Oxidation-responsive cationic water-soluble pillararene, and preparation method and use

Abstract

Disclosed are an oxidation-responsive cationic water-soluble pillararene, and a preparation method and a use. The oxidation-responsive water-soluble cationic pillararene is a cyclic small-molecule nucleic acid carrier and is obtained by carrying out quaternization reaction on 4-methyl borate and a tertiary amine modified pillararene. A delivery carrier having a high positive charge density is obtained by means of small molecule synthesis, the delivery carrier can tightly bind with a negatively-charged nucleic acid substance to form a nanocomposite, and after entering cells, in an oxidative environment, positive charges of the carrier fall off and the nucleic acid substance is released to efficiently express nucleic acid information.

Description

Oxidation-responsive cationic water-soluble pillar aromatic hydrocarbon, preparation method and application Technical Field

[0001] The present invention belongs to the field of organic synthetic biomedical science, and specifically relates to an oxidation-responsive cationic water-soluble pillararomatic hydrocarbon, a preparation method and an application thereof as a nucleic acid delivery carrier. Background Art

[0002] Among the many cancer treatments currently available, gene therapy, by directly repairing and improving genetic defects, can achieve therapeutic effects that are difficult to achieve with traditional drugs. Thanks to the development and completion of the Human Genome Project, the identification of numerous disease-causing genes has provided a broader range of approaches and methods for gene therapy, enabling researchers to more specifically explore innovative and effective gene therapy strategies. The research and development of gene delivery vectors is a key component of this approach.

[0003] Gene delivery technologies primarily fall into three categories: viral vectors, non-viral vectors, and physical transfection techniques. Compared to the other two gene delivery methods, non-viral vectors are more easily able to overcome inherent defects such as pathogenicity and immunogenicity, offer high biosafety, and are low-cost, thus demonstrating significant advantages. Currently, the main research challenges facing the development of non-viral vectors include: 1. Improving the in vivo circulation stability of the gene delivery system; 2. Efficient lesion targeting; and 3. Overcoming the bottleneck of low transfection efficiency. Therefore, improving the performance of the gene delivery system through innovative optimization of the vector structure is undoubtedly a powerful approach to overcoming the challenges of non-viral gene vector research.

[0004] Host-guest recognition based on macrocyclic molecules is one of the most promising areas for research and development in supramolecular chemistry. This host-guest recognition between macrocyclic molecules and guest molecules, achieved through the synergistic action of multiple weak interactions, provides researchers with new, simpler, and more efficient approaches and methods for constructing stimuli-responsive and innovative materials. Pillarene, due to its unique structural properties, has shown promising application prospects and has become a leader among the new generation of macrocyclic hosts. The repeating units of pillararenes, 1,4-dimethoxybenzene, are linked at the 2 and 5 positions by methylene bridges, forming a symmetrical columnar structure. This gives pillararenes unique advantages and features: (a) an electron-rich molecular cavity, facilitating the formation of novel host-guest complexes; (b) the molecular structure is amenable to chemical synthesis and modification, allowing pillararenes of desired structures to be obtained through simple and efficient chemical reactions; and (c) a rich stimuli-responsiveness, which can be achieved not only through dynamic and reversible host-guest complex interactions but also by introducing specific responsive modifying groups. These characteristics are key to the widespread application and success of pillararene-based functional materials in various fields.

[0005] Therefore, the present invention provides a method for preparing an oxidation-responsive cationic water-soluble pillararene, which can tightly bind to negatively charged nucleic acids through electrostatic interactions. At the same time, it combines with the physiological microenvironment characteristics of cancer, injury, inflammation and other lesions, which have a large number of reactive oxygen free radicals (ROS). The pillararene can effectively load nucleic acid drugs and, after reaching the lesion site, remove the positive charge of the pillararene by oxidatively shedding the boric acid ester and releasing the nucleic acid drug, thereby improving the efficient delivery and release of nucleic acid drugs in gene therapy.

[0006] In summary, the oxidation-responsive cationic water-soluble pillararenes of the present invention utilize the ROS microenvironment within diseased cells to trigger charge changes and release the loaded nucleic acid drug. Currently, there are no reports of such oxidation-responsive decationized pillararenes, nor are there reports on their preparation methods and applications.

