Use of a charge cross-linking carrier as a tumor chemotherapy drug

Abstract

The application discloses a kind of nano-carrier based on charge crosslinking and its preparation method and as the application of tumor chemotherapy drug.Therein, nano-carrier is assembled by the electrostatic interaction of two polymer chains with positive and negative charges, and the organic ligand is released under exogenous or endogenous stimulation by the pre-ligand on polymer chain B, and metal ions on positive charge polymer chain A are competed, and metal complex is generated in situ, while the electrostatic interaction of nano-carrier is weakened, leading to the dissociation of nano-carrier, and the release of polymer chain B containing organic ligand, playing the role of tumor chemotherapy or immunoregulation.The method is simple in operation, short in time consumption and mild in reaction conditions;The material has good biocompatibility and high safety.In some embodiments, pharmacodynamic experiments show that the nano-carrier can effectively kill breast cancer and induce cell immunogenic death, while inhibiting the metastasis of tumor cells, providing a new idea and means for the current urgent clinical problem of breast cancer lung metastasis.

Description

Technical Field

[0001] This invention belongs to the field of nanomaterials and their preparation, specifically relating to a charge crosslinking carrier, its preparation method, and its application as a tumor therapeutic drug. Background Technology

[0002] Malignant tumors are one of the leading diseases threatening the health of Chinese residents. Data from China's cause-of-death surveillance shows that malignant tumors account for nearly a quarter of all deaths. With the aging population, the burden of major chronic diseases, including malignant tumors, is on the rise. Currently, traditional cancer treatments include surgery, chemotherapy, and radiotherapy. However, due to limitations in surgical resection, immunosuppression caused by radiotherapy, and drug resistance resulting from chemotherapy, these treatment modalities often have poor efficacy. Currently, clinical cancer treatment recommends combination therapy, using synergistic effects across multiple mechanisms, combining radiotherapy and chemotherapy, or combining chemotherapy and immunotherapy, to improve efficacy and reduce adverse reactions.

[0003] Drug carrier design is a crucial step in the development of drug delivery systems, directly impacting drug efficacy, targeting, and safety. In recent years, much research has focused on developing drug carriers for highly effective cancer therapy. However, most drug carriers involve complex synthesis and preparation processes, resulting in high production costs, poor controllability, and weak reproducibility. Furthermore, drug carriers also face off-target effects; ensuring drug stability during in vivo circulation and selectively delivering drugs to tumor cells for effective release remain pressing issues.

[0004] Therefore, it is necessary to develop a simple and efficient drug carrier for loading multiple drugs. Based on this, a design pattern for a charge-crosslinked drug carrier is proposed. This type of carrier is simple to manufacture, controllable, and reproducible; in vivo, it can respond to the tumor microenvironment, release drugs synchronously, avoid side effects caused by metabolic differences between drugs, and improve therapeutic efficacy through drug synergy. Summary of the Invention

[0005] The purpose of this invention is to provide a charge-crosslinked carrier based on dual polymer chains for tumor treatment. A positively charged polymer chain is prepared by coordination of metal ions with organic ligands on polymer chain A, and then uniformly mixed with a negatively charged polymer chain B loaded with a prodrug through electrostatic interactions.

[0006] The purpose of this invention is to overcome two major difficulties in existing treatment modalities: first, the complexity of drug carrier design, which often involves complex synthesis and preparation processes, resulting in poor stability, reproducibility, and unstable efficacy; second, the problem of multi-drug delivery in tumor treatment, where differences in drug metabolism and off-target effects lead to uneven spatiotemporal distribution of drugs during combined administration, poor tumor cell selectivity, poor efficacy, and significant side effects.

[0007] The objective of this invention can be achieved through the following technical solutions:

[0008] A metal coordination-mediated charge crosslinking carrier is characterized by preparing a positively charged polymer chain through the coordination interaction between metal ions and ligands on polymer chain A, and uniformly mixing it with a negatively charged polymer chain B loaded with a prodrug through electrostatic interaction to obtain a uniform drug carrier that can be dispersed in an aqueous solution.

