyz-pym) in the present invention has the effect of inhibiting tumor growth and inhibiting bacterial growth, especially in the presence of Zn 2+ Under such circumstances, the effects of inhibiting tumor growth and inhibiting bacterial growth are more significant, thereby completing the present invention.

[0007] The first object of the present invention is to provide an ion transport molecule that has the effect of inhibiting tumor growth and inhibiting bacterial growth.

[0008] The second object of the present invention is to provide a pharmaceutical composition comprising the aforementioned ion transport molecule and a pharmaceutically acceptable carrier.

[0009] The third object of the present invention is to provide the use of the above-mentioned ion transport molecules in the preparation of drugs for treating tumors or for treating bacterial infections.

[0010] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0011] The first aspect of the present invention provides an ion transport molecule having the structure shown below:

[0012]

[0013] Where x is 1, 2, 3, 4, or 5;

[0014] R1 and R2 are each independently a crown ether group, an azacrown ether group, a cyclodextrin group, a thiourea group, an amide ring group, a benzamide ring group, an aryltriazole group, a cholic acid group, a cholesterol group, a CH3CH2(OCH2CH2) group, k -O-, hydrogen, or alkyl, wherein k = 0 to 10;

[0015] Furthermore, R1 and R2 are not hydrogen or alkyl at the same time;

[0016] R3 and R4 are each independently a linking group, an alkylene group, an alkyleneoxy group, an alkyleneacyl group, an alkyleneamino group, an alkylenetriazolyl group, an alkynylene group, an iminoalkylacyl group, a sulfinylalkylacyl group, a phosphinylalkylacyl group, an arylene group, or a heteroarylene group.

[0017] The second aspect of the present invention provides a pharmaceutical composition comprising the aforementioned ion transport molecule and a pharmaceutically acceptable carrier.

[0018] The third aspect of the present invention provides the use of the aforementioned ion transport molecules in the preparation of drugs for treating cancer or drugs for treating bacterial infections.

[0019] The two ends of the ion transport molecule of the present invention can be inserted into the phospholipid bilayer, and the ion balance inside and outside the cancer cell or bacteria can be changed by transport to induce apoptosis, thereby inhibiting the growth of cancer cells or inhibiting the growth of bacteria to obtain the effect of treating cancer or treating bacterial infection. In particular, the ion transport molecule of the present invention has a repeated pyrimidine-hydrazone-pyrimidine structural unit, which in the presence of Zn 2+ Folding can occur in the case of Zn 2+ Its ion transmission capacity is further enhanced in the case of 2+To regulate the folding / unfolding of the structure of ion transport molecules and thus regulate the transmembrane transport activity of ion transport molecules. DETAILED DESCRIPTION

[0020] In order to more clearly illustrate the present invention, the above contents of the present invention are further described in detail below by way of examples. It should be understood by those skilled in the art that the contents specifically described below are illustrative rather than restrictive, and should not be used to limit the scope of protection of the present invention.

[0021] The experimental materials used in the examples of the present invention, unless otherwise specified, are all commercially available. The chemical reagents are all analytically pure and do not require further purification. The cells used in the present invention were purchased from the Cell Bank of the Chinese Academy of Sciences.

[0022] The ion transport molecule of the present invention has the structure shown in the following formula:

[0023]

[0024] Where x = 1, 2, 3, 4, or 5;

[0025] R1 and R2 are each independently a crown ether group, an azacrown ether group, a cyclodextrin group, a thiourea group, an amide ring group, a benzamide ring group, an aryltriazole group, a cholic acid group, a cholesterol group, a CH3CH2(OCH2CH2) group, k -O-, hydrogen, or alkyl, wherein k = 0 to 10;

[0026] R3 and R4 are each independently a linking group, an alkylene group, an alkyleneoxy group, an alkyleneacyl group, an alkyleneamino group, an alkylenetriazolyl group, an alkynylene group, an iminoalkylacyl group, a sulfinylalkylacyl group, a phosphinylalkylacyl group, an arylene group, or a heteroarylene group.

[0027] Among the groups represented by the aforementioned R1 and R2, the crown ether group, azacrown ether group, and cyclodextrin group have cation recognition ability, the thiourea group, amide ring group, benzamide ring group, and aryltriazole group have anion recognition ability, and the cholic acid group, cholesterol group, CH3CH2(OCH2CH2) k -O- (where k = 0 to 10) has a membrane anchoring effect. By utilizing the cation recognition ability, anion recognition ability or membrane anchoring effect of these groups, the aforementioned ion transport molecules can be further modified with cations or anions, or anchored to a membrane with a functional group. As the aforementioned cations, for example, there are Na + , Li + , K + Mg 2+ , Ca 2+ As the aforementioned anion, for example, there are Cl - , HCO- 、NO3 - Examples of the functional group carried by the aforementioned membrane carrying a functional group include a bile acid group, a cholesterol group, a cholesterol group, and a fatty chain group.

[0028] In some embodiments of the ion transport molecule, the aforementioned R1 and R2 are preferably each independently selected from any one of the group consisting of the following structural formulas.

[0029]

[0030]

[0031] In some embodiments of the ion transport molecule, R3 and R4 are each independently an alkylene group having 1 to 5 carbon atoms, an alkyleneoxy group having 1 to 5 carbon atoms, an alkyleneacyl group having 1 to 5 carbon atoms, or an alkyleneamino group having 1 to 5 carbon atoms.

[0032] Examples of the alkylene group having 1 to 5 carbon atoms include a methylene group, an ethylene group, a propylene group, a butylene group, and a pentylene group.

[0033] Examples of the alkyleneoxy group having 1 to 5 carbon atoms include a methyleneoxy group, an ethyleneoxy group, a propyleneoxy group, a butyleneoxy group, and a pentyleneoxy group.

[0034] Examples of the alkylene acyl group having 1 to 5 carbon atoms include a methylene acyl group, an ethylene acyl group, a propylene acyl group, a butylene acyl group, and a pentylene acyl group.

[0035] Examples of the alkyleneamino group having 1 to 5 carbon atoms include a methyleneamino group, an ethyleneamino group, a propyleneamino group, a butyleneamino group, and a pentyleneamino group.

[0036] In some embodiments of the ion transport molecule, the aforementioned ion transport molecule is selected from the group consisting of the following compounds A to K.

[0037]

[0038]

[0039] The aforementioned ion transport molecules can be inserted into the phospholipid bilayer, and by transporting ions across the membrane, they disrupt the ion balance inside and outside the cell, have significant anti-cancer cell activity, and inhibit the growth of tumor cells. They can be used alone to prepare drugs for treating cancer, or combined or conjugated with other cancer treatment compounds for cancer treatment. In particular, the aforementioned ion transport molecules have a significant effect on divalent metal ions (such as Zn 2+ , Fe 2+ , Cu2+ , Pb 2+ The molecule has a specific response to divalent metal ions, and its ion transmembrane transport activity is greatly increased by coordination with these divalent metal ions, making the molecule gated. For example, in the presence of 0.8-8.0 mM of the aforementioned divalent metal ions, especially Zn 2+ In this case, the aforementioned ion transport molecules change their molecular structure by folding, thereby having a significantly enhanced killing effect on tumor cells, IC 50 (Half inhibition rate) is reduced to 1 / 3 to 1 / 20 of the original.

[0040] The aforementioned cancers include, for example, cervical cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, hepatic cancer, leukemia, and the like.

[0041] In addition, the aforementioned ion transport molecules can disrupt the ion balance inside and outside the cell by transporting ions across the membrane, and thus have a broad spectrum of antibacterial properties. In particular, the aforementioned ion transport molecules have a wide range of antibacterial properties for divalent metal ions (such as Zn 2+ , Fe 2+ , Cu 2+ , Pb 2+ The ion transport activity of the membrane is greatly increased by the coordination with these divalent metal ions. For example, in the presence of 0.8 to 8.0 mM of the aforementioned divalent metal ions, especially Zn 2+ In the case of , the aforementioned ion transport molecules change their molecular structure by folding, thereby having a significantly enhanced killing effect on bacterial cells, thereby significantly reducing the ability of bacteria to grow and proliferate. Therefore, the aforementioned ion transport molecules can be used alone or in combination or conjugated with other drugs that inhibit bacterial growth to prepare drugs for treating bacterial infections.

