Supramolecular Complex, Preparation Method Thereof and Application Thereof

Abstract

The supramolecular complex provided by the present disclosure, its preparation method and application. The structural formula of the supramolecular complex is CB-EG x -L-Guest, where CB is the host molecule, and cucurbituril is selected; EG x is a class of linking segments; L is a dynamic covalent bond; Guest is the guest molecule. The preparation method includes: synthesizing a conjugate molecule of Guest-activated ester; synthesizing a conjugate molecule of Guest-L; synthesizing a conjugate molecule of Guest-L-activated ester; synthesizing a conjugate molecule of CB-EG x ; mixing the conjugate molecule of CB-EG x and the conjugate molecule of Guest-L-activated ester to obtain a crude product containing the supramolecular complex; and obtaining the supramolecular complex after purification. The supramolecular complex of the present disclosure has a self-inclusion property and can achieve stable encapsulation and efficient release of guest molecules. The preparation method is simple and rapid, and has broad application prospects in the field of anti-tumor drugs.

Description

Technical Field

[0001] The present disclosure belongs to the field of biomedicine, and particularly relates to a supramolecular complex, a preparation method thereof, and an application thereof. Background Art

[0002] Chemotherapy is one of the common treatment regimens for cancer. However, due to the lack of certain selectivity of chemotherapy drugs, the toxic and side effects on patients are relatively obvious during the treatment process. In recent years, both experimental studies and clinical manifestations have shown that using a nanocarrier to encapsulate chemotherapy drugs can effectively reduce the toxic and side effects of the drugs and improve the quality of life of patients. However, the disadvantage is that the nanocarrier is not stable enough, manifested as unstable components, unstable drug loading capacity, and unstable driving force for drug loading. The uncontrollable chemical composition between batches and the premature drug leakage in vivo make the drug efficacy of the nanocarrier not stable enough. In contrast, the supramolecular drug system constructed by host-guest chemistry has characteristics such as a clear structure and stimulus response. The strategy of using supramolecular drugs is expected to achieve the precision and intelligence of the drug delivery system, providing a method for precision medicine.

[0003] In the construction of supramolecular drugs, the core lies in the design of the molecular structure, aiming to encapsulate the drug molecules in the cavity of the host molecule under normal physiological conditions. On the one hand, it encapsulates the drug active sites, reduces its anti-tumor activity, and improves safety. On the other hand, it also protects the drug molecules from being adsorbed by in vivo proteins and improves the drug utilization rate. When reaching the tumor site, the supramolecular drug can release the drug molecules and restore the anti-tumor activity of the drug to kill cancer cells. However, in the existing supramolecular drug design, the drug molecules and the host molecule adopt intermolecular binding, and the formed supramolecular drug will dissociate as the concentration decreases, resulting in premature drug leakage and making the efficacy and toxicity reduction effect of the supramolecular drug not significant enough.

[0004] Therefore, how to further design a supramolecular drug with both stable encapsulation and efficient release has important clinical application prospects and medical significance. Summary of the Invention

[0005] The present disclosure aims to solve at least one of the technical problems in the related art to some extent.

[0006] To this end, a supramolecular complex with self-inclusion characteristics and capable of achieving stable encapsulation and efficient release of guest molecules provided in the first aspect embodiment of the present disclosure has a structural formula of CB-EG x -L-Guest, wherein:

[0007] CB is a host molecule, and cucurbituril is selected;

[0008] EG x is a linking segment;

[0009] L is a dynamic covalent bond;

[0010] Guest is the guest molecule.

[0011] In some embodiments, the host molecule is selected from cucurbit[6]uril, cucurbit[7]uril or cucurbit[8]uril.

[0012] In some embodiments, the guest molecule is selected from anti-tumor drug molecules.

[0013] In some embodiments, the anti-tumor drug molecule is selected from chlorambucil, cisplatin, paclitaxel, camptothecin, hydroxycamptothecin or irinotecan.

[0014] In some embodiments, the linking segment is selected from oxyethyl, x is the number of repeating units, and x ranges from 1 to 5.

[0015] In some embodiments, the dynamic covalent bond is selected from disulfide bond, diselenide bond, phenylboronic ester or Schiff base.

[0016] In some embodiments, the supramolecular complex formed undergoes intramolecular binding, that is, the formed molecular structure is a self-inclusion structure.

[0017] The supramolecular complex provided by the first aspect embodiment of the present disclosure has the following characteristics and beneficial effects:

[0018] The cucurbituril used in the embodiments of the present disclosure, especially cucurbit[7]uril (CB[7]), has good water solubility and biocompatibility, and it can form host-guest complexes with many chemotherapy drug molecules, which is the basis for constructing supramolecular drugs. In addition, designing the molecular structure to enable self-inclusion of the drug-host conjugate molecule is one of the key points of the present disclosure. Self-inclusion is an intramolecular host-guest binding. On the one hand, the binding constant of self-inclusion is dimensionless, so the binding rate of the drug to cucurbituril will not decrease with the dilution of the sample; on the other hand, after the host-guest conjugate molecule is formed between cucurbituril and the drug molecule, the local concentration is increased in the spatial structure, and the entropy change of the host-guest binding is also more favorable, thus greatly enhancing the binding ability of the drug to cucurbituril. Therefore, the constructed self-inclusion supramolecular drug can stably encapsulate drug molecules at various concentrations, prevent the premature leakage of the drug, and increase the drug safety and drug utilization rate. The reducible-responsive cleavable dynamic covalent bond enables the self-inclusion supramolecular drug to break the conjugate molecule in the tumor-reducing microenvironment, and the drug molecule and cucurbituril will change from the intramolecular self-inclusion binding mode to the intermolecular binding, so that the drug molecule is efficiently released.

[0019] The present disclosure first proposes and synthesizes a self-inclusion supramolecular drug, realizing the stable encapsulation and efficient release of anti-tumor drugs at any concentration, and showing a highly efficient and low-toxic anti-tumor effect in organisms.

[0020] In summary, the supramolecular complex provided by the present disclosure has the advantages of precise structure, high encapsulation efficiency, stable encapsulation and efficient release compared with the traditional drug-loading system, and has good clinical application prospects in the field of cancer treatment.

[0021] The preparation method of the supramolecular complex provided by the second aspect of the present disclosure includes:

[0022] Adding the guest molecule into a first solvent, and synthesizing a conjugate molecule of the guest molecule-activated ester through an activated ester reaction;

[0023] Adding the conjugate molecule of the guest molecule-activated ester and the dynamic covalent bond molecule into a second solvent, and synthesizing a conjugate molecule of the guest molecule-dynamic covalent bond through a bimolecular nucleophilic substitution reaction;

[0024] Adding the conjugate molecule of the guest molecule-dynamic covalent bond into a third solvent, and synthesizing a conjugate molecule of the guest molecule-dynamic covalent bond-activated ester through an activated ester reaction;

[0025] Adding the host molecule and the azide-containing linker segment into a fourth solvent, and synthesizing a conjugate molecule of the host molecule-linker segment through a click chemical reaction catalyzed by cuprous ions;

[0026] Mixing the conjugate molecule of the host molecule-linker segment and the conjugate molecule of the guest molecule-dynamic covalent bond-activated ester in a fifth solvent to obtain a crude product containing the supramolecular complex;

[0027] Purifying the crude product to obtain the supramolecular complex.