[0007] Summary of the Invention

[0008] This invention addresses the use of an oxidation-responsive cationic water-soluble pillararene as a nucleic acid carrier in gene therapy. This formulation exhibits advantages such as efficient loading and release, excellent biocompatibility, and the ability to form nanocomplexes with DNA, RNA, and other short-chain nucleic acids, demonstrating high transfection efficiency.

[0009] An oxidation-responsive cationic water-soluble pillar aromatic hydrocarbon comprises the following structure:

[0010] In the above formula:

[0011] X is an integer from 1 to 4, R1 and R2 are each independently the following fragment, wherein R3 and R4 are each independently H, a C1-C6 alkyl group or an acyl group;

[0012] R5, R6, R7, and R8 are independently H, C1-C6 alkyl or aromatic groups;

[0013] R7 and R9 are independently H, C1-C20 alkyl or aromatic groups;

[0014] The anion is bromide or chloride.

[0015] Preferably, the copolymerized pillar[5]arene is obtained by reacting the monomers 1-(2-haloethoxy)-4-methoxybenzene and 4-alkoxymethyl(ethyl)oxybenzene in the presence of a Lewis acid catalyst in a monomer ratio of 2:3 and 1:4, and then reacting with dimethylamine and diethylamine to obtain a copolymerized pillar[5]arene substituted with a tertiary amine, and then reacting with boric acid benzylidene bromide, boric acid ester benzylidene bromide, boric acid benzylidene chloride or boric acid ester benzylidene chloride to obtain the copolymerized pillar[5]arene.

[0016] Preferably, the pillar aromatics are copolymerized pillar[5] aromatics, and the copolymerization ratio is 1:4, i.e., x=1 or 4, and 2:3, i.e., x=2 or 3;

[0017] Preferably, the copolymerized pillar[5]arene includes pillar[5]arene obtained by copolymerization and ring-forming reaction with a reaction monomer molar ratio of 2:3, where the product x=2 or 3; a reaction monomer molar ratio of 1:4, where the product x=1 or 4.

[0018] Preferably, R3 and R4 are boric acid, methyl borate, ethyl borate or pinacol borate.

[0019] Preferably, R5, R6, R7, and R8 are methyl or ethyl.

[0020] As a preference, the above R9 is methyl or ethyl, R 10 It is an alkyl group or an aromatic group having more than 6 carbon atoms, and more preferably an n-alkyl group.

[0021] Preferably, the above compound can be prepared by the following method:

[0022] 1-(2-haloethoxy)-4-methoxybenzene and 4-alkoxymethyl(ethyl)oxybenzene are reacted together under Lewis acid catalyst conditions to obtain a copolymerized pillar[5]arene copolymerized in a monomer ratio of 1:4, which is then reacted with dimethylamine and diethylamine to obtain a copolymerized pillar[5]arene substituted with a tertiary amine, which is then reacted with boric acid benzyl bromide, boric acid ester benzyl bromide, boric acid benzyl chloride or boric acid ester benzyl chloride to obtain the copolymerized pillar[5]arene.

[0023] Its preparation method is as follows:

[0024] The oxidation-responsive cationic water-soluble pillararenes of the present invention can drop the positive charge of the quaternary amine salt through an oxidation reaction with active oxygen species such as hydrogen peroxide, and transform into an electrically neutral tertiary amine. The reaction formula of this process is shown below:

[0025] The present invention also provides an application of the oxidation-responsive cationic water-soluble pillar aromatic hydrocarbon in the transport of DNA, RNA and other short-chain nucleic acids.

[0026] Compared with the existing technology, the present invention has the following beneficial effects:

[0027] (1) The oxidation-responsive cationic water-soluble pillar aromatic hydrocarbon prepared by the present invention is a small molecule gene delivery vector. It not only has the characteristics of high charge density and high nucleic acid loading efficiency, but also has a simple structure and is cheap to synthesize. Compared with traditional polymer-type nucleic acid vectors, it also has higher stability and reproducibility.

[0028] (2) The oxidation-responsive cationic water-soluble columnar aromatic hydrocarbon nucleic acid carrier prepared by the present invention can achieve effective nucleic acid release by reacting with active oxygen species, avoiding the problem of decreased transfection efficiency caused by traditional quaternary ammonium carriers being too tightly bound to nucleic acids and difficult to relay.