[0042] As the aforementioned bacterium, for example, gram-positive bacteria or gram-negative bacteria can be used. As the aforementioned gram-positive bacteria, for example, Staphylococcus aureus can be listed. As the aforementioned gram-negative bacteria, for example, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, etc. can be listed.

[0043] Example

[0044] Example 1

[0045]

[0046] (1) Synthesis of Compound 1: Methylhydrazine sulfate (20 g, 138.7 mmol) was dissolved in 100 mL of methanol, and then 4,6-dichloropyrimidine (5 g, 33.5 mmol) was added dropwise. After the solution was stable, anhydrous potassium carbonate (24 g, 173.6 mmol) was added and the temperature was raised to 80°C and heated under argon protection for 24 h. After the reaction was completed, the reaction solution was filtered and washed with 100 mL of chloroform. The filtrate was removed from the solvent under vacuum, and the obtained solid residue was dissolved in 150 mL of chloroform solution and stirred for 10 min. The obtained mixture was washed with water three times (30 mL). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by reverse phase preparative HPLC (MeOH:H2O, from 30:70 to 100:0, 20 min) to obtain Compound 1.

[0047] The physical parameters of compound 1 are: 1 H NMR (400MHz, DMSO-d6) δ7.92 (d, J = 1.0 Hz, 1H), 6.53 (d, J = 1.0 Hz, 1H), 4.47 (s, 4H), 3.14 (s, 6H). 13 C NMR(151MHz,DMSO-d6)δ165.35,156.72,81.82,39.86.MS(ESI):m / z:Calcd.For C6H 13 N6 + [M+H] + :169.1;found:169.1.

[0048] (2) Synthesis of compound 2: 4,6-dichloropyrimidine (2 g, 13.4 mmol) and tributyl (1-ethoxyethylene) tin (11 g, 30.4 mmol) were dissolved in 100 mL of DMF, and then (Ph3)2PdCl2 (0.47 g, 0.66 mmol) was added. The mixture was heated at 80°C under argon protection for 12 h. After the reaction was completed, the reaction solution was poured into a saturated KF solution (100 mL) to produce a brown precipitate. The filter cake was filtered and washed with ether (80 mL × 4). The resulting mixture solution was washed with a saturated NaCl solution (50 mL × 3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel chromatography (petroleum ether: dichloromethane = 3:1 to 1:2) to obtain a yellow solid. The obtained yellow solid was dissolved in acetone (80 mL), and a 2 mol / L hydrochloric acid solution (15 mL) was added, and the mixture was stirred at room temperature for 6 h. The solvent was removed under vacuum to obtain a white solid residue, which was dissolved in dichloromethane (200 mL), washed with a saturated sodium bicarbonate solution (60 mL), and the aqueous layer was washed three times with dichloromethane (100 mL×3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel chromatography (petroleum ether: dichloromethane = 3:1 to 1:2) to obtain compound 2.

[0049] The physical parameters of compound 2 are: 1 H NMR (400MHz, CDCl3) δ9.50 (d, J = 1.4Hz, 1H), 8.44 (d, J = 1.4Hz, 1H), 2.76 (s, 6H). 13 C NMR(151MHz, CDCl3)δ198.47,160.75,159.03,113.35,77.26,77.05,76.84,25.67.MS(ESI):m / z:Calcd.For C8H9N2O2 + [M+H] + :165.1;found:165.1.

[0050] (3) Synthesis of compound A-1: ​​Cholic acid (0.50 g, 1.2 mmol) was dissolved in dry DMF (10 mL), and HBTU (0.48 g, 1.3 mmol) and NMM (0.12 g, 1.2 mmol) were added. The mixture was stirred at room temperature for 0.5 h, and the reaction system turned bright yellow. Then, the aforementioned compound 1 (0.31 g, 1.8 mmol) and NMM (0.12 g, 1.2 mmol) were added to the reaction mixture. The solution was stirred at room temperature for 48 h. After the raw material was completely consumed (detected by TLC), the solvent was removed under vacuum, and the reaction mixture was diluted with brine (30 mL), and then extracted with DCM (30 mL×3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound A-1.

[0051] The physical parameters of compound A-1 are: 1 H NMR (400MHz, DMSO-d6) δ10.03(s,1H),8.03(d,J=0.9Hz,1H),6.21(d,J=1.0H z,1H),4.52(s,2H),4.32(d,J=4.3Hz,1H),4.17–3.98(m,4H),3.80(d,J=3.5H z,1H),3.67–3.56(m,1H),3.23–3.13(m,12H),2.32–1.96(m,6H),1.91–1.61 (m,7H),1.53–1.08(m,14H),0.97(d,J=6.2Hz,5H),0.81(s,4H),0.60(s,3H). 13 C NMR(101MHz,DMSO-d6)δ172.17,165.49,164.03,157.01,81.61,71.48,70. 90,66.71,46.51,46.22,41.97,41.85,40.62,40.57,40.36,40.16,39.95,3 9.74,39.53,39.32,37.33,35.77,35.64,35.36,34.86,31.86,30.86,29.0 3,27.74,26.69,23.29,23.10,17.55,12.82,0.58.MS(ESI):m / z:Calcd.For C 30 H 51 N6O4 + [M+H] + :559.4;found:559.4.

[0052] (4) Synthesis of Compound A: Compound A-1 (0.72 g, 1.2 mmol) was dissolved in EtOH (5 mL), and the aforementioned compound 2 (0.10 g, 0.6 mmol) was added. The mixture was stirred at 90°C for 12 h. The mixture was cooled to room temperature, and the solvent was removed under vacuum. After concentration, it was purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound 4.

[0053] The physical parameters of compound A are: 1 H NMR (600MHz, DMSO-d6) δ10.11(s,2H),9.39(d,J=1.3Hz,1H),8.55(s,1H),8.30(d,J=1.0Hz,2H),4. 32(d,J=4.3Hz,2H),4.15–3.97(m,5H),3.59(t,J=3.2Hz,4H),3.45(s,6H),3.17(d,J=5.0Hz,8H),2 .46(s,6H),2.25–2.07(m,5H),2.01–1.86(m,6H),1.83–1.76(m,2H),1.70–1.58(m,5H),1.57–1.19 (m,26H),0.99(dd,J=49.0,14.2Hz,5H),0.81(s,12H),0.76–0.67(m,6H).MS(ESI):m / z:Calcd.forC 68 H 105 N 14 O8 + [M+H] + :1245.8234; found:1245.8239.

[0054] Example 2

[0055]

[0056] (1) Synthesis of compound B-1: To a solution of 4-(4-aminophenyl)butyric acid (0.80 g, 4.67 mmol) in dichloromethane was added isothiocyanate (4.67 mmol), stirred overnight at room temperature, dried by rotary evaporation, and recrystallized (dichloromethane: petroleum ether = 1:1) to obtain a white solid compound B-1.

[0057] The physical parameters of compound B-1 are: 1H NMR(600MHz, CDCl3)δ11.93(s,1H),9.32(s,1H),7.86(s,1H),7.55–7.21(m,2H),7.06 (dt,J=7.5,1.0Hz,2H),3.04(s,3H),2.53(t,J=1.0Hz,2H),2.26(m,2H),1.76(m,2H). 13 C NMR(100MHz, CDCl3)δ181.45,177.33,139.32,134.58,127.69,117.53,34.33,33.50,30.05,26.63.LR-MS(ESI)(m / z):[M] + Calculate for C 12 H 16 N2O2S:252.09,found:252.09.

[0058] (2) Synthesis of compound B-2: B-1 (0.14 g, 1.4 mmol) was dissolved in dry DMF (10 mL), and HBTU (0.55 g, 1.5 mmol) and NMM (0.15 g, 1.5 mmol) were added. The mixture was stirred at room temperature for 0.5 h, and the reaction system turned bright yellow. Compound 1 (0.34 g, 2.0 mmol) and NMM (0.15 g, 1.5 mmol) were then added to the reaction mixture. The solution was stirred at room temperature for 48 h. After the raw material was completely consumed (detected by TLC), the solvent was removed under vacuum, and the reaction mixture was diluted with brine (30 mL), and then extracted with DCM (30 mL×3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound B-2.