[0028] In some embodiments, the first solvent is selected from any one or a mixture of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide, and dichloromethane is preferably selected.

[0029] In some embodiments, the second solvent is selected from any one or a mixture of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide, and dichloromethane is preferably selected.

[0030] In some embodiments, the third solvent is selected from any one or a mixture of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide, and dichloromethane is preferably selected.

[0031] In some embodiments, the fourth solvent is selected from any one or a mixture of ultrapure water, deionized water, dimethyl sulfoxide and acetonitrile, and deionized water is preferably selected.

[0032] In some embodiments, the fifth solvent is selected from any one or a mixture of methanol, trifluoroacetic acid, ultrapure water, deionized water, dimethyl sulfoxide, and acetonitrile, and dimethyl sulfoxide is preferably selected.

[0033] In some embodiments, the purification of the crude product includes the following steps:

[0034] The crude product is successively subjected to sedimentation, washing, and recrystallization with a sixth solvent to obtain a preliminarily purified product, and the supramolecular complex is obtained by column chromatography of the preliminarily purified product.

[0035] In some embodiments, the sixth solvent is selected from any one or a mixture of acetone, ether, methanol, dichloromethane, chloroform, and deionized water.

[0036] The preparation method of the supramolecular complex provided by the second aspect embodiment of the present disclosure has the following characteristics and beneficial effects: The preparation is modular, and a variety of drug molecules can be replaced; The reactions involved in the process are all characterized by mild reaction, fast rate, and high efficiency; The self-inclusion structure of the host-guest conjugate molecule is different from that of the supramolecular polymer, which is convenient for the purification of the product. It is suitable for mass production.

[0037] In the application of the supramolecular complex provided by the third aspect embodiment of the present disclosure in the field of anti-tumor drugs, in the normal physiological environment, the drug guest molecule in the supramolecular complex of the embodiment of the present disclosure is self-included in the host molecule to form a host-guest complex with low drug toxicity; In the reducing microenvironment of the tumor, the dynamic covalent bond of the supramolecular complex of the embodiment of the present disclosure is broken, the effect of self-inclusion enhancing guest binding disappears, the host-guest complex dissociates, and the released drug guest molecule can restore its anti-cancer effect and precisely kill cancer cells. Description of the Drawings

[0038] Figure 1 It is a schematic diagram of the supramolecular complex stably loading drugs and efficiently releasing drugs proposed by the embodiment of the present disclosure.

[0039] Figure 2 It is the nuclear magnetic resonance hydrogen spectrum of the supramolecular complex prepared in Example 1 of the present disclosure.

[0040] Figure 3 It is the electrospray ionization mass spectrum of the supramolecular complex prepared in Example 1 of the present disclosure.

[0041] Figure 4 It is the characterization of the host-guest composite structure of the supramolecular complex prepared in Example 1 of the present disclosure by nuclear magnetic resonance hydrogen spectrum.

[0042] Figure 5The trapped ion mobility spectrometry of the supramolecular complex prepared in Example 1 of the present disclosure at different concentrations and the demonstration of its self-inclusion structure.

[0043] Figure 6 It is the result of the competitive isothermal titration calorimetry experiment of the supramolecular complex prepared in Example 1 of the present disclosure.

[0044] Figure 7 a) - c) of is the drug release curve of the supramolecular complex prepared in Example 1 of the present disclosure under different concentrations of glutathione (GSH).

[0045] Figure 8 It is the result of cell viability detected by CCK-8 kit staining and microplate reader after co-incubating the supramolecular complex prepared in Example 1 of the present disclosure with human cervical cancer cells (Hela) at different concentrations for 48 h.

[0046] Figure 9 a) - c) of is the adsorption result of human serum albumin on camptothecin solution (CPT), cucurbit[7]uril-camptothecin (CPT@CB[7]) and the supramolecular complex (CB-EG 1 -SS-CPT) prepared in Example 1 of the present disclosure detected by fluorescence spectrometer.

[0047] Figure 10 a), b) of are the change curves of tumor volume and body weight of nude mice with tumors over time after tail vein injection of phosphate buffer solution (PBS), camptothecin solution (CPT), self-inclusion supramolecular drug (CB-EG 3 -CPT) solution and the supramolecular complex (CB-EG 1 -SS-CPT) solution prepared in Example 1 of the present disclosure. Detailed implementation manners

[0048] In order to more clearly describe the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. The drawings in the following description are only some embodiments of the present invention. If not clearly stated, all reagents used in the following embodiments are commercially available, or can be synthesized according to the reported methods or the methods in this article, which are easily obtained for those skilled in the art, and other drawings can also be derived from these drawings.

[0049] Example 1:

[0050] Based on EG 1 The preparation of the supramolecular complex CB-EG 1 -SS-CPT containing disulfide bond and camptothecin includes the following steps:

[0051] 1) Disperse 1.4 g of (S)-(+)-camptothecin (CPT) into 100 mL of deoxygenated anhydrous dichloromethane. Add 0.65 g of 4-dimethylaminopyridine under an ice-water bath, and then slowly add triphosgene (600 mg). Stir at room temperature for 4 hours under nitrogen protection. That is, a conjugate molecule of CPT-activated ester is generated.

[0052] 2) Extract 3.85 g of bis(2-hydroxyethyl) disulfide with dichloromethane. After the reaction solution of the above CPT-activated ester conjugate molecule becomes clear, drop it into the disulfide substrate and stir at room temperature overnight under nitrogen. Then spin-dry the reaction solution, wash it three times with 10 mL of deionized water, and then wash it twice with 10 mL of methanol. Vacuum dry to obtain a pale yellow powder. That is, a conjugate molecule of drug-dynamic covalent bond, and its structural formula is:

[0053]

[0054] 3) Dissolve 260 mg of the above conjugate molecule of drug-dynamic covalent bond into 20 mL of anhydrous dichloromethane, and successively add 50 mg of triethylamine and 10 mg of 4-dimethylaminopyridine under an ice-water bath to make a reaction solution. Dissolve 120 mg of 4-nitrophenyl chloroformate into 5 mL of anhydrous dichloromethane, and add it to the reaction solution under an ice-water bath. React at room temperature for 1 hour under nitrogen. After the reaction is completed, rotary evaporate and concentrate the reaction solution to about 10 mL, add it to 200 mL of anhydrous ether to precipitate for 30 minutes, then filter to obtain the filter cake, and wash it twice with anhydrous ether to obtain a pale yellow powder, that is, a conjugate molecule of CPT-SS-activated ester, and its structural formula is:

[0055]

[0056] 4) Dissolve mono-alkynyl-modified cucurbit[7]uril C8bim@CB[7]-OA (192 mg) and azido-monoethylene glycol-amine (52.0 mg) in 2 mL of ultrapure water. Then add CuAAC catalyst (2.0 mg), use a double-tube to displace the atmosphere in the system with nitrogen, and then stir at 55 °C for 18 hours. After the reaction is completed, add 500 mg of KBr and dissolve it by ultrasonic wave, and then freeze-dry the reaction solution. Wash the above freeze-dried powder twice with anhydrous dichloromethane and anhydrous methanol respectively, and then vacuum dry the solid overnight to obtain a white powder, that is, the conjugate molecule of cucurbit[7]uril-EG 1 with the structural formula:

[0057]

[0058] 5) Dissolve 140 mg of the conjugate molecule of CPT-SS-activated ester into 5 mL of ultra-dry dimethyl sulfoxide (DMSO) to make reaction solution 1. Take 128 mg of cucurbit[7]uril-EG 1The conjugate molecules were dispersed into 5 mL of ultradry DMSO, and 15.0 mg of N,N-diisopropylethylamine was added. After ultrasonic homogenization, reaction solution 2 was prepared. Reaction solution 2 was added dropwise to reaction solution 1, and the mixture was stirred at room temperature for 0.5 - 4 hours until the solution became clear and orange-red, indicating the end point of the reaction. The orange-red reaction solution was dropped into 50 mL of anhydrous ether and allowed to precipitate in an ice-water bath for 2 hours. Then, the precipitate was filtered, and the filter cake was washed twice with ice-cold anhydrous ether. Subsequently, the filter cake was washed ultrasonically once with 5 mL of dichloromethane, dried in vacuo, washed once with 5 mL of methanol, dried in vacuo, and finally washed once with 5 mL of deionized water and centrifuged to obtain a pale yellow solid, namely CB-EG 1 -SS-CPT crude product.

[0059] 6) After activating the normal-phase column packing with methanol for half an hour, the activated packing was obtained by centrifugation. The packing was dispersed with a methanol / water mixture to fill the chromatography column, and the mobile phase of methanol / water was gradually converted to ultrapure water by washing the chromatography column. CB-EG 1 -SS-CPT crude product was dissolved in an acetonitrile / water solution containing trifluoroacetic acid, and column chromatography was performed. The eluent was a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. During the process, a hand-held ultraviolet lamp with 365 nm excitation light was used to monitor the bands. The second yellow-emitting band was the product. The product fraction solution was collected, the acetonitrile was removed by rotary evaporation, and the product was freeze-dried to obtain a yellow powder, namely CB-EG 1 -SS-CPT, and its structural formula is:

[0060]

[0061] The supramolecular complex CB-EG 1 -SS-CPT prepared in this example can spontaneously form a self-inclusion structure in aqueous solution, and its structural formula is:

[0062]

[0063] Example 2:

[0064] Based on EG 2 、dithiol bond and camptothecin supramolecular complex CB-EG 2 -SS-CPT preparation.

[0065] The difference between this example and Example 1 is that the number x of ethoxy groups is different, which is manifested as the difference in steps 4) to 6). The remaining steps are the same as those in Example 1 and will not be elaborated here. Now, steps 4) to 6) of Example 2 are described in detail:

[0066] 4) Weigh the mono-alkynyl-modified cucurbit[7]uril C8bim@CB[7]-OA (211 mg) and 2-[2-(2-azidoethoxy)ethoxy]ethylamine (95.0 mg) and dissolve them in 2 mL of ultrapure water. Then add the CuAAC catalyst (2.0 mg), use a double-tube to displace the atmosphere in the system with nitrogen, and then stir at 55 °C for 18 hours. After the reaction is completed, add 500 mg of KBr and dissolve it by ultrasonic treatment, and then freeze-dry the reaction solution. Wash the above freeze-dried powder twice with anhydrous dichloromethane and anhydrous methanol respectively, and then vacuum-dry the solid overnight to obtain a white powder, namely cucurbit[7]uril-EG 2 conjugate molecule.

[0067] 5) Dissolve 120 mg of the conjugate molecule of CPT-SS-activated ester in 5 mL of ultradry DMSO to prepare reaction solution 1. Disperse 137 mg of the conjugate molecule of cucurbit[7]uril-EG 2 in 5 mL of ultradry DMSO, add 15.0 mg of N,N-diisopropylethylamine, and after ultrasonic homogenization, prepare reaction solution 2. Dropwise add reaction solution 2 into reaction solution 1, stir at room temperature for 0.5 - 4 hours, and when the solution becomes clear and orange-red, it is the end point of the reaction. Drop the orange-red reaction solution into 50 mL of anhydrous ether, precipitate in an ice-water bath for 2 hours, then filter and collect the filter cake, and wash it twice with ice-cold anhydrous ether. Then, wash the filter cake once with 5 mL of dichloromethane by ultrasonic treatment, vacuum-dry it, wash it once with 5 mL of methanol, vacuum-dry it, and finally wash it once with 5 mL of deionized water and centrifuge to obtain a pale yellow solid, namely the crude product of CB-EG 2 -SS-CPT.

[0068] 6) After activating the normal-phase column packing with methanol for half an hour, centrifuge to obtain the activated packing, disperse the packing with a methanol / water mixture to fill the chromatography column, and wash the chromatography column to gradually convert the mobile phase of methanol / water to ultrapure water. Dissolve the CB-EG 2 -SS-CPT crude product with an acetonitrile / water solution containing trifluoroacetic acid, perform column chromatography, and the eluent is a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. During the process, monitor the band with a handheld ultraviolet lamp with an excitation light of 365 nm, collect the product component solution, rotary evaporate to remove acetonitrile, and freeze-dry to obtain a pale yellow powder, namely CB-EG 2 -SS-CPT.

[0069] Example 3:

[0070] Preparation of the supramolecular complex CB-EG 3 based on EG 3 , disulfide bond and camptothecin.

[0071] The difference between this example and Example 1 lies in the different number x of ethoxy groups, which is manifested as the differences in steps 4) to 6). The remaining steps are the same as those in Example 1 and will not be elaborated here. Now, steps 4) to 6) of Example 3 will be described in detail:

[0072] 4) Weigh mono-alkynyl-modified cucurbit[7]uril C8bim@CB[7]-OA (208 mg) and 11-azido-3,6,9-trioxaundecan-1-amine (105 mg) and dissolve them in 2 mL of water. Then add CuAAC catalyst (2.0 mg), and use a double-tube to displace the atmosphere in the system with nitrogen. Then stir at 55 °C for 18 hours. After the reaction is completed, add 500 mg of KBr and dissolve it by ultrasonic treatment. Subsequently, lyophilize the reaction solution. Wash the above lyophilized powder twice with anhydrous dichloromethane and anhydrous methanol respectively, and then dry the solid under vacuum overnight to obtain a white powder, namely the conjugate molecule of cucurbit[7]uril-EG 3 of the conjugate molecule.

