Ion transport molecules, pharmaceutical compositions and uses

Abstract

This invention relates to ion transport molecules, pharmaceutical compositions, and their uses. The ion transport molecule has the structure shown in Formula I, wherein 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 aryltriazolyl group, a cholic acid group, a cholesterol group, or CH3CH2(OCH2CH2). k -O-, hydrogen, or alkyl, wherein k = 0-10; R3 and R4 are each independently a linker, alkylene, alkyleneoxy, alkyleneacyl, alkyleneamino, alkylenetriazole, alkynyl, iminoalkylacyl, sulfinylalkylacyl, phosphonoalkylacyl, aryl, or heteroaryl. The ion transport molecule of this invention can insert into a phospholipid bilayer to inhibit the growth of cancer cells or bacteria, thereby achieving therapeutic effects for cancer or bacterial infections.

Description

Technical Field

[0001] This invention relates to the field of artificial ion transport technology, and more specifically to an ion transport molecule with activity in treating bacterial infections and cancer. Background Technology

[0002] As the basic building block of living organisms, cells need to exchange nutrients and transmit biological signals with their surrounding environment to ensure the orderly conduct of various life activities. Maintaining the ion balance inside and outside the cell is achieved by ion transport proteins embedded on the cell surface, which are crucial for the realization of various cellular functions. Abnormalities in ion transport protein function can lead to many diseases. For example, many ion channel disorders are related to the transmembrane transport of chloride ions, such as cystic fibrosis (CF), Thomsen's ankylosing spondylitis, and hereditary kidney stones.

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

[0004] To date, many artificial ion transporters have exhibited 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 the requirements for medical applications.

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

[0006] After conducting in-depth research on the above-mentioned problems, this invention discovered that ion transport molecules with a pyrimidine-hydrazone-pyrimidine (pym-hyz-pym) structure have the effect of inhibiting tumor growth and bacterial growth, especially in the presence of Zn. 2+ The inhibitory effects on tumor growth and bacterial growth are more significant under certain conditions, thus completing this invention.

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

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

[0009] A third object of the present invention is to provide the use of the above-mentioned ion transport molecule in the preparation of a medicament for treating tumors or in the preparation of a medicament for treating bacterial infections.

[0010] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0011] A 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 cyclic group, a benzamide cyclic group, an aryltriazolyl group, a cholic acid group, a cholesterol group, or CH3CH2(OCH2CH2). k -O-, hydrogen, or alkyl, where k = 0 to 10;

[0015] Furthermore, R1 and R2 are not both hydrogen or alkyl groups;

[0016] R3 and R4 are each independently a linker, alkylene, alkyleneoxy, alkyleneacyl, alkyleneamino, alkylenetriazolyl, alkynyl, iminoalkylacyl, sulfinylalkylacyl, phosphonoalkylacyl, aryl, or heteroaryl.

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

[0018] A third aspect of the present invention provides the use of the aforementioned ion transport molecule in the preparation of medicaments for treating cancer or for treating bacterial infections.

[0019] The ion transport molecule of this invention can insert into the phospholipid bilayer at both ends, altering the ion balance inside and outside cancer cells or bacteria through transport to induce apoptosis, thereby inhibiting the growth of cancer cells or bacteria, and achieving therapeutic effects for cancer or bacterial infections. In particular, the ion transport molecule of this invention has repeating pyrimidine-hydrazone-pyrimidine structural units, which, in the presence of Zn... 2+ Under certain conditions, folding can occur, thus allowing Zn to exist. 2+ Its ion transport capability is further enhanced under certain conditions. Utilizing this property, ion transport can be achieved through Zn... 2+This can be used to regulate the folding / unfolding of ion transport molecules, thereby regulating their transmembrane transport activity. Detailed Implementation

[0020] To more clearly illustrate the present invention, the following examples provide a more detailed description of the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive and should not be construed as limiting the scope of protection of the present invention.

[0021] Unless otherwise specified, all experimental materials used in the embodiments of this invention are commercially available. All chemical reagents are of analytical grade and require no further purification. The cells used in this 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 cyclic group, a benzamide cyclic group, an aryltriazolyl group, a cholic acid group, a cholesterol group, or CH3CH2(OCH2CH2). k -O-, hydrogen, or alkyl, where k = 0 to 10;

[0026] R3 and R4 are each independently a linker, alkylene, alkyleneoxy, alkyleneacyl, alkyleneamino, alkylenetriazolyl, alkynyl, iminoalkylacyl, sulfinylalkylacyl, phosphonoalkylacyl, aryl, or heteroaryl.

