CN114796514B - Covalent nanometer bionic drug delivery system of water-soluble positive ion type supermolecule organic framework - Google Patents

Covalent nanometer bionic drug delivery system of water-soluble positive ion type supermolecule organic framework Download PDF

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CN114796514B
CN114796514B CN202210345116.3A CN202210345116A CN114796514B CN 114796514 B CN114796514 B CN 114796514B CN 202210345116 A CN202210345116 A CN 202210345116A CN 114796514 B CN114796514 B CN 114796514B
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sof
inno206
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organic framework
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CN114796514A (en
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黎占亭
刘月阳
张丹维
周伟
王辉
王泽坤
余尚博
刘传志
刘亚敏
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Fudan University
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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a covalent nano bionic drug delivery system of a water-soluble positive ion type supermolecule organic framework. The drug delivery system takes a supermolecule organic framework which is self-assembled in a water phase through the action of a host and a guest as a carrier, and a drug or a prodrug is connected to the carrier through a covalent bond to obtain a nanoparticle drug delivery system; the system can concentrate the targeting of the drug or the prodrug to tumor tissues and release the drug so as to improve the therapeutic effect of the drug; in particular, the anti-tumor drug is doxorubicin prodrug INNO206, which overcomes the multidrug resistance of tumor cells, and animal experiments prove that the anti-tumor drug can inhibit tumor growth and reduce the damage to normal cells and tissues.

Description

Covalent nanometer bionic drug delivery system of water-soluble positive ion type supermolecule organic framework
Technical Field
The invention belongs to the technical field of biological medicine, and particularly relates to a covalent nano bionic drug delivery system of a water-soluble positive ion type supermolecule organic framework.
Background
Cancer is commonly used to describe a range of diseases in which cells grow abnormally. The world health organization WHO subordinate international cancer research institute IARC issued "ARC Biennial Report 2020-2021" to suggest that new cancer cases worldwide reach 1929 ten thousand in 2020, and that the number of 3020 ten thousand is expected to reach 2040; the global cancer death number in 2020 is 996 ten thousand, and the cancer death number in 2040 year can reach 1640 ten thousand. The incidence and mortality of cancer continue to increase year by year, and cancer treatment is a medical problem that humans have long needed to overcome. One report on cancer issued by WHO in month 2020 shows that about $4580 billion are spent annually in cancer diagnosis, prevention and treatment. Despite significant advances in therapy, diagnosis and prevention, cancer remains a high mortality disease, with about one sixth of deaths worldwide being caused by cancer. At present, cancer treatment mainly comprises means such as surgery, chemotherapy, radiotherapy and the like. Chemotherapy is one of the most effective means of treating cancer, and tumor multidrug resistance is the leading cause of chemotherapy failure.
The transporter P-glycoprotein (P-gp) is capable of transporting drugs in the cell membrane out of the cell and thereby reducing intracellular drug concentration. The currently widely accepted action mechanism is that P-gp has a similar function of a 'hydrophobic vacuum aspirator', and after ATP is combined, hydrophobic substances embedded in the inner layer of a cell membrane phosphoester bilayer can be discharged out of the membrane, so that the concentration of the intracellular medicine is reduced, and the cell generates drug resistance. Studies have reported that P-gp may be involved in the development of multidrug resistance through some cellular pathways. The existing commonly used hydrophobic chemotherapeutics comprise vinblastine, vincristine, doxorubicin, daunorubicin, actinomycin D, etoposide, teniposide, taxol and the like, and the hydrophobic chemotherapeutics can enter cells through free diffusion without a carrier, so that the possibility of being pumped by P-gp is increased to a certain extent, and the problem can be effectively solved by using nano particles to enter cells through an endocytic way.
Human serum albumin is a natural nano-drug delivery system with multiple functions. Albumin is one of the most abundant and important proteins in the body, accounting for about 60% of all proteins in the blood. It has low toxicity, is a highly water-soluble globular protein, has a molecular weight of 67kDa and an average half-life of 19 days. It exhibits stability in the pH range of 4-9 and can be heated at 60℃for 10 hours. It plays an important role in maintaining intravascular gel osmotic pressure, neutralizing toxins, and transporting therapeutic agents. Albumin has high binding capacity and relatively long half-life for both hydrophobic and hydrophilic drugs, and can specifically target inflammatory sites. In addition to the targeted binding of the drug to albumin, the drug may also be bound by covalent coupling. In many tumors, albumin binding proteins, such as SPARC and gp60, and the like, often appear to be overexpressed to transport albumin as an amino acid and energy source for rapidly growing cancer cells, which also makes it possible for albumin to provide active targeting without the use of external ligands.