[0009]

[0010] In some specific embodiments of the present invention, the metal ion in polymer chain A is preferably Cu. 2+ Cu + Fe 3+ ,Fe 2+ Zn 2+ ;

[0011] More preferably Cu 2+ Cu + Zn 2+ ;

[0012] In some specific embodiments of the present invention, the metal ion is Cu. 2+ .

[0013] The organic ligands in polymer chain A include, but are not limited to, biologically active molecules; the molecules may be molecules with tumor chemotherapy or immunomodulatory effects, including but not limited to praxavir (AMD3100), paclitaxel, and sotolas.

[0014] In some specific embodiments of the present invention, the organic ligand of polymer chain A is preferably plexafor (AMD3100).

[0015] In some embodiments, polymer chain A is preferably hyaluronic acid, chitosan, or chondroitin sulfate; the molecular weight of the polymer is preferably 2k-10w.

[0016] In some embodiments, the prodrug in the negatively charged polymer chain B may be a proligand constituting a metal complex; the proligand constituting the metal complex includes, but is not limited to, prodrug molecules of dithiocarbamic acid derivatives.

[0017] In some embodiments, the negatively charged polymer is preferably chondroitin sulfate, a sulfated polysaccharide.

[0018] In some embodiments, the negatively charged polymer chain B modified with the prodrug is a polymer compound modified with a dithiocarbamic acid prodrug as shown in general formula I:

[0019] General Formula I:

[0020] in, It is a dithiocarbamic acid group.

[0021] N represents the negatively charged polymer chain B mentioned above;

[0022] In some of these embodiments, R1 and R2 are preferably independent C2-C5 short-chain aliphatic hydrocarbons or combined into C3-C8 cycloalkanes;

[0023] M is an exogenous / endogenous stimulus-responsive group. In some embodiments of the present invention, ROS-sensitive bonds such as phenylboronic acid esters, thioclase bonds, GSH-sensitive disulfide bonds, light-sensitive groups such as o-nitrobenzyl alcohol, enzyme-specific cleaved chemical bonds such as the sequence of FAP-α enzyme, and X-ray-sensitive bonds such as fluoroaryl azides are preferred.

[0024] In some embodiments of the present invention, the ROS-sensitive bond is more preferably a phenylboronic ester bond, with the following structure:

[0025]

[0026] L is a linker connecting the prodrug and the negatively charged polymer chain B. In some embodiments of the present invention, it is preferably adipic acid dihydrazide, with the following structure:

[0027]

[0028] Under exogenous or endogenous stimulation, the proligand on polymer chain B releases free ligands and competes for metal ions on positively charged polymer chain A, generating metal complexes in situ. At the same time, the electrostatic effect of the carrier weakens, leading to the dissociation of the carrier and the release of polymer chain B containing metal ligands, which exerts the effects of tumor chemotherapy or immunomodulation.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The preparation of this charge crosslinking carrier is simple, time-saving, and under mild reaction conditions; in vivo, it can respond to the tumor microenvironment and release drugs synchronously, avoiding side effects caused by metabolic differences between drugs, and improving the therapeutic effect through drug synergy.

[0031] In some specific embodiments, the carrier overcomes the problem that dithiocarbamate derivative drugs and copper cannot be synchronously distributed at the tumor site to exert an efficient anti-tumor effect; secondly, in view of the characteristics of easy tumor metastasis, it proposes a two-pronged strategy of in situ killing tumor cells and activating immunogenic death and inhibiting cell invasion signals to inhibit tumor cell growth and metastasis. Attached Figure Description

[0032] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings, wherein:

[0033] Figure 1 : The 1H NMR spectra of the prodrug BpDTC and the negative control pDTC in Example 1 of this invention.

[0034] Figure 2 Example 2 of the present invention: 1H NMR spectra of the carrier polymer chains CBD, CD and HA-AMD.