[0073] 5) Take 152 mg of the conjugate molecule of CPT-SS-activated ester and dissolve it in 5 mL of ultra-dry DMSO to prepare reaction solution 1. Take 166 mg of the conjugate molecule of cucurbit[7]uril-EG 3 of the conjugate molecule and disperse it in 6 mL of ultra-dry DMSO. Add 18.0 mg of N,N-diisopropylethylamine, and after ultrasonic homogenization, prepare reaction solution 2. Dropwise add reaction solution 2 into reaction solution 1, and stir at room temperature for 0.5 - 4 hours until the solution becomes clear and orange-red, which is the end point of the reaction. Drop the orange-red reaction solution into 50 mL of anhydrous ether, precipitate in an ice-water bath for 2 hours, then filter and collect the filter cake, and wash it twice with ice-cold anhydrous ether. Then, wash the filter cake once by ultrasonic treatment with 5 mL of dichloromethane, dry it under vacuum, wash it once with 5 mL of methanol, dry it under vacuum, and finally wash it once with 5 mL of deionized water and centrifuge to obtain a pale yellow solid, namely the crude product of CB-EG 3 -SS-CPT.

[0074] 6) After activating the normal-phase column packing with methanol for half an hour, centrifuge to obtain the activated packing. Disperse the packing with a methanol / water mixed solution to pack the chromatography column, and wash the chromatography column to gradually convert the mobile phase of methanol / water into ultrapure water. Dissolve the CB-EG3-SS-CPT crude product with an acetonitrile / water solution containing trifluoroacetic acid, perform column chromatography, and the eluent is a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. During the process, monitor the band with a handheld UV lamp with 365 nm excitation light, collect the product component solution, rotary evaporate to remove acetonitrile, and lyophilize to obtain a pale yellow powder, namely CB-EG 3 -SS-CPT.

[0075] Example 4:

[0076] Based on EG 5、Supramolecular complex CB-EG of disulfide bond and camptothecin 5 - Preparation of -SS-CPT

[0077] The difference between this example and Example 1 lies in the different number x of ethoxy groups, and cucurbit[8]uril is used in this example, which is manifested as the differences in steps 4) to 6). The remaining steps are the same as those in Example 1 and will not be elaborated here. Now, steps 4) to 6) of Example 4 are described in detail:

[0078] 4) Weigh monoynyl-modified cucurbit[8]uril C8bim@CB[8]-OA (85 mg) and 17-azido-3,6,9,12,15-pentaoxaheptadecane-1-amine (68 mg) and dissolve them in 2 mL of ultrapure water. Then add CuAAC catalyst (2.0 mg), use a double-tube to displace the atmosphere in the system with nitrogen, and then stir at 55 °C for 18 hours. After the reaction is completed, lyophilize the reaction solution. Subsequently, dissolve ammonium hexafluorophosphate in anhydrous dichloromethane and anhydrous methanol, and perform Soxhlet extraction on the above lyophilized powder for 6 h. Then vacuum dry the solid overnight to obtain a white powder, which is the conjugate molecule of cucurbit[8]uril-EG 5

[0079] 5) Take 75 mg of the conjugate molecule of CPT-SS-activated ester and dissolve it in 5 mL of ultradry DMSO to prepare reaction solution 1. Take 58 mg of the conjugate molecule of cucurbit[8]uril-EG 5 and disperse it in 6 mL of ultradry DMSO, add 5.0 mg of N,N-diisopropylethylamine, and after ultrasonic homogenization, prepare reaction solution 2. Slowly add reaction solution 2 dropwise to reaction solution 1, stir at room temperature for 0.5 - 4 hours, and when the solution becomes clear and orange-red, it is the end point of the reaction. Drop the orange-red reaction solution into 50 mL of anhydrous ether, precipitate in an ice-water bath for 2 hours, then filter and collect the filter cake, and wash it twice with ice-cold anhydrous ether. Then, wash the filter cake once by ultrasonic treatment with 5 mL of dichloromethane, vacuum dry it, wash it once with 5 mL of methanol, vacuum dry it, and finally wash it once with 5 mL of deionized water, and centrifuge to obtain a pale yellow solid, which is the crude product of CB-EG 5 -SS-CPT

[0080] 6) After activating the normal-phase column packing with methanol for half an hour, centrifuge to obtain the activated packing, disperse the packing with a methanol / water mixture to fill the chromatography column, and wash the chromatography column to gradually convert the mobile phase of methanol / water to ultrapure water. Dissolve the CB-EG 5 -SS-CPT crude product in an acetonitrile / water solution containing trifluoroacetic acid, perform column chromatography, and the eluent is a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. During the process, monitor the band with a handheld UV lamp with 365 nm excitation light, collect the product component solution, rotary evaporate to remove acetonitrile, and lyophilize to obtain a pale yellow powder, which is CB-EG​5 -SS-CPT.

[0081] Example 5:

[0082] Based on EG 1 、biselenide bond and camptothecin supramolecular complex CB-EG 1 -SeSe-CPT preparation.

[0083] The difference between this example and Example 1 lies in the different dynamic covalent bonds. Steps 1) and 4) of this example are the same as Steps 1) and 4) of Example 1 respectively, which will not be elaborated here. Now, the remaining steps of Example 4 will be described in detail:

[0084] 2) Extract 9.3 g of bis(2-hydroxyethyl) diselenide with dichloromethane. When the reaction solution of the CPT-activated ester conjugate molecule becomes clear, it is dropped into the diselenide substrate, and stirred at room temperature overnight under nitrogen. Then the reaction solution is rotary evaporated to dryness, washed three times with 15 mL of deionized water, and then washed twice with 15 mL of methanol. The yellow powder obtained by vacuum drying is the CPT-activated ester conjugate molecule, and its structural formula is:

[0085]

[0086] 3) Take 100 mg of the CPT-activated ester conjugate molecule prepared in step 2), dissolve it in 20 mL of anhydrous dichloromethane, and successively add 40 mg of triethylamine and 8.0 mg of 4-dimethylaminopyridine in an ice-water bath. Take 100 mg of 4-nitrophenyl chloroformate and dissolve it in 5 mL of anhydrous dichloromethane, and add it to the reaction solution in an ice-water bath, and react at room temperature for 1 hour under nitrogen. After the reaction, the reaction solution is rotary evaporated and concentrated to about 10 mL, added to 150 mL of anhydrous ether for sedimentation for 30 minutes, then filtered to obtain the filter cake, and washed twice with anhydrous ether to obtain a yellow powder, which is the CPT-SeSe-activated ester conjugate molecule, and its structural formula is:

[0087]

[0088] 5) Take 140 mg of the CPT-SeSe-activated ester conjugate molecule and dissolve it in 5 mL of ultradry DMSO to prepare reaction solution 1. Take 155 mg of cucurbit[7]uril-EG 1The conjugate molecules were dispersed in 5 mL of ultra-dry DMSO, and 15.0 mg of N,N-diisopropylethylamine was added. After ultrasonic homogenization, reaction solution 2 was prepared. Reaction solution 2 was added dropwise to reaction solution 1, and the mixture was stirred at room temperature for 4 hours until the solution became clear and orange-red, which was the end point of the reaction. The orange-red reaction solution was dropped into 50 mL of anhydrous ether and allowed to precipitate in an ice-water bath for 2 hours. Then, the filter cake was washed ultrasonically once with 5 mL of dichloromethane, dried in vacuo, washed once with 5 mL of methanol, dried in vacuo, and finally washed once with 5 mL of deionized water and centrifuged to obtain a pale yellow solid, namely CB-EG 1 The crude product of -SeSe-CPT.

[0089] 6) After activating the normal-phase column packing with methanol for half an hour, the activated packing was obtained by centrifugation. The packing was dispersed with a methanol / water mixture to fill the chromatography column, and the chromatography column was washed to gradually convert the mobile phase of methanol / water to ultrapure water. The crude product was dissolved in an acetonitrile / water solution containing trifluoroacetic acid for column chromatography, and the eluent was a gradient solution of ultrapure water and acetonitrile containing trifluoroacetic acid. The product fraction solution was collected, the acetonitrile was removed by rotary evaporation, and the residue was freeze-dried to obtain a yellow powder, namely CB-EG 1 -SeSe-CPT, and its structural formula is:

[0090]

[0091] Example 6:

[0092] Based on EG 1 The supramolecular complex CB-EG 1 -SS-Iron containing disulfide bonds and irinotecan was prepared, including the following steps:

[0093] 1) Irinotecan hydrochloride (1.6 g) was dispersed in 100 mL of deoxygenated anhydrous dichloromethane. Under an ice-water bath, 0.70 g of 4-dimethylaminopyridine was added, and then triphosgene (620 mg) was slowly added. The mixture was stirred at room temperature for 4 hours under nitrogen protection.

[0094] 2) 4.1 g of bis(2-hydroxyethyl) disulfide was extracted with dichloromethane. When the irinotecan reaction solution became clear, it was dropped into the disulfide substrate, and the mixture was stirred at room temperature overnight under nitrogen. Then, the reaction solution was evaporated to dryness, washed three times with 15 mL of deionized water and twice with 15 mL of methanol, and dried in vacuo to obtain a pale yellow powder, namely the conjugate molecules of Iron-dynamic covalent bonds, and its structural formula is:

[0095]

[0096] 3) Take 120 mg of the conjugate molecule of Iron-activated ester prepared in step 1), dissolve it in 20 mL of anhydrous dichloromethane, and successively add 40 mg of triethylamine and 8.0 mg of 4-dimethylaminopyridine under an ice-water bath. Take 100 mg of 4-nitrophenyl chloroformate and dissolve it in 5 mL of anhydrous dichloromethane, and add it to the reaction solution under an ice-water bath. React at room temperature for 1 hour under a nitrogen atmosphere. After the reaction is completed, rotary evaporate and concentrate the reaction solution to about 10 mL, add it to 150 mL of anhydrous ether for sedimentation for 30 minutes, then filter to obtain the filter cake, and wash it twice with anhydrous ether to obtain a yellow powder, which is the conjugate molecule of Iron-SS-activated ester, and its structural formula is:

[0097]

[0098] 4) Take the mono-alkynyl-modified cucurbit[7]uril C8bim@CB[7]-OA (192 mg) and azido-monoethylene glycol-amine (52.0 mg) and dissolve them in 2 mL of ultrapure water. Then add the CuAAC catalyst (2.0 mg), use a double-tube to displace the atmosphere in the system with nitrogen, and then stir at 55 °C for 18 hours. After the reaction is completed, add 500 mg of KBr and dissolve it by ultrasonic treatment, and then lyophilize the reaction solution. Wash the above lyophilized powder twice with anhydrous dichloromethane and anhydrous methanol respectively, and then vacuum-dry the solid overnight to obtain a white powder, which is the conjugate molecule of cucurbit[7]uril-EG 1 of the conjugate molecule.

[0099] 5) Take 150 mg of the conjugate molecule of Iron-SS-activated ester and dissolve it in 5 mL of ultradry DMSO to prepare reaction solution 1. Take 152 mg of the conjugate molecule of cucurbit[7]uril-EG 1 and disperse it in 5 mL of ultradry DMSO, add 15.0 mg of N,N-diisopropylethylamine, and make it uniform by ultrasonic treatment to prepare reaction solution 2. Add reaction solution 2 dropwise to reaction solution 1, stir at room temperature for 0.5 - 4 hours, and when the solution becomes clear and orange-red, it is the end point of the reaction. Drop the orange-red reaction solution into 50 mL of anhydrous ether, sediment it in an ice-water bath for 2 hours, then wash the filter cake once with 5 mL of dichloromethane by ultrasonic treatment, vacuum-dry it, wash it once with 5 mL of methanol, vacuum-dry it, and finally wash it once with 5 mL of deionized water and centrifuge to obtain a pale yellow solid, which is the crude product of CB-EG 1 -SS-Iron.

[0100] 6) After activating the normal-phase column packing with methanol for half an hour, centrifuge to obtain the activated packing. Disperse the packing with a mixed solution of methanol / water to fill the chromatography column, and wash the chromatography column to gradually convert the mobile phase of methanol / water into ultrapure water. Dissolve the crude product with an acetonitrile / water solution containing trifluoroacetic acid, perform column chromatography, and use a gradient solution of ultrapure water and acetonitrile containing trifluoroacetic acid as the eluent. Collect the product fraction solution, rotary evaporate to remove acetonitrile, and freeze-dry to obtain a pale yellow powder, which is CB-EG 1 -SS-Iron, and its structural formula is:

[0101]

[0102] Example 7:

[0103] Based on EG 1 、supramolecular complex CB-EG 1 -SS-Cbl is prepared, including the following steps:

[0104] 1) Add a solution of 800 mg of 4-nitrophenol, 2.5 g of chlorambucil, 1.8 g of dicyclohexylcarbodiimide, and 50 mg of 4-dimethylaminopyridine to 100 mL of dichloromethane in sequence, and then stir at room temperature for 24 h. After suction filtration, wash the filtrate with a mixed solution of glacial acetic acid and water, dry it over anhydrous sodium sulfate, concentrate it, and separate the product Cbl-activated ester conjugate molecule by column chromatography. Its structural formula is:

[0105]

[0106] 2) Take 1.2 g of the product Cbl-activated ester conjugate molecule and 1.7 g of aminoethyl-disulfide-ethanol in 50 mL of dichloromethane, mix them in an ice-water bath, gradually add 330 mg of triethylamine, react for 30 min, then rotary evaporate and concentrate to 10 mL, and precipitate with hydrochloric acid-ether solution to obtain 220 mg of the precipitated product, that is, the drug-dynamic covalent bond conjugate molecule. Its structural formula is:

[0107]

[0108] 3) Dissolve the above drug-dynamic covalent bond conjugate molecule in 20 mL of anhydrous dichloromethane, take 150 mg of 4-nitrophenyl chloroformate and dissolve it in 8 mL of anhydrous dichloromethane, add it to the above reaction solution in an ice-water bath, and then add 60 mg of triethylamine and 10 mg of 4-dimethylaminopyridine in sequence. React at room temperature for 1 hour under nitrogen. After the reaction, rotary evaporate and concentrate the reaction solution to about 10 mL, add it to 150 mL of anhydrous ether to precipitate for 30 minutes, then suction filter to collect the filter cake, and wash it twice with anhydrous ether to obtain a white powder, that is, the Cbl-SS-activated ester conjugate molecule. Its structural formula is:

[0109]

[0110] 4) The mono-alkynyl modified cucurbit[7]uril C8bim@CB[7]-OA (192 mg) and azido-monoethylene glycol-amine (52.0 mg) were dissolved in 2 mL of ultrapure water. Then, CuAAC catalyst (2.0 mg) was added, and the atmosphere in the system was replaced with nitrogen using a double-tube. Subsequently, the mixture was stirred at 55 °C for 18 hours. After the reaction, 500 mg of KBr was added and dissolved by ultrasound, and then the reaction solution was lyophilized. The resulting lyophilized powder was washed twice with anhydrous dichloromethane and anhydrous methanol respectively, and then the solid was dried under vacuum overnight to obtain a white powder, namely the conjugate molecule of cucurbit[7]uril-EG. 1 of the conjugate molecule.

[0111] 5) 100 mg of the conjugate molecule of Cbl-SS-activated ester was dissolved in 5 mL of ultradry DMSO to prepare reaction solution 1. 100 mg of the conjugate molecule of cucurbit[7]uril-EG 1 of the conjugate molecule was dispersed in 5 mL of ultradry DMSO and sonicated until homogeneous to prepare reaction solution 2. Reaction solution 2 was added dropwise to reaction solution 1, and the mixture was stirred at room temperature for 0.5 - 4 hours. The reaction was terminated when the solution became clear. The clear reaction solution was dropped into 50 mL of anhydrous ether and allowed to precipitate in an ice-water bath for 2 hours. Then, the filter cake was washed once with 5 mL of dichloromethane by ultrasound, dried under vacuum, washed once with 5 mL of methanol, dried under vacuum, and finally washed once with 5 mL of deionized water and centrifuged to obtain a white solid, namely the crude product of CB-EG 1 -SS-Cbl.

[0112] 6) After activating the normal-phase column packing with methanol for half an hour, the activated packing was obtained by centrifugation. The packing was dispersed with a methanol / water mixture to fill the chromatography column, and the mobile phase of methanol / water was gradually converted to ultrapure water by washing the chromatography column. The crude product was dissolved in an acetonitrile / water solution containing trifluoroacetic acid for column chromatography, and the eluent was a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. The product fraction solution was collected, the acetonitrile was removed by rotary evaporation, and the residue was lyophilized to obtain a white powder, namely CB-EG 1 -SS-Cbl, and its structural formula is:

[0113]

[0114] Example 8:

[0115] Based on the supramolecular complex CB-EG 3 of EG 3 and camptothecin was prepared.

[0116] The difference between this example and Example 3 is that there is no dynamic covalent bond. Now, the steps of Example 8 are described in detail: including the following steps:

[0117] 1) Dissolve 3.0 g of (S)-(+)-camptothecin in 200 mL of anhydrous dichloromethane, add 6.32 g of 4-dimethylaminopyridine, and stir for half an hour. Weigh 5.20 g of 4-nitrophenyl chloroformate and dissolve it in 40 mL of anhydrous dichloromethane. Then, dropwise add it to the reaction solution of (S)-(+)-camptothecin under an ice-water bath condition and stir at room temperature overnight. After completion, rotary evaporate and concentrate the reaction solution to 20 mL, add 200 mL of anhydrous ether and ultrasonicate to form a turbid solution, and let it settle in an ice-water bath for 2 hours. Filter out the solid, dissolve it with a dichloromethane and methanol solution, add column chromatography silica gel, rotary evaporate the solvent, perform dry loading, and use column chromatography to separate and purify it. The mobile phase is a gradient of petroleum ether - dichloromethane. During this period, monitor the band with a handheld ultraviolet lamp with 365 nm excitation light. The first blue fluorescent band is the product. Collect the product fraction solution and rotary evaporate it to dryness to obtain a pale yellow powder, which is the conjugate molecule of CPT-activated ester, and its structural formula is:

[0118]

[0119] 2) Weigh the mono-alkynyl modified cucurbit[7]uril C8bim@CB[7]-OA (208 mg) and 11-azido-3,6,9-trioxaundecan-1-amine (105 mg) and dissolve them in 2 mL of water. Then add the CuAAC catalyst (2.0 mg), use a double-tube to displace the atmosphere in the system with nitrogen, and then stir at 55 °C for 18 hours. After the reaction is completed, add 500 mg of KBr and ultrasonically dissolve it, and then lyophilize the reaction solution. Wash the above lyophilized powder twice with anhydrous dichloromethane and anhydrous methanol respectively, and then vacuum dry the solid overnight to obtain a white powder, which is the conjugate molecule of cucurbit[7]uril-EG 3 of the conjugate molecule.

[0120] 3) Weigh 100 mg of the conjugate molecule of CPT-activated ester prepared in step 1) and dissolve it in 5 mL of ultra-dry DMSO to prepare reaction solution 1. Take 122 mg of the conjugate molecule of cucurbit[7]uril-EG 3 of the conjugate molecule, disperse it in 5 mL of ultra-dry DMSO, add 13.3 mg of N,N-diisopropylethylamine, ultrasonically mix evenly, and then prepare reaction solution 2. Dropwise add reaction solution 2 to reaction solution 1 and stir at room temperature for 0.5 - 4 hours. When the solution becomes clear and orange-red, it is the end point of the reaction. Drop the orange-red reaction solution into 50 mL of anhydrous ether and let it settle in an ice-water bath for 2 hours. Then, wash the filter cake once with 5 mL of dichloromethane by ultrasonication, vacuum dry it, wash it once with 5 mL of methanol, vacuum dry it, and finally wash it once with 5 mL of deionized water and centrifuge to obtain a white solid, which is the CB-EG 3 -CPT crude product.