To improve the cardiotoxicity of the broad spectrum antitumor drug doxorubicin, researchers developed the acid-sensitive doxorubicin prodrug INNO206 (doxorubicin-maleimide derivative DOXO-EMCH). INNO206 is rapidly combined with human serum albumin after intravenous infusion, the conjugate is preferentially retained in tumor tissues and is taken up by tumor cells, hydrolysis occurs in the acidic environment of endocytic lysosomes, and doxorubicin is released to achieve the therapeutic purpose.
Disclosure of Invention
The invention aims to overcome the problems of multi-drug resistance of cancer cells, low drug and nanoparticle loading and weak combination, and provides a covalent bionic drug delivery system of a water-soluble positive ion type supermolecule organic framework.
The water-soluble supermolecular organic frame is used as a carrier to deliver the prodrug to the tumor site and then release the drug to improve the drug treatment effect, and the nanoparticle bionic drug delivery system for inhibiting the growth of multi-drug resistant breast cancer tumor is provided.
The water-soluble positive ion type supermolecule organic frame covalent bionic drug delivery system provided by the invention takes the water-soluble supermolecule organic frame nano material as a carrier, and the loaded drug or prodrug is enriched in tumor tissues in a targeting way, and then the drug is released, so that the drug treatment effect is improved; specifically, taking a supermolecule organic framework obtained by self-assembly through the action of a host and a guest in a water phase as a carrier, and marking the carrier as SOF-SH; the drug or prodrug is denoted Y; the drug or prodrug is covalently linked to a carrier to give a nanoparticle delivery system, designated y@sof-SH.
In the invention, the drug-loading rate of the water-soluble supermolecule organic frame nano material is 4-12% of the mass of the organic frame nano material.
For example, the doxorubicin prodrug INNO206 is covalently linked to a carrier, resulting in a nanoparticle delivery system, designated INNO206@SOF-SH; the amount of INNO206 is 4-12% of the mass of SOF-SH nanoparticles.
In the invention, the supermolecule organic framework nano material has a mesoporous structure, and the hydration particle size is 90-120nm; the shape is spherical, blocky or flaky.
The terminal sulfhydryl of the supermolecule organic frame nano material can be in covalent bond connection with a drug or prodrug to realize drug loading.
The water-soluble supermolecule organic framework nano material is multivalent positive ions, is obtained by assembling in a water phase, and has good water solubility.
In the present invention, the pharmaceutical assembly further comprises a pharmaceutically acceptable additive.
In the present invention, the drug is an anticancer drug, for example, a broad-spectrum cancer therapeutic drug.
In the present invention, the drug is a multidrug resistant cancer therapeutic drug model, such as an doxorubicin prodrug.
In the nano bionic drug delivery system, 2 preparation routes exist for the supermolecule organic framework nanomaterial serving as a carrier:
Figure BDA0003576097260000031
SOF-SH1 and SOF-SH2 correspond to two supermolecule organic frame materials respectively.
The water-soluble positive ion type supermolecule organic framework material provided by the invention has two construction methods:
(1) Dissolving a cysteine modified pyridylphenol derivative and tetra-p-bromomethyl phenyl methane in a dry N, N-dimethylformamide solution according to a stoichiometric ratio of 6:1, reacting for 12 hours at 100 ℃ to obtain a protected tetrahedral precursor compound, reacting for 12 hours at 40 ℃ with hexafluoroisopropanol solution (HFIP) containing 0.1N hydrochloric acid to obtain a positive ionic tetrahedral compound with a mercapto substituent at the tail end, which is marked as T1, and then self-assembling T1 and CB 8 in an aqueous phase according to a mass ratio of 1:2-1:2.5 to obtain a supramolecular framework with a cysteine structure at the tail end of a side chain, which is marked as SOF-SH1;
(2) Dissolving a pyridylphenol derivative and tetra-p-bromomethyl phenyl methane in a dry N, N-dimethylformamide solution according to a stoichiometric ratio of 6:1, reacting for 12 hours at 100 ℃ to obtain an amino-protected tetrahedral compound, reacting for 12 hours at 40 ℃ with a hexafluoroisopropanol solution containing 0.1N hydrochloric acid to synthesize a positive ionic tetrahedral compound with a terminal amino group, which is denoted as T2, then self-assembling T2 and CB 8 in an aqueous phase according to a mass ratio of 1:2-1:2.5 to obtain a framework structure, reacting the obtained supramolecular framework with 2-iminothiolane hydrochloride (Traut reagent) to obtain a supramolecular framework with a terminal mercapto group loaded on a side chain, which is denoted as SOF-SH2.