[0035] Figure 3 Example 3 of the present invention: Dynamic light scattering particle size measurement diagram

[0036] Figure 4 The response and release of the carrier and its individual components to H2O2 in Example 4 of this invention.

[0037] Figure 5 : Cytotoxicity diagram of the vector in Example 5 of this invention.

[0038] Figure 6 Example 6 of this invention: Vector-induced reactive oxygen species generation diagram.

[0039] Figure 7 Example 7 of this invention: Vector-induced proteasome accumulation diagram.

[0040] Figure 8 Example 8 of the present invention: Anti-migration effect of carrier component HA-AMD Detailed Implementation

[0041] The present invention will be further described below with reference to the embodiments, but the present invention is not limited to the following embodiments. For example, the concentration and volume of the solution used can be adjusted as needed.

[0042] The CXCL12 (SDF-1) / CXCR4 biological axis consists of the chemokine CXCL12 and its specific receptor CXCR4. Studies have shown that this signaling axis is highly activated under tumor pathological conditions and is associated with tumor growth, invasion, and prognosis. Currently, the only clinically available small molecule inhibitor of CXCR4, praxavir (AMD3100), is used for autologous transplantation in patients with non-Hodgkin's lymphoma and multiple myeloma. Furthermore, studies have shown that AMD3100 can effectively disrupt the CXCL12 / CXCR4 axis, inhibiting tumor development and metastasis. Simultaneously, AMD3100 has a macrocyclic structure, enabling the delivery of metal ions to the tumor region, facilitating the in-situ formation of metal complexes.

[0043] Example 1: Preparation of ROS-sensitive diethyldithiocarbamate prodrug BpDTC and its negative control pDTC

[0044] (1) The preparation process of BpDTC compound is shown below:

[0045]

[0046] 1) Preparation of intermediate 2: p-Bromophenylhydroxyacetic acid (5 g, 1 eq) was dissolved in methanol (50 mL), and concentrated sulfuric acid (1.73 mL, 1.5 eq) was dissolved in methanol and slowly added dropwise to the above solution. The reaction was carried out at 65 °C for 2 h, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 8:1). After the reaction was completed, the mixture was cooled to room temperature, the solvent was evaporated under reduced pressure, the mixture was reconstituted with ethyl acetate, and washed three times with saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to remove the solvent, yielding intermediate 2.

[0047] 2) Preparation of Intermediate 3: Intermediate 2 (5.348 g, 1 eq), bis-pinacolborate (11.08 g, 2 eq), [1,1′-bis(diphenylphosphine)ferrocene]palladium dichloride dichloromethane complex (893 mg, 0.05 eq), and potassium acetate (6.42 g, 3 eq) were dissolved in 100 mL of dioxane. The reaction was carried out at 80°C under nitrogen protection, and the reaction was monitored by TLC (petroleum ether:ethyl acetate = 10:1). After the reaction was completed, the solvent was evaporated under reduced pressure, and the solution was redissolved in dichloromethane. The filtrate was collected by suction filtration. Intermediate 3 was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 20:1).

[0048] 3) Preparation of intermediate 4: Intermediate 3 (5 g, 1 eq) was dissolved in 10 mL of dichloromethane. Thionyl chloride (26.5 mL, 20 eq) was slowly added to the reaction system under ice bath conditions. Two drops of DMF were added to catalyze the reaction. The mixture was refluxed at 50 °C under nitrogen protection, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 8:1). After the reaction was complete, the mixture was quenched with ice water and extracted multiple times with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered to obtain intermediate 4.

[0049] 4) Preparation of Intermediate 5: Intermediate 4 (4.88 g, 1 eq) was dissolved in 20 mL of acetonitrile, and sodium diethyldithiocarbamate (2.83 g, 1.05 eq) was dissolved in 20 mL of acetonitrile and gradually added to the above system. The reaction was carried out at room temperature, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 20:1). After the reaction was completed, the solvent was evaporated under reduced pressure, reconstituted with ethyl acetate, and washed with water and saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and filtered. Intermediate 5 was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 30:1).