[0059] The physical parameters of compound B-2 are: 1 H NMR (600MHz, DMSO-d6) δ9.44(d,J=0.7Hz,1H),8.16(d,J=1.4Hz,0H),7.77(q,J=4.3Hz,1H),7.48–7.39(m,1H),7.10(dt,J=7.9,1.0Hz,1H),5.39 (dd,J=37.0,8.6Hz,1H),3.42(s,1H),3.25(s,1H),3.01(d,J=4.4Hz,1H),2.63(tt,J=8.2,1.0Hz,1H),2.33(t,J=8.9Hz,1H),1.95–1.70(m,1H). 13C NMR (125MHz, DMSO-d6) δ180.36,171.44,160.49,157.37,154.81,137.93,135.89,129. 21,123.51,88.77,39.84,39.06,34.84,34.79,31.26,25.16.MS(ESI):m / z:Calcd.For C 18 H 27 N8OS + [M+H] + :403.2;found:403.2.

[0060] (4) Synthesis of Compound B: Compound B-2 (0.84 g, 2.1 mmol) was dissolved in EtOH (5 mL), and Compound 2 (0.11 g, 0.6 mmol) was added. The mixture was stirred at 90°C for 12 h. The mixture was cooled to room temperature, and the solvent was removed under vacuum. After concentration, compound B was purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min).

[0061] The physical parameters of compound B are: 11H NMR (600 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.56 (d, J = 1.4 Hz, 1H), 9.45 (d, J = 13.0 Hz, 3H), 8.25 (d, J = 1.4 Hz, 1H), 8.22 (d, J = 1.5 Hz, 2H), 7.77 (q, J = 4.3 Hz, 2H), 7.46 (d, J = 1.4 Hz, 2H), 7.45 (d, J = 1.2 Hz, 3H), 5.83 (d, J = 1.4 Hz, 1H), 5.77 (d, J = 1.4 Hz, 1H), 3.79 (s, 3H), 3.55 (s, 3H), 3.34 (s, 3H), 3.25 (s, 3H), 3.01 (d, J = 4.4 Hz, 7H), 2.85 (td, J = 8.3, 0.9 Hz, 5H), 2.58–2.55 (m, 2H), 2.55–2.51 (m, 2H), 2.33 (s, 6H). 13C NMR (125 MHz, DMSO-d6) δ 181.11, 171.12, 171.09, 158.89, 158.66, 158.64, 158.55, 157.94, 157.54, 156.92, 156.09, 156.07, 155.95, 155.90, 155.49, 155.47, 148.46, 148.43, 148.30, 148.23, 137.92, 134.37, 134.33, 129.23, 123.35, 114.82, 85.48, 84.79, 40.00, 39.94, 38.77, 38.74, 38.54, 38.48, 36.59, 31.33, 30.95, 30.82, 14.90, 14.87, 14.85. MS (ESI): m / z: Calcd. For C 44 H 57 N 18 O2S2 + [M + H] + : 933.4348; found: 933.4340.

[0062] Example 3

[0063]

[0064] (1) Synthesis of compound C-1: 4-dimethylaminopyridine (DMAP, 0.20 g, 1.70 mmol) was added to a dichloromethane solution (50 mL) of 4-carboxybenzo-18-crown-6 (1.03 g, 2.89 mmol) and 4-butyn-1-ol (0.49 g, 5.78 mmol). The mixture was stirred at 0°C for 10 min and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 0.54 g, 2.89 mmol) was added. The reaction was carried out at room temperature for 36 h, the solvent was removed under vacuum, 100 mL of dichloromethane was added to dissolve the product, and it was washed with saturated sodium chloride solution (100 mL×3), the organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel column chromatography (dichloromethane: methanol = 99:1) to obtain a white solid compound C-1.

[0065] The physical parameters of compound C-1 are: 1 H NMR(500MHz,Chloroform-d)δ7.65(dd,J=8.6,1.8Hz,1H),7.51(d,J=2.0Hz,1H),6.93(d,J=8.7Hz,1H),4.28(t,J=6.2Hz,2H),4.19(t ,J=5.0Hz,4H),3.83–3.77(m,4H),3.66(s,7H),3.66(s,4H),2.57(td,J=6.5,3.0Hz,2H),2.04(t,J=3.0Hz,1H),1.96(p,J=6.3Hz,2H). 13 C NMR(125MHz,Chloroform-d)δ166.30,153.28,148.61,125.30,123.79,114.36,113.64,83.46,70.74,70.73, 70.59,70.58,70.48,70.45,69.70,69.68,69.14,69.11,68.91,65.07,28.03,15.35.MS(ESI):m / z:Calcd.For C 22 H 31 8Na + [M+Na] + :445.2; found:445.2.

[0066] (2) Synthesis of compound C-2: 3-Hydroxypropionic acid (0.11 g, 1.2 mmol) was dissolved in dry DMF (10 mL), and HBTU (0.48 g, 1.3 mmol) and NMM (0.12 g, 1.2 mmol) were added. The mixture was stirred at room temperature for 0.5 h, and the reaction system turned yellow. Compound 1 (0.31 g, 1.8 mmol) and NMM (0.12 g, 1.2 mmol) were then added to the reaction mixture. The solution was stirred at room temperature for 48 h. After the raw material was completely consumed (detected by TLC), the solvent was removed under vacuum, and the reaction mixture was diluted with brine (30 mL), and then extracted with DCM (30 mL×3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound C-2.

[0067] The physical parameters of compound C-2 are: 1 H NMR (600MHz, DMSO-d6) δ9.77(s,1H),8.15(d,J=1.4Hz,1H),6.41(d,J=1.4Hz,1H),4.82–4.73(m ,1H),4.64–4.47(m,2H),3.72(q,J=5.8Hz,2H),3.34(s,3H),3.23(s,3H),2.44(t,J=5.7Hz,3H). 13 C NMR(125MHz,DMSO-d6)δ171.28,159.18,157.37,155.23,84.48,57.91,40.84,39.89,37.53.MS(ESI):m / z:Calcd.For C9H 17 O6N2 + [M+H] + :241.1; found:241.2.

[0068] (3) Synthesis of compound C-3: Compound C-2 (0.50 g, 2.1 mmol) was dissolved in EtOH (5 mL), and compound 2 (0.11 g, 0.6 mmol) was added. The mixture was stirred at 90°C for 12 h. The mixture was cooled to room temperature, and the solvent was removed under vacuum. After concentration, it was purified by reverse phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound C-3.

[0069] The physical parameters of compound C-3 are: 1H NMR (600MHz, DMSO-d6) δ9.77 (s, 1H), 9.56 (d, J = 1.4Hz, 1H), 9.47 (s, 1H), 8.25 ( d,J=1.4Hz,1H),8.22(d,J=1.5Hz,2H),5.83(d,J=1.4Hz,1H),5.77(d,J=1.4Hz ,1H),4.77(td,J=6.0,0.7Hz,2H),3.79(s,2H),3.70(dq,J=12.6,5.8Hz,4H),3 .55(s,2H),3.34(s,3H),3.25(s,2H),2.44(td,J=5.7,1.3Hz,4H),2.33(s,6H). 13 CNMR(125MHz,DMSO-d6)δ171.28,171.13,158.89,158.64,157.94,157.24,156.94,156.07,155.90,155.49,14 8.43,148.23,114.82,84.79,57.91,39.98,39.91,38.74,38.48,37.93,37.53,14.85.MS(ESI):m / z:Calcd.For C 26 H 37 N 14 O4 + [M+H] + :609.3; found:609.3.

[0070] (4) Synthesis of compound C-4: 4-Toluenesulfonyl chloride (2.0 g, 10.5 mmol) and DMAP (catalytic amount) were added to a dichloromethane solution of compound C-3 (1.27 g, 2.1 mmol). Triethylamine (1.5 mL, 11.0 mmol) was added under ice bath conditions. The reaction was continued for 18 hours, and the resulting yellow solution was washed with water (40 mL × 3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel column chromatography (petroleum ether: dichloromethane = 2:8) to obtain a light yellow oily compound C-4.