[0029] (3) The oxidation-responsive cationic water-soluble columnar aromatic hydrocarbon nucleic acid carrier prepared by the present invention exhibits a higher transfection efficiency in cells, has lower cytotoxicity than the gold standard PEI for gene transfection, and has good biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] FIG1 is a graph showing the reaction of compound 1 under oxidative conditions in Examples 1, 2 and 3. 1 HNMR spectrum

[0031] FIG2 shows the particle size and Zeta potential of nanocomplexes with different N / P ratios formed by compound 1 and plasmid DNA in Example 4 of the present invention;

[0032] FIG3 is a gel retardation electrophoresis diagram of nanocomplexes with different N / P ratios formed by compound 1 and plasmid DNA in Example 4 of the present invention;

[0033] FIG4 is a gel retardation electrophoresis diagram of a nanocomplex formed by compound 1 and plasmid DNA in Example 4 of the present invention under oxidative conditions;

[0034] FIG5 is a graph showing changes in particle size and zeta potential of a nanocomplex formed by compound 1 and plasmid DNA in Example 4 of the present invention under oxidative conditions;

[0035] FIG6 is a transmission electron micrograph of a nanocomplex formed by compound 1 and plasmid DNA in Example 4 of the present invention;

[0036] FIG7 is a graph showing the cytotoxicity of compound 1 in Example 5 of the present invention at different concentrations;

[0037] FIG8 is a diagram showing the cell transfection effect of the nanocomplex formed by compound 1 and plasmid DNA in Example 6 of the present invention under different N / P conditions;

[0038] FIG9 is a diagram showing the cell transfection effect of the nanocomplex formed by compound 1 and mRNA in Example 7 of the present invention under different N / P conditions;

[0039] FIG10 is a diagram showing the cell transfection effect of the compound 1 / DNA nanocomplex in Example 8 of the present invention under oxidative conditions; DETAILED DESCRIPTION

[0040] The present invention provides some specific implementation examples, but the present invention is not limited to these embodiments.

[0041] Embodiment 1:

[0042] Synthesis of compound 1

[0043] 1-Methoxy-4-hexadecylbenzene (1.74 g, 5.00 mmol) and 4-bis(2-bromoethoxy)benzene (6.48 g, 20.0 mmol) were placed in 80 mL of 1,2-dichloroethane, and then boron trifluoride etherate (3.20 mL, 25 mM) was added and stirred at room temperature for 2 h. After the reaction, the reaction solution was poured into methanol to precipitate a large amount of white solid. The precipitate was filtered and dissolved in dichloromethane. The insoluble matter was removed by filtration again. The organic phase was washed twice with dichloromethane solution to obtain the organic phase, which was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was chromatographed by column chromatography with a mobile phase of petroleum ether / ethyl acetate = 50:1 (R f =0.50), the product compound 3 was obtained by spin drying as a white powdery solid (0.85 g, 10%).

[0044] Compound 3 (1.64 g, 1.00 mmol) and excess diethylamine (7.50 g, 100 mmol) were added to 100 ml of anhydrous ethanol and heated with stirring under reflux at 80 ° C for 24 h. After the reaction, the solvent was removed by rotary evaporation, and 200 mL of 1 M sodium hydroxide solution was poured into it and stirred for 1 h. The reaction solution was then fully extracted with ethyl acetate, and the organic phase was dried to obtain dark yellow oily compound 2 (1.55 g, 98%).

[0045] Compound 2 (0.49 g, 0.30 mM) and 4-bromomethylphenylboronic acid pinacol ester (0.78 g, 2.64 mmol) were dissolved in 25 mL of acetonitrile and heated with stirring at 75°C under reflux for 24 hours. After the reaction, the mixed solution was concentrated to 2.0 mL and poured into excess ether. The precipitated white precipitate was collected by filtration and washed thoroughly with ether. The obtained white precipitate was dried in a vacuum oven to obtain the product compound 1 (0.98 g, 81.6%).