[0027] Among the groups represented by R1 and R2 mentioned above, crown ether, azacrown ether, and cyclodextrin groups have cation recognition capabilities; thiourea, amide ring, benzamide ring, and aryltriazole groups have anion recognition capabilities; and cholic acid, cholesterol, and CH3CH2(OCH2CH2) groups have anion recognition capabilities. k -O- (where k = 0–10) has membrane anchoring properties. 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 membranes containing functional groups. Examples of the aforementioned cations include Na. + Li + K + Mg 2+ Ca 2+ And so on. For example, Cl- is an anion mentioned above. - HCO- NO3 - Examples of functional groups on membranes that are functionalized include, for instance, bile acid groups, cholesterol groups, and fatty acid chain groups.

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

[0029]

[0030]

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

[0032] The aforementioned alkylene groups having 1 to 5 carbon atoms can be, for example, methylene, ethylene, propylene, butylene, or pentylene.

[0033] The aforementioned alkeneoxy group having 1 to 5 carbon atoms can be, for example, methyleneoxy, ethyloxy, propyloxy, butyloxy, or pentyleoxy.

[0034] The aforementioned alkylene acyl group having 1 to 5 carbon atoms can be, for example, methylene acyl, ethylene acyl, propylene acyl, butylene acyl, or pentylene acyl.

[0035] The aforementioned alkylamino group having 1 to 5 carbon atoms can be, for example, methyleneamino, ethylamino, propylamino, butylamino, or pentyleminamino.

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

[0037]

[0038]

[0039] The aforementioned ion transport molecules can all insert into the phospholipid bilayer, disrupting the intracellular and extracellular ion balance through transmembrane ion transport. They exhibit significant anti-cancer cell activity and inhibit tumor cell growth, and can be used alone to prepare drugs for cancer treatment, or combined or conjugated with other cancer-treating compounds for cancer therapy. In particular, these ion transport molecules are effective against divalent metal ions (such as Zn). 2+ Fe 2+ Cu2+ Pb 2+ These molecules exhibit specific responses, significantly increasing their transmembrane ion transport activity through coordination with these divalent metal ions, thus giving the molecules gating properties. For example, the presence of the aforementioned divalent metal ions, especially Zn, at concentrations of 0.8–8.0 mM in the environment... 2+ In this context, the aforementioned ion transport molecules alter their molecular structure through folding, thereby exhibiting a significantly enhanced killing effect on tumor cells. 50 The half-inhibition rate decreased to 1 / 3 to 1 / 20 of its original value.

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

[0041] Furthermore, the aforementioned ion transport molecules disrupt the intracellular and extracellular ion balance by transporting ions across the membrane, thus exhibiting broad-spectrum inhibitory properties against bacterial infections. In particular, these ion transport molecules are effective against divalent metal ions (such as Zn). 2+ Fe 2+ Cu 2+ Pb 2+ (e.g., Zn) exhibits specific responses, significantly increasing its transmembrane ion transport activity through coordination with these divalent metal ions. For example, in the presence of the aforementioned divalent metal ions, especially Zn, at concentrations of 0.8–8.0 mM in the environment... 2+ In such cases, the aforementioned ion transport molecules alter their molecular structure through folding, thereby significantly enhancing their killing effect on bacterial cells and substantially reducing their ability to grow and proliferate. Therefore, these ion transport molecules can be used alone or in combination with other drugs that inhibit bacterial growth to prepare drugs for treating bacterial infections.

[0042] The aforementioned bacteria can be, for example, Gram-positive or Gram-negative bacteria. Examples of Gram-positive bacteria include Staphylococcus aureus. Examples of Gram-negative bacteria include Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii.