[0050] 5) Preparation of pDTC: Intermediate 5 (3.2 g, 1 eq) was dissolved in 5 mL of tetrahydrofuran, and 5 mL of NaOH solution (605 mg, 2 eq) was added. The reaction was carried out at room temperature, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 6:1). After the reaction was completed, the mixture was extracted with ethyl acetate, and the aqueous layer was collected. The pH was adjusted to 5, at which point a white precipitate formed. The precipitate was extracted multiple times with dichloromethane, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the final product BpDTC.

[0051] (2) The preparation process of the pDTC compound is shown below:

[0052]

[0053] 1) Preparation of intermediate 6: Methyl bromophenylacetate (1.2 g, 1 eq) was dissolved in 10 mL of acetonitrile, and sodium diethyldithiocarbamate (898 mg, 1 eq) was dissolved in 10 mL of acetonitrile and slowly added to the above system. The reaction was carried out at room temperature, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 6:1). After the reaction was completed, the filtrate was filtered, the solvent was evaporated under reduced pressure, and the intermediate 6 was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1).

[0054] 2) Preparation of pDTC: Intermediate 6 (1.1 g, 1 eq) was dissolved in 5 mL of THF, and 5 mL of NaOH solution (443 mg, 3 eq) was added. The reaction was carried out at room temperature, and the reaction was monitored by TLC (petroleum ether: ethyl acetate = 6:1). After the reaction was completed, the mixture was extracted with ethyl acetate, and the aqueous layer was collected. The pH was adjusted to 5, at which point a white precipitate formed. This precipitate was extracted multiple times with dichloromethane. The organic layer was collected, dried over anhydrous sodium sulfate, and filtered to obtain the final product pDTC.

[0055] Example 2: Chondroitin A polymer chains modified with prodrugs, CBD and CD, and hyaluronic acid modified with AMD3100. Preparation of polymer chain HA-AMD

[0056] (1) The preparation process of CBD and CD polymers is shown below:

[0057]

[0058] 1) Preparation of CS-ADH: 2 g of chondroitin sulfate A (CS-A) was dissolved in 20 mL of distilled water and stirred until dissolved. 4 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and 0.585 g of N-hydroxysuccinimide (NHS) were added. The pH was adjusted to 6-7 with hydrochloric acid, and the mixture was stirred for approximately 15 min to activate the carboxyl groups. 8.8 g of adipic acid dihydrazide (ADH) was added to the system, and the pH was adjusted to 6-7 with NaOH. The mixture was stirred at room temperature for 24 h, dialyzed for three days (Mw=3500), and then freeze-dried to obtain CS-ADH. The grafting rate was calculated using 1H NMR spectroscopy.

[0059] 2) Preparation of CBD: BpDTC (0.6 g, 1 eq) was dissolved in 10 mL DMSO, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) (0.232 g, 0.6 eq) was dissolved in 7.5 mL water and added to the above system. The mixture was activated for 0.5 h. CS-ADH (0.188 g, 0.72 eq) was dissolved in 7.5 mL water and added to the above system. The mixture was stirred at room temperature for 24 h. After the reaction was complete, the mixture was dialyzed for three days (Mw=3500) and lyophilized to obtain CBD. The grafting rate was calculated using 1H NMR spectroscopy.

[0060] 3) Preparation of CD: pDTC (0.6 g, 1 eq) was dissolved in 10 mL DMSO, and 4-(4,6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) (0.165 g, 0.6 eq) was dissolved in 7.5 mL water and added to the above system. Activation was carried out for 0.5 h. CS-ADH (0.188 g, 0.72 eq) was dissolved in 7.5 mL water and added to the above system. The mixture was stirred at room temperature for 24 h. After the reaction was completed, the mixture was dialyzed for three days (Mw=3500) and lyophilized to obtain CD. The grafting rate was calculated using 1H NMR spectroscopy.

