CN113648423B - Amphiphilic conjugate anti-tumor nano-drug, preparation method thereof, nano-assembly and application - Google Patents

Amphiphilic conjugate anti-tumor nano-drug, preparation method thereof, nano-assembly and application Download PDF

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CN113648423B
CN113648423B CN202110828637.XA CN202110828637A CN113648423B CN 113648423 B CN113648423 B CN 113648423B CN 202110828637 A CN202110828637 A CN 202110828637A CN 113648423 B CN113648423 B CN 113648423B
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CN113648423A (en
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汪枭睿
李阳
王钰
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Southern Medical University
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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
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Abstract

The invention provides an amphiphilic conjugate anti-tumor nano-drug, which comprises an anti-tumor inhibitor, an anti-tumor prodrug and a hydrophilic segment which are connected through covalent bonds, wherein the anti-tumor prodrug comprises at least one of a cisplatin precursor, a carboplatin precursor and an oxaliplatin precursor. The amphiphilic conjugate antitumor nano-drug disclosed by the invention can be assembled into a uniform nano-structure in water due to a simple chemical structure, the damage to normal cells is greatly reduced by reducing the cytotoxicity of the antitumor drug outside tumor cells, and the blood circulation time in a human body is prolonged due to the EPR effect of the uniform nano-structure in vivo. Compared with small molecular antitumor drugs, the conjugate can prolong the blood circulation time in a human body and has a passive targeting function, so that the enrichment of the conjugate in a tumor part is greatly improved.

Description

Amphiphilic conjugate anti-tumor nano-drug, preparation method thereof, nano-assembly and application
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to an amphiphilic conjugate anti-tumor nano-drug, a preparation method thereof, a nano-assembly and application thereof.
Background
Tumors are one of the main causes of human death in the world at present, and chemotherapy is the most important treatment means in clinical tumor treatment. However, the traditional small molecule anticancer drugs have poor single-drug chemotherapy selectivity, low efficiency, damage to normal cells and poor drug resistance, which severely limits the efficacy of the traditional small molecule anticancer drugs in clinical tumor treatment. In order to overcome the defects, the mode of multi-drug synergistic chemotherapy is carried out and is rapidly becoming the hot point of tumor treatment. However, conventional multi-drug chemotherapy has limitations of low practical range, cross-resistance and cytotoxicity, low cellular uptake, and increased efflux, which can lead to treatment failure.
With the development of nanotechnology, the nano drug-loaded system improves the defect that the anti-cancer drugs are quickly removed in a human body. The intrinsic EPR Effect (Enhanced Permeability and Retention Effect, EPR, characteristic of nanocarriers in tumor tissues) of nanocarrier systems can enhance the circulation time of anticancer drugs in vivo by their "passive targeting" properties (Brent Weinberg, et al, angelwald, chemie.2004,116, 6483-6487). In addition, amphiphilic drug conjugates formed by non-covalent linkage of hydrophobic antitumor drugs and hydrophilic antitumor-like drugs have also begun to show the angle. (Deyue Yan, et al.J.am.chem.Soc.2014,136,33, 11748-11756). Compared with a nano drug-carrying system, the nano particles formed by the amphiphilic drug conjugate can complete the delivery of the antitumor drug, so that the side effect caused by the nano carrier is avoided.
However, the high cytotoxicity of amphiphilic drug conjugates severely limits their application in clinical tumor therapy (Xinyuan Zhu, et al.j.am.chem.soc.2018,140,28, 8797-8806). In order to reduce the high toxicity of antitumor drugs outside tumor cells, several methods have been developed: introducing a targeting motif over-expressed by tumor cells on a drug molecule, or introducing the targeting motif on a nano-carrier, and the like. These above strategies either require the addition of some substances, with complicated manufacturing processes; or the enrichment of the nano drug-loaded system at the tumor part is limited due to the low drug-loaded rate and the drug-loaded efficiency of the nano drug-loaded system, so that the curative effect of the anti-tumor drug is greatly reduced.
On the other hand, the nano drug-carrying system has few and few applications in clinical tumor treatment due to inherent heterogeneity, complex chemical structure and other factors. Recent studies show that the low tumor cell penetration capability and low tumor lesion site enrichment of the nano drug-loaded system severely limit the anti-tumor efficacy (Warren. Chan, et al. ACS. Nano.2018,12,8, 8423-8435).