[0046] The structural detection data of compound 1 are as follows:

[0047] 1H NMR(400MHz,D2O,298K)δ(ppm):7.80–7.73(m,16H),7.54–7.45(m,16H),6.97–6.63(m,10H),4.79–4.58(m,16H),4.48–4.38(m,16H),4.30–4 .25(t,2H),3.86–3.77(m,16H),3.68(s,3H),3.65–3.50(s,10H),3.50 –3.42(m,32H),1.46–1.39(m,48H),1.23(s,96H),0.71–0.41(m,31H). 13 C NMR (600MHz, CD3OD, 298K) δ (ppm): 151.45, 136.54, 135.77, 133.44, 133.01, 131.50, 117.47, 85.54, 75. 81,70.92,63.47,58.05,55.57,33.05,30.79,30.46,25.27,25.03,23.72,14.47,8.96.HR-MS:[M–8Br] 8+ m / z determined to be 422.8757, [M–7Br] 7+ The m / z was determined to be 494.5405. The melting point was 162.4-162.9°C.

[0048] Example 2:

[0049] H2O2 responsiveness of compounds

[0050] A certain amount of compound 1 was weighed and dissolved in D2O (1mM), and a small amount of hydrogen peroxide was added to make the final concentration 10mM. Under the oxidative conditions, compound 1 quickly reacted and produced quinone, which was converted into p-hydroxybenzyl alcohol in water. The process changes through 1 The corresponding proton peaks a, b, and c can be observed in H NMR, indicating that compound 1 undergoes a redox response. The detection results are shown in Figure 1.

[0051] Example 4:

[0052] Preparation and characterization of compound 1 and plasmid DNA nanocomplex

[0053] A certain amount of compound 1 was dissolved in HEPES buffer solution (pH = 7.4, 10mM) at a concentration of 2.0mg / ml. At the same time, plasmid DNA was also diluted with HEPES buffer solution to a concentration of 40μg / ml. After the compound was diluted to the corresponding concentration according to the corresponding N / P molar ratio, it was quickly added to the plasmid DNA solution at a 1:1 volume ratio, vortexed for 30s, and then allowed to stand for 30min to obtain a series of nanocomplexes with different N / P ratios.

[0054] Nanocomposite Particle Size and Zeta Potential: Appropriate amounts of the prepared nanocomposites with varying N / P ratios were placed in a sample cell. Dynamic light scattering was used to measure the particle size and zeta potential of the nanocomposite solutions. Three replicates were performed for each sample, and the average value was taken. As shown in Figure 2, the nanocomposite particle size was approximately 70-100 nm, and the zeta potential was 20-30 mV.

[0055] Nanocomplex Gel Retardation Experiment: A 1.0% agarose gel (containing 2 μg / ml of gelred) was prepared and placed in 1×TAE buffer. 20 μL of the different N / P nanocomplexes to be tested and 20 μL of pure plasmid DNA of the same concentration as a control were added to the gel wells. Electrophoresis was performed at 120 mV for 30 minutes. After electrophoresis, the gel was placed in a gel imaging system and photographed. The results are shown in Figure 3. After wrapping DNA, compound 1 can effectively block the migration of DNA, thereby effectively protecting DNA during gene delivery.

[0056] Gel retardation experiment of nanocomplexes under oxidative conditions: Take a nanocomplex with an N / P ratio of 15 and incubate it in H2O2 solutions of different concentrations at 37°C for 30 minutes. Then, similarly, prepare 1.0% agarose gel (containing 2μg / ml of gelred) and place it in 1×TAE buffer. Add 20μL of the incubated nanocomplex to the gel wells and 20μL of pure plasmid DNA of the same concentration as a control. Apply a voltage of 120mV and electrophoresis for 30 minutes. After the end, place the gel in a gel imaging system and take pictures. The results are shown in Figure 4. The results show that under oxidative conditions, the nanocomplex can effectively release DNA.

[0057] Changes in the particle size and potential of the nanocomplex under oxidative conditions: Nanocomplexes with an N / P ratio of 15 were incubated in H2O2 solutions of varying concentrations at 37°C for 30 min. Appropriate amounts were placed in sample cells and the particle size of the nanocomplex solutions with varying N / P ratios was measured using dynamic light scattering. Three replicates were performed for each sample and the average value was calculated. Nanocomplexes with an N / P ratio of 15 were incubated in H2O2 solutions of 1.0 mM at 37°C. Appropriate amounts were placed in sample cells at different time points and the particle size was measured using dynamic light scattering. Three replicates were performed for each sample and the average value was calculated. As shown in Figure 5, the nanocomplex particle size gradually increased with increasing hydrogen peroxide concentration, and the tightly bound nanocomplexes dissociated. Over time, the surface potential of the nanocomplexes shifted from positive to negative, and the electrostatic attraction with the negatively charged DNA disappeared.