[0061] (2) The preparation process of HA-AMD polymer is shown below:

[0062]

[0063] 1) Synthesis of HA-MA: 3g of HA (1w-10w) was dissolved in 50mL of water. The reaction solution was placed at 0℃, and 5.734g of methacrylic anhydride was slowly added dropwise. The pH was adjusted to 8-9 with 5M NaOH, and the reaction was continued at 0℃ with stirring for 8h (the pH was maintained at 8-9 during the reaction). After the reaction was completed, the reaction solution was added dropwise to 500mL of anhydrous ethanol. After stirring for 10min, the precipitate was collected and washed three times with anhydrous ethanol (500mL×3). The precipitate was collected by filtration and then dried under vacuum to obtain HA-MA. The grafting rate was calculated using 1H NMR spectroscopy.

[0064] 2) Synthesis of HA-AMD: HA-MA (100 mg, 1 eq) was dissolved in a mixture of 0.5 mL DMSO and 2 mL water. Triethylamine (15 mg, 1 eq) was added, and plerixafor (AMD3100) (100 mg, 1.5 eq) was added to methanol and slowly added dropwise to the above system. The reaction was carried out under nitrogen protection at room temperature for 24 h in the dark. After the reaction was completed, the mixture was dialyzed for three days (Mw=3500) and lyophilized to obtain HA-AMD. The grafting rate was calculated using 1H NMR spectroscopy.

[0065] Example 3: Construction of charge-crosslinked nanocarriers and determination of dynamic light scattering particle size

[0066] A stock solution of HA-AMD (containing 5 mM AMD3100, all subsequent concentrations refer to AMD3100 concentration) was prepared using ultrapure water. A stock solution of prodrug-modified CBD polymer chains (containing 1 mM BpDTC, all subsequent concentrations refer to BpDTC concentration) was prepared using DMSO:ulpure water at a 1:1 ratio. A 20 mM copper chloride solution was prepared using ultrapure water. The copper-coordinated HA-AMD-Cu (1 mM) was obtained by mixing HA-AMD and the copper solution. A 1 mM HA-AMD-Cu aqueous solution was added dropwise to a 1 mM CBD solution at a ratio of 1:8, and the mixture was continuously magnetically stirred (650 r / min) for 60 min to allow HA-AMD-Cu and CBD to complex via electrostatic interaction. The resulting nanocarrier (CBD-HM-Cu) was obtained by ultrasonication and extrusion, and stored at 4°C.

[0067] The corresponding negative nanocarrier CD-HM-Cu replaces the CBD chain with a CD chain, while maintaining the same other methods.

[0068] Figure 3 Dynamic light scattering particle size analysis showed that the obtained nanocarriers had uniform particle size, approximately 230 nm, and a PDI of 0.176.

[0069] Example 4: Response and release of nanocarriers and their individual components to H2O2

[0070] HPLC was used to determine the sensitivity of the selected nanocarrier and its individual components to different concentrations of H2O2 and the activity of the active metal complex Cu(DTC)2. A C8 column was used, with a mobile phase of CH3OH:H2O = 90%:10% (0.05% HCOOH, v / v). The characteristic UV absorption peak of Cu(DTC)2 was selected at 432 nm.

[0071] The nanocarrier and individual components were mixed with H2O2 (50 mM) or without H2O2 and dissolved in a solution containing 5% DMSO and 95% H2O (v / v), and detected after incubation for 2 h.

[0072] Figure 4 The results showed that, under a concentration of 50 mM H2O2, the nanocarrier group CBD-HM-Cu with responsive activation ability and the prodrug-modified polymer chain CBD-Cu alone produced significant Cu(DTC)2, while no Cu(DTC)2 was detected in other experimental groups without responsive activation ability.