Therefore, how to enhance the curative effect of the antitumor drugs and ensure the antitumor inhibition and the effective release of the chemotherapeutic drugs, and ensure the uniformity, the concise chemical structure and the low toxic and side effects of the antitumor drugs is still a serious challenge and an urgent problem to be solved. Recently, there has been a lot of research on the internal connection between the pathway of growth factors specifically expressed by tumor cells and the pathway of DNA damage signals of tumor cells, and on this basis, two drugs with different mechanisms for anti-tumor, i.e., various tumor-related inhibitors and chemotherapeutic drugs, can be screened for better therapeutic effect and reduced toxic and side effects.
Disclosure of Invention
The invention aims to design an anti-tumor system which introduces a new anti-tumor inhibitor element and has a simple and effective chemical structure, and particularly provides an amphiphilic conjugate anti-tumor nano-drug, a preparation method thereof, a nano-assembly and application thereof.
The technical scheme adopted by the invention is as follows:
in a first aspect of the present invention, there is provided:
an amphiphilic conjugate anti-tumor nano-drug, which comprises an anti-tumor inhibitor fragment, an anti-tumor prodrug fragment and a substance containing a hydrophilic segment, wherein the anti-tumor prodrug is connected by covalent bonds, and comprises at least one of a cisplatin precursor, a carboplatin precursor and an oxaliplatin precursor.
Preferably, the antitumor prodrug is at least one of a carboxyl-modified cisplatin precursor, a carboplatin precursor, and an oxaliplatin precursor.
Preferably, the antitumor inhibitor is an antitumor drug modified with at least one selected from the group consisting of a carboxyl group, a hydroxyl group, an amino group, and a thiol group.
Preferably, the anti-tumor drug is further modified by a fluorescent probe motif.
Preferably, the antitumor drug is at least one selected from the group consisting of a tyrosine kinase inhibitor, a beta-lactamase inhibitor, and a Bromodomain (Bromodomain) protein small molecule inhibitor.
Preferably, the antitumor drug is at least one selected from sunitinib, erlotinib, gefitinib, clavulanic acid, (4S) -6- (4-chlorophenyl) -N-ethyl-8-methoxy-1-methyl-4H- [1,2,4] triazolo [4,3-A ] [1,4] benzodiazepine-4-acetamide (I-BET 762) and abeta-tone (RVX-208).
Preferably, the hydrophilic segment material includes at least one of polyethylene glycol, mono-substituted polyethylene glycol, polyacrylic acid, poly (N-isopropylacrylamide), poly (oligoethylene glycol monomethyl ether), and poly (N, N-dimethylaminoethyl methacrylate).
Preferably, the degree of polymerization of the hydrophilic segment containing substance moiety is 2 to 200; preferably 10 to 50.
In a second aspect of the present invention, there is provided:
an amphiphilic conjugate anti-tumor nano-drug combination comprises the amphiphilic conjugate anti-tumor nano-drug and at least one chemotherapeutic drug bonded with the amphiphilic conjugate anti-tumor nano-drug by chemical bonds.
Preferably, the chemotherapeutic drug comprises at least one of camptothecin, doxorubicin, paclitaxel, coumarin and podophyllotoxin.
In a third aspect of the present invention, there is provided:
a preparation method of the amphiphilic conjugate anti-tumor nano-drug comprises the following steps:
(1) The antitumor prodrug and dibasic acid anhydride are substituted by carboxyl;
(2) The antitumor drug is modified by at least one of carboxyl, hydroxyl, amino and sulfhydryl;
(3) The carboxyl-substituted antitumor prodrug reacts with the antitumor drug modified by carboxyl, hydroxyl, amino and sulfydryl to obtain a covalent bond antitumor inhibitor-antitumor prodrug;
(4) The antitumor inhibitor-antitumor prodrug reacts with a substance containing a hydrophilic segment to obtain the antitumor nano-drug of the amphiphilic conjugate of the antitumor inhibitor-antitumor prodrug-hydrophilic segment which is bonded by covalent bonds.
Preferably, in the step (2), the antitumor drug is subjected to amino modification with at least one of ethylamine, triethylamine, and tetraethyl amine.