[0058] Transmission electron microscopy observation experiment of nanocomplexes: A nanocomplex with an N / P ratio of 15 was taken and dropped onto a 300-mesh copper grid. It was negatively stained with phosphotungstic acid, and the liquid was washed off with the edge of filter paper and naturally dried at room temperature. The nanocomplex sample on the copper grid was then observed by transmission electron microscopy. As shown in Figure 6, compound 1 and plasmid DNA complexed to form nearly spherical nanoparticles with regular morphology and a particle size of approximately 80 nm, which is basically consistent with the dynamic light scattering detection results.

[0059] Example 5:

[0060] Cytotoxicity experiment of compound 1: The cytotoxicity evaluation of compound 1 was characterized using a CCK8 kit. The cells were cultured in different concentrations of compound 1 carriers, and the traditional polymer gene carrier PEI was used as a control. The cells were incubated for 48 hours. After the culture was completed, the culture medium was discarded and the diluted CCK8 reagent was added. The cells were incubated for 1-2 hours. The light absorbance at a wavelength of 450nm was measured with a microplate reader. The cell survival rate was calculated by comparing it with the control group. The results are shown in Figure 7. As the concentration increases, the survival rate of cells treated with PEI decreases sharply, while the survival rate of cells treated with compound 1 decreases slowly and is significantly higher than that of PEI, proving that compound 1 has higher biosafety within the measured concentration range.

[0061] Example 6:

[0062] Luciferase gene transfection assay: A549 cells were cultured in 96-well plates at a density of 15,000 cells per well in 200 μL of culture medium. The cells were incubated at 37°C in an incubator with 5% CO₂ and 95% humidity for 24 h. The medium was then removed and replaced with fresh serum-free medium. Compound 1 was then added to the luciferase gene plasmid to form nanocomplexes with varying N / P ratios. The complexes were then added to the culture medium and incubated at 37°C for 4 h. The medium was again removed and replaced with fresh medium for an additional 48 h. Following incubation, the medium was removed and 20 μL of 1× cell lysis buffer was added. After lysis for 10 min, 5 μL of the supernatant was added to 20 μL of luciferase substrate. The chemiluminescence intensity was measured using a chemiluminescence detector. Protein concentration was determined using a Bradford protein assay kit. Each data set was measured in triplicate and the average value was calculated. The chemiluminescence intensity was normalized by protein concentration to obtain the luminescence intensity per milligram of protein (RLU / mg protein). The results are shown in FIG8 . Compared with the commonly used non-viral gene vector PEI, the transfection efficiency of compound 1 is increased by 1-2 orders of magnitude, indicating that compound 1 is a more efficient gene vector.

[0063] Example 7:

[0064] Green fluorescent protein mRNA transfection experiment: RAW264.7 cells were cultured in 15mm radius glass bottom culture dishes with a cell density of 25,000 cells per dish. 1.5mL of culture medium was added and the cells were cultured in a 37°C incubator containing 5% CO2 and 95% humidity for 24 hours. Subsequently, the culture medium in the culture dish was replaced with serum-free medium, and the prepared complex of compound 1 and green fluorescent protein mRNA was added. The cells were incubated at 37°C for 4 hours. The culture medium was discarded again and replaced with fresh culture medium for a further 48 hours. After the culture was completed, the expression of green fluorescent protein was observed using a laser confocal microscope with an excitation wavelength of 488nm and an emission wavelength of 510-540nm. All photos were taken under a 10x objective lens and the light intensity was kept consistent for all samples. The results are shown in Figure 9. Compared with PEI, compound 1 loaded with mRNA can produce higher transfection efficiency.