[0073] Example 5: Cytotoxicity of Nanocarriers

[0074] MTT assay: 4T1 cells were seeded in 96-well plates (5 × 10⁶ cells / well). 4 Cells / well were incubated for 24 h. Then, they were treated with CBD-HM-Cu and CD-HM-Cu nanocarriers at concentrations of 0.625 μM, 1.25 μM, 2.5 μM, 5 μM, 10 μM, and 20 μM (referring to the drug concentration in the CBD or CD polymer chains). After co-incubating each group at 37°C for 24 h, 5 mg / mL LMT solution was added to each well, and after another 4 hours of incubation, the culture medium was replaced with 100 μL DMSO. The optical density (OD) of each well was measured at 490 nm using a microplate reader.

[0075] Using the group without drug treatment as the control group and the group without cell pores as the blank group, the cell viability was calculated according to formula 2-1:

[0076]

[0077] Figure 5 The results showed that after 24 h of co-incubation, the CBD-HM-Cu vector group exhibited a significant tumor cell killing effect with increasing concentration. At a concentration of 20 μM, the cell survival rate was only 5%, while the unresponsive CD-HM-Cu group showed no killing effect even at high concentrations. This indicates that CBD-HM-Cu can be activated in response to H2O2 within tumor cells, generating Cu(DTC)2 to exert its effect.

[0078] Example 6: Nanocarrier-induced generation of reactive oxygen species

[0079] Using DCFH-DA as a ROS probe, intracellular ROS generation was detected by FCM. The specific sample processing method is as follows:

[0080] 4T1 cells were loaded at 5 × 10 4Cells were seeded at a density of cells / well in 12-well plates and incubated at 37°C with 5% CO2 for 24 h. After complete cell adhesion, the original culture medium was aspirated, and 1 mL of DMEM containing the following drug-treated medium was added: Cu(DTC)2 (2.5 μM), CBD (20 μM), CBD+HM (CBD 20 μM, HM 2.5 μM), CD-HM-Cu NPs (CD 20 μM, HM 2.5 μM, copper 5 μM), and CBD-HM-Cu NPs (CBD 20 μM, HM 2.5 μM, copper 5 μM). Each group was incubated at 37°C for 7 h. After incubation, the drug-treated DMEM solution was aspirated, and the cells were washed three times with PBS. Then, DMEM medium containing the DCFH-DA probe (10 μM) was added, and the plates were incubated for 30 min. After incubation, the DMEM solution containing DCFH-DA was aspirated, the cells were washed three times with PBS, digested with trypsin, and the cells were collected for FCM detection.

[0081] Figure 6 The results showed that after incubation with 4T1 cells, the Cu(DTC)2 group and the CBD-HM-Cu NPs group exhibited significantly enhanced DCFH-DA fluorescence signals compared to the CT group, indicating an increase in intracellular ROS levels.

[0082] Example 7: Nanocarrier-induced proteasome accumulation diagram

[0083] 4T1 cells were seeded in 6-well plates (10 5 Cells were incubated for 24 h with the cells / well group from Example 6. Cells were then incubated with each group for another 10 h (Cu(DTC)2 incubation time was 5 h). Cells were washed three times with PBS and collected by centrifugation at 1500 rpm for 5 min. Cells were lysed on ice for 30 min with RIPA lysis buffer. Proteins in the supernatant were obtained by centrifugation at 12000 rpm for 20 min and quantified using a BCA kit. Proteins were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a PVDF membrane. After blocking in 5% skim milk at room temperature for 2 h, the PVDF membrane was incubated overnight at 4°C with ubiquitin and β-Actin primary antibodies, respectively. The PVDF membrane was then incubated with HRP-bound secondary antibodies at room temperature for 2 h, and finally observed by chemiluminescent autoradiography.

[0084] Figure 7 The results show that Cu(DTC)2 and CBD-HM-Cu NPs can effectively ubiquitinate the accumulation of proteins.