[0071] The physical parameters of compound C-4 are: 1H NMR(600MHz,DMSO-d6)δ9.56(d,J=1.4Hz,1H),8.23(dd,J=14.9,1.4Hz,3H),7.77–7.74(m,4H),7.40–7.36(m,4H),5.83(d,J=1.4Hz ,1H),5.77(d,J=1.4Hz,1H),4.19(t,J=6.5Hz,4H),3.79(s,3H),3.55(s,3H),3.34(s,3H),2.69(td,J=6.5,1.4Hz,4H),2.33(s,6H). 13 C NMR (125MHz, DMSO-d6) δ170.07,169.97,158.89,158.64,157.95,157.42,156.94,156.07,155.90,155.49,148.43,148.23,142.72,13 4.28,129.76,127.76,114.82,84.79,65.22,65.18,39.98,39.91,38.74,38.48,35.58,35.56,21.50,14.85.MS(ESI):m / z:Calcd.For C 40 H 49 N 14 O8S2 + [M+H] + :916.3; found:916.3.

[0072] (5) Synthesis of compound C-5: Azide trimethylsilane (0.12 g, 1.00 mmol) was added to a DMF (15 mL) solution of compound C-4 (0.55 g, 0.60 mmol), and the mixture was stirred at 80°C overnight. After cooling to room temperature, dichloromethane was added to the mixture, and then washed with water (30 mL×3). The organic phase was dried over anhydrous Na2SO4 and concentrated to obtain a light yellow solid compound C-5.

[0073] The physical parameters of compound C-5 are: 1 H NMR (600MHz, DMSO-d6) δ9.56(d,J=1.4Hz,1H),8.60(s,2H),8.25(d,J=1.4Hz,1H),8.20(d,J=1.6Hz, 2H), 7.01 (d, J = 1.4Hz, 2H), 3.51 (s, 6H), 3.31 (t, J = 5.8Hz, 10H), 2.66 (t, J = 5.8Hz, 4H), 2.39 (s, 6H). 13C NMR (125MHz, DMSO-d6) δ172.10,157.95,157.34,156.65,155.47,155.41,147. 43,116.94,84.82,47.16,40.17,39.36,33.16,15.27.MS(ESI):m / z:Calcd.For C 40 H 49 N 14 O8S2 + [M+H] + :659.3246; found:659.3240.

[0074] (6) Synthesis of compound C: Compound C-1 (84.4 mg, 0.20 mmol) and compound C-5 (65.8 mg, 0.10 mmol) were dissolved in a mixed solvent of dichloromethane / acetonitrile (4:1, 3 mL), and Cu(CH3CN)4PF6 (55.6 mg, 0.15 mmol) was added, and stirred at room temperature for 12 h. The solvent was removed in vacuo, and the residue was purified by reverse phase high performance liquid chromatography (mobile phase: MeOH / H2O=50 / 50-100 / 0, 15 min) to obtain yellow solid compound C-5.

[0075] The physical parameters of compound C are: 1 H NMR (600MHz, DMSO-d6) δ9.71(s,1H),9.56(d,J=1.5Hz,1H),9.52(s,1H),8.25(d,J=1.5Hz,1H),8.22( d,J=1.5Hz,2H),7.65(dd,J=8.7,2.0Hz,2H),7.51–7.46(m,4H),6.94(d,J=8.7Hz,2H),5.83(d,J=1.5 Hz,1H),5.77(d,J=1.5Hz,1H),4.39–4.29(m,8H),4.17(t,J=5.0Hz,8H),3.83–3.77(m,11H),3.63(d, J=1.8Hz,24H),3.55(s,3H),3.34(s,3H),2.84–2.75(m,8H),2.33(s,6H),2.23(tt,J=8.6,6.7Hz,4H). 13C NMR(125MHz,DMSO-d6)δ169.28,169.10,165.94,157.90,157.81,157.41,157.30,156.95,1 56.15,155.92,155.15,152.70,149.57,149.27,148.15,147.33,125.30,125.07,123.79,1 16.26,114.20,113.96,88.84,70.69,70.54,70.40,69.67,69.65,68.91,68.81,64.97,45. 68,39.06,38.95,37.89,37.43,34.33,34.31,27.25,22.72,14.96.MS(ESI):m / z:Calcd.For C 70 H 95 N 20 O 18 + [M+H] + :1503.7128; found:1503.7115.

[0076] Example 4

[0077]

[0078] (1) Synthesis of compound D-1: Dissolve tetraethylene glycol (4 mL, 23 mmol) in dichloromethane, add triethylamine (30 mL) in an ice bath, add a dichloromethane solution (50 mL) of p-toluenesulfonyl chloride (12 g, 64 mmol), and react at room temperature for 4 hours. Remove the solvent under vacuum, and extract with water (30 mL×5). Collect the organic phase, and wash it three times with 240 mL of 2 mol / L hydrochloric acid aqueous solution, 5% NaHCO3 aqueous solution (150 mL), and 150 mL of water, respectively. Then, dry the collected organic phase with anhydrous Na2SO4, concentrate, and purify it on a silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain compound D-1.

[0079] The physical parameters of compound D-1 are: 1 H NMR (400MHz, CDCl3) δ: 2.45 (s, 6H), 3.57 (s, 8H), 3.68 (t, J = 4.7Hz, 4H), 4.16 (t, J = 4.7Hz, 4H), 7.35 (d, J = 8.2Hz, 4H), 7.80 (d, J = 8.2Hz, 4H). 13C NMR (100MHz, CDCl3) δ: 144.8, 133.0, 129.8, 128.0, 70.7, 70.5, 69.3, 68.7, 21.6. LR-MS (ESI-TOF): Calcd.For C 22 H 30 O9S2[M]+:502.1.Found:502.1.

[0080] (2) Synthesis of compound D-2: K2CO3 (7.05 g, 50.00 mmol) was added to an acetonitrile (260 mL) solution containing p-hydroxybenzonitrile (2.24 g, 18.80 mmol) and compound D-1 (4.64 g, 9.25 mmol), and the resulting mixture was heated to reflux for 3 days. The mixture was filtered and the filtrate was purified by silica gel column chromatography (dichloromethane:ethyl acetate=9:1) to obtain a white solid D-2.

[0081] The physical parameters of compound D-2 are: 1 H NMR(400MHz, CDCl3)δ:7.62-7.53(m,4H),7.00-6.92(m,4H),4.20-4.13(m,4H),3.90-3.83(m,4H),3.77-3.64(m,8H). 13 C NMR(100MHz, CDCl3)δ:162.08,133.97,119.17,115.32,104.17,70.91,70.66,69.44,67.76.LR-MS(ESI-TOF):Calcd.For C 22 H 24 N2O5Na[M+Na] + :419.2.Found:419.3.

[0082] (3) Synthesis of compound D-3: A 1M LiAlH4 solution in THF (75 mL, 75 mmol) was added dropwise to a THF solution (70 mL) of compound D-2 (4.72 g, 15.83 mmol) under ice bath conditions. The reaction solution was stirred until it turned yellow and heated to reflux for 4 hours. The reaction solution was cooled to room temperature, and 2.88 mL of H2O, 2.88 mL of 15% NaOH aqueous solution and 8.58 mL of H2O were slowly added respectively. The filtrate was filtered and concentrated to obtain a white solid D-3.

[0083] The physical parameters of compound D-3 are: 1H NMR (400MHz, CDCl3) δ: 7.23-7.16 (m, 4H), 6.89-6.84 (m, 4H), 4.10 (dd, J = 5.7, 4.1Hz, 4H), 3.84 (dd, J = 5.8, 4.0Hz, 4H), 3.75-3.65 (m, 8H). 13 CNMR(100MHz, CDCl3)δ:157.74,135.52,128.27,114.69,70.82,70.67,69.76,67.48,45.81.LR-MS(ESI-TOF):Calcd.For C 22 H 33 N2O5[M+H] + :405.2.Found:405.2.

[0084] (4) Synthesis of compound D-4: Anhydrous potassium carbonate (40.00 g, 289.8 mmol), 4-hydroxy-2,6-dicarboxypyridine (15.00 g, 81.96 mmol), benzyl bromide (30.00 mL, 262.2 mmol) were dissolved in 250 mL of acetone and refluxed at 70°C for 12 h under argon protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and the filtrate was taken. After removing the solvent under vacuum, it was dissolved in dichloromethane. The resulting mixture solution was washed with 5% potassium hydroxide aqueous solution (350 mL) and saturated brine (200 mL), respectively, and the organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel chromatography (petroleum ether: ethyl acetate = 9:1) to obtain a light yellow solid compound D-4.