In a fourth aspect of the present invention, there is provided:
an amphiphilic conjugate anti-tumor nano-drug has a structure shown in formula I:
Figure BDA0003174612100000031
wherein R is an anti-tumor inhibitor; n =2 to 6; p =2 to 6; q =2 to 200.
Preferably, formula I above is selected from the group consisting of:
Figure BDA0003174612100000032
Figure BDA0003174612100000041
wherein R1 is an antitumor drug; m =2 to 4; n =2 to 6; p =2 to 6; q =2 to 200.
Preferably, the anti-tumor drug is an anti-tumor drug modified by a fluorescent probe motif.
Preferably, R1 is selected from the group consisting of:
Figure BDA0003174612100000042
in a fifth aspect of the present invention, there is provided:
an amphiphilic conjugate anti-tumor nano-drug combination comprises the amphiphilic conjugate anti-tumor nano-drug and at least one chemotherapeutic drug bonded with the amphiphilic conjugate anti-tumor nano-drug.
Preferably, the chemotherapeutic drug comprises at least one of camptothecin, doxorubicin, paclitaxel, coumarin and podophyllotoxin.
In a sixth aspect of the present invention, there is provided:
the amphiphilic conjugate antitumor nanometer medicine is prepared with the compound in the formula I and through reversible addition-fragmentation transfer polymerization or atom transfer radical polymerization.
In a seventh aspect of the present invention, there is provided:
an amphiphilic conjugate anti-tumor nano-drug nano-assembly at least comprises one amphiphilic conjugate anti-tumor nano-drug.
Preferably, the nano-assembly includes at least one of a nanoparticle, a vesicle, a nanorod, and a composite micelle.
An eighth aspect of the present invention provides:
the application of the amphiphilic conjugate anti-tumor nano-medicament in preparing medicaments for treating tumors, microbial infection and inflammation is disclosed, wherein the amphiphilic conjugate anti-tumor nano-medicament is the amphiphilic conjugate anti-tumor nano-medicament or is prepared by the preparation method.
In a ninth aspect of the present invention, there is provided:
the application of the amphiphilic conjugate anti-tumor nano-drug composition in the preparation of drugs for treating tumors, microbial infections and inflammations is disclosed.
In a tenth aspect of the present invention, there is provided:
an application of an amphiphilic conjugate anti-tumor nano-drug assembly in preparation of drugs for treating tumors, microbial infections and inflammations is disclosed.
The beneficial effects of the invention are:
1. the conjugate of the invention changes bivalent cisplatin technical material with high cytotoxicity as an anticancer drug into tetravalent cisplatin prodrug with low cytotoxicity through exquisite chemical design, thereby greatly reducing the toxicity of the anticancer drug outside tumor cells. Only when the conjugate enters tumor cells, the tetravalent cisplatin prodrug is converted into chemotherapeutic cisplatin with antitumor activity under the reductive condition in the tumor cells, and an antitumor inhibitor motif is released, so that the curative effect is generated.
2. The amphiphilic conjugate antitumor nano-drug has the property of simultaneously releasing the chemotherapeutic drug and the antitumor inhibitor in response to tumor microenvironment, skillfully overcomes the problems of different administration time, low cell uptake, cross drug resistance and the like of the conventional multi-drug combination chemotherapy, and greatly improves the antitumor activity of the drug.
3. The amphiphilic conjugate antitumor nano-drug disclosed by the invention can be assembled into a uniform nano-structure in water due to a simple chemical structure, the damage to normal cells is greatly reduced by reducing the cytotoxicity of the antitumor drug outside tumor cells, and the blood circulation time in a human body is prolonged due to the EPR effect of the uniform nano-structure in vivo. Compared with small molecular antitumor drugs, the conjugate can prolong the blood circulation time in a human body and has a passive targeting function, so that the enrichment of the conjugate in a tumor part is greatly improved.
4. The amphiphilic conjugate anti-tumor nano-drug can be used for realizing the cooperative treatment of tumors by additionally modifying a fluorescent probe element on the conjugate to track the drug in real time or modifying another anti-tumor drug, and has important biological medicine application prospect.
5. The invention realizes the adjustable composition, controllable molecular weight and narrow molecular weight distribution of the conjugate by adjusting the molecular weight of each component in the conjugate, and expands the application range of the amphiphilic conjugate antitumor drug.
Drawings
FIG. 1 is a scheme showing the synthesis scheme of Sunitinib-DCP-PEG.