[0121] 4) After activating the normal-phase column packing with methanol for half an hour, centrifuge to obtain the activated packing. Disperse the packing with a methanol / water mixture to fill the chromatography column, and wash the chromatography column to gradually convert the methanol / water mobile phase into ultrapure water. Dissolve the crude product with an acetonitrile / water solution containing trifluoroacetic acid, perform column chromatography, and the eluent is a gradient solution of ultrapure water containing trifluoroacetic acid and acetonitrile. Collect the product fraction solution, rotary evaporate to remove acetonitrile, and finally obtain a yellow powder, which is the product CB-EG3-CPT, and its structural formula is:

[0122]

[0123] The effectiveness of the embodiments of the present disclosure is verified as follows:

[0124] 1. Structural characterization of the supramolecular complex

[0125] The structural characterization is divided into the molecular structure characterization and the characterization of its self-inclusion conformation. Dissolve the prepared complex in a deuterated nuclear magnetic resonance solvent and characterize it by nuclear magnetic resonance hydrogen spectrum, and then combine electrospray ionization mass spectrometry to characterize the molecular structure. Then, for the self-inclusion structure of the self-inclusion supramolecular drug, the nuclear magnetic resonance hydrogen spectrum of guest competition, two-dimensional diffusion ordered spectroscopy, and trapped ion mobility spectrometry can be used. The characterization results of Example 1 are as Figures 2 to 5 shown. In the nuclear magnetic resonance hydrogen spectrum, after adding the competing guest, the peaks in the aromatic region of CPT shifted downfield, which indicates that CB-EG 1 -SS-CPT underwent host-guest complexation, and CPT was encapsulated in the cavity of CB[7]. Moreover, the sharp and clear aromatic region peaks also indicate that no supramolecular polymer was formed in this system, suggesting that the host-guest complexation in this system may occur in a self-inclusion manner; in addition, the trapped ion mobility spectrometry standard shows that after adding the competing guest, the volume of CB-EG 1 -SS-CPT increased after the host-guest binding was released, which means that CB-EG 1 -SS-CPT in the original solution has a self-inclusion structure. The above results prove that the target self-inclusion supramolecular drug can be obtained through the relevant synthesis steps disclosed in the present invention.

[0126] 2. Characterization of the self-inclusion binding constant of the supramolecular complex

[0127] The experimental instrument for Isothermal Titration Calorimetry (ITC) is a GE MicroCal VP-ITC. The temperature of the titration experiment is 298.1K, and the experimental buffer solution used is phosphate buffer solution. The experiment adopts a competitive titration method. The competing guest is a solution of p-trimethyl-p-phthalic diamine as the titrant, and the supramolecular complex CB-EG 1-SS-CPT was used as the titrant, and the titration time interval was set to 400 s. The function model used was the ITCone set of site function. The characterization results are as Figure 6 shown. By calculation, the binding constant of the supramolecular complex can be obtained, which is approximately 2.31×10 3 , that is, the encapsulation efficiency of the guest molecule is greater than 99%.

[0128] 3. Characterization of the controlled release of guest molecules in the supramolecular complex

[0129] The drug release process of the self-inclusive supramolecular drug was monitored by high-performance liquid chromatography under the simulated tumor microenvironment of reduced glutathione. The high-performance liquid chromatograph used was composed of a binary pump, an ultraviolet-visible detector, a column oven, and an autoinjector connected in series. The mobile phase was divided into two phases: a high-polarity phase and a low-polarity phase. The high-polarity phase was an ultrapure water phase, and the low-polarity phase was an acetonitrile phase. 0.1% formic acid was added to both phases of the mobile phase. The monitoring wavelength was 365 nm, and the baseline needed to be balanced with the ultrapure water phase before each experiment. In terms of experimental operation, the reaction stock solution of the supramolecular drug prepared with phosphate buffer was used, and the reaction stock solution of reduced glutathione with different concentrations was also prepared. The two were quickly mixed in a volume ratio of 1:1 to form reaction solution 2, and the reaction was carried out at room temperature. This time point was recorded as the reaction start point. During the reaction process, 80 μL of the reaction solution was taken every once in a while and added to 400 μL of the ultrapure aqueous solution containing 0.1% formic acid to quench the reaction and make a test solution, and then the high-performance liquid chromatography test was carried out. The results are as Figure 7 shown in a) - c). The drug molecules in the self-inclusive supramolecular drug can achieve controlled release of the drug according to the different concentrations of glutathione. When the concentration of glutathione is low, the drug molecules will be stably encapsulated in the cavity of the host molecule due to self-inclusion, while when in an environment with a high concentration of glutathione (such as the tumor site), they can be released quickly and efficiently. The release rate exceeds 97%.

[0130] The above results show that the supramolecular complex of Example 1 can not only stably encapsulate drug molecules at low concentrations, but also efficiently release drug molecules in the tumor microenvironment.

[0131] 4. Apoptosis test of the supramolecular complex

[0132] The inhibitory effect of the supramolecular complex of Example 1 on cell growth was studied by using a CCK-8 kit in combination with an enzyme-labeled instrument. The cell line involved in this experiment was human cervical cancer cells (Hela). There were two experimental groups, namely the camptothecin group (CPT) and the supramolecular complex group (CB-EG 1 -SS-CPT). The drugs in each group were co-cultured with Hela cells at different concentrations for 48 h. The experimental results are shown in Figure 8, the supramolecular complex has a concentration-dependent cytotoxicity on Hela cells similar to that of the CPT original drug, and the IC 50 is relatively close, indicating that the self-included supramolecular drug can efficiently release the chemotherapeutic drug under cancer cell conditions, thus obtaining antitumor activity equivalent to that of the original drug and effectively killing cancer cells.

[0133] 5. Test of the supramolecular complex with human serum albumin

[0134] Human serum albumin (HSA) is the most abundant protein in blood, with a concentration of about 30 - 50 mg / mL. It often uses the hydrophobic cavity between domains to transport small molecules such as amino acids, fatty acids, or therapeutic drugs. The dynamic equilibrium of the binding between the drug and albumin determines the drug concentration at the lesion site, and thus determines the drug efficacy.