[0085] The physical parameters of compound D-4 are: 1 H NMR(400MHz, CDCl3)δ:7.84(s,2H),7.47-7.45(m,5H),7.42-7.29(m,10H),5.43(s,4H),5.17(s,2H). 13 C NMR(100MHz, CDCl3)δ:166.52,164.39,150.04,135.04,130.28,124.56,114.81,69.25.LR-MS(ESI-TOF):Calcd.ForC 28 H3NO5Na[M+Na] + :476.1.Found:476.2.

[0086] (5) Synthesis of compound D-5: Compound D-4 (3.0 g, 6.6 mmol) was dissolved in ethanol (200 mL) solution, KOH (3.0 g, 53.5 mmol) was added, and the mixture was stirred and refluxed at 90°C overnight. After the reaction was completed, the mixture was cooled to room temperature, and the reaction mixture was filtered, and the filter cake was washed with ethanol (50 mL × 3). The filter cake was dissolved in 100 mL of water again, and trifluoroacetic acid was added dropwise to the solution until a white solid was completely precipitated. The white solid was collected by suction filtration and dried to obtain a white solid product D-5.

[0087] The physical parameters of compound D-5 are: 1 H NMR(400MHz, CDCl3)δ:7.77(s,2H),7.58-7.28(m,5H),5.41(s,2H). 13 C NMR (100MHz, CDCl3) δ: 164.3, 162.1, 150.2, 135.9, 129.0, 128.7, 128.3, 113.6, 70.8. LR-MS (ESI-TOF): Calcd.For C 14 H 11 NO5K[M+K] + :312.0.Found:312.0.

[0088] (6) Synthesis of compound D-6: Compound D-5 (3.0 g, 7.44 mmol) and triethylamine (2.5 mL) dissolved in dichloromethane solution (1500 mL) were added to a dichloromethane solution (15 mL) of compound D-3 (2.05 g, 7.44 mmol) under ice bath conditions. The reaction was carried out in an anhydrous environment. The reaction was carried out at room temperature for 12 hours, and the solvent was removed under vacuum. The crude product was purified by silica gel column chromatography (dichloromethane: methanol = 96:4) to obtain white solid compound D-6.

[0089] The physical parameters of compound D-6 are: 1 H NMR (400MHz, CDCl3) δ: 7.99 (s, 2H), 7.89 (s, 2H), 7.50-, 7.32 (m, 5H), 7.11 (d, J = 8.5Hz, 4H), 6.76 (d, J = 8.4Hz ,4H),5.28(s,2H),4.56(d,J=,5.5Hz,4H),4.08(dd,J=5.8,3.5Hz,4H),3.94-3.84(m,4H),3.77-3.64(m,8H). 13C NMR(100MHz, CDCl3)δ:163.13,158.28,150.65,135.15,129.90,128.84,128.57,127.77, ,114.72,111.64,77.23,70.90,70.63,69.66,67.64,43.00.LR-MS(ESI-TOF):Calcd.ForC 36 H 39 N3O 8, Na[M+Na] + :664.3.Found:664.4.

[0090] (7) Synthesis of compound D-7: Compound D-6 (1.28 g, 2.00 mmol) was dissolved in a mixed solvent of methanol (30 mL) and dichloromethane (30 mL), and then Pd / C (10%, 0.10 g) was added. The reaction solution was reacted at room temperature under a hydrogen pressure of 70 atm for 15 hours, and the palladium carbon catalyst was filtered off and rinsed with tetrahydrofuran. The filtrate was dried to obtain a white solid compound D-7.

[0091] The physical parameters of compound D-7 are: 1 H NMR(400MHz,DMSO-d6,)δ:11.44(s,1H),9.69(t,J=,6.3Hz,2H),7.58(s,2H),7.21(d,J=8.3Hz,4H),6 .89(d,J=8.4Hz,4H),4.54(d,J=6.4Hz,,4H),4.09-3.96(m,4H),3.78-3.67(m,4H),3.58-3.48(m,8H). 13 C NMR(100MHz,DMSO-d6,)δ:,163.03,157.38,150.72,131.20,128.11,114.20,111.45,69.89,68.83,67.16,40.97.LR-,MS(ESI-TOF):Calcd.For C 29 H 33 N3O8Na[M+Na] + :574.2.Found:574.2.

[0092] (8) Synthesis of compound D-8: Compound D-7 (1.0 g, 1.8 mmol) was dissolved in DMF (100 mL), methyl-3-bromopropionate (0.3 g, 1.8 mmol) and anhydrous potassium carbonate (2.5 g, 18 mmol) were added, and the mixture was stirred at 90°C for 12 h under argon protection. After the reaction was completed, the mixture was filtered, and the filter cake was rinsed with dichloromethane (100 mL). The solvent was removed under vacuum, and the mixture was purified by silica gel column chromatography (dichloromethane: methanol = 97:3) to obtain a white solid compound D-8.

[0093] The physical parameters of compound D-8 are: 1 H NMR (600MHz, DMSO-d6) δ8.30(t,J=6.0Hz,2H),7.85(s,2H),7.23(dt,J=8.8,1.1Hz,4H),6.82–6.76(m,4H),4.62(dt,J=6.0,0.9Hz,4H ),4.27(t,J=6.6Hz,2H),4.16–4.09(m,6H),3.72(t,J=5.0Hz,4H),3.63(d,J=3.3Hz,8H),2.73(t,J=6.6Hz,2H),1.22(t,J=6.6Hz,3H). 13 C NMR (125MHz, DMSO-d6) δ170.63,165.50,164.13,158.23,146.81,132.94,128.39,115.00,109. 81,70.65,70.41,69.83,67.92,63.34,60.45,44.32,34.83,14.15.LC-MS(ESI-TOF):Calcd.For C 34 H 41 N3O 10 Na + [M+Na] + :674.3.Found:674.2.

[0094] (9) Synthesis of compound D-9: Compound D-8 (1.0 g, 1.6 mmol) was dissolved in methanol (20 mL), 10 mL of 2 M NaOH solution was added, and the mixture was stirred for 1 h. Then, 6 M HCl solution (20 mL) was added and stirred for 1 h. After the reaction was completed, the solvent was removed in vacuo. The remaining white solid mixture was dissolved in dichloromethane (50 mL) and extracted with water (15 mL×3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel chromatography (dichloromethane: methanol = 95:5) to obtain a light yellow oily compound D-9.

[0095] The physical parameters of compound D-9 are:1 H NMR (600MHz, DMSO-d6) δ8.30(t,J=6.0Hz,2H),7.85(s,2H),7.23(dt,J=8.8,1.1Hz,4H),6.82–6.76(m,4H),4.62(dt,J= 6.0,0.9Hz,4H),4.27(s,2H),4.11(t,J=5.0Hz,4H),3.72(t,J=5.0Hz,4H),3.63(d,J=3.3Hz,8H),2.70(t,J=6.3Hz,2H). 13 C NMR(125MHz,DMSO-d6)δ172.41,165.53,164.13,158.23,146.81,132.94,128.39,115.0 0,109.81,70.65,70.41,69.83,67.92,63.60,44.32,34.20.LC-MS(ESI-TOF):Calcd.For C 32 H 37 N3O 10 Na + [M+Na] + :646.2371.Found:646.2362.

[0096] (10) Synthesis of Compound D-10: Compound A-1 was synthesized by following the method described in Example 1.

[0097] The physical parameters of compound D-10 are: 1 H NMR(500MHz,DMSO-d6)δ9.75(s,1H),8.30(t,J=6.0Hz,2H),8.15(d,J=1.5Hz,1H), 7.85(s,2H),7.23(dt,J=8.8,1.1Hz,4H),6.82–6.76(m,4H),6.41(d,J=1.5Hz,1H) ,4.65–4.58(m,5H),4.55(d,J=8.6Hz,1H),4.27–4.21(m,2H),4.11(t,J=5.0Hz,4H ),3.72(t,J=5.0Hz,4H),3.63(d,J=1.7Hz,8H),3.34(s,3H),2.62(t,J=6.0Hz,2H). 13C NMR (125MHz, DMSO-d6) δ170.36,165.70,164.13,159.18,158.23,157.40,155.23,146.81,132.94,128.39,115. 00,109.62,84.48,70.65,70.41,69.83,67.92,63.81,44.32,40.84,39.92,36.12.LC-MS(ESI-TOF):Calcd.For C 38 H 47 N9O9Na + [M+Na] + :796.3389.Found:796.3378.