[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 stabilized, anhydrous potassium carbonate (24 g, 173.6 mmol) was added, and the temperature was raised to 80 °C and heated under argon protection for reflux 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 resulting solid residue was dissolved in 150 mL of chloroform solution and stirred for 10 min. The resulting mixture was washed three times with water (30 mL). The organic phase was dried with anhydrous Na2SO4, concentrated, and purified by reversed-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 LMF, followed by the addition of (Ph3)2PdCl2 (0.47 g, 0.66 mmol). The mixture was heated at 80 °C under argon protection for 12 h. After the reaction was complete, the reaction solution was poured into saturated KF solution (100 mL), resulting in a brown precipitate. The precipitate was collected by filtration and washed with diethyl ether (80 mL × 4). The resulting mixture was washed with saturated NaCl solution (50 mL × 3). The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel column 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 2 mol / L hydrochloric acid solution (15 mL) was added. The mixture was stirred at room temperature for 6 h, and the solvent was removed under vacuum to obtain a white solid residue. The residue was dissolved in dichloromethane (200 mL), washed with 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 column 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 starting material was completely consumed (as detected by TLC), the solvent was removed under vacuum, 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 reversed-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 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, compound 4 was obtained by reversed-phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min).

[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: Isothiocyanate (4.67 mmol) was added to dichloromethane containing 4-(4-aminophenyl)butyric acid (0.80 g, 4.67 mmol), stirred overnight at room temperature, evaporated to dryness, and recrystallized (dichloromethane: petroleum ether = 1:1) to obtain 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] + calcd forC 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. Then, compound 1 (0.34 g, 2.0 mmol) and NMM (0.15 g, 1.5 mmol) were added to the reaction mixture. The solution was stirred at room temperature for 48 h. After the starting material was completely consumed (as detected by TLC), the solvent was removed under vacuum, 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 reversed-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 reversed-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, and the product was dissolved in 100 mL of dichloromethane. The product was washed with saturated sodium chloride solution (100 mL × 3), 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 O8Na + [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. Then, 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 starting material was completely consumed (as detected by TLC), the solvent was removed under vacuum, 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 reversed-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, compound C-3 was purified by reversed-phase preparative HPLC (MeOH:H2O, from 60:40 to 100:0, 15 min).

[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 allowed to proceed for 18 hours. 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 pale 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: Trimethylsilane azide (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 overnight at 80 °C. After cooling to room temperature, dichloromethane was added to the mixture, and then it was washed with water (30 mL × 3). The organic phase was dried over anhydrous Na2SO4 and concentrated to give a pale 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 dichloromethane / acetonitrile (4:1, 3 mL) mixed solvent, and Cu(CH3CN)4PF6 (55.6 mg, 0.15 mmol) was added. The mixture was stirred at room temperature for 12 h. The solvent was removed under vacuum, and the residue was purified by reversed-phase high-performance liquid chromatography (mobile phase: MeOH / H2O = 50 / 50-100 / 0, 15 min) to obtain a 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: Tetraethylene glycol (4 mL, 23 mmol) was dissolved in dichloromethane. Triethylamine (30 mL) was added in an ice bath, followed by a dichloromethane solution (50 mL) of p-toluenesulfonyl chloride (12 g, 64 mmol). The reaction was carried out at room temperature for 4 hours. The solvent was removed under vacuum, and the mixture was extracted with water (30 mL × 5). The organic phase was collected and washed three times, successively with 240 mL of 2 mol / L hydrochloric acid aqueous solution, 150 mL of 5% NaHCO3 aqueous solution, and 150 mL of water. The collected organic phase was then dried over anhydrous Na2SO4, concentrated, and purified by silica gel column chromatography (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). 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 THF solution (75 mL, 75 mmol) was added dropwise to a 70 mL THF solution of compound D-2 (4.72 g, 15.83 mmol) under ice bath conditions. The reaction mixture was stirred until it turned yellow, then heated to reflux for 4 hours. The reaction mixture 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. The mixture was filtered, and the filtrate was 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), and 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 collected. The solvent was removed under vacuum, and the filtrate was dissolved in dichloromethane. The resulting mixture was washed with 350 mL of 5% potassium hydroxide aqueous solution and 200 mL of saturated brine. The organic phase was dried over anhydrous Na2SO4, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 9:1) to obtain a pale 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), and KOH (3.0 g, 53.5 mmol) was added. The mixture was stirred and refluxed overnight at 90 °C. After the reaction was completed, the mixture was cooled to room temperature, filtered, and the filter cake was washed with ethanol (50 mL × 3). The resulting filter cake was then dissolved again in 100 mL of water, and trifluoroacetic acid was added dropwise until a white solid completely precipitated. The mixture was filtered, the white solid was collected, dried, and the white solid product D-5 was obtained.