Figure 2 is the nuclear magnetic hydrogen spectrum of sunitinib aminotriafluoroacetate.
FIG. 3 shows the nuclear magnetic hydrogen spectrum of Sunitinib-DCP.
FIG. 4 shows the nuclear magnetic hydrogen spectrum of Sunitinib-DCP-PEG.
FIG. 5 shows the nuclear magnetic hydrogen spectrum of Sunitinib-DCP-PEG-OH.
FIG. 6 shows the NMR spectrum of Sunitinib-DCP-PEG.
FIG. 7 shows the particle size distribution of the assembly of Sunitinib-DCP-PEG.
FIG. 8 is a TEM image of the assembly formed by Sunitinib-DCP-PEG.
FIG. 9 is a comparison graph of cytotoxicity of the assembly formed by Sunitinib-DCP-PEG, sunitinib prodrug and cisplatin prodrug in HeLa cells.
FIG. 10 is a comparison of cytotoxicity of the assembly formed by Sunitinib-DCP-PEG, sunitinib prodrug and cisplatin in A549 cells.
Detailed Description
In order to make the objects, technical solutions and technical effects of the present invention more clear, the present invention will be described in further detail with reference to specific embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.
The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and biomaterials, if not specifically indicated, are commercially available.
FIG. 1 shows a synthesis route of Sunitinib-DCP-PEG, and the preparation method of other amphiphilic conjugate antitumor nano-drugs of the present invention can refer to the synthesis route of Sunitinib-DCP-PEG.
Example 1: preparation of Sunitiib-DCP-PEG
(1) 0.3g (1 mmol) of cisplatin original drug was dissolved in 6 to 60mL of ultrapure water, and the reaction system was stirred at 10 to 100 ℃ for 30 minutes, followed by adding 10 to 100mL of 30% aqueous hydrogen peroxide solution thereto. After reacting for 4 hours at 10-100 ℃, the reaction system is restored to room temperature to generate light yellow crystals. The crude product was then filtered with suction and dried in vacuo to yield 0.210g of pure platinum prodrug DHP as a pale yellow solid (67% yield).
(2) 0.165g (0.5 mmol) of DHP and 0.5 (5 mmol) of succinic anhydride are dissolved in 4-40 ml of DMF, and stirred in an oil bath at 10-100 ℃ for 24 hours under the protection of sufficient nitrogen. After the reaction was terminated, the DMF solvent was spin-dried, dissolved in methanol and precipitated in anhydrous ether, precipitated three times, and dried in a vacuum oven to give carboxyl-bearing platinum prodrug DCP as a pale yellow solid in a yield of 0.216g (80.2% yield).
(3) 0.108g (0.2 mmol) of DCP and 0.30g (1 mmol) of TBTU (O-benzotriazole-N, N, N ', N' -tetramethylurea tetrafluoroborate) are dissolved in 4-40 mL of DMF, 0.035mL of triethylamine is added under the protection of sufficient nitrogen, and the mixture is stirred for half an hour at 10-100 ℃. 0.045g of Sunitinib (Sunitinib) aminotrifluoroacetate was then added and stirred at room temperature for 12 hours. After the reaction was terminated, the DMF solvent was spin-dried, dissolved in a small amount of DMF and precipitated in ultrapure water, which was then suction-filtered, washed with water three times and dried in a vacuum oven to give Sunitinib-DCP-COOH as a yellow solid in a yield of 0.101g (74.1% yield).
(4) 0.85g (0.01 mmol) of Sunitinib-DCP-COOH and 0.2g (300-50000) (0.01 mmol) of PEG were dissolved in 2-20 ml of DMF, and 0.02g (0.1 mmol) of DCC and 0.2mg (0.002 mmol) of DMAP were added under sufficient nitrogen protection, followed by stirring at 10-100 ℃ for 48 hours. After the reaction was terminated, the DMF solvent was spin-dried, dissolved with a small amount of DMF and precipitated in anhydrous ether, and repeated three times. After suction filtration, the precipitate was dissolved in a small amount of THF, dialyzed against ice water using a dialysis membrane with a molecular weight of 3500, and lyophilized to give Sunitinib-DCP-PEG as a yellow solid at a yield of 0.191g (67.0%).