Self-assembly of the tetrahedral compounds with CB [8] can be referred to as Nat.Commun.2014,5,5574.
The invention provides a water-soluble positive ion type supermolecule organic frame nano material, which is characterized in that a sulfhydryl functional group is loaded at the end position of the supermolecule organic frame. The supermolecule organic framework has the advantages of low toxicity, good stability, better water solubility and the like, and utilizes the conjugated addition of the sulfydryl loaded at the end position of SOF-SH and an anticancer drug, such as a maleimide structural unit of INNO206, to generate a novel covalently-bonded bionic nano drug delivery system. The drug delivery mode has high permeation long retention effect (EPR effect) and pH responsiveness, can aggregate nano particles loaded with drugs to tumor tissues and release the drugs under the weak acidic condition, and effectively avoids the generation of multi-drug resistance.
The supermolecule organic framework material provided by the invention can obviously improve the capability of medicines entering cancer cells and inhibiting tumor growth. The specific operation is as follows: the nano-drug assembly with a certain concentration is injected into the mouse body through tail vein every three days.
The invention carries out dynamic light scattering experiments, dialysis experiments, hemolysis experiments, cytotoxicity experiments, laser confocal experiments and mouse tumor inhibition experiments on the nano-drug assembly. Dynamic light scattering experiments show that the nano-drug assembly has stability and can exist stably in various buffer systems; dialysis experiments show that the nano-drug assembly has pH sensitivity and can rapidly release drugs under acidic conditions; the hemolysis experiment shows that the nano-drug assembly has high biological safety; cytotoxicity experiments show that the nano-drug assembly has good capability of overcoming the proliferation of multi-drug resistant cancer cells; laser confocal experiments show that the nano-drug assembly can promote drugs to enter drug-resistant breast cancer cells; the mouse tumor inhibition experiment shows that the nano-drug assembly can inhibit the growth of drug-resistant tumors, the proliferation of cancer cells can be observed to be inhibited by slicing, the apoptosis of the cancer cells is promoted, meanwhile, the animal weight is not reduced in the experiment process, obvious lesions are not seen on organ slices, and the toxicity of the assembly to the animal is small.
The experimental results show that the nano-drug assembly has the characteristics of stable structure, good biocompatibility and pH-dependent release, has proliferation inhibition effect on multidrug resistant cancer cells, can promote the drug to enter the drug resistant breast cancer cells, and inhibit the growth of drug resistant tumors, and has low living toxicity, so that the covalent bionic nano-drug delivery system of the water-soluble positive ion type supermolecular organic framework has wide application prospect.
Drawings
FIG. 1 is a graph of the hydrated particle size of SOF-SH1 at various concentrations.
FIG. 2 is a graph of the hydrated particle size of SOF-SH2 at various concentrations.
FIG. 3 is a graph of the hydrated particle size of T1 and SOF-SH1 at various times.
FIG. 4 is a graph of the hydrated particle size of T2 and SOF-SH2 at various times.
FIG. 5 is a graph showing the hydrated particle size of SOF-SH1 and SOF-SH2 before and after drug loading.
FIG. 6 is a graph of hydration particle size of INNO206@SOF-SH1 and INNO206@SOF-SH2 in different buffer systems.
FIG. 7 shows the in vitro cytotoxicity of SOF-SH1 and SOF-SH2 on H9C2 cells, as measured by the cell counting reagent CCK-8.
FIG. 8 shows the in vitro cytotoxicity of SOF-SH1 and SOF-SH2 on L02 cells, as measured by the cell counting reagent CCK-8.