[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: A dichloromethane solution (1500 mL) containing compound D-5 (3.0 g, 7.44 mmol) and triethylamine (2.5 mL) was added to a dichloromethane solution (15 mL) containing compound D-3 (2.05 g, 7.44 mmol) under ice bath conditions. An anhydrous environment was maintained during the reaction. The reaction was carried out at room temperature for 12 hours. The solvent was removed under vacuum, and the crude product was purified by silica gel column chromatography (dichloromethane:methanol = 96:4) to obtain a 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 hydrogen pressure of 70 atm for 15 hours. The palladium catalyst on carbon was removed by filtration, and the solution was washed with tetrahydrofuran. The filtrate was evaporated to dryness 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), and methyl-3-bromopropionate (0.3 g, 1.8 mmol) and anhydrous potassium carbonate (2.5 g, 18 mmol) were added. 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 washed 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 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 under vacuum. The remaining white solid mixture was dissolved in dichloromethane (50 mL), extracted with water (15 mL × 3), and the organic phase was dried over anhydrous Na2SO4 and concentrated. The mixture was purified by silica gel chromatography (dichloromethane:methanol = 95:5) to obtain a pale 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) Compound D-10 was synthesized according to the method of compound A-1 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) Compound D was synthesized using the same method as compound A 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 according 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 concentration, the system was purified by reversed-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) The synthesis of compound F is performed according to the synthesis method of compound A in Example 1.

[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 performed in accordance with 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 follows the same method as the synthesis 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 was performed in accordance with 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 is based on 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] Example 11

[0127]

[0128] The synthesis of compound K was performed in accordance with 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 assay of ion transport molecules

[0131] The CCK-8 assay was used to determine the inhibitory effects of the ion transport molecules A-K prepared in the aforementioned examples on the proliferation of 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 testing procedure is as follows:

[0132] (1) Sample solution preparation:

[0133] Folded body group: Ion transport molecules A to K prepared in the examples and the commercial anticancer drug doxorubicin 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.

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

[0135]

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

[0137] (2) Single-cell suspensions with a concentration of 5000 cells / mL were prepared for the above tumor cell lines using the corresponding culture media. 200 μL of the cell suspension was seeded into 96-well culture plates and cultured in a CO2 incubator (37℃, 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% penicillin 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% penicillin antibody); and human breast cancer cells (MCF-7) were cultured in MEM medium (containing 10% newborn calf serum and 1% penicillin antibody).

[0138] (3) After culturing the cells for 24 hours, remove the culture medium from the 96-well culture plate and add the culture medium solutions of the ion transport molecules A to K prepared in (1) and the commercial anticancer drug doxorubicin, as well as the culture medium solution containing zinc ions, respectively, and culture for 12 hours.

[0139] (4) Then, add 10 μL of CCK-8 (Cell Counting Kits-8) reagent to each well, incubate at 37°C for 1.5 hours, and measure the absorbance at 450 nm using a Biotek multi-microplate reader. Cell viability is represented by the cell survival rate obtained by the ratio of the number of viable cells after treatment to the number of untreated viable cells (as shown in the following formula). Take the average value of the results of 4 parallel experiments.

[0140]

[0141] Among them, OD experiment OD represents the absorbance of cells treated with ion transport molecules. control OD0 represents the absorbance of cells without any treatment, while OD0 represents the absorbance of a blank sample containing no cells.

[0142] The concentration of ion transport molecules required to achieve a cell viability of 50%, as determined by the above experiments, is called IC50. 50 The results are shown in Tables 1 and 2 below.

[0143] It should be noted that the presence of 0.1% DMSO and the corresponding Zn 2+ The presence of it has no cytotoxic effect on any of the selected cells.

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

[0145]

[0146] As can be seen from Table 1, the ion transport molecules prepared in Examples 1-11, without Zn 2+ Under all conditions, they showed general inhibitory effects on cancer cells, while Zn... 2+ In the presence of [a specific substance], its anticancer activity is greatly increased. For example, the ion transport molecule F [is present] in the absence of Zn. 2+ Under these conditions, for various types of cancer cells, IC 50 The concentration was 20.2–26.3 μM, after adding Zn 2+ After unfolding, for cancer cells IC 50 When the concentration is reduced to 5.9–10.1 μM, the lethality increases by 2–4 times, and the lethal effect is closer to the anticancer effect of commercially available doxorubicin hydrochloride.