[0085] Example 8: Anti-migration effect of the nanocarrier component HA-AMD

[0086] Three horizontal scratches were made in each well of a 6-well plate beforehand. 4T1 cells were then seeded into the 6-well plates and incubated until 90% confluence. Using a sterile 20 μL pipette tip, three vertical scratches were made on the cells. Dead cells were washed away with PBS. 2 mL of drug-containing DMEM medium (HA-AMD (2.5 μM) and AMD (5 μM)) was added to the experimental wells, while drug-free medium was added to the control wells. Images were taken at 0 h and 24 h after scratching. Three intersection points of the scratches in each well were photographed, and the area of ​​the empty space in the center was measured using ImageJ software.

[0087] Migration rate calculation method: Migration rate 24 h (%) = (area) 0 h -area 24 h ) / area 0 h x100%.

[0088] Figure 8 The results showed that both AMD and HA-AMD can inhibit the migration of tumor cells.

Claims

1. A metal coordination mediated charge crosslinked carrier, characterized in that, The carrier is prepared by uniformly mixing a positively charged polymer chain A with a negatively charged polymer chain B loaded with a prodrug through electrostatic interaction; the positively charged polymer chain A is prepared by coordination between metal ions and organic ligands on polymer chain A; wherein the metal ion includes, but is not limited to, Cu 2+ , Cu + , Fe 3+ , Fe 2+ , Mn 2+ ; The polymer chain A includes, but is not limited to, hyaluronic acid, chitosan, and chondroitin sulfate, with a molecular weight of 2k-50w. The organic ligand on polymer chain A is a biologically active molecule containing a coordinating group selected from carboxyl, hydroxyl, N electron donor, N and O electron donor or N, O and S electron donor. The biologically active molecule includes, but is not limited to, praxavir, abametapir, doxorubicin, paclitaxel, sotolas, or bortezomib. The negatively charged polymer chain B includes, but is not limited to, chondroitin sulfate or sulfated polysaccharides; The prodrug is a dithiocarbamic acid prodrug represented by general formula I: Formula I: wherein is a dithio carbamic group, Wherein, N is the aforementioned negatively charged polymer chain B; R1 and R2 can be different or the same C1-C10 alkyl chains, C2-C10 olefin chains or alkyne chains, or they can be aliphatic chains mono- or poly-substituted by hydroxyl, carboxyl, cyano, nitro or halogen atoms in the middle or at the end, or they can be combined into 3-8 membered aliphatic rings, aliphatic aromatic rings and heterocycles containing nitrogen, sulfur, oxygen and boron; M is an exogenous or endogenous stimulus-responsive group, including but not limited to ROS-sensitive bonds, thioketone bonds, GSH-sensitive bonds, photosensitive groups, enzyme-specific cleaving chemical bonds or X-ray-sensitive bonds; L is a linker connecting the prodrug and the negatively charged polymer chain B, including but not limited to adipic acid dihydrazide, short-chain PEG, C2-C10 saturated or unsaturated aliphatic chains or short chains linked by triazoles.

2. The metal coordination mediated charge crosslinking carrier of claim 1, wherein, The metal ion is Cu 2+ ; the high molecular chain A is hyaluronic acid; and the organic ligand is plerixafor.

3. The metal coordination mediated charge crosslinked carrier according to claim 1, wherein, The negatively charged polymer chain B is chondroitin sulfate; in general formula I, M includes, but is not limited to, phenylboronic acid esters, disulfide bonds, o-nitrobenzyl alcohol, FAP-α enzyme or MMP enzyme corresponding sequences, and fluoroaryl azides.

4. The metal coordination-mediated charge crosslinking support according to claim 1, characterized in that, The prodrug is a ROS-sensitive diethyldithiocarbamate prodrug.

5. The metal coordination-mediated charge crosslinking carrier according to any one of claims 1-4, characterized in that, Under exogenous or endogenous stimulation, the proligand on polymer chain B releases free ligands and competes for metal ions on positively charged polymer chain A, forming metal complexes in situ. At the same time, the electrostatic effect of the carrier weakens, leading to the dissociation of the carrier and the release of polymer chain B containing organic ligands.

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