FIG. 9 shows the in vitro cytotoxicity of SOF-SH1 and SOF-SH2 on Ana-1 cells, as measured by the cell counting reagent CCK-8.
FIG. 10 is a graph of experiments on hemolysis of human and murine erythrocytes by SOF-SH1 and INNO206@SOF-SH 1.
FIG. 11 is a graph of experiments on hemolysis of human and murine erythrocytes with SOF-SH2 and INNO206@SOF-SH2.
FIG. 12 shows the drug release profile of INNO206@SOF-SH1 at different pH.
FIG. 13 is a graph showing the drug release profile of INNO206@SOF-SH2 at different pH.
FIG. 14 is a graph showing the time-dependent drug uptake of INNO206 in MCF-7 cells.
FIG. 15 is a graph of drug uptake in MCF-7 cells as a function of time for INNO206@SOF-SH 1.
FIG. 16 is a graph of drug uptake of INNO206@SOF-SH2 in MCF-7 cells over time.
FIG. 17 is a graph of INNO206 drug uptake in MCF-7/ADR cells over time.
FIG. 18 is a graph of the time-dependent drug uptake of INNO206@SOF-SH1 in MCF-7/ADR cells.
FIG. 19 is a graph of the time dependent drug uptake of INNO206@SOF-SH2 in MCF-7/ADR cells.
FIG. 20 is a graph showing survival curves of MCF-7/ADR cells in the presence of Doxorubicin (DOX), INNO206, SOF-SH1, SOF-SH2, INNO206@SOF-SH1 and INNO206@SOF-SH2.
Figure 21 is a graph of the change in body weight of mice over the course of 4 injections of drug at the animal level.
Figure 22 is a graph of tumor volume change in mice over the course of 4 injections of drug at the animal level.
FIG. 23 is a normal organ slice of mice at the animal level after completion of drug treatment.
Figure 24 is a graph of tumor weights of mice at the animal level after completion of drug treatment.
Figure 25 is a schematic representation of all mice tumors at the animal level after completion of drug treatment.
Fig. 26 is a view of tumor sections of mice at the animal level after completion of drug treatment.
Detailed Description
The invention is further described by the following examples, which should not be construed as limiting the invention.
Example 1: preparation of the supramolecular organic framework drug delivery System INNO206@SOF-SH.
The invention provides a preparation method of a water-soluble positive ion type supermolecule organic framework material, which is referred to as Nat.Commun.2014,5,5574.
Preparation of SOF-SH 1: and dispersing tetrahedral monomer T1 and CB 8 in the stoichiometric ratio of 1 to 2 ultrasonically, heating to dissolve, and cooling to room temperature to obtain SOF-SH1.
Preparation of SOF-SH 2: heating tetrahedral molecules T2 and CB 8 in the chemical dosage ratio of 1 to 2 to dissolve, and naturally cooling to room temperature. 2-iminothiolane hydrochloride (Traut reagent) is added and reacted for 18 hours at 70 ℃ to obtain the mercapto-modified framework compound SOF-SH2.
INNO206@SOF-SH preparation: taking the loading of 8% as an example, adjusting the pH value of the aqueous solution of SOF-SH1 or SOF-SH2 to 6.5-7.0, adding INNO206 with the mass ratio of 8% in the frame, and carrying out light-shielding reaction at room temperature for 18 hours to obtain the corresponding INNO206@SOF-SH without further treatment.
Example 2: particle size characterization of SOF-SH1 and SOF-SH2 and INNO206@SOF-SH1 and INNO206@SOF-SH2 in solution.
Dynamic light scattering experiments: 1.5mM of SOF-SH1 and SOF-SH2 aqueous solutions were prepared, respectively, and the particle size obtained by the test was between 90 and 110nm, and the particle size did not become smaller with decreasing concentration, as shown in FIGS. 1-2. The difference in particle size between SOF-SH1 and SOF-SH2, which are both around 1nm for both tetrahedral monomer molecules, and the corresponding monomer particles also indicates the formation of an assembly (FIGS. 3-4).