[0147] As can be seen from the results in Table 1 above, without the addition of Zn 2+ Under the conditions specified, the ion transport molecules prepared in each embodiment exhibit certain inhibitory cell activity, especially with the addition of Zn. 2+ Under certain conditions, it exhibits higher cell-inhibiting activity, demonstrating a significant effect in killing tumor cells and inhibiting their growth. Furthermore, compared to normal cells, it exhibits higher cytotoxicity against cancer cells, selectively targeting and killing them.

[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 with doxorubicin

[0150]

[0151] As can be seen from Table 2, the ion transport molecule F prepared in the examples, in the absence of Zn 2+ Under certain conditions, its cytotoxicity is lower than that of the commercially available anticancer drug molecule doxorubicin, but it exhibits lower cytotoxicity in the presence of Zn. 2+ Under these conditions, its cytotoxicity increased significantly, and its anticancer activity approached that of doxorubicin, exhibiting significant Zn activity. 2+ The lethal behavior of responding cancer cells.

[0152] Assay for the inhibitory activity of ion transport molecules against bacteria

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

[0154] (1) Sample solution preparation:

[0155] Folded body group: Ion transport molecules A to K and broad-spectrum antibacterial molecule chitosan prepared in the examples were dissolved in DMSO, 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: Ion transport molecules A to K prepared in the examples were dissolved in DMSO. A series of culture medium solutions with ion transport molecule concentrations of 2.50, 5.00, 10.00, 15.00, 30.00, and 50.00 μM (final concentration, 0.1% DMSO) were prepared in TSB medium, and 10 μL of ZnCl2 aqueous solution (final concentration is 10 eq of the response ion transport molecule concentration) was added.

[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 shaker (150 RPM). Every 1 hour, 1 mL of medium was taken to determine the OD of the bacterial solution. 600 To determine the bacterial growth curve, take 2 mL of bacterial culture in the exponential growth phase, centrifuge at 4°C (3000 rpm) for 10 minutes, remove the supernatant, wash three times with sterile PBS, and then prepare a solution with a concentration of 10. 6 The bacterial suspension at CFU / mL was prepared for use. The bacterial concentration was determined using a standard curve.

[0158] (3) Antibacterial activity test: 200 μL of bacterial solution was spread evenly on a TSA agar plate, and sample solutions prepared in step (1) above with different concentration gradients were added. The plates were then incubated at 37°C for 24 h in a biochemical incubator. After incubation, the bacterial counts were recorded. The lowest sample concentration at which no bacterial growth was observed was taken as the minimum inhibitory concentration (MIC) and recorded in Table 3 below. At the same time, a blank control without any sample and a solvent sample with added DMSO (<5%) served as control groups.

[0159] Table 3: Ion transport molecules A-K in the absence of Zn 2+ And the existence of Zn 2+ Data on antibacterial activity under the specified conditions

[0160]

[0161] As shown in Table 3, regardless of the presence or absence of Zn 2+ The ion transport molecules A through K prepared in the examples all exhibited inhibitory effects on the activity of Staphylococcus aureus and Escherichia coli. Among them, in the presence of Zn... 2+ In this case, the MIC value further decreased, indicating that in Zn 2+ The presence of [a substance] further enhances the inhibition of bacterial activity.

[0162] The results of the tests on the antibacterial activity of ion transport molecule F prepared in Example 6 and chitosan are shown in Table 4 below. Table 4: Test results on the antibacterial activity of 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 is 9.9 μg / mL. -1 It exhibits a much higher inhibitory effect on bacterial activity than chitosan, demonstrating a better ability to inhibit bacterial activity.

[0165] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. An ion transport molecule having the structure shown in the following formula: in, x = 1 or 2; R3 and R4 are each independently an alkylene group having 1 to 5 carbon atoms; R1 and R2 are each independently selected from any of the following substituents:

2. An ion transport molecule selected from the group consisting of compounds A to K:

3. A pharmaceutical composition comprising the ion transport molecule of claim 1 or 2 and a pharmaceutically acceptable carrier.

4. Use of the ion transport molecule according to claim 1 or 2 in the preparation of a medicament for treating cancer or a medicament for treating bacterial infections, wherein, The cancer is cervical cancer, liver cancer, lung cancer, breast cancer, or prostate cancer; the bacteria is Staphylococcus aureus or Escherichia coli.

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