[0098] (11) Synthesis of Compound D: Compound A was synthesized by following the method described in Example 1.

[0099] The physical parameters of compound D are: 1 H NMR (600MHz, DMSO-d6) δ9.75(s,1H),9.59(s,1H),9.56(d,J=1.5Hz,1H),8.30(t,J=6.0Hz,4H),8.25(d,J=1.5 Hz,1H),8.22(d,J=1.5Hz,2H),7.85(s,4H),7.23(dt,J=8.8,1.1Hz,8H),6.84–6.76(m,8H),5.83(d,J=1.5Hz, 1H),5.77(d,J=1.5Hz,1H),4.62(dt,J=6.1,1.0Hz,8H),4.24(td,J=5.9,1.1Hz,4H),4.11(t,J=5.0Hz,8H),3. 79(s,3H),3.72(t,J=5.0Hz,8H),3.63(d,J=1.7Hz,16H),3.55(s,3H),3.34(s,3H),2.62(s,4H),2.33(s,6H). 13C NMR(125MHz,DMSO-d6)δ169.36,169.32,165.36,162.65,158.16,157.90,157.81,1 57.56,157.30,156.98,156.15,155.92,155.15,149.57,149.27,148.79,133.77,1 28.61,116.26,115.00,107.85,88.84,70.64,70.40,69.94,67.70,63.40,63.35,4 4.53,39.04,38.96,37.89,37.43,34.93,34.59,14.96.LC-MS(ESI-TOF):Calcd.For C 84 H 98 N 20 O 18 Na + [M+Na] + :1699.7401.Found:1699.7421.

[0100] Example 5

[0101]

[0102] (1) Synthesis of Compound E: Compound E was synthesized by referring to the method of Compound A in Example 1.

[0103] The physical parameters of compound E are: 1 H NMR(500MHz,Chloroform-d)δ9.45(d,J=1.4Hz,1H),8.79(s,1H),8.71(s,1H),8.34(dd,J=4.2,1.5Hz,2H),7.74( d,J=8.8Hz,2H),7.26(t,J=5.9Hz,2H),6.87(d,J=1.4Hz,2H),6.30(t,J=6.0Hz,2H),6.05(t,J=5.8Hz,4H),4.31(d td,J=8.6,6.0,4.9Hz,2H),3.89–3.80(m,16H),3.78(s,3H),3.66(s,3H),3.57(s,3H),3.40(s,3H),2.44(s,5H), 2.27(td,J=8.4,0.9Hz,4H),1.81(td,J=8.8,6.0Hz,2H),1.75–1.60(m,4H),1.60–1.55(m,1H),1.55–1.34(m,4H). 13C NMR (125MHz, DMSO-d6) δ172.84,172.53,172.40,171.55,170.93,170.88,170.84,170. 44,159.91,159.69,158.31,158.05,158.03,156.38,156.28,155.97,148.45,148.20,1 16.15,85.03,53.65,53.64,43.44,43.06,42.72,42.63,42.54,40.18,40.03,39.53,3 9.28,35.50,35.48,30.82,30.76,24.35,24.00,15.16,15.11.MS(ESI):m / z:Calcd.For C 50 H 71 N 24 O 12 + [M+H] + :1199.5678; found:1199.5670.

[0104] Example 6

[0105]

[0106] (1) Synthesis of compound F-1: A-1 (0.24 g, 0.4 mmol) was dissolved in EtOH (5 mL), and then 2 (0.10 g, 0.6 mmol) was added. The mixture was stirred at 90 °C for 12 h. The mixture was cooled to room temperature and the solvent was removed under vacuum. After the system was concentrated, it was purified by reverse phase preparative chromatography (MeOH:H2O, from 60:40 to 100:0, 15 min) to obtain compound F-1.

[0107] The physical parameters of compound F-1 are: 1H NMR (400MHz, DMSO-d6) δ10.24(s,1H),9.50(s,1H),8.36(d,J=25.3Hz,2H),6.12(s,1H),4.36(d,J=4 .2Hz,1H),4.04(dd,J=11.5,3.4Hz,2H),3.61(d,J=16.1Hz,2H),3.51(s,3H),3.23–3.15(m,5H),2.7 0(s,3H),2.49(s,2H),2.28–1.97(m,5H),1.91(q,J=5.2Hz,1H),1.78(dt,J=8.4,4.0Hz,1H),1.67–1 .60(m,2H),1.54–1.23(m,12H),1.12–0.98(m,2H),0.81(s,5H),0.76(d,J=6.2Hz,3H),0.45(s,3H). 13 CNMR(101MHz,DMSO-d6)δ199.30,172.35,164.73,164.20,164.05,159.30,158.89,157.32,112 .62,84.73,71.37,70.90,66.67,49.07,46.05,45.97,41.96,41.71,40.56,40.35,40.14,39.9 3,39.73,39.52,39.31,38.86,37.35,36.26,35.77,35.36,35.22,34.84,31.65,31.24,30.85, 30.35,28.97,27.40,26.64,26.20,23.19,23.07,17.23,16.55,12.68.MS(ESI):m / z:Calcd.For C 38 H 57 N8O5 + [M+H] + :705.4454; found:705.4451.

[0108] (2) Synthesis of Compound F The synthesis method of Compound A in Example 1 was used as reference.

[0109] The physical parameters of compound F are: 11H NMR (600 MHz, DMSO-d6) δ 10.25–10.09 (m, 2H), 9.40 (t, J = 1.3 Hz, 1H), 9.00 (dd, J = 11.0, 1.3 Hz, 1H), 8.36–8.26 (m, 3H), 6.04 (d, J = 38.7 Hz, 2H), 5.79 (d, J = 3.3 Hz, 1H), 5.38–5.29 (m, 1H), 4.31 (t, J = 4.0 Hz, 2H), 4.05–3.98 (m, 4H), 3.64 (dd, J = 42.1, 19.2 Hz, 5H), 3.51–3.42 (m, 6H), 3.23–3.14 (m, 9H), 2.48–2.39 (m, 6H), 2.20 (q, J = 12.3 Hz, 2H), 2.10 (d, J = 11.5 Hz, 4H), 2.00 (dt, J = 12.5, 6.7 Hz, 3H), 1.91 (d, J = 5.0 Hz, 2H), 1.78 (d, J = 5.2 Hz, 2H), 1.70–1.20 (m, 40H), 1.11–1.04 (m, 2H), 0.97 (d, J = 6.8 Hz, 1H), 0.89–0.69 (m, 19H). 13 13C NMR (125 MHz, DMSO-d6). δ 171.72, 158.41, 158.19, 157.90, 157.88, 157.38, 157.30, 156.15, 155.92, 155.18, 155.15, 149.57, 149.27, 116.26, 116.22, 88.84, 87.08, 74.12, 70.04, 68.81, 68.79, 48.86, 46.96, 44.16, 41.08, 40.28, 38.98, 37.89, 37.63, 37.43, 36.30, 35.70, 35.53, 34.93, 34.39, 32.91, 30.94, 30.80, 30.08, 30.04, 27.42, 24.71, 20.38, 18.42, 14.96, 13.84, 13.33. MS (ESI): m / z: Calcd. for C 82 H 121 N 22 O8 + [M + H] + : 1541.9733; found: 1541.9734.

[0110] Example 7

[0111]

[0112] The synthesis of compound G was carried out according to the synthesis of compound C in Example 3.