1.5mM INNO206@SOF-SH1 and INNO206@SOF-SH2 aqueous solutions were prepared, respectively, and the results of the tests were shown in FIG. 5, in which the particle size of the drug assembly was not different from that before the drug was not loaded, and the loading of the drug did not affect the self-structure of the frame. 1.5mM PBS buffer, FBS solution and aqueous solution of INNO206@SOF-SH1 and INNO206@SOF-SH2, respectively, were formulated to simulate in vivo environment, and as shown in FIG. 6, the particle size of the nano-drug assembly was not changed, and it was inferred that INNO206@SOF-SH1 and INNO206@SOF-SH2 could also maintain stable structures in vivo without being destroyed in vivo circulation.
Example 3: toxicity experiments of SOF-SH nanoparticles on normal cells.
By using the cell counting kit Cell Counting Kit-8 (CCK-8), we studied the in vitro cytotoxicity of the framework structures using human normal hepatocytes (L02 cells), mouse macrophages (Ana-1 cells) and rat cardiomyocytes (H9C 2 cells), respectively, as shown in FIGS. 7-9. The results show that the SOF-SH1 and SOF-SH2 can keep good survival rate within the range of 200 mug/mL, and the results also show that the SOF-SH1 and SOF-SH2 have lower toxicity and less damage to normal cells, and can be used as a carrier safely used in vivo.
Example 4: biosafety studies of SOF-SH1 and SOF-SH2.
We performed experimental studies of hemolysis on SOF-SH1 and SOF-SH2, respectively. As shown in FIGS. 10 to 11, SOF-SH1 and SOF-SH2 did not undergo hemolysis in both human erythrocytes and rat erythrocytes at the concentrations used, indicating that SOF-SH1 and SOF-SH2 have high safety.
Example 5: drug loading studies of INNO206@SOF-SH1 and INNO206@SOF-SH2.
Since INNO206 has an acid-sensitive hydrazone linkage unit, the literature reports that it can rapidly release doxorubicin under weakly acidic conditions, and we therefore indirectly determine drug loading by testing the released doxorubicin content. The drug concentration of the exotic solution was calculated by measuring the UV absorbance of the exotic dialysis solution and substituting the standard UV absorbance curve of doxorubicin at three pH values, by placing 1.5mM INNO206@SOF-SH1 and INNO206@SOF-SH2 glucose solutions in dialysis bags (molecular weight cut-off 1000 Da) respectively immersed in neutral (pH=7.4, simulated physiological environment), weakly acidic (pH=6.5 and 6.0, simulated tumor tissue environment) and acidic (pH=5.0, simulated in-vivo lysosome environment) solutions, thereby obtaining the drug amount loaded on SOF-SH material (FIGS. 12-13). The results show that the drug permeation of both nano-drug assemblies is less than 5% under neutral conditions, which means that the INNO206 is stably covalently attached to the SOF-SH material over 72 hours. At pH 6.5, both frameworks were able to load 65% and 40% respectively, and the loading was reduced with the acidic enhancement drug. The drug was released 44% and 59% in the lysosomal environment at 1 hour, respectively, reaching 99% and 95% of maximum release at 72 hours. Based on the feature that the drug is hardly released under neutral conditions, INNO206@SOF-SH1 and INNO206@SOF-SH2 can be used directly without further purification.
Example 6: laser confocal experiments behavioural studies of INNO206@SOF-SH1 and INNO206@SOF-SH2 in resistant and non-resistant cancer cells.
We selected human breast cancer cells (MCF-7 cells) and doxorubicin-resistant human breast cancer cells (MCF-7/ADR cells) for studies of INNO206@SOF-SH1 and INNO206@SOF-SH2 against tumor cell multidrug resistance using MCF-7/ADR cells at a drug resistance of 1000ng/mL. First we stained nuclei and lysosomes with Hoechst 33342 and Lysotracker Green DND-26 dyes, respectively, as shown in fig. 14-16, with INNO206, INNO206@sof-SH1 and INNO206@sof-SH2 both partially entering MCF-7 cells at 0.5 hours and almost fully entering cells at 2 hours, demonstrating therapeutic effect on both non-drug resistant cell small molecule drugs and nano-drug assemblies. For MCF-7/ADR cells, INNO206 has little drug content entering cells even if the cells are incubated for 2 hours, INNO206@SOF-SH1 and INNO206@SOF-SH2 show that the cells enter the cells in 0.5 hour, and the cell entering efficiency reaches the maximum after 2 hours of incubation (figures 17-19), which shows that INNO206@SOF-SH1 and INNO206@SOF-SH2 can effectively promote the drugs to enter drug-resistant cells, thereby achieving the aim of overcoming drug resistance.