Example 2: preparation of Sunitiib-DCP-PEG nanoparticles
1mg of Sunitinib-DCP-PEG prepared in example 1 was dissolved in 1mL of DMSO, and 9mL of ultrapure water was rapidly added thereto under high-speed stirring. The obtained nanoparticle solution was placed in a dialysis bag and dialyzed against water until the organic solvent was removed. Thus obtaining nanoparticles with a diameter size of about 70 nm.
Example 3: preparation of DOX-wrapped Sunitiib-DCP-PEG nano-assembly
10mg of Sunitinib-DCP-PEG obtained in example 1 and 1mg of DOX were dissolved in 1mL of DMSO, and 9mL of ultrapure water was rapidly added thereto under high-speed stirring. The obtained nanoparticle solution was placed in a dialysis bag and dialyzed against water until the organic solvent was removed.
Test example 1: material nuclear magnetic hydrogen spectrum and nuclear magnetic carbon spectrum test in preparation process of Sunitiib-DCP-PEG
The materials, intermediates and products involved in the preparation of Sunitinib-DCP-PEG in example 1 were subjected to nmr hydrogen spectroscopy and nmr carbon spectroscopy, and the results are shown in fig. 2 to 6. Wherein, fig. 2 is nuclear magnetic hydrogen spectrum test of reaction material Sunitinib amino trifluoroacetate, fig. 3 is nuclear magnetic hydrogen spectrum of intermediate Sunitinib-DCP, fig. 4 is nuclear magnetic hydrogen spectrum of product Sunitinib-DCP-PEG, fig. 5 is nuclear magnetic hydrogen spectrum of hydroxyl-containing product Sunitinib-DCP-PEG-OH, and fig. 6 is nuclear magnetic carbon spectrum of product Sunitinib-DCP-PEG.
Test example 2: sunitinib-DCP-PEG particle size determination and TEM determination
The test method comprises the following steps: taking the aqueous solution of the Sunitinib-DCP-PEG nano-assembly prepared in example 2, placing 1ml of the solution in an unopened dynamic light scattering culture dish, performing a particle size test on the solution, dropping a small amount of the solution on a copper mesh special for TEM, and starting the TEM test after the copper mesh is completely dried. The results are shown in FIGS. 7 and 8.
The results show that: the Sunitinib-DCP-PEG nano-assembly is nano-particles with uniform size and diameter of about 70 nm.
Test example 3: toxicity of Sunitinib-DCP-PEG on HeLa cells
The test method comprises the following steps: the cytotoxicity test was performed on Sunitinib-DCP-PEG, and cisplatin and Sunitinib as a comparative two prodrugs. The cells used were HeLa cells using standard MTT assay methods. First, heLa cells were cultured on a 96-well cell culture plate for 24 hours. After culturing for 24 hours, the cell culture medium is replaced, and Sunitinib-DCP-PEG, cisplatin and Sunitinib are added to ensure that the concentration of the drug in the HeLa cell culture solution reaches the corresponding concentration. After 24 hours of addition, the 96-well cell culture plate was removed and subjected to cytotoxicity assay according to MTT standard protocol, and the results are shown in fig. 9.
The results show that: compared with two technical products of Sunitinib-DCP-PEG, cisplatin and Sunitinib, the toxic and side effects of Sunitinib-DCP-PEG and cisplatin at the same concentration are greatly reduced. Meanwhile, the inhibition effect of the Sunitinib-DCP-PEG under a certain concentration on tumor cells is equivalent to that of two technical products of cisplatin and Sunitinib. This shows that the Sunitinib-DCP-PEG can inhibit tumor without generating large toxic and side effects on normal cells.
Test example 4: toxicity of Sunitiib-DCP-PEG to cisplatin-resistant cells
The test method comprises the following steps: first, heLa cells were cultured in a cisplatin-containing medium, and cells that survived the culture for a corresponding period of time were cisplatin-resistant HeLa cells. First, cisplatin-resistant HeLa cells were cultured on 96-well cell culture plates for 24 hours. After culturing for 24 hours, the cell culture medium is replaced, and Sunitinib-DCP-PEG, cisplatin and Sunitinib are added to ensure that the drug concentration in the HeLa cell culture solution reaches the corresponding concentration. After 24 hours of addition, the 96-well cell culture plate was removed and subjected to cytotoxicity assay according to MTT standard protocol, and the results are shown in fig. 10.