[0135] Prepare three groups of experiments with phosphate buffer, namely: 1) Camptothecin group (CPT), prepare camptothecin solution and camptothecin + human serum albumin solution respectively; 2) Host-guest complex group with a molar ratio of camptothecin to cucurbit[7]uril of 1:1 (CPT@CB[7]), prepare camptothecin-cucurbit[7]uril solution and camptothecin-cucurbit[7]uril + human serum albumin solution respectively; 3) Solution of the supramolecular complex (CB-EG 1 -SS-CPT) prepared in Example 1 and solution of the self-included supramolecular drug + human serum albumin. Place the above solutions in a 37 °C water bath and stir for 2 hours. After completion, transfer each group of solutions to an ultrafiltration tube with a molecular weight cut-off of 10k, centrifuge at 3500 rpm for 2 minutes, and take the filtrate. The drug concentration in this filtrate is the concentration of the free drug in the original solution. Then use an Agilent Cary Eclipse fluorescence spectrometer to measure the fluorescence value of the filtrate. The test parameters are as follows: excitation wavelength 360 nm, and both the excitation light slit width and the emission light slit width are 20 nm. By calculating the fluorescence ratio before and after adding human serum albumin, the proportion of the free drug can be obtained. See a) - c) of Figure 9 for the experimental results. When human serum albumin is added, although the fluorescence value of the filtrate in all groups decreases, only the self-included supramolecular drug group still has 29.5% remaining, indicating that human serum albumin and cucurbit[7]uril are competing for the camptothecin fragment, and the strong binding ability of self-inclusion enables camptothecin to remain in the cucurbit[7]uril cavity, reducing the interference of human serum albumin to a certain extent. The results show that the self-included supramolecular drug can better resist the encapsulation of human serum albumin compared with the camptothecin molecule itself, improving the blood drug concentration and drug utilization rate in vivo.

[0136] 6. Antitumor experiment of the supramolecular complex on tumor-bearing nude mice

[0137] The human tumor cells used in tumor-bearing experiments were Hela cells used in cell experiments. Each mouse was inoculated subcutaneously in the flank. After the tumor volume reached the standard, the drug administration experiment was carried out. The four experimental groups were: biological phosphate buffer group; (S)-(+)-camptothecin group; supramolecular complex CB-EG 3 -CPT group prepared in Example 8 and supramolecular complex CB-EG 1 -SS-CPT group prepared in Example 1. Tail vein injection was used, and the treatment lasted for two weeks in total. After each treatment, the tumor size and body weight of the nude mice were measured. After the injection treatment was completed, observation continued for two weeks, and the tumor size and body weight were recorded.

[0138] See the experimental results in Figure 10 a) and b). The experimental results show that due to the existence of self-inclusion and disulfide bonds in the self-inclusion supramolecular drug CB-EG3-CPT group, the drug molecules can be stably loaded and have the characteristics of efficient release at the same time. While significantly inhibiting the growth of nude mouse tumors, it also has safety.

[0139] In summary, the present disclosure improves the construction method of existing supramolecular complexes, especially supramolecular drugs. Based on the principle of host-guest intramolecular self-inclusion independent of concentration, through the regulation of molecular structure, a variety of supramolecular complexes that can undergo self-inclusion are designed. The self-inclusion supramolecular complexes provided by the present disclosure can achieve stable loading and efficient release at any drug concentration. On this basis, the present disclosure also develops its medical applications, indicating that the self-inclusion supramolecules can improve the anti-tumor effect of the original drugs, providing a potential solution for the design of anti-tumor drugs.

[0140] In the description of this specification, the content not described in detail belongs to the well-known technology of those skilled in the art. The description with reference to terms such as "one embodiment", "some embodiments", "schematic embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present disclosure. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0141] Although the embodiments of the present disclosure have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and purposes of the present disclosure. The scope of the present disclosure is defined by the claims and their equivalents.

Claims

1. A supramolecular complex, characterized in that, The structural formula of the supramolecular complex is CB-EG x -L-Guest, where: CB is the host molecule, and cucurbit[6]uril, cucurbit[7]uril or cucurbit[8]uril is selected; EG x is a type of linking segment; L is a dynamic covalent bond; Guest is the guest molecule, and camptothecin or irinotecan is selected.

2. The supramolecular complex according to claim 1, characterized in that, the connecting segment is selected as oxyethyl, and the number of repeating units is 1 to 5; and / or the dynamic covalent bond is selected as a disulfide bond, a diselenide bond, a phenylborate ester or a Schiff base.

3. The supramolecular complex according to claim 1, characterized in that, the supramolecular complexation that occurs in the supramolecular complex is intramolecular binding, that is, the formed molecular structure is a self-inclusion structure.

4. A preparation method of the supramolecular complex according to any one of claims 1 to 3, characterized in that, comprising: adding the guest molecule into a first solvent, and synthesizing a conjugate molecule of the guest molecule-activating ester through an activated ester reaction; adding the conjugate molecule of the guest molecule-activating ester and the dynamic covalent bond molecule into a second solvent, and synthesizing a conjugate molecule of the guest molecule-dynamic covalent bond through a bimolecular nucleophilic substitution reaction; adding the conjugate molecule of the guest molecule-dynamic covalent bond into a third solvent, and synthesizing a conjugate molecule of the guest molecule-dynamic covalent bond-activating ester through an activated ester reaction; adding the host molecule and the azide-containing connecting segment in a fourth solvent, and synthesizing a conjugate molecule of the host molecule-connecting segment through a click chemical reaction catalyzed by cuprous ions; mixing the conjugate molecule of the host molecule-connecting segment and the conjugate molecule of the guest molecule-dynamic covalent bond-activating ester in a fifth solvent to obtain a crude product containing the supramolecular complex; purifying the crude product to obtain the supramolecular complex.

5. The preparation method according to claim 4, characterized in that, the first solvent is selected as any one or a mixture of more than one of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide; and / or the second solvent is selected as any one or a mixture of more than one of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide; and / or the third solvent is selected as any one or a mixture of more than one of dichloromethane, chloroform, acetonitrile, tetrahydrofuran and dimethyl sulfoxide; and / or the fourth solvent is selected as any one or a mixture of more than one of ultrapure water, deionized water, dimethyl sulfoxide and acetonitrile; and / or the fifth solvent is selected as any one or a mixture of more than one of methanol, trifluoroacetic acid, ultrapure water, deionized water, dimethyl sulfoxide and acetonitrile.

6. The preparation method according to claim 5, characterized in that, the fifth solvent is selected as dimethyl sulfoxide.

7. The preparation method according to claim 4, characterized in that, purifying the crude product includes the following steps: performing sedimentation, washing and recrystallization on the crude product through a sixth solvent to obtain a preliminary purified product, and obtaining the supramolecular complex through column chromatography of the preliminary purified product.

8. The preparation method according to claim 7, characterized in that, The sixth solvent is selected from any one or a mixture of more than one of acetone, ether, methanol, dichloromethane, chloroform and deionized water.

9. Use of the supramolecular complex according to any one of claims 1 to 3 in the field of preparation of anti-tumor drugs.

10. Use of the supramolecular complex prepared by the preparation method according to any one of claims 4 to 8 in the field of preparation of anti-tumor drugs.

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