[0113] The physical parameters of compound G are: 1 H NMR(500MHz,DMSO-d6)δ9.71(s,2H),9.56(d,J=1.5Hz,2H),8.27–8.20(m,5H),7.65(dd,J=8.7 ,2.0Hz,2H),7.51–7.46(m,4H),6.94(d,J=8.7Hz,2H),6.68(d,J=1.7Hz,1H),5.83(d,J=1.5Hz, 2H),4.34(dt,J=21.2,6.8Hz,8H),4.17(t,J=5.0Hz,8H),3.83–3.77(m,11H),3.63(d,J=1.8Hz ,24H),3.55(s,9H),3.34(s,5H),2.84–2.75(m,8H),2.33(s,11H),2.23(tt,J=8.6,6.7Hz,4H). 13 C NMR(125MHz,DMSO-d6)δ169.10,165.94,158.41,158.19,157.90,157.88,157.41,157.30,156 .15,155.92,155.18,155.15,152.70,149.57,149.27,148.15,147.33,125.30,125.07,123.79 ,116.26,116.22,114.20,113.96,88.84,87.08,70.69,70.54,70.40,69.67,69.65,68.91,68 .81,64.97,45.68,38.95,37.89,37.43,34.31,27.25,22.72,14.96.MS(ESI):m / z:Calcd.forC 84 H 111 N 28 O 18 + [M+H] + :1799.8626; found:1799.8621.

[0114] Example 8

[0115]

[0116] The synthesis of compound H was carried out according to the synthesis method of compound B in Example 2.

[0117] The physical parameters of compound H are:1 H NMR(600MHz,DMSO-d6)δ9.77(s,2H),9.56(d,J=1.4Hz,2H),9.44(s,2H),8.27–8.20 (m,5H),7.77(q,J=4.3Hz,2H),7.44–7.38(m,4H),7.10(dt,J=7.9,1.0Hz,4H),6.68( d,J=1.6Hz,1H),5.83(d,J=1.4Hz,2H),3.79(s,3H),3.55(s,9H),3.34(s,6H),3.01( d,J=4.4Hz,6H),2.63(tt,J=8.2,1.1Hz,4H),2.37–2.29(m,16H),1.89–1.79(m,4H). 13 C NMR(125MHz,DMSO-d6)δ181.11,171.82,158.66,158.64,158.55,157.94,157.93 ,157.24,156.09,155.95,155.90,155.49,155.47,148.46,148.30,148.23,137.8 6,134.80,128.84,123.38,114.82,114.76,85.48,84.79,39.94,38.77,38.54,3 8.48,35.03,34.76,31.33,26.49,14.90,14.87,14.85.MS(ESI):m / z:Calcd.forC 58 H 73 N 26 O2S2 + [M+H] + :1229.5846; found:1229.5838.

[0118] Example 9

[0119]

[0120] The synthesis of compound I refers to compound D in Example 4.

[0121] The physical parameters of compound I are: 1H NMR(600MHz,DMSO-d6)δ9.75(s,2H),9.56(d,J=1.5Hz,2H),8.30(t,J=6.0Hz,4H),8.27–8.20(m,5 H),7.85(s,4H),7.23(dt,J=8.8,1.1Hz,8H),6.82–6.76(m,8H),6.68(d,J=1.6Hz,1H),5.83(d,J=1 .5Hz,2H),4.62(dt,J=6.1,1.0Hz,8H),4.24(s,4H),4.11(t,J=5.0Hz,8H),3.79(s,3H),3.72(t,J= 5.0Hz,8H),3.63(d,J=1.7Hz,16H),3.55(s,9H),3.34(s,5H),2.62(t,J=5.9Hz,4H),2.33(s,12H). 13 C NMR(125MHz,DMSO-d6)δ169.36,165.36,162.65,158.41,158.19,158.16,157.90,1 57.88,157.56,157.30,156.15,155.92,155.18,155.15,149.57,149.27,148.79,1 33.77,128.61,116.26,116.22,115.00,107.85,88.84,87.08,70.64,70.40,69.94 ,67.70,63.40,44.53,38.96,37.89,37.43,34.93,14.96.MS(ESI):m / z:Calcd.for C 98 H 115 N 28 O 18 + [M+H] + :1972.8973; found:1972.8977.

[0122] Example 10

[0123]

[0124] The synthesis of compound J refers to compound A in Example 1.

[0125] The physical parameters of compound J are: 1H NMR (600MHz, DMSO-d6) δ9.77(s,2H),9.56(d,J=1.4Hz,1H),8.23(dd,J=15.0,1.4Hz,3H),5.83(d,J=1.4Hz,2H),3.55(s, 6H),3.34(s,6H),2.33(s,6H),2.27(t,J=8.8Hz,4H),1.58(tt,J=8.5,7.5Hz,4H),1.35–1.24(m,17H),0.94–0.85(m,6H). 13 C NMR(125MHz,DMSO-d6)δ171.98,158.64,157.93,157.24,155.90,155.49,148.23,114.76,84.79 ,39.94,38.48,35.16,31.17,28.71,28.42,24.95,22.62,14.85,14.05.MS(ESI):m / z:Calcd.for C 36 H 57 N 14 O2 + [M+H] + :717.4783; found:717.4775.

[0126] Embodiment 11

[0127]

[0128] The synthesis of compound K refers to compound F in Example 6.

[0129] The physical parameters of compound J are: 1 H NMR(500MHz,DMSO-d6)δ9.75(s,2H),9.56(d,J=1.5Hz,2H),8.27–8.20(m,5H),6.68(d,J=1.6Hz,1H),5.83(d,J=1.5Hz,2 H),3.79(s,3H),3.65(t,J=5.9Hz,4H),3.61–3.50(m,34H),3.35(d,J=15.0Hz,12H),2.54(t,J=6.0Hz,4H),2.33(s,12H). 13C NMR (125MHz, DMSO-d6) δ170.60,158.66,158.64,158.55,157.94,157.93,157.2 4,156.09,155.95,155.90,155.49,155.47,148.46,148.30,148.23,114.82,11 4.76,85.48,84.79,71.77,70.62,70.55,70.54,70.52,69.56,65.96,59.02,39 .91,38.77,38.54,38.48,36.22,14.90,14.87,14.85.MS(ESI):m / z:Calcd.for C 54 H 81 N 22 O 11 + [M+H] + :1197.6501; found:1197.6522.

[0130] Cytotoxicity assays of ion transport molecules

[0131] The CCK-8 method was used to determine the proliferation inhibition effects of the ion transport molecules A to K prepared in the above examples on human cervical cancer cells (HeLa), human lung cancer cells (A549), human liver cancer cells (HepG2), human breast cancer cells (MCF-7) and human prostate cancer cells (PC-3). The specific test process is as follows:

[0132] (1) Sample solution preparation:

[0133] Folding body group: DMSO was used to dissolve the ion transport molecules A~K prepared in the example and the commercial anticancer drug doxorubicin, and then a series of culture medium solutions with concentrations of 2.50, 5.00, 10.00, 15.00, 30.00, and 50.00 μM (final concentration, 0.1% DMSO) were prepared using culture medium.

[0134] The structure of the aforementioned doxorubicin is shown below:

[0135]

[0136] Zn 2+Regulated unfolding group: The ion transport molecules A to K prepared in the example were dissolved in DMSO respectively, and a series of culture medium solutions with concentrations of 2.50, 5.00, 10.00, 15.00, 30.00, and 50.00 μM (final concentration, 0.1% DMSO) of the ion transport molecules were prepared in culture medium, and 10 μL of ZnCl2 aqueous solution was added (the final concentration was 10 eq of the concentration of the ion transport molecules).

[0137] (2) The above tumor cell lines were prepared into single cell suspensions with a concentration of 5000 cells / mL using the corresponding culture medium, and 200 μL of the cell suspension was inoculated into a 96-well culture plate and placed in a CO2 incubator (37°C, 5% CO2, 95% air) for 24 h. Human cervical cancer cells (HeLa), human liver cancer cells (HepG2), mouse embryonic fibroblasts (NIH3T3), human umbilical vein endothelial cells (HUVEC), and mouse fibroblasts (L929) were cultured in DMEM high-glucose medium (containing 10% newborn calf serum and 1% double antibody); human prostate cancer cells (PC-3) and human lung cancer cells (A549) were cultured in 1640 medium (containing 10% newborn calf serum and 1% double antibody); human breast cancer cells (MCF-7) were cultured in MEM medium (containing 10% newborn calf serum and 1% double antibody).