The results show that: compared with two technical products of Sunitinib-DCP-PEG, cisplatin and Sunitinib, the toxic and side effects of Sunitinib-DCP-PEG and cisplatin at the same concentration are greatly reduced. Meanwhile, the inhibition effect of the Sunitinib-DCP-PEG under a certain concentration on the cisplatin-resistant tumor cells is equivalent to that of Sunitinib raw drug and stronger than that of the cisplatin raw drug. This shows that the Sunitinib-DCP-PEG can overcome the resistance of tumor cells to cis-platinum and does not generate larger toxic and side effects on normal cells.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.

Claims (10)

1. An amphiphilic conjugate anti-tumor nano-drug nano-assembly is characterized in that: the nano assembly at least comprises an amphiphilic conjugate anti-tumor nano drug, the drug comprises an anti-tumor inhibitor fragment, an anti-tumor prodrug fragment and a substance containing a hydrophilic segment which are connected through covalent bonds, the anti-tumor prodrug comprises a cis-platinum precursor, and the amphiphilic conjugate anti-tumor nano drug has the structure shown in formula I:
Figure FDA0003783952410000011
wherein R is an antitumor inhibitor selected from hydroxyl or amino modified antitumor drugs, and the antitumor drugs are selected from at least one of sunitinib, erlotinib, clavulanic acid and Abetatone; n =2 to 6; p =2 to 6; q =2 to 200;
the nano assembly is at least one of nano particles, vesicles, nano rods and composite micelles.
2. The amphiphilic conjugate anti-tumor nano-drug nano-assembly according to claim 1, characterized in that: the anti-tumor medicine is further modified by a fluorescent probe motif.
3. The amphiphilic conjugate anti-tumor nano-drug nano-assembly according to claim 1, characterized in that: the preparation method of the amphiphilic conjugate anti-tumor nano-drug comprises the following steps:
(1) The antitumor prodrug and dibasic acid anhydride are substituted by carboxyl;
(2) Modifying hydroxyl or amino of the antitumor drug;
(3) The carboxyl-substituted antitumor prodrug reacts with the antitumor drug modified by hydroxyl or amino to obtain a covalent bond antitumor inhibitor-antitumor prodrug;
(4) The antitumor inhibitor-antitumor prodrug reacts with a substance containing a hydrophilic segment to obtain the antitumor nano-drug of the amphiphilic conjugate of the antitumor inhibitor-antitumor prodrug-hydrophilic segment which is bonded by covalent bonds.
4. The amphiphilic conjugate anti-tumor nano-drug nano-assembly according to claim 1, characterized in that: the formula I is selected from the group consisting of:
Figure FDA0003783952410000012
Figure FDA0003783952410000021
wherein R1 is an antitumor drug; m =2 to 4; n =2 to 6; p =2 to 6; q =2 to 200.
5. The amphiphilic conjugate anti-tumor nano-drug nano-assembly according to claim 4, characterized in that:
the R1 is selected from the following group:
Figure FDA0003783952410000022
6. an amphiphilic conjugate anti-tumor nano-drug composition is characterized in that: comprising the amphiphilic conjugate anti-tumor nano-drug nano-assembly of any one of claims 1 to 5 and at least one chemotherapeutic drug chemically bonded thereto.
7. The amphiphilic conjugate anti-tumor nano-drug composition according to claim 6, characterized in that: the chemotherapy medicine is at least one of camptothecin, adriamycin, paclitaxel, coumarin and podophyllotoxin.
8. The amphiphilic conjugate anti-tumor nano-drug nano-assembly according to claim 1, characterized in that: the compound of formula I is prepared by reversible addition-fragmentation transfer polymerization or atom transfer radical polymerization.
9. The application of the amphiphilic conjugate anti-tumor nano-drug composition in the preparation of drugs for treating tumors, microbial infections or inflammations is characterized in that: the amphiphilic conjugate anti-tumor nano-drug composition is the amphiphilic conjugate anti-tumor nano-drug composition of claim 6 or 7.
10. An application of amphiphilic conjugate anti-tumor nano-drug assembly in preparing drugs for treating tumors, microbial infections or inflammations is characterized in that: the amphiphilic conjugate anti-tumor nano-drug nano-assembly is the amphiphilic conjugate anti-tumor nano-drug nano-assembly according to any one of claims 1 to 5 and 8.
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