Example 7: the cytotoxicity experiment is used for comparing the killing effect of the small molecular medicine and the supermolecular organic framework nano medicine delivery system on the drug-resistant breast cancer cells.
After determining that the supermolecule organic framework nano drug delivery system can load drugs into drug-resistant breast cancer cells, the treatment effect of the supermolecule organic framework nano drug delivery system on the multidrug-resistant breast cancer cells is researched through cytotoxicity experiments. It was found that the median lethal concentration of drug IC of MCF-7/ADR cells by DOX or INNO206 alone 50 150.0 μg/mL and 124.0 μg/mL, respectively, while IC to MCF-7/ADR cells was administered by INNO206@SOF-SH1 and INNO206@SOF-SH2 50 The values were 12.3. Mu.g/mL and 5.1. Mu.g/mL, respectively (FIG. 20). The results show that compared with the killing capacity of doxorubicin on drug-resistant cancer cells, the effect of INNO206 is slightly better than that of doxorubicin. Drug-loaded SOF-SH material IC 50 Value compared to INNO206 alone, IC 50 The improvement is 12 times and 30 times respectively. This suggests that INNO206 is covalently linked to SOF-SH material to enhance therapeutic effects on drug-resistant cells. And under the same concentration condition, the cell survival rate is far higher than that of the corresponding INNO206 load material only when the SOF-SH material exists, which indicates that the killing effect on MCF-7/ADR cells is not from the SOF-SH material, and INNO206@SOF-SH plays a vital role in the apoptosis of doxorubicin-resistant human breast cancer cells.
Example 8: SOF-SH material is used as carrier to treat drug-resistant tumor under living condition.
In vitro experiments show that INNO206@SOF-SH1 and INNO206@SOF-SH2 have good killing ability to drug-resistant cancer cells, and the in vivo therapeutic effect of the INNO206@SOF-SH2 is further explored. Female nude mice with the weight of about 16g and the age of 5-6 weeks are selected as animal models, the forelimbs of the nude mice are inoculated with A549/ADR tumor, and the tumor survives in vivo after one week of inoculation. Tumor-bearing mice were divided into 7 groups of 4, each injected with physiological saline, DOX, INNO206, SOF-SH1, SOF-SH2, INNO206@SOF-SH1 and INNO206@SOF-SH2, respectively. Experiments show that mice survive well when the dosage is 5mg/kg (animal body weight), no death occurs, and literature data show that the drug dosage has clinical reference value, so that the later experiments adopt the dosage, and the SOF-SH dosage is 50mg/kg (animal body weight). On the day of injection we measured the body weight and tumor size of the mice and the calculation formula for tumor volume was: v=1/2 (length of tumor) x (width of tumor) 2 . The drug was then injected every three days and the body weight and tumor size were measured (fig. 21 and 22). Tumor volumes of mice injected with normal saline groups after four injections reach the sacrifice standard, all mice are euthanized according to ethical requirements of animal experiments, and tissue sections are taken from the heart, liver, spleen, lung and kidney. H&E staining results showed that the mice injected with INNO206@SOF-SH1 and INNO206@SOF-SH2 had no obvious tissue lesions, and SOF-SH1 and SOF-SH2 did not damage normal organs as well (FIG. 23), which also demonstrates that SOF-SH material can be used in vivo as a safe carrier.