[0065] Example 8:

[0066] Cell transfection experiment of nanocomplexes under oxidative conditions: A549 cells were cultured in 96-well plates at a density of 15,000 cells per well in 200 μL of culture medium and incubated at 37°C in an incubator with 5% CO2 and 95% humidity for 24 hours. The culture medium was discarded and replaced with fresh culture medium. H2O2 was added to the culture medium to prepare a highly oxidative environment with different concentrations (5 μM, 10 μM, 20 μM, 50 μM, and 100 μM) to simulate the hyperoxidative microenvironment of the tumor. Compound 1 was then added to the luciferase gene plasmid to form nanocomplexes with different N / P ratios. The complexes were added to the culture medium and incubated at 37°C for 4 hours. The culture medium was then discarded again and replaced with fresh culture medium for a further 48 hours. After the incubation period, the culture medium was discarded and 20 μL of 1× cell lysis buffer was added. After lysis for 10 minutes, 5 μL of the supernatant was added to 20 μL of luciferase substrate. The chemiluminescence intensity was measured using a chemiluminescence detector, and the protein concentration was determined using a Bradford protein assay kit. Each set of data was measured in parallel for three replicate wells and the average value was taken. The chemiluminescence intensity was normalized by protein concentration to obtain the luminescence intensity per milligram of protein (RLU / mg protein). The results are shown in Figure 10. In an oxidative environment, the expression efficiency of the plasmid DNA loaded with compound 1 was improved.

Claims

1. An oxidation-responsive cationic water-soluble pillararene, Characterized in that, It includes the following structure: In the above formula: X is an integer from 1 to 4, R 1 , R 2 are each independently the following fragments, where R 3 , R 4 are each independently H, C1-C6 alkyl or acyl; R 5 、R 6 and R 7 、R 8 are each independently an alkyl or aryl group having 1 to 6 carbon atoms; R 9 、R 10 are each independently H, a C1-C20 alkyl group or an aryl group; The anion is bromide or chloride ion.

2. The oxidation-responsive cationic water-soluble pillararene according to claim 1, Characterized in that, The pillararene is prepared by reacting a pillararene containing primary, secondary amino groups or tertiary amine with a benzyl group containing boronic acid group or benzyl borate group.

3. The oxidation-responsive cationic water-soluble pillararene according to claim 2, Characterized in that, The pillararene is a copolymerized pillar[5]arene, and the copolymerization ratio is 1:4, that is, x = 1 or 4, 2:3, that is, x = 2 or 3.

4. The oxidation-responsive cationic water-soluble pillararene according to claim 1 or 3, Characterized in that, The copolymerized pillar[5]arene includes the pillar[5]arene obtained by copolymerization cyclization reaction according to the molar ratio of reaction monomers 2:3, and the product x = 2 or 3; according to the molar ratio of reaction monomers 1:4, the product x = 1 or 4.

5. The oxidation-responsive cationic water-soluble pillararene according to claim 1, 3 or 4, Characterized in that: R 5 、R 6 and R 7 、R 8 are methyl or ethyl, R 9 is methyl or ethyl, R 10 is an alkyl or aryl group having 6 to 20 carbon atoms.

6. The preparation method of the oxidation-responsive cationic water-soluble pillararene according to claim 1, 2, 3 or 4, Characterized in that: The pillararene compound is specifically as follows: 1-(2-haloethoxy)-4-methoxybenzene and 4-alkoxymethoxy(ethoxy)benzene react together under the condition of Lewis acid catalyst to obtain copolymerized pillar[5]arene copolymerized according to the molar ratio of 2:3 and 1:4, and then react with dimethylamine and diethylamine to obtain a copolymerized pillar[5]arene substituted with tertiary amine, and then react with benzyl bromide with boronic acid group, benzyl borate bromide, benzyl chloride with boronic acid group or benzyl borate chloride.

7. The application of an oxidation-responsive cationic water-soluble pillararene in transporting nucleic acid substances, including DNA, RNA and other short-chain nucleic acids.

8. The preparation method of the oxidation-responsive cationic water-soluble pillararene according to claim 2, Characterized in that: The preparation method of the pillararene compound is specifically as follows: 1-(2-haloethoxy)-4-methoxybenzene reacts with paraformaldehyde or trioxane under the condition of Lewis acid catalyst to obtain cyclic pillar[n]arene, where n = 5-15, and then reacts with dimethylamine and diethylamine to obtain a pillar[n]arene substituted with tertiary amine, and then reacts with benzyl bromide with boronic acid group, benzyl borate bromide, benzyl chloride with boronic acid group or benzyl borate chloride.

9. The preparation method of the oxidation-responsive cationic water-soluble pillararene according to claim 2, Characterized in that: The structure of the water-soluble pillararene prepared by this method is as follows:

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