[0138] (3) After culturing the cells for 24 hours, the culture medium in the 96-well culture plate was removed, and culture medium solutions of different concentrations of the ion transport molecules A to K prepared in the above (1) and the commercial anticancer drug doxorubicin, as well as a culture medium solution further containing zinc ions, were added, and the cells were cultured for 12 hours.

[0139] (4) Then, 10 μL of CCK-8 (Cell Counting Kits-8) reagent was added to each well. After incubation at 37°C for 1.5 h, the absorbance at 450 nm was measured using a Biotek multi-function microplate reader. The cell viability was represented by the ratio of the number of live cells after treatment to the number of live cells without treatment (as shown in the following formula). The average of the results of four parallel experiments was taken.

[0140]

[0141] Among them, OD experiment represents the absorbance of cells treated with ion transport molecules, OD control OD0 represents the absorbance of cells without any treatment, and OD1 represents the absorbance of a blank sample without cells.

[0142] The concentration of ion transport molecules required for the cell survival rate to be 50% obtained from the above experiment is IC 50 The results are shown in Tables 1 and 2 described below.

[0143] It should be noted that the presence of 0.1% DMSO and the corresponding Zn 2+ The presence of was not cytotoxic to all selected cells.

[0144] Table 1 Ion transport molecules A~K in the absence of Zn 2+ And the presence of Zn 2+ Cytotoxicity test data under the conditions of

[0145]

[0146] It can be seen from Table 1 that the ion transport molecules prepared in Examples 1 to 11 have no Zn 2+ Under the conditions of Zn 2+ For example, the ion transport molecule F has a strong anticancer activity in the absence of Zn 2+ Under the condition of 50 20.2~26.3μM, when Zn was added 2+ After unfolding, IC 50 When it drops to 5.9-10.1 μM, the killing power increases by 2-4 times, and the killing effect is closer to the anti-cancer effect of commercially available doxorubicin hydrochloride.

[0147] From the results in Table 1 above, it can be seen that without adding Zn 2+ Under the conditions of 2+ It shows higher inhibitory activity in the case of , and has obvious effects of killing tumor cells and inhibiting tumor cell growth. Moreover, compared with normal cells, it has higher cytotoxicity to cancer cells and can selectively target and kill cancer cells.

[0148] The test results of the ion transport molecule F prepared in Example 6 and the commercial anticancer drug doxorubicin are shown in Table 3 below.

[0149] Table 2: Cytotoxicity test of ion transport molecule F and doxorubicin

[0150]

[0151] It can be seen from Table 2 that the ion transport molecule F prepared in Example 1 has a high 2+ The cytotoxicity of the drug was lower than that of the commercial anticancer drug doxorubicin under the condition of Zn 2+ Under the condition of Zn 2+ Responding cancer cell lethal behavior.

[0152] Antibacterial activity test of ion transport molecules

[0153] Test method: Agar dilution method was used to determine the ion transport molecules A-K and broad-spectrum antibacterial molecule chitosan prepared in the example in the absence of Zn 2+ Under the conditions and in the presence of Zn 2+ The minimum inhibitory concentration (MIC) of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli under the conditions of . The specific test process is as follows:

[0154] (1) Sample solution preparation:

[0155] Folding body group: The ion transport molecules A~K and the broad-spectrum antibacterial molecule chitosan prepared in the example were dissolved in DMSO respectively, and then a series of culture medium solutions with concentrations of 2.50, 5.00, 10.00, 15.00, 30.00, and 50.00 μM (final concentration, 0.1% DMSO) were prepared in tryptic soy broth (TSB).

[0156] Zn 2+ Regulated unfolding group: The ion transport molecules A to K prepared in the example were dissolved in DMSO respectively, and a series of culture medium solutions with concentrations of 2.50, 5.00, 10.00, 15.00, 30.00, and 50.00 μM (final concentration, 0.1% DMSO) of the ion transport molecules were prepared in TSB culture medium, and 10 μL of ZnCl2 aqueous solution was added (the final concentration was 10 eq of the concentration of the response ion transport molecule).

[0157] (2) Bacterial culture: Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli were used as bacterial strains for the bacterial inhibition activity test. The revived strains were inoculated into TSB medium and cultured in a 37°C constant temperature shaker (150 RPM). Every 1 hour, 1 mL of the culture medium was taken to measure the OD value of the bacterial solution. 600 Take 2 mL of bacterial culture in the exponential growth phase and centrifuge at 4°C (3000 rpm) for 10 minutes. After removing the supernatant, wash three times with sterile PBS and prepare a concentration of 10 6 The bacterial solution with CFU / mL is ready for use. The bacterial concentration is determined by the standard curve.

[0158] (3) Antibacterial activity test: Take 200 μL of bacterial solution onto TSA agar plate, spread evenly, add different concentration gradients of the sample solution prepared in (1) above, and culture in a biochemical incubator at 37°C for 24 h. Count after the culture. The lowest sample concentration at which no bacterial growth is observed is taken as the minimum inhibitory concentration (MIC) and recorded in Table 3 described below. At the same time, a blank control without any sample and a solvent sample with DMSO (﹤5%) added are used as the control group.

[0159] Table 3: Ion transport molecules A~K in the absence of Zn 2+ And the presence of Zn 2+ Test data on antibacterial activity under the conditions of

[0160]

[0161] As shown in Table 3, whether there is Zn 2+ The ion transport molecules A to K prepared in the examples all showed inhibitory effects on the activity of Staphylococcus aureus and Escherichia coli. 2+ In the case of Zn 2+ The inhibition of bacterial activity was further enhanced in the presence of

[0162] The test results of the antibacterial activity of the ion transport molecule F and chitosan prepared in Example 6 are shown in Table 4 below. Table 4: Test results of the antibacterial activity of the ion transport molecule F and chitosan

[0163]

[0164] As shown in Table 4, the MIC of molecule F prepared in Example 6 against Staphylococcus aureus is 10.3 μg mL -1 The MIC for Escherichia coli was 9.9 μg mL -1 The inhibitory effect on bacterial activity is much higher than that of chitosan, showing a better ability to inhibit bacterial activity.

[0165] The above is only a preferred embodiment of the present invention, and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment, it is not used to limit the present invention. Any technician familiar with this patent can make some changes or modify the technical contents suggested above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the solution of the present invention.

Claims

1. An ion transport molecule having a structure shown in the following formula: in, x = 1, 2, 3, 4, or 5; R1 and R2 are each independently a crown ether group, an azacrown ether group, a cyclodextrin group, a thiourea group, an amide ring group, a benzamide ring group, an aryltriazole group, a cholic acid group, a cholesterol group, a CH3CH2(OCH2CH2) group, k -O-, hydrogen, or alkyl, wherein k = 0 to 10; R3 and R4 are each independently a linking group, an alkylene group, an alkyleneoxy group, an alkyleneacyl group, an alkyleneamino group, an alkylenetriazolyl group, an alkynylene group, an iminoalkylacyl group, a sulfinylalkylacyl group, a phosphinylalkylacyl group, an arylene group, or a heteroarylene group.

2. The ion transport molecule according to claim 1, wherein The R1 and R2 are each independently selected from any one of the following substituents:

3. The ion transport molecule according to claim 1, wherein The R3 and R4 are each independently a linking group, an alkylene group having 1 to 5 carbon atoms, an alkyleneoxy group having 1 to 5 carbon atoms, an alkyleneacyl group having 1 to 5 carbon atoms, or an alkyleneamino group having 1 to 5 carbon atoms.

4. The ion transport molecule according to claim 1, which is selected from the group consisting of the following compounds A to K: 5 . A pharmaceutical composition comprising the ion transport molecule according to claim 1 and a pharmaceutically acceptable carrier.

6. Use of the ion transport molecule according to any one of claims 1 to 4 in the preparation of a drug for treating cancer or a drug for treating bacterial infection.

7. The use according to claim 6, wherein The cancer is cervical cancer, liver cancer, lung cancer, colorectal cancer, breast cancer, prostate cancer, hepatocellular carcinoma or leukemia.

8. The use according to claim 6, wherein The bacteria are Gram-positive bacteria or Gram-negative bacteria.

9. The use according to claim 8, wherein The Gram-positive bacteria is Staphylococcus aureus.

10. The use according to claim 8, wherein The Gram-negative bacteria is Escherichia coli.

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