In the reported animal experiments, the tumor volumes of mice injected with the same dose of doxorubicin and INNO206 showed comparable effects in inhibiting growth during the treatment period, but the treatment effect was better with increasing dose of INNO206; in clinical experiments in the third period, it has been reported that INNO206 has no significant difference between the therapeutic effect on tumors and doxorubicin, but shows more excellent effects on the indexes of the maximum drug tolerance dose, the median survival time, the total survival time and the like, so that the INNO206 has better application prospect. It was found in the experiments that both groups of INNO206 and doxorubicin injected mice also gave consistent results, the tumor volumes of INNO206 and doxorubicin injected mice were therapeutically effective compared to the blank group, but both groups of INNO206@SOF-SH1 and INNO206@SOF-SH2 injected showed more pronounced tumor inhibition (FIG. 22), and the weight and size of the tumors also indicated that the tumors were smaller and better inhibition after treatment with the supermolecular organic frame nano-drug delivery system (FIGS. 24 and 25). Tumor tissue section results showed that tumor tissue proliferation was better inhibited in mice treated with INNO206@SOF-SH1 and INNO206@SOF-SH2, and tumor cell apoptosis was more severe (FIG. 26), indicating that INNO206@SOF-SH1 and INNO206@SOF-SH2 also have the effect of inhibiting drug-resistant tumor growth in vivo. The SOF-SH has no inhibition effect on tumor growth, which indicates that the anticancer activity of the supermolecular organic framework nano-drug delivery system is not caused by a supermolecular organic framework carrier, and the combination of laser confocal experiment results shows that the supermolecular organic framework nano-drug delivery system can effectively promote drugs to enter multi-drug resistant cells so as to better inhibit tumor cell proliferation, promote tumor cell apoptosis and achieve the effect of inhibiting tumor tissue growth.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the particular embodiments disclosed, but on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Claims (3)

1. The covalent bionic drug delivery system of the water-soluble positive ion type supermolecule organic framework is characterized in that the water-soluble supermolecule organic framework nano material is used as a carrier, the loaded drug is targeted and enriched to tumor tissues, and then the drug is released, so that the drug treatment effect is improved; specifically, a water-soluble supermolecule organic framework nano material obtained by self-assembly through the action of a host and a guest in a water phase is taken as a carrier and is marked as SOF-SH; the drug is connected to a carrier through a covalent bond to obtain a nanoparticle drug delivery system; wherein:
the drug is doxorubicin prodrug INNO206 of a multi-drug resistant cancer treatment drug model;
the water-soluble supermolecule organic framework nanomaterial is prepared by the following two construction methods:
(1) Firstly, dissolving a cysteine modified pyridylphenol derivative and tetra-p-bromomethyl phenyl methane in a dry ratio of 6:1 in stoichiometric ratioN,NReacting in dimethylformamide solution at 100deg.C for 12 hours to obtain protected tetrahedral precursor compound, reacting in hexafluoroisopropanol solution containing 0.1N hydrochloric acid at 40deg.C for 12 hours to obtain terminal mercapto-substituted positive ionic tetrahedral compound, designated as T1, and then reacting T1 and CB [8]]Self-assembling in a water phase according to the mass ratio of 1:2-1:2.5 to obtain a supramolecular framework with a cysteine structure at the tail end of a side chain, which is marked as SOF-SH1; the drug delivery system was designated INNO206@SOF-SH1;
cysteine modified pyridylphenol derivatives have the following structure:
Figure QLYQS_1
；
(2) Pyridinylphenol derivativesThe tetra-p-bromomethyl phenyl methane is dissolved in dry state according to the stoichiometric ratio of 6:1N,NReacting in dimethylformamide solution at 100deg.C for 12 hr to obtain amino-protected tetrahedral compound, reacting in hexafluoroisopropanol solution containing 0.1N hydrochloric acid at 40deg.C for 12 hr to obtain terminal amino-terminated positive ion tetrahedral compound, denoted as T2, and reacting T2 and CB [8]]Self-assembling in a water phase according to the mass ratio of 1:2-1:2.5 to obtain a framework structure, reacting the obtained supermolecular framework with 2-iminothiolane hydrochloride to obtain a supermolecular framework with a side chain terminal loaded with mercapto, and marking the supermolecular framework as SOF-SH2; the drug delivery system was designated INNO206@SOF-SH2;
the structure of the pyridylphenol derivative is as follows:
Figure QLYQS_2
；
the sulfydryl loaded at the SOF-SH terminal is conjugated and added with the maleimide structural unit of the anticancer drug INNO206 to generate a covalently linked bionic nano drug delivery system.
2. The supramolecular organic framework covalent biomimetic drug delivery system of claim 1, wherein the drug loading of the water-soluble supramolecular organic framework nanomaterial is 4-12% of the mass of the organic framework nanomaterial.
3. The supramolecular organic framework covalent biomimetic drug delivery system of claim 1, wherein the water-soluble supramolecular organic framework nanomaterial has a mesoporous structure with a hydrated particle size of 90-120nm; the shape is spherical, blocky or flaky.
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