Ros / gsh-responsive hyperbranched zwitterionic micelles, and preparation method and application thereof
By preparing ROS/GSH-responsive hyperbranched zwitterionic micelles, the problems of nanodrug enrichment and limited penetration in vascular normalization therapy were solved, realizing the combined treatment of tumor vascular normalization and tumor stem cell killing, thus improving the treatment effect and survival.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- CHINA PHARM UNIV
- Filing Date
- 2023-07-19
- Publication Date
- 2026-07-24
AI Technical Summary
In existing technologies, when vascular normalization therapy is combined with nanomedicine therapy, conventional chemotherapy drugs are highly hydrophobic, have short half-lives in vivo, and have significant toxic side effects. Furthermore, abnormal tumor blood vessels limit the accumulation and penetration of nanomedicines at the tumor site, making it impossible to effectively inhibit tumor growth.
ROS/GSH-responsive hyperbranched zwitterionic micelles were used to prepare small-sized micelles via acylation and Michael addition reactions. These micelles were loaded with anti-angiogenic drugs and anti-tumor stem cell drugs, and the drugs were released in response to ROS and GSH in the tumor microenvironment to achieve combined therapy of vascular normalization and tumor stem cell killing.
This approach improves drug stability, prolongs in vivo circulation time, reduces toxic side effects, and significantly enhances therapeutic efficacy through deep penetration and targeted release of small-sized nanomedicines at tumor sites, combining treatment of tumor angiogenesis and tumor stem cells.
Smart Images
Figure CN117281773B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to a hyperbranched zwitterionic micelle drug delivery system, particularly to a ROS / GSH-responsive hyperbranched zwitterionic micelle drug delivery system, and also to the preparation method and application of the above-mentioned zwitterionic micelles. Background Technology
[0002] Angiogenesis is an important process in tumor progression. Newly formed blood vessels not only provide channels for tumor cell metastasis, but also create abnormal microenvironments such as hypoxia, acidosis, and increased interstitial pressure (IFP) in tumor tissue, thereby affecting the therapeutic effect of cytotoxic drugs.
[0003] Angiogenesis inhibitors can suppress tumor angiogenesis, promote tumor vascular "normalization," and subsequently restore vascular structure and function, providing a favorable environment for other drug treatments and achieving synergistic therapeutic effects. Peter et al., through intraperitoneal injection of sunitinib into nude mice with SF126 gliomas, found that the number of blood vessels in tumor tissue decreased, but the density of functional vessels within the tumor increased, the tumor perfusion index increased, and the blood flow velocity in microvessels increased tenfold. Doxorubicin accumulation at the tumor site was significantly enhanced. The combination of vascular normalization and conventional chemotherapy small molecule drugs showed a significant synergistic tumor-suppressing effect. However, the high hydrophobicity, short half-life, and significant toxic side effects of conventional chemotherapy drugs still limit the widespread application of vascular normalization therapy.
[0004] Nanomedicines can effectively improve drug stability, prolong in vivo circulation time, and reduce drug toxicity. They can also achieve the co-encapsulation and delivery of multiple drugs, such as anti-angiogenic inhibitors and chemotherapeutic drugs, enhancing retention and penetration at the tumor site and providing a good means for efficient synergistic tumor treatment. Tumor angiogenesis normalization therapy increases the number of functional blood vessels within the tumor, but it also reduces the gaps between vascular endothelial cells and decreases vascular permeability, posing higher requirements for the design and application of nanomedicine systems. Results of angiogenesis normalization combined with nanomedicine therapy show that small-sized Abraxane (~10nm) is more effective in inhibiting tumor growth than large-sized Doxil (~100nm). Repairing abnormal tumor blood vessels significantly improves the delivery of smaller-scale nanomedicines, but hinders the retention and penetration of larger nanomedicines, failing to achieve a good inhibitory effect. Summary of the Invention
[0005] Objectives of the invention: The first objective is to provide a ROS / GSH-responsive zwitterionic micelle drug delivery system; the second objective is to provide a method for preparing the above-mentioned small-sized zwitterionic micelle drug delivery system; and the third objective is to provide the application of the above-mentioned zwitterionic micelles in the preparation of drugs for combined therapy that promotes vascular normalization and tumor stem cell killing.
[0006] Technical solution: The ROS / GSH responsive zwitterionic drug delivery system of the present invention comprises: acylation reaction of hyperbranched zwitterionic polycarbonate and acrylate oxaloyl chloride to obtain ROS responsive hyperbranched zwitterionic polycarbonate; and Michael addition reaction of acrylate functional hyperbranched zwitterionic polycarbonate with dimercapto compounds and cystamine bisacrylamide to obtain GSH responsive hyperbranched zwitterionic polycarbonate.
[0007] ROS-responsive and GSH-responsive hyperbranched zwitterionic polycarbonate were self-assembled and photocrosslinked in aqueous solution to obtain ROS-responsive and GSH-responsive zwitterionic micelles.
[0008] By loading anti-angiogenic drugs onto ROS-responsive zwitterions and anti-tumor stem cell drugs onto GSH-responsive zwitterions, a ROS / GSH-responsive zwitterion drug delivery system for combined therapy of vascular normalization and tumor stem cell killing is obtained.
[0009] Preferably, the structure of the hyperbranched zwitterionic polycarbonate is shown in general formula (I), and the structure of the acrylate functionalized hyperbranched zwitterionic polycarbonate is shown in general formula (II), wherein R1 is selected from H or CH3, and R2 is selected from C2-C4 alkyl or C4-C8 aryl:
[0010]
[0011] Preferably, the general structural formula of the acrylate oxaloyl chloride is as follows: R is selected from C2-C6 alkyl and C2-C6 heteroalkyl.
[0012] Preferably, the dithiol compound is selected from compounds with the following structures:
[0013] Where n = 1 - 50.
[0014] Preferably, the anti-angiogenic drug is selected from tyrosine kinase inhibitors of the tyrosine class, and the anti-tumor stem cell drug is salinomycin, thioridazine, all-trans retinoic acid, or curcumin.
[0015] The ROS / GSH-responsive zwitterionic micelles are small micelles ranging from 5 to 25 nm in size.
[0016] This invention also provides a method for preparing the above-mentioned ROS / GSH-responsive zwitterionic drug delivery system, characterized by comprising the following steps:
[0017] (1) ROS-responsive hyperbranched zwitterionic polycarbonate was obtained by acylation reaction of hyperbranched zwitterionic polycarbonate and acrylate oxaloyl chloride; GSH-responsive hyperbranched zwitterionic polycarbonate was obtained by Michael addition reaction of acrylate functional hyperbranched zwitterionic polycarbonate with dimercapto compound and cystamine bisacrylamide in sequence.
[0018] (2) The ROS-responsive hyperbranched zwitterionic polycarbonate and the GSH-responsive hyperbranched zwitterionic polycarbonate are self-assembled and photocrosslinked in aqueous solution, respectively, to obtain the ROS-responsive zwitterionic micelles and the GSH-responsive zwitterionic micelles.
[0019] (3) Loading anti-angiogenic drugs onto ROS-responsive zwitterions and anti-tumor stem cell drugs onto GSH-responsive zwitterions yields the ROS / GSH-responsive zwitterion drug delivery system.
[0020] Preferably, the reaction formula for the synthesis of the ROS-responsive hyperbranched zwitterionic polycarbonate is as follows:
[0021] Preferably, the reaction formula for the synthesis of the GSH-responsive hyperbranched zwitterionic polycarbonate is as follows:
[0022] This invention also provides the application of the above-mentioned ROS / GSH-responsive zwitterionic drug delivery system in the preparation of drugs for combined therapy that promotes vascular normalization and tumor stem cell killing.
[0023] Invention Principle: Tumor stem cells possess self-renewal capacity, multi-lineage differentiation potential, and high tumorigenicity. They are often located within hypoxic and high-IFP tumor regions. Conventional treatments result in very low drug concentrations in tumor stem cell (CSC) regions, making effective killing difficult and leading to tumor recurrence and metastasis. Vascular normalization can regulate the tumor microenvironment and reduce tumor IFP, effectively promoting the tumor enrichment and penetration of small-sized nanomedicines, increasing drug concentrations near tumor stem cells, and thus achieving a highly efficient combined therapy of vascular normalization and tumor stem cell killing. This invention utilizes small-sized, tumor extracellular / intracellular microenvironment intelligently responsive (ROS / GSH) nanomicelles to load anti-angiogenic drugs and tumor stem cell killing drugs, applying them in combination for vascular normalization therapy and tumor stem cell killing.
[0024] ROS / GSH-responsive hyperbranched zwitterionic polycarbonate can be directly dissolved in high-purity water and self-assembled, then cross-linked under ultraviolet light to obtain small-sized nanomicelles. Adding a DMSO solution containing an anti-angiogenic drug tyrosine kinase inhibitor under ultrasound can yield ROS-responsive nanomedicines for tumor microenvironment normalization; adding a DMSO solution containing a tumor stem cell killing drug under ultrasound can yield GSH-responsive nanomedicines for eliminating tumor stem cells within tumor cells.
[0025] ROS / GSH-responsive nanomedicines can effectively improve drug stability, prolong in vivo circulation time, reduce drug toxicity and side effects, and achieve effective drug encapsulation and targeted release, such as the extracellular matrix release of anti-angiogenic inhibitors and the intracellular release of chemotherapeutic drugs, providing a good means for efficient synergistic treatment of tumors. ROS-responsive nanomedicines release tyrosine kinase inhibitors near tumor blood vessels to inhibit angiogenesis and normalize blood vessels, thereby regulating the tumor microenvironment, reducing interstitial pressure in tumor tissue, and promoting the deep penetration of subsequent small-sized GSH-responsive nanomedicines into tumor tissue. The small-sized carrier is less affected by the narrowing of intercellular spaces and the pore size of the extracellular matrix of tumor cells, thus accumulating more at the tumor site and having strong tumor penetration ability, effectively reaching the tumor stem cell region inside the tumor, and releasing tumor stem cell drugs under high GSH concentrations within tumor cells to kill tumor stem cells. Therefore, the drug delivery system of the present invention avoids the tumor enrichment limitation of conventional nanomedicines (size above 100nm) by vascular normalization therapy due to the small size effect of amphoteric ionized micelles. It integrates multiple treatment strategies, modulates the tumor microenvironment by pruning blood vessels, and can also kill tumor stem cells to reduce the risk of tumor metastasis, significantly enhance the treatment effect and improve survival.
[0026] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) The ROS / GSH responsive zwitterionic micelle drug delivery system of the present invention can effectively solve the limitation of the narrowing of the intercellular space of vascular endothelial cells caused by vascular normalization on the subsequent enrichment and penetration of nanomedicines in the tumor site while realizing vascular normalization. It can also make full use of the change in intercellular pressure of tumor tissue brought about by vascular normalization to kill tumor stem cells, effectively realizing the combined treatment of vascular normalization therapy and tumor stem cell therapy; (2) The preparation method of the ionic micelles of the present invention is simple and convenient, and the reaction conditions are mild. Attached Figure Description
[0027] Figure 1 The hydrogen nuclear magnetic resonance spectrum of ROS-responsive zwitterionic hyperbranched polycarbonate (hPC-CB-OBA) in Example 1;
[0028] Figure 2The hydrogen NMR spectrum of the mercapto-zwitterionic hyperbranched polycarbonate (hPC-CB-SH) in Example 2;
[0029] Figure 3 The hydrogen NMR spectrum of the GSH-responsive hyperbranched zwitterionic polycarbonate (hPC-CB-CBA) in Example 2;
[0030] Figure 4 The particle size distributions of small-sized ROS-responsive micelles (RM) and small-sized GSH-responsive micelles (GM) in Example 3 are shown.
[0031] Figure 5 The results of the ROS-responsive drug-loaded micelles (RM@Sun) on endothelial cell migration inhibition in Example 4 are shown.
[0032] Figure 6 The particle size distribution of the large-size GSH-responsive micelles (BGM) in Example 5 is shown.
[0033] Figure 7 The results of the large and small-sized GSH-responsive drug-loaded micelles killing tumor stem cell spheres in Example 5;
[0034] Figure 8 This is an in vitro tumor image and tumor weight analysis of 4T1 tumor-bearing mice after treatment with the combined therapy of tumor angiogenesis normalization and tumor stem cell killing in Example 6. Detailed Implementation
[0035] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.
[0036] Example 1
[0037] Synthesis of ROS-responsive hyperbranched zwitterionic polycarbonate hPC-CB-OBA
[0038] Oxaloyl chloride (0.57 g, 4.50 mmol) was dissolved in 10 mL of dichloromethane (DCM), and hydroxyethyl acrylate (0.10 g, 0.90 mmol) and triethylamine (0.09 g, 0.90 mmol) were slowly added dropwise with stirring. After reacting in an ice bath for 4 h, unreacted hydroxyethyl acrylate was removed by rotary evaporation to obtain ROS-responsive crosslinking linker oxaloyl acrylate. The oxaloyl acrylate dimethyl sulfoxide solution was added dropwise to hPC-CB hyperbranched polycarbonate (2 g, 2.25 mmol hydroxyl groups), and the reaction was allowed to proceed overnight. The reaction solution was then dialyzed in high-purity water to remove unreacted small molecules. After dialysis, the liquid in the dialysis bag was freeze-dried to obtain the product, ROS-responsive zwitterionic hyperbranched polycarbonate. NMR results showed that the hyperbranched zwitterionic polymer modified with 10% oxaloyl acrylate had the structure designated hPC-CB-OBA. (NMR results are attached.) Figure 1 .
[0039] Example 2
[0040] Synthesis of GSH-responsive hyperbranched zwitterionic polycarbonate
[0041] (1) Synthesis of thiolized hyperbranched zwitterionic polycarbonate hPC-CB-SH
[0042] 3,6-Dioxa-1,8-octanedithiol (0.50 g, 2.76 mmol) was dissolved in 10 mL of methanol. Under stirring, hPC-AC-CB (2 g, 0.69 mmol acrylate unit) and a catalytic amount of triethylamine, dissolved in a mixed solvent of dimethylformamide (DMF) and methanol (MeOH), were slowly added dropwise. The reaction was carried out overnight at room temperature. After the reaction was complete, the reaction solution was dialyzed in high-purity water to remove excess 3,6-dioxa-1,8-octanedithiol and other small molecules. After dialysis, the liquid in the dialysis bag was lyophilized to obtain the product, a thiolized zwitterionic hyperbranched polycarbonate. NMR results showed that the characteristic peak of the AC unit disappeared, indicating complete substitution by thiol groups. Its structure is labeled hPC-CB-SH. Its NMR spectra are attached. Figure 2 .
[0043] (2) Synthesis of GSH-responsive hyperbranched zwitterionic polycarbonate hPC-CB-CBA
[0044] GSH-sensitive small molecule cystamine bisacrylamide (CBA) (0.50 g, 2.76 mmol) was dissolved in 10 mL of methanol. Under nitrogen protection, a mixture of DMF and methanol containing 2 g (0.69 mmol) of thiol-modified zwitterionic hyperbranched polycarbonate hPC-CB-SH and triethylamine was slowly added dropwise. After the reaction, the reaction solution was dialyzed in high-purity water to remove excess CBA and triethylamine. After dialysis, the liquid in the dialysis bag was freeze-dried to obtain the product, GSH-responsive zwitterionic hyperbranched polycarbonate. NMR results showed that the zwitterionic hyperbranched polymer modified with 10% cystamine bisacrylamide had the structure designated hPC-CB-CBA. (NMR results are attached.) Figure 3 .
[0045] Example 3
[0046] Preparation and drug loading of small-sized ROS / GSH responsive micelles
[0047] Under ultrasonic conditions, 6 mg of hPC-CB-OBA polymer was added to 2 mL of purified water and sonicated for half an hour. Then, a catalytic amount of the photoinitiator 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone (I2959) was added to the micelle solution, and the mixture was crosslinked under ultraviolet light for 20 min. After the light irradiation, small-sized ROS-responsive micelles (RM) were obtained. The average particle size of the micelles was measured to be 9.8 nm using dynamic light scattering. The particle size distribution is shown in the attached figure. Figure 4 Freshly prepared ROS-responsive micelles were slowly added to a DMSO solution of sunitinib (Sun) (50 mg / mL) with stirring. After stirring for half an hour, the unloaded drug and organic solvent were removed by dialysis with high-purity water to obtain small-sized ROS-responsive micelles (RM@Sun) loaded with anti-angiogenic drugs.
[0048] The preparation of small-sized GSH-responsive micelles was similar to the above process. Under ultrasonic conditions, 6 mg of hPC-CB-CBA polymer was added to 2 ml of purified water and sonicated for half an hour. Then, photoinitiator I2959 was added to the micelle solution, and the mixture was crosslinked under ultraviolet light for 20 min. After the irradiation, small-sized GSH-responsive micelles (GM) were obtained. The average particle size of the micelles was measured to be 11.2 nm using a dynamic light scattering instrument. The particle size distribution is shown in the attached figure. Figure 4 Freshly prepared GSH-responsive micelles were slowly added to a DMSO solution of salinomycin (50 mg / mL) with stirring. After stirring for half an hour, the unloaded drug and organic solvent were removed by dialysis with high-purity water to obtain small-sized GSH-responsive micelles (GM@Sal) loaded with anti-tumor stem cell drugs.
[0049] Example 4
[0050] Inhibition of vascular endothelial cell migration by ROS-responsive micelles loaded with anti-angiogenic drugs (RM@Sun):
[0051] Wound healing assays were performed using human umbilical vein endothelial cells (HUVECs) cultured in 12-well plates at 37°C and 5% CO2 in DMEM medium containing 10% serum, at a cell density of 30,000 cells / well. When the cell density reached 90%, uniform cell scratches were made using a pipette tip, and the width was photographed and recorded. The original medium was then replaced with serum-free medium containing freeSun, RM@Sun, and RM@Sun + H2O2 (100 μM). The width was photographed again after 48 hours.
[0052] like Figure 5 As shown, