A photosensitive chloride transporter based on azobenzene semi-aza bamboo ring, its preparation method and application

Abstract

The present invention provides a photo-controlled chloride ion transporter based on azobenzene semi-aza-calixarene, and its preparation method and application, belonging to the technical field of artificial ion transporters. The photo-controlled chloride ion transporter based on azobenzene semi-aza-calixarene provided by the present invention is composed of an azobenzene unit and a calixarene skeleton; the calixarene is a macrocyclic molecule composed of six N,N-dimethylglycoluril units, and the methylene group facing the inside of the ring effectively binds and transports anions through the ion-dipole interaction of the C-H bond with the anion, and both ends of the ring are covalently connected to the azobenzene through carbon chains of different lengths. The semi-aza-calixarene chloride ion transporter provided by the present invention has high chloride ion transport efficiency and high photo-responsive characteristics, and can efficiently achieve E / Z isomerization under the switching stimulation of ultraviolet / visible light in the external environment, realizing effective photo-controlled anion transmembrane transport.

Description

Technical Field

[0001] The present invention relates to the technical field of artificial ion transporters, and particularly to a light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring, and its preparation method and application. Background Art

[0002] Ion transport proteins are composed of special proteins produced by cells and are widely present in various cells. These ion transport proteins control the entry and exit of various physiologically relevant ions and other polar substances into and out of cells, and play a crucial role in maintaining cellular ion homeostasis and basic life activities. However, structural mutations of natural ion transport proteins can lead to severe ion channel diseases. In order to explore the pathogenic mechanisms of these diseases and discover specific therapeutic drugs, people have begun to analyze natural ion transport proteins and design artificially synthesized ion transporters to mimic the functions of natural transporters, hoping to use artificially synthesized transporters to replace mutant channel proteins to treat ion channel diseases. Research has shown that artificial chloride ion transporters play a complementary role with commercially available drugs in the treatment of cystic fibrosis related to chloride ion channels. In addition, artificial ion transporters can also disrupt cellular ion homeostasis and induce apoptosis by mediating ion flow, and have great application prospects in anti-cancer and other aspects.

[0003] The analysis of the structure of natural ion channels provides important ideas for the design and development of artificial ion channels. Natural ion channels usually have gating properties. For example, rhodopsin is a natural light-responsive transport protein. However, currently reported artificial chloride ion transmembrane transport systems with light-controlled properties mainly rely on the photolysis properties of o-nitrobenzylamine and its derivatives, and the irreversibility of photolysis limits their applications to a certain extent.

[0004] Therefore, it is urgent to develop a chloride ion transporter with reversible light-responsive properties, which is of great significance for studying the transmembrane transport behavior of ion channels and developing new drugs. Summary of the Invention

[0005] The purpose of the present invention is to provide a light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring with reversible light-responsive properties, and its preparation method and application.

[0006] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0007] The present invention provides a light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring, which is characterized in that it has a chemical structure as shown in Formula I:

[0008]

[0009] In the said Formula I, n is an integer from 1 to 6.

[0010] The present invention also provides a method for preparing the light-controlled chloride ion transporter based on the azobenzene semiaza bamboo ring described in the above technical solution, comprising the following steps:

[0011] (1) mixing an N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, 4-aminoazobenzene and a solvent, and performing a condensation reaction to obtain an azobenzene intermediate;

[0012] The structural formula of the N-Boc-amino acid derivative is shown in Formula II: Wherein, n is an integer from 1 to 6;

[0013] (2) mixing the azobenzene intermediate obtained in step (1) with trifluoroacetic acid, and performing a protonation reaction to obtain a protonated azobenzene intermediate;

[0014] (3) mixing the protonated azobenzene intermediate obtained in step (2) with a methylated semi-thio bamboo ring, triethylamine, and solvent 2, and performing a substitution reaction to obtain a protonated azobenzene semi-aza bamboo ring;

[0015] (4) dissolving the protonated azobenzene semiazabamboo ring obtained in step (3) in an alcohol solvent to obtain a protonated azobenzene semiazabamboo ring solution, mixing the protonated azobenzene semiazabamboo ring solution with an ion exchange resin, and performing a deprotonation reaction to obtain a light-controlled chloride ion transporter based on the azobenzene semiazabamboo ring.

[0016] Preferably, in step (1), the molar ratio of the N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine and 4-aminoazobenzene is 1:(1.2-5):(1.5-10):(0.5-1.5).

[0017] Preferably, the solvent in step (1) comprises dichloromethane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide or acetonitrile.

[0018] Preferably, the temperature of the condensation reaction in step (1) is 25 to 40° C.; and the time of the condensation reaction is 24 to 72 hours.

[0019] Preferably, the molar ratio of the azobenzene intermediate to trifluoroacetic acid in step (2) is 1:(10-1000).

[0020] Preferably, the molar ratio of the protonated azobenzene intermediate, methylated semi-thiophane, and triethylamine in step (3) is (10-20):(1-2):(100-200).

[0021] Preferably, the temperature of the substitution reaction in step (3) is 65-100 °C; the time of the substitution reaction is 8-24 h.

[0022] Preferably, the temperature of the deprotonation reaction in step (4) is 25-40 °C; the time of the deprotonation reaction is 3-24 h.

[0023] The present invention also provides the use of the azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter described in the above technical solution or the azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter prepared by the preparation method described in the above technical solution in the preparation of anti-cancer drugs.

[0024] The present invention provides a light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring, having a chemical structure as shown in Formula I:

[0025]

[0026] In Formula I, n is an integer from 1 to 6. The light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring provided by the present invention is composed of an azobenzene unit and a bamboo ring skeleton; the bamboo ring is a macrocyclic molecule composed of six N,N-dimethylglycoluril units, and the methylene group facing the inside of the ring effectively binds and transports anions through the ion-dipole interaction of the C-H bond with the anion. The two ends of the ring are covalently connected to azobenzene through carbon chains of different lengths, and it is a bamboo ring chloride ion transporter with light-controlled properties; moreover, the azobenzene bamboo ring, as a single molecular channel, has better stability. The semi-aza bamboo ring chloride ion transporter provided by the present invention has high chloride ion transport efficiency and high light response characteristics, and can efficiently achieve E / Z isomerization under the switching stimulation of external ultraviolet / visible light. Among them, the (E)-isomer can effectively carry out ion transport on the bilayer membrane, while the (Z)-isomer cannot carry out ion transport. The results of the examples show that the light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring provided by the present invention has good anion transport activity, and its ion transport EC 50 value is 0.34 μM. By switching the ultraviolet / visible light irradiation, the isomerization of azobenzene is driven, and it has an extremely fast light response switch characteristic and high ion transport activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a schematic diagram of the light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring provided by the present invention for controlling the transmembrane transport of chloride ions;

[0028] Figure 2 1H NMR spectrum of azobenzene intermediate 3a prepared in Example 1 of the present invention 1 ;

[0029] Figure 3 1H NMR spectrum of azobenzene intermediate 3a prepared in Example 1 of the present invention 13 13C NMR spectrum;

[0030] Figure 4 1H NMR spectrum of protonated azobenzene semi-aza-bambus[6]arene 4a prepared in Example 1 of the present invention 1 ;

[0031] Figure 5 1H NMR spectrum of azobenzene semi-aza-bambus[6]arene-based light-controlled chloride transporter 5a prepared in Example 1 of the present invention 1 ;

[0032] Figure 6 1H NMR spectrum of azobenzene intermediate 3b prepared in Example 2 of the present invention 1 ;

[0033] Figure 7 1H NMR spectrum of azobenzene intermediate 3b prepared in Example 2 of the present invention 13 13C NMR spectrum;

[0034] Figure 8 1H NMR spectrum of protonated azobenzene semi-aza-bambus[6]arene 4b prepared in Example 2 of the present invention 1 ;

[0035] Figure 9 1H NMR spectrum of azobenzene semi-aza-bambus[6]arene-based light-controlled chloride transporter 5b prepared in Example 2 of the present invention 1 ;

[0036] Figure 10 1H NMR spectrum of azobenzene intermediate 3c prepared in Example 3 of the present invention 1 ;

[0037] Figure 11 1H NMR spectrum of azobenzene intermediate 3c prepared in Example 3 of the present invention 13 13C NMR spectrum;

[0038] Figure 12 1H NMR spectrum of protonated azobenzene semi-aza-bambus[6]arene 4c prepared in Example 3 of the present invention 1 ;

[0039] Figure 13 1H NMR spectrum of azobenzene semi-aza-bambus[6]arene-based light-controlled chloride transporter 5c prepared in Example 3 of the present invention 1 ;

[0040] Figure 14 13C NMR spectrum of the light-controlled chloride ion transporter 5c based on azobenzene semi-aza-bamboo ring prepared in Example 3 of the present invention 13 13C NMR spectrum

[0041] Figure 15 1H NMR spectrum of the azobenzene intermediate 3d prepared in Example 4 of the present invention 1 1H NMR spectrum

[0042] Figure 16 13C NMR spectrum of the azobenzene intermediate 3d prepared in Example 4 of the present invention 13 13C NMR spectrum

[0043] Figure 17 1H NMR spectrum of the protonated azobenzene semi-aza-bamboo ring 4d prepared in Example 4 of the present invention 1 1H NMR spectrum

[0044] Figure 18 1H NMR spectrum of the light-controlled chloride ion transporter 5d based on azobenzene semi-aza-bamboo ring prepared in Example 4 of the present invention 1 1H NMR spectrum

[0045] Figure 19 13C NMR spectrum of the light-controlled chloride ion transporter 5d based on azobenzene semi-aza-bamboo ring prepared in Example 4 of the present invention 13 13C NMR spectrum

[0046] Figure 20 1H NMR spectrum of the azobenzene intermediate 3e prepared in Example 5 of the present invention 1 1H NMR spectrum

[0047] Figure 21 13C NMR spectrum of the azobenzene intermediate 3e prepared in Example 5 of the present invention 13 13C NMR spectrum

[0048] Figure 22 1H NMR spectrum of the protonated azobenzene semi-aza-bamboo ring 4e prepared in Example 5 of the present invention 1 1H NMR spectrum

[0049] Figure 23 1H NMR spectrum of the light-controlled chloride ion transporter 5e based on azobenzene semi-aza-bamboo ring prepared in Example 5 of the present invention 1 1H NMR spectrum

[0050] Figure 24 13C NMR spectrum of the light-controlled chloride ion transporter 5e based on azobenzene semi-aza-bamboo ring prepared in Example 5 of the present invention 13 13C NMR spectrum

[0051] Figure 25 1H NMR spectrum of the azobenzene intermediate 3f prepared in Example 6 of the present invention 1 1H NMR spectrum

[0052] Figure 26 13C NMR spectrum of the azobenzene intermediate 3f prepared in Example 6 of the present invention 13 13C NMR spectrum;

[0053] Figure 27 1H NMR spectrum of the protonated azobenzene semi-aza-bamboo ring 4f prepared in Example 6 of the present invention 1 1H NMR spectrum;

[0054] Figure 28 1H NMR spectrum of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter 5f prepared in Example 6 of the present invention 1 1H NMR spectrum;

[0055] Figure 29 13C NMR spectrum of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter 5f prepared in Example 6 of the present invention 13 13C NMR spectrum;

[0056] Figure 30 Ion transport activity of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter prepared in Examples 1-6 of the present invention

[0057] Figure 31 Light-controlled ion transport activity of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter 5b prepared in Example 2 of the present invention

[0058] Figure 32 Experimental results of the in vitro anti-cancer activity of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter 5b prepared in Example 2 of the present invention

[0059] Figure 33 CCK8 experimental results of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter 5b prepared in Example 2 of the present invention Detailed implementation mode

[0060] The present invention provides a light-controlled chloride ion transporter based on an azobenzene semi-aza-bamboo ring, which is characterized in that it has a chemical structure as shown in Formula I:

[0061]

[0062] In the present invention, in Formula I, n is an integer from 1 to 6.

[0063] The schematic diagram of the azobenzene semi-aza-bamboo ring-based light-controlled chloride ion transporter provided by the present invention for controlling the transmembrane transport of chloride ions is preferably as Figure 1 shown. From Figure 1It can be seen that the semi-aza-bamboo-ring chloride ion transporter provided by the present invention has high chloride ion transport efficiency and high light response characteristics, and can efficiently achieve E / Z isomerization under the switching stimulation of ultraviolet / visible light in the external environment. Among them, the (E)-isomer can effectively transport ions on the bilayer membrane, while the (Z)-isomer cannot transport ions, so the effect of controlling the transmembrane transport of chloride ions by a light-controlled chloride ion transporter can be achieved.

[0064] The present invention also provides a preparation method of the light-controlled chloride ion transporter based on azobenzene semi-aza-bamboo-ring described in the above technical solution, including the following steps:

[0065] (1) Mix an N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, 4-aminoazobenzene and a solvent I, and carry out a condensation reaction to obtain an azobenzene intermediate;

[0066] The structural formula of the N-Boc-amino acid derivative is shown in Formula II:

[0067] Among them, n is an integer from 1 to 6;

[0068] (2) Mix the azobenzene intermediate obtained in the step (1) with trifluoroacetic acid, and carry out a protonation reaction to obtain a protonated azobenzene intermediate;

[0069] (3) Mix the protonated azobenzene intermediate obtained in the step (2) with methylated semi-thia-bamboo-ring, triethylamine and a solvent II, and carry out a substitution reaction to obtain a protonated azobenzene semi-aza-bamboo-ring;

[0070] (4) Dissolve the protonated azobenzene semi-aza-bamboo-ring obtained in the step (3) in an alcohol solvent to obtain a protonated azobenzene semi-aza-bamboo-ring solution, and mix the protonated azobenzene semi-aza-bamboo-ring solution with an ion exchange resin to carry out a deprotonation reaction to obtain a light-controlled chloride ion transporter based on azobenzene semi-aza-bamboo-ring.

[0071] In the present invention, unless otherwise specified, the chemical reagents used in the present invention are all commercially available products well-known to those skilled in the art.

[0072] The present invention mixes an N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, 4-aminoazobenzene and a solvent I, and carries out a condensation reaction to obtain an azobenzene intermediate.

[0073] In the present invention, the structural formula of the N-Boc-amino acid derivative is shown in Formula II:

[0074] Among them, n is an integer from 1 to 6. In the present invention, the above N-Boc-amino acid derivative is used as the starting material, and the value of n is the same as the value of n in the chemical structure of the azobenzene semi-azabicyclic photocontrolled chloride transporter.

[0075] In the present invention, the molar ratio of the N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, and 4-aminoazobenzene is preferably 1:(1.2 - 5):(1.5 - 10):(0.5 - 1.5), more preferably 1:(1.2 - 2):(2 - 5):(1 - 1.2). By controlling the molar ratio of the N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, and 4-aminoazobenzene within the above range in the present invention, the condensation reaction between the N-Boc-amino acid derivative and 4-aminoazobenzene can be carried out sufficiently to obtain an azobenzene intermediate.

[0076] In the present invention, the first solvent preferably includes dichloromethane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, or acetonitrile, more preferably dichloromethane. The present invention uses the first solvent to provide a suitable reaction medium for the condensation reaction.

[0077] In the present invention, the molar ratio of the amount of the N-Boc-amino acid derivative to the volume of the first solvent is preferably 1 - 10 mmol:1 - 100 mL, more preferably 5 mmol:5 mL. By controlling the molar ratio of the amount of the N-Boc-amino acid derivative to the volume of the first solvent within the above range in the present invention, the N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, and 4-aminoazobenzene can be completely dissolved in the first solvent, which is more conducive to promoting the condensation reaction more fully.

[0078] In the present invention, the method of mixing the N-Boc-amino acid derivative, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine, 4-aminoazobenzene and Solvent 1 is preferably to stir and mix the N-Boc-amino acid derivative, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N',N'-diisopropylethylamine and Solvent 1 to obtain a mixed solution, and then mix 4-aminoazobenzene with the mixed solution. In the present invention, stirring and mixing is used to promote the dissolution of each component in Solvent 1. The present invention has no special limitation on the rotation speed and time of the stirring and mixing, and it can be adjusted according to the dissolution situation of each component. In the examples of the present invention, the stirring and mixing is carried out at room temperature, and the time of the stirring and mixing can be 30 min.

[0079] In the present invention, the temperature of the condensation reaction is preferably 25-40 °C, more preferably 25-30 °C; the time of the condensation reaction is preferably 24-72 h, more preferably 48 h. In the present invention, the above temperature and time are more conducive to promoting the condensation reaction more fully.

[0080] The present invention preferably washes, dries and purifies the mixed system obtained by the condensation reaction by silica gel column chromatography to obtain an azobenzene intermediate. The present invention has no special limitation on the specific operation methods of the washing, drying and silica gel column chromatography purification, and the conventional operation methods of washing, drying and silica gel column chromatography purification can be used, as long as the impurities in the azobenzene intermediate can be sufficiently removed.

[0081] In the present invention, the washing reagent is preferably distilled water. The present invention has no special limitation on the dosage of the distilled water, and it can be adjusted according to the dosage of Solvent 1, as long as the water-soluble impurities in the mixed system can be sufficiently removed. In the examples of the present invention, the volume ratio of Solvent 1 to distilled water can be 1:1.

[0082] In the present invention, the drying method is preferably: mixing the organic phase obtained after washing with anhydrous sodium sulfate. The present invention has no special limitation on the dosage of the anhydrous sodium sulfate, and it can be carried out according to the conventional dosage, as long as the water in the organic phase can be sufficiently removed.

[0083] The present invention preferably filters the system obtained after drying, and purifies the obtained filtrate by silica gel column chromatography. In the present invention, anhydrous sodium sulfate is removed by filtration.

[0084] In the present invention, the mobile phase for the silica gel column chromatography purification is preferably petroleum ether and acetic acid, and the volume ratio of the petroleum ether to the acetic acid can be 10:1.

[0085] After obtaining the azobenzene intermediate, in the present invention, the azobenzene intermediate is mixed with trifluoroacetic acid and then subjected to a protonation reaction to obtain a protonated azobenzene intermediate.

[0086] In the present invention, the molar ratio of the azobenzene intermediate to trifluoroacetic acid is preferably 1:(10 - 1000), more preferably 1:(10 - 100). The present invention uses trifluoroacetic acid to provide protons, which combine with the basic groups in the azobenzene intermediate to form a protonated intermediate; by controlling the molar ratio of the azobenzene intermediate to trifluoroacetic acid within the above range, the protonation treatment of the azobenzene intermediate by trifluoroacetic acid can be more sufficient.

[0087] In the present invention, the protonation reaction occurs immediately after the azobenzene intermediate is mixed with trifluoroacetic acid. In the present invention, the temperature of the protonation reaction is preferably 0 - 60°C, more preferably 25 - 40°C. The present invention has no special limitation on the time of the protonation reaction, which is adjusted according to the amount of trifluoroacetic acid added so that the azobenzene intermediate is completely consumed. In the present invention, the protonation reaction is preferably carried out under stirring. The present invention has no special limitation on the rotation speed of the stirring, which is adjusted according to the experimental needs to promote the uniform mixing of the azobenzene intermediate and trifluoroacetic acid.

[0088] The present invention preferably neutralizes, washes and dries the system obtained from the protonation reaction to obtain a protonated azobenzene intermediate.

[0089] In the present invention, the reagents for neutralization are preferably dichloromethane and saturated sodium bicarbonate solution. In the examples of the present invention, the volume ratio of dichloromethane to saturated sodium bicarbonate solution can be 1:1. The present invention obtains an organic phase and an aqueous phase through neutralization. The present invention has no special limitation on the amount of the neutralization reagents, which is adjusted according to the needs to ensure that the acid in the system obtained from the protonation reaction is fully neutralized.

[0090] In the present invention, the reagent for washing is preferably dichloromethane. By using dichloromethane to wash the aqueous phase obtained from neutralization, the protonated azobenzene intermediate in the aqueous phase obtained from neutralization can be extracted into the organic phase.

[0091] In the present invention, the drying method is preferably to mix the organic phase obtained from washing with anhydrous sodium sulfate, followed by filtration and solvent removal to obtain a protonated azobenzene intermediate. The present invention has no special limitation on the amount of anhydrous sodium sulfate, which is added according to conventional operations to fully remove the moisture in the organic phase. The present invention has no special limitation on the method of solvent removal, and any conventional solvent removal method can be used.

[0092] After obtaining the protonated azobenzene intermediate, the present invention mixes the protonated azobenzene intermediate with methylated semi-thio-bamboo ring, triethylamine and solvent II, and conducts a substitution reaction to obtain protonated azobenzene semi-aza-bamboo ring.

[0093] In the present invention, the preparation method of the methylated semi-thio-bamboo ring is preferably Chem. Eur. J., 2016, 22, 8848 - 8854.

[0094] In the present invention, the synthesis route of the methylated semi-thio-bamboo ring is preferably as shown in Formula III:

[0095]

[0096] In the present invention, the molar ratio of the protonated azobenzene intermediate, methylated semi-thio-bamboo ring and triethylamine is preferably (10 - 20):(1 - 2):(100 - 200), more preferably 10:1:100. By controlling the molar ratio of the protonated azobenzene intermediate, methylated semi-thio-bamboo ring and triethylamine within the above range, the present invention can enable the protonated azobenzene intermediate and methylated semi-thio-bamboo ring to fully react to obtain protonated azobenzene semi-aza-bamboo ring.

[0097] In the present invention, the solvent II preferably includes tetrahydrofuran, acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide or N,N-di-methylacetamide, more preferably tetrahydrofuran. Using the above solvent II in the present invention can provide a suitable reaction environment for the substitution reaction.

[0098] In the present invention, the molar ratio of the protonated azobenzene intermediate to the volume of the solvent II is preferably (0.24 - 1.2) mmol:50 mL, more preferably (0.48 - 1.0) mmol:50 mL.

[0099] In the present invention, the method of mixing the protonated azobenzene intermediate with methylated semi-thio-bamboo ring, triethylamine and solvent II is preferably: dissolving the protonated azobenzene intermediate and methylated semi-thio-bamboo ring in the solvent II to obtain a mixed solution, and then mixing triethylamine with the mixed solution.

[0100] In the present invention, the temperature of the substitution reaction is preferably 65 - 100 °C, more preferably 70 - 80 °C; the time of the substitution reaction is preferably 8 - 24 h, more preferably 12 h. In the present invention, the substitution reaction is preferably carried out under refluxing. Within the above temperature and time range, the present invention can promote the substitution reaction to be sufficient and improve the yield of protonated azobenzene semi-aza-bamboo ring.

[0101] Preferably, the product obtained from the substitution reaction is successively evaporated to dryness of the solvent, washed, and dried to obtain protonated azobenzene semi-aza bamboo ring. There are no special limitations on the specific operations of evaporating the solvent to dryness, washing, and drying in the present invention. By using conventional operation methods, it is sufficient to dry the protonated azobenzene semi-aza bamboo ring. In the examples of the present invention, the washing reagent can be diethyl ether.

[0102] After obtaining the protonated azobenzene semi-aza bamboo ring, the present invention dissolves the protonated azobenzene semi-aza bamboo ring in an alcohol solvent to obtain a protonated azobenzene semi-aza bamboo ring solution, and mixes the protonated azobenzene semi-aza bamboo ring solution with an ion exchange resin to carry out a deprotonation reaction to obtain a light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring.

[0103] In the present invention, the alcohol solvent preferably includes methanol, ethanol, tert-butanol, or isopropanol.

[0104] In the present invention, the molar ratio of the protonated azobenzene semi-aza bamboo ring to the volume of the alcohol solvent is preferably (0.005 - 0.1) mmol: 50 mL, more preferably (0.005 - 0.05) mmol: 50 mL. By controlling the molar ratio of the protonated azobenzene semi-aza bamboo ring to the volume of the alcohol solvent within the above range in the present invention, the protonated azobenzene semi-aza bamboo ring can be completely dissolved.

[0105] In the present invention, the ion exchange resin is preferably Amberlyst A26 resin or Ambersep 900 resin, more preferably Amberlyst A26 resin. There are no special limitations on the source of the ion exchange resin in the present invention, and conventional commercially available products can be used.

[0106] In the present invention, the mass ratio of the protonated azobenzene semi-aza bamboo ring to the ion exchange resin is preferably (1 - 2):(1 - 20), more preferably 1:1. By controlling the molar ratio of the protonated azobenzene semi-aza bamboo ring to the mass of the ion exchange resin within the above range in the present invention, the deprotonation reaction can be made more complete.

[0107] In the present invention, the temperature of the deprotonation reaction is preferably 25 - 40 °C, more preferably 25 - 30 °C; the time of the deprotonation reaction is preferably 3 - 24 h, more preferably 8 - 12 h. Under the above temperature and time in the present invention, the deprotonation reaction can be promoted to be sufficient, and the yield of the light-controlled chloride ion transporter based on azobenzene semi-aza bamboo ring can be increased.

[0108] The present invention preferably evaporates the solvent from the system obtained by the deprotonation reaction to obtain a photoswitchable chloride ion transporter based on azobenzene semi-aza-bambus[6]arene. The present invention has no special limitation on the operation method and temperature for evaporating the solvent, which can be adjusted according to the alcohol solvent used, and the solvent can be fully removed.

[0109] The synthetic route of the photoswitchable chloride ion transporter based on azobenzene semi-aza-bambus[6]arene provided by the present invention is preferably shown in Formula IV:

[0110]

[0111] The preparation method provided by the present invention is simple and controllable. By controlling the structure of the photoswitchable chloride ion transporter based on azobenzene semi-aza-bambus[6]arene, a photoswitchable chloride ion transporter composed of an azobenzene unit and a bambus[6]arene skeleton can be obtained.

[0112] The present invention also provides an application of the photoswitchable chloride ion transporter based on azobenzene semi-aza-bambus[6]arene described in the above technical solution in the preparation of anticancer drugs.

[0113] In the present invention, the anticancer drug is preferably one or more of an anti-glioma drug, an anti-pigmented tumor drug, and an anti-lung cancer drug.

[0114] The photoswitchable chloride ion transporter based on azobenzene semi-aza-bambus[6]arene provided by the present invention can drive the isomerization of azobenzene through the switching of ultraviolet / visible light irradiation, and finally realize effective photoswitchable anion transmembrane transport. Therefore, it can be used as a substitute for natural transport proteins or developed into the above types of anticancer drugs.

[0115] Next, the technical solutions in the present invention will be clearly and completely described in conjunction with the examples in the present invention. Obviously, the described examples are only a part of the examples of the present invention, rather than all the examples. Based on the examples in the present invention, all other examples obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0116] The reference for the preparation method of methylated semi-thiobambus[6]arene used in the examples of the present invention is Chem. Eur. J., 2016, 22, 8848 - 8854. The characterization of the obtained methylated semi-thiobambus[6]arene is as follows: 1 H NMR(500MHz, DMSO-d6)δ5.94(s, 12H), 5.26(s, 12H), 3.45(s, 36H), 2.76(s, 18H); 13 C NMR(126MHz, DMSO-d6)δ172.81, 158.03, 74.47, 49.18, 36.26, 16.31.

[0117] Example 1

[0118] A light-controlled chloride transporter based on azobenzene semi-aza calixarene has a chemical structure as shown in the following formula:

[0119] where n = 1.

[0120] The preparation method of the light-controlled chloride transporter based on azobenzene semi-aza calixarene comprises the following steps:

[0121] (1) In a 100 mL round-bottom flask, add 5 mmol of N-Boc-amino acid derivative 2a

[0122] 1.2 equivalents of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 2 equivalents of N',N'-diisopropylethylamine (DIPEA), and add 50 mL of dichloromethane as a solvent and stir at room temperature for 30 min. Subsequently, add 1.2 equivalents of 4-aminoazobenzene 1a and carry out a condensation reaction at 25 °C for 48 h; after the reaction is completed, wash the organic phase with 50 mL of distilled water, dry the organic phase with anhydrous sodium sulfate after washing, filter, and then remove the solvent under reduced pressure. Purify by silica gel column chromatography to obtain an orange-red solid, which is azobenzene intermediate 3a (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0123] The 1 1H NMR spectrum of 3a is as Figure 2 shown, and the 13 13C NMR spectrum of 3a is as Figure 3 shown: 1 1H NMR (500 MHz, CDCl3) δ 8.67 (s, 1H), 7.93 - 7.87 (m, 4H), 7.72 - 7.67 (m, 2H), 7.52 - 7.48 (m, 2H), 7.46 (d, J = 7.1 Hz, 1H), 5.45 (s, 1H), 3.98 (d, J = 6.0 Hz, 2H), 1.49 (s, 9H); 13 13C NMR (126 MHz, CDCl3) δ 168.10, 156.74, 152.66, 149.13, 140.10, 130.78, 129.06, 123.98, 122.75, 119.94, 80.99, 45.69, 28.32; HRMS (ESI) m / z: [M+Na] + Calculated for C 19 H 22 N4O3Na 377.1584, Found 377.1581.

[0124] (2) Dissolve the azobenzene intermediate 3a obtained in step (1) in 50 mL of dichloromethane, add 10 equivalents of trifluoroacetic acid, and stir at room temperature until the raw materials are completely consumed. Then, completely evaporate the solvent under reduced pressure; add 50 mL of dichloromethane and 50 mL of saturated sodium bicarbonate solution. After stirring the mixed solution at room temperature for 30 min, collect the obtained organic phase, and wash the aqueous phase with dichloromethane; the obtained organic phase is dried over anhydrous sodium sulfate, washed, and then the solvent is evaporated under reduced pressure to obtain an orange-red powder solid, which is the protonated azobenzene intermediate;

[0125] (3) Dissolve the protonated azobenzene intermediate (0.48 mmol) obtained in step (2) and methylated thiaazacalix[5] (0.048 mmol) in 50 mL of tetrahydrofuran, add triethylamine (4.8 mmol), and reflux for 12 h. Then, evaporate the solvent to obtain a crude product, wash it with diethyl ether and dry it to obtain protonated azobenzene hemi-azacalix[5] 4a;

[0126] For 4a, 1 the 1H NMR spectrum is as Figure 4 follows: 1 1H NMR (500 MHz, DMSO-d6) δ 10.46 (s, 6H), 8.15 (s, 6H), 7.93 - 7.78 (m, 36H), 7.57 (dt, J = 17.0, 7.3 Hz, 18H), 5.68 (s, 12H), 5.06 (s, 12H), 3.86 - 3.66 (m, 12H), 3.31 (s, 36H), 2.84 - 2.70 (m, 12H).

[0127] (4) Dissolve 100 mg of the protonated azobenzene hemi-azacalix[5] obtained in step (3) in 50 mL of methanol to obtain a protonated azobenzene hemi-azacalix[5] solution. Mix the protonated azobenzene hemi-azacalix[5] solution with 100 mg of ion exchange resin and carry out a deprotonation reaction at 25 °C for 4 h. Evaporate the solvent to obtain a photoresponsive chloride ion transporter 5a based on azobenzene hemi-azacalix[5];

[0128] For 5a, 1 the 1H NMR spectrum is as Figure 5 follows: 1 1H NMR (500 MHz, DMSO-d6) δ 10.38 (s, 6H), 7.84 (dt, J = 19.0, 8.9 Hz, 36H), 7.61 - 7.49 (m, 18H), 5.35 (s, 12H), 4.98 (s, 12H), 3.60 (d, J = 87.2 Hz, 12H), 3.10 (s, 12H), 2.95 (s, 36H); 1313C NMR (126 MHz, DMSO-d6) δ 171.03, 161.84, 158.59, 152.50, 148.16, 141.73, 131.48, 129.85, 124.16, 122.80, 119.88, 79.79, 52.13, 46.21, 34.71.

[0129] Example 2

[0130] A photoswitchable chloride transporter based on azobenzene semi-aza calixarene has the chemical structure shown in the following formula:

[0131] wherein, n = 2.

[0132] The preparation method of the photoswitchable chloride transporter based on azobenzene semi-aza calixarene is different from that of Example 1 in that:

[0133] The N-Boc-amino acid derivative 2b in step (1) is An orange solid was obtained as the azobenzene intermediate 3b (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0134] 1H NMR spectrum of 3b is as 1 shown, and the 13C NMR spectrum of 3b is as Figure 6 shown: 13 1H NMR (500 MHz, CDCl3) δ 8.34 (s, 1H), 7.94 - 7.87 (m, 4H), 7.74 (d, J = 8.5 Hz, 2H), 7.53 - 7.48 (m, 2H), 7.48 - 7.43 (m, 1H), 5.25 (s, 1H), 3.53 (q, J = 6.1 Hz, 2H), 2.66 (t, J = 5.9 Hz, 2H), 1.45 (s, 9H); Figure 7 13C NMR (126 MHz, CDCl3) δ 169.89, 156.63, 152.67, 148.98, 140.55, 130.75, 129.06, 123.97, 122.73, 119.78, 80.00, 37.98, 34.45, 28.41; HRMS (ESI) m / z: [M+Na]+ 1 Calculated for C 13 H + N4O3Na 391.1741, Found 391.1722. 20 H 24 N4O3Na 391.1741, Found 391.1722.

[0135] (2) is the same as that in Example 1;

[0136] (3) The same as in Example 1, the protonated azobenzene semi-aza bamboo ring 4b was obtained;

[0137] The 1 1H NMR spectrum of 4b is as Figure 8 follows: 1 1H NMR (500 MHz, DMSO-d6) δ 10.46 (s, 6H), 8.15 (s, 6H), 7.93 - 7.78 (m, 36H), 7.57 (dt, J = 17.0, 7.3 Hz, 18H), 5.68 (s, 12H), 5.06 (s, 12H), 3.86 - 3.66 (m, 12H), 3.31 (s, 36H), 2.84 - 2.70 (m, 12H);

[0138] (4) The same as in Example 1, the azobenzene semi-aza bamboo ring-based photocontrolled chloride ion transporter 5b was obtained;

[0139] The 1 1H NMR spectrum of 5b is as Figure 9 follows: 1 1H NMR (500 MHz, DMSO-d6) δ 10.38 (s, 6H), 7.84 (dt, J = 19.0, 8.9 Hz, 36H), 7.61 - 7.49 (m, 18H), 5.35 (s, 12H), 4.98 (s, 12H), 3.60 (d, J = 87.2 Hz, 12H), 3.10 (s, 12H), 2.95 (s, 36H); 13 13C NMR (126 MHz, DMSO) δ 171.03, 161.84, 158.59, 152.50, 148.16, 141.73, 131.48, 129.85, 124.16, 122.80, 119.88, 74.31, 52.13, 46.21, 38.52, 34.71.

[0140] Example 3

[0141] A photocontrolled chloride ion transporter based on azobenzene semi-aza bamboo ring has the chemical structure shown in the following formula:

[0142] where n = 3.

[0143] The preparation method of the azobenzene semi-aza bamboo ring-based photocontrolled chloride ion transporter is as follows:

[0144] (1) Different from Example 1 in that N-Boc-amino acid derivative 2c gave an orange-red solid, which is azobenzene intermediate 3c (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0145] 3c's 1 1H NMR spectrum is as follows Figure 10 shown, for 3c 13 13C NMR spectrum is as follows Figure 11 shown: 1 1H NMR (500 MHz, CDCl3) δ 9.29 (s, 1H), 7.94 - 7.87 (m, 4H), 7.82 - 7.78 (m, 2H), 7.50 (dd, J = 8.3, 6.6 Hz, 2H), 7.46 - 7.43 (m, 1H), 4.86 (t, J = 6.5 Hz, 1H), 3.28 (d, J = 6.3 Hz, 2H), 2.43 (t, J = 6.4 Hz, 2H), 1.93 - 1.88 (m, 2H), 1.48 (s, 9H); 13 13C NMR (126 MHz, CDCl3) δ 186.47, 152.70, 148.84, 140.82, 130.69, 129.05, 128.90, 123.95, 122.70, 119.67, 79.45, 39.22, 36.89, 29.56, 28.45, 22.49; HRMS (ESI) m / z: [M+Na] + Calculated for C 21 H 26 N4O3Na 405.1897, Found 405.1891.

[0146] (2) The same as Example 1, an orange - red powdered solid was obtained, which is the protonated azobenzene intermediate;

[0147] (3) The same as Example 1, protonated azobenzene semi - azabamboo ring 4c was obtained;

[0148] 1H NMR spectrum of 4c 1 is as follows Figure 12 shown: 1 1H NMR (500 MHz, DMSO - d6) δ 10.68 (s, 6H), 8.52 (s, 6H), 7.92 - 7.82 (m, 36H), 7.60 - 7.51 (m, 18H), 5.64 (s, 12H), 5.07 (s, 12H), 3.49 (s, 12H), 3.37 (s, 36H), 3.32 (s, 12H), 1.94 (s, 12H).

[0149] (4) The same as Example 1, a light - controlled chloride ion transporter 5c based on azobenzene semi - azabamboo ring was obtained;

[0150] 1H NMR spectrum of 5c 1 is as follows Figure 13As shown, for 5c 13 The 13C NMR spectrum is as Figure 14 shown: 1 1H NMR (500 MHz, DMSO-d6) δ 10.21 (s, 6H), 7.85 (q, J = 7.1, 4.8 Hz, 36H), 7.59 - 7.50 (m, 18H), 5.33 (s, 12H), 4.94 (s, 12H), 3.17 (s, 12H), 2.92 (s, 36H), 2.47 - 2.38 (m, 12H), 1.75 (s, 12H); 13 13C NMR (126 MHz, DMSO-d6) δ 181.91, 172.61, 170.94, 152.51, 147.72, 143.07, 131.38, 129.84, 124.11, 122.75, 119.57, 85.46, 65.42, 44.40, 35.07, 28.71.

[0151] Example 4

[0152] A photoswitchable chloride transporter based on azobenzene semi-aza bambusuril has a chemical structure as shown in the following formula:

[0153] where n = 4.

[0154] The preparation method of the photoswitchable chloride transporter based on azobenzene semi-aza bambusuril is as follows:

[0155] (1) Different from Example 1 in that N-Boc-amino acid derivative 2d gave an orange-red solid, which is azobenzene intermediate 3d (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0156] The 1H NMR spectrum of 3d 1 is as Figure 15 shown, and the 13C NMR spectrum of 3d 13 is as Figure 16 shown: 1 1H NMR (500 MHz, CDCl3) δ 7.97 - 7.85 (m, 5H), 7.73 (d, J = 8.4 Hz, 2H), 7.50 (dd, J = 8.3, 6.6 Hz, 2H), 7.47 - 7.43 (m, 1H), 4.70 (s, 1H), 3.20 (q, J = 6.5 Hz, 2H), 2.45 (t, J = 7.6 Hz, 2H), 1.79 (p, J = 7.3 Hz, 2H), 1.61 (s, 2H), 1.45 (s, 9H); HRMS(ESI) m / z: [M+Na] + Calculated for C22 H 28 N4O3Na 419.2054, Found 419.2036.

[0157] (2) The same as in Example 1, an orange - red powdered solid was obtained, which is the protonated azobenzene intermediate;

[0158] (3) The same as in Example 1, protonated azobenzene semi - azabamboo ring 4d was obtained;

[0159] 1H NMR spectrum of 4d is as 1 follows: Figure 17 shown: 1 1H NMR (500 MHz, DMSO - d6) δ 10.29 (s, 6H), 8.05 (s, 6H), 7.91 - 7.81 (m, 36H), 7.56 (dt, J = 23.1, 6.9 Hz, 18H), 5.69 (s, 12H), 5.12 (s, 12H), 3.44 - 3.37 (m, 12H), 3.29 (s, 36H), 2.42 (t, J = 6.6 Hz, 12H), 1.65 (s, 24H).

[0160] (4) The same as in Example 1, a light - controlled chloride ion transporter 5d based on azobenzene semi - azabamboo ring was obtained;

[0161] 1H NMR spectrum of 5d is as 1 follows, and 13C NMR spectrum of 5d is as Figure 18 shown, and 13C NMR spectrum of 5d is as 13 follows: Figure 19 shown: 1 1H MMR (500 MHz, DMSO - d6) 6 10.24 (s, 6H), 7.90 - 7.80 (m, 36H), 7.59 - 7.50 (m, 18H), 5.43 (s, 12H), 4.98 (s, 12H), 3.13 (d, J = 58.7 Hz, 12H), 2.97 (s, 36H), 2.37 (t, J = 7.3 Hz, 12H), 1.67 (q, J = 7.9 Hz, 12H), 1.54 - 1.38 (m, 12H); 13 13C NMR (126 MHz, DMSO - d6) δ 184.55, 172.40, 165.55, 152.50, 147.80, 142.95, 131.41, 129.84, 124.12, 122.75, 119.62, 86.68, 47.39, 40.85, 36.95, 32.63, 23.44.

[0162] Example 5

[0163] A light-controlled chloride ion transporter based on azobenzene semi-azabamboo ring has a chemical structure as shown in the following formula:

[0164] where n = 5.

[0165] The preparation method of the light-controlled chloride ion transporter based on azobenzene semi-azabamboo ring is as follows:

[0166] (1) Different from Example 1 in that N-Boc-amino acid derivative 2e An orange solid is obtained, which is azobenzene intermediate 3e (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0167] The 1 1H NMR spectrum of 3e is as Figure 20 shown, and the 13 13C NMR spectrum of 3e is as Figure 21 shown: 1 1H NMR (500 MHz, CDCl3) δ 8.03 (s, 1H), 7.92 - 7.87 (m, 4H), 7.72 (d, J = 8.5 Hz, 2H), 7.52 - 7.48 (m, 2H), 7.47 - 7.42 (m, 1H), 4.67 (s, 1H), 3.12 (q, J = 6.7 Hz, 2H), 2.38 (t, J = 7.5 Hz, 2H), 1.75 (p, J = 7.6 Hz, 2H), 1.52 (p, J = 7.3 Hz, 2H), 1.44 (s, 9H), 1.41 - 1.35 (m, 2H); 13 13C NMR (126 MHz, CDCl3) δ 171.77, 156.22, 152.67, 148.78, 140.91, 130.69, 129.06, 123.93, 123.90, 119.75, 79.24, 40.23, 37.45, 29.69, 28.4, 26.23, 25.00; HRMS (ESI) m / z: [M+Na] + Calculated for C 23 H 30 N4O3Na 433.2210, Found 433.2189.

[0168] (2) The same as Example 1, an orange powder solid is obtained, which is the protonated azobenzene intermediate;

[0169] (3) The same as Example 1, the protonated azobenzene semi-azabamboo ring 4e is obtained;

[0170] The 1 1H NMR spectrum of 4e is as Figure 22 shown:1 1H NMR (500 MHz, DMSO-d6) δ 10.27 (s, 6H), 8.06 (s, 6H), 7.90 - 7.79 (m, 36H), 7.56 (dt, J = 16.1, 7.3 Hz, 18H), 5.67 (s, 12H), 5.11 (s, 12H), 3.43 - 3.35 (m, 12H), 3.31 (d, J = 29.1 Hz, 36H), 2.38 (q, J = 7.6 Hz, 12H), 1.63 (dt, J = 17.2, 8.2 Hz, 24H), 1.35 (q, J = 7.7 Hz, 12H).

[0171] (4) Similar to Example 1, a photoswitchable chloride transporter 5e based on azobenzene semi-aza-calixarene was obtained;

[0172] For 5e, 1 the 1H NMR spectrum is as ​ shown, and the 13C NMR spectrum of 5e is as 13 shown: ​ 1 1H NMR (500 MHz, DMSO-d6) δ 10.25 (s, 6H), 7.85 (d, J = 11.1 Hz, 36H), 7.55 (dt, J = 19.3, 8.2 Hz, 18H), 5.32 (s, 12H), 4.92 (s, 12H), 3.11 (d, J = 31.5 Hz, 12H), 2.88 (s, 36H), 2.35 (s, 12H), 1.62 (s, 12H), 1.41 (d, J = 40.3 Hz, 24H); 13 13C NMR (126 MHz, DMSO-d6) δ 172.35, 158.51, 152.50, 147.80, 142.97, 131.38, 129.82, 124.10, 122.75, 119.62, 99.98, 86.82, 68.36, 47.71, 37.15, 32.87, 26.97, 25.53.

[0173] Example 6

[0174] A photoswitchable chloride transporter based on azobenzene semi-aza-calixarene has a chemical structure shown in the following formula:

[0175] where n = 6.

[0176] The preparation method of the photoswitchable chloride transporter based on azobenzene semi-aza-calixarene is as follows:

[0177] (1) Different from Example 1 in that N-Boc-amino acid derivative 2f ​The orange-red solid obtained is the azobenzene intermediate 3f (mobile phase: petroleum ether and ethyl acetate with a volume ratio of 10:1);

[0178] The 1 1H NMR spectrum of 3f is as ​ shown, and the 13 13C NMR spectrum of 3f is as ​ shown: 1 1H NMR (500 MHz, CDCl3) δ 8.09 (s, 1H), 7.90 (td, J = 8.5, 1.5 Hz, 4H), 7.75 (d, J = 8.4 Hz, 2H), 7.50 (dd, J = 8.3, 6.6 Hz, 2H), 7.47 - 7.43 (m, 1H), 4.61 (s, 1H), 3.13 (dd, J = 13.7, 7.0 Hz, 2H), 2.38 (t, J = 7.3 Hz, 2H), 1.74 (p, J = 7.2 Hz, 2H), 1.45 (s, 9H), 1.42 - 1.35 (m, 6H); 13 13C NMR (126 MHz, CDCl3) δ 171.93, 156.27, 152.69, 148.82, 140.93, 130.70, 129.06, 123.94, 122.70, 119.74, 79.25, 43.59, 40.18, 37.38, 29.94, 28.46, 26.03, 25.38; HRMS (ESI) m / z: [M+Na] + Calculated for C 24 H 32 N4O3Na 447.2367, Found 447.2348.

[0179] (2) The same as in Example 1, an orange-red powdered solid is obtained, which is the protonated azobenzene intermediate;

[0180] (3) The same as in Example 1, the protonated azobenzene semi-azabicyclic ring 4f is obtained;

[0181] The 1 1H NMR spectrum of 4f is as ​ shown: 11H NMR (500 MHz, DMSO-d6) δ 10.25 (s, 6H), 8.04 (s, 6H), 7.85 (dt, J = 16.0, 8.5 Hz, 36H), 7.61 - 7.51 (m, 18H), 5.67 (s, 12H), 5.11 (s, 12H), 3.41 - 3.35 (m, 12H), 3.28 (s, 36H), 2.37 (t, J = 6.9 Hz, 12H), 1.66 - 1.53 (m, 24H), 1.38 - 1.28 (m, 24H).

[0182] (4) Similar to Example 1, a photoswitchable chloride transporter 5f based on azobenzene semi-aza-bamboo ring was obtained;

[0183] For 5f, 1 the 1H NMR spectrum is as ​ shown. For 5f, 13 the 13C NMR spectrum is as ​ shown: 1 1H NMR (500 MHz, DMSO-d6) δ 10.22 (s, 6H), 7.85 (q, J = 9.8 Hz, 36H), 7.54 (dt, J = 25.8, 7.3 Hz, 18H), 5.33 (s, 12H), 4.95 (s, 12H), 3.14 (s, 12H), 2.91 (s, 36H), 2.34 (t, J = 7.4 Hz, 12H), 1.65 - 1.55 (m, 12H), 1.37 (d, J = 44.7 Hz, 36H); 13 13C NMR (126 MHz, DMSO-d6) δ 180.08, 172.31, 152.50, 147.80, 142.94, 131.39, 129.83, 124.11, 122.75, 119.62, 99.98, 96.46, 88.73, 67.89, 47.74, 37.03, 32.94, 29.07, 27.03, 25.59.

[0184] Test Example

[0185] (1) Tests on transport activity and light-controlled ion transport behavior: The artificial lipid vesicles (LUVs) were used to test the anion transport activity. The method for preparing vesicles is as follows: 25 mg of egg yolk phosphatidylcholine (EYPC) was completely dissolved in 1 mL of chloroform to form a homogeneous solution. Then, the phospholipid solution was placed in a high-vacuum environment for 4 h. After the solvent was evaporated, the dissolved phospholipids formed a thin film on the bottle wall. Subsequently, 1 mL of NaNO3 solution containing the chloride ion fluorescent probe 6-methoxy-N-(3-sulfopropyl) quinolinium inner salt (SPQ) (0.5 mM SPQ, 200 mM NaNO3) was added. The resulting mixed solution was hydrated at 40 °C for 3 h, and then repeatedly frozen with liquid nitrogen and thawed in a 40 °C water bath for 10 times to form vesicles with uniform size. Then, it was filtered 10 times through a 0.22 μm aqueous filter membrane. Finally, the uncoated fluorescent probe was removed using a Sephadex G-50 column to obtain a suspension solution of LUVs coated with SPQ probes.

[0186] The fluorescence test method is as follows: In a clean fluorescence cuvette, 2 mL of 200 mM NaCl solution and 50 μL of the above LUVs were added. Due to the chloride ion concentration difference inside and outside the membrane of LUVs, the chloride ion transporter can mediate the influx of chloride ions, resulting in fluorescence quenching. This method was used to evaluate the chloride ion transport efficiency, and the final concentration of liposomes was approximately 102 μM. Subsequently, 10 μL of dimethyl sulfoxide (DMSO) solution containing the transporter molecule was added to the cuvette and mixed evenly. The fluorescence intensity change of the system was continuously monitored for 300 seconds using a fluorescence spectrometer (PerkinElmer FL6500) (excitation wavelength: 360 nm, emission wavelength: 430 nm). Subsequently, 10 μL of 10% Triton X-100 aqueous solution was added to lyse the vesicles to correct the fluorescence intensity to obtain the minimum value.

[0187] The transport activity was calculated using the following formula: I f =(F t -F ∞ ) / (F0 - F ∞ ). Among them, F0 and F ∞ represent the initial and final fluorescence intensities.

[0188] As ​ shown, it is the ion transport activity of the light-controlled chloride ion transporters based on azobenzene semi-aza calixarene prepared in Examples 1 - 6. In ​ , a is the schematic diagram of the ion transport process based on SPQ, b is the comparison diagram of the ion transport activities of different azobenzene semi-aza calixarenes at a final concentration of 1.46 μM, c is the concentration-dependent ion transport test of azobenzene semi-aza calixarene 5b, and d is the Hill analysis of the ion transport of 5b and the fitting of the half-maximal effective concentration. From ​It can be seen that among the azobenzene semi-aza-bamboo-ring molecules with different carbon chain lengths, the transport activity of 5b prepared in Example 2 is the best, and its transport activity EC 50 value at 300 seconds is 0.34 μM, indicating its high transport activity.

[0189] (2) Photo-controlled anion transport experiment:

[0190] As ​ shown is the photo-controlled ion transport activity of the photo-controlled chloride ion transporter 5b based on azobenzene semi-aza-bamboo-ring prepared in Example 2. In ​ , a is the ion transport activity of 5b in different states. 5b E : The ion transport activity of 5b without illumination; 5b Z : 5b E : The ion transport activity of 5b after being irradiated with 365 nm ultraviolet light for 5 seconds to 30 seconds; 5b E PSS: The ion transport activity of 5b z after being irradiated with 450 nm blue light for 5 seconds to 30 seconds, b is the schematic diagram of the change in the ion transport activity of 5b during the multiple cycle switching irradiation of ultraviolet / visible light, c is the "off-on-off-on-off-on" process of the activity under in-situ conditions by switching irradiation of ultraviolet / visible light, d is the "on-off-on-off-on-off" process of the activity under in-situ conditions by switching irradiation of ultraviolet / visible light. It can be seen from ​ that after completing the test of the (E)-isomer of azobenzene semi-aza-bamboo-ring, the azobenzene semi-aza-bamboo-ring molecule 5b is irradiated with 365 nm ultraviolet light for three minutes to obtain the (Z)-isomer. Subsequently, 10 μL of the DMSO solution of the (Z)-isomer is added to the test system and mixed evenly. The (Z)-isomer has almost no ion transport activity. Subsequently, the molecule is irradiated with 450 nm visible light for one minute to obtain the (E)-isomer and then tested. This process is cycled multiple times without a significant decrease in the transport efficiency. Subsequently, an in-situ photo-controlled experiment on the chloride ion transport activity of azobenzene semi-aza-bamboo-ring is carried out, which demonstrates the application potential of azobenzene semi-aza-bamboo-ring.

[0191] Application Example 1

[0192] The photo-controlled chloride ion transporter 5b based on azobenzene semi-aza-bamboo-ring prepared in Example 2, which showed the best performance in the LUVs test, was selected as the research object, and its anti-cancer activity was further studied. ​ are the results of the in vitro anti-cancer activity experiment of 5b. From ​It can be seen that after 5b was co-incubated with human glioblastoma cells (U87MG), mouse melanoma cells (B16), and human non-small cell lung cancer cells (A549) for 24 h respectively, it showed significant cytotoxicity to the three cancer cell lines, and its half-maximal inhibitory concentration (IC50) was 5.65 μM, 7.09 μM, and 10.14 μM respectively.

[0193] The results of the CCK8 experiment of 5b are as ​ shown. As ​ can be seen, 5b can effectively kill cancer cells. Based on its light-controlled characteristics in the LUVs experiment, we speculated that 5b might exhibit similar light-controlled responses at the cellular level. Therefore, we performed CCK8 cytotoxicity tests on 5b E and 5b z respectively. The results were consistent with the LUVs experiment: 5b z failed to induce apoptosis in cancer cells, which proved that 5b has the ability to achieve switch regulation in the cellular environment.

[0194] As can be seen from the above results, the light-controlled chloride transporter based on azobenzene semi-aza-bamboo-ring provided by the present invention has high chloride transport efficiency and high light-responsive characteristics, and can efficiently achieve E / Z isomerization under the switching stimulation of ultraviolet / visible light in the external environment.

[0195] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A photosensitive chloride transporter based on azobenzene semi-aza bamboo ring, characterized in that, It has a chemical structure as shown in Formula I: In the said Formula I, n is an integer from 1 to 6.

2. The preparation method of the azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter according to claim 1, comprising the following steps: (1) Mix an N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine, 4-aminoazobenzene and a first solvent, and carry out a condensation reaction to obtain an azobenzene intermediate; The structural formula of the said N-Boc-amino acid derivative is as shown in Formula II: wherein, n is an integer from 1 to 6; (2) After mixing the azobenzene intermediate obtained in the said step (1) with trifluoroacetic acid, carry out a protonation reaction to obtain a protonated azobenzene intermediate; (3) Mix the protonated azobenzene intermediate obtained in the said step (2) with methylated semi-thia bamboo ring, triethylamine and a second solvent, and carry out a substitution reaction to obtain a protonated azobenzene semi-aza bamboo ring; (4) Dissolve the protonated azobenzene semi-aza bamboo ring obtained in the said step (3) in an alcohol solvent to obtain a protonated azobenzene semi-aza bamboo ring solution, mix the protonated azobenzene semi-aza bamboo ring solution with an ion exchange resin, and carry out a deprotonation reaction to obtain an azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter.

3. The preparation method according to claim 2, characterized in that, In the said step (1), the molar ratio of the N-Boc-amino acid derivative, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine and 4-aminoazobenzene is 1∶(1.2 - 5)∶(1.5 - 10)∶(0.5 - 1.5).

4. The preparation method according to claim 2, characterized in that, The first solvent in the said step (1) includes dichloromethane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide or acetonitrile.

5. The preparation method according to claim 2, wherein The temperature of the condensation reaction in the said step (1) is 25 - 40 °C; the time of the condensation reaction is 24 - 72 h.

6. The preparation method according to claim 2, characterized in that, In the said step (2), the molar ratio of the azobenzene intermediate to trifluoroacetic acid is 1∶(10 - 1000).

7. The preparation method according to claim 2, characterized in that, In the said step (3), the molar ratio of the protonated azobenzene intermediate, methylated semi-thia bamboo ring and triethylamine is (10 - 20)∶(1 - 2)∶(100 - 200).

8. The preparation method according to claim 2, characterized in that, The temperature of the substitution reaction in the said step (3) is 65 - 100 °C; the time of the substitution reaction is 8 - 24 h.

9. The preparation method according to claim 2, characterized in that, The temperature of the deprotonation reaction in the said step (4) is 25 - 40 °C; the time of the deprotonation reaction is 3 - 24 h.

10. The application of the azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter according to claim 1 or the azobenzene semi-aza bamboo ring-based light-controlled chloride ion transporter prepared by the preparation method according to any one of claims 2 - 9 in the preparation of anticancer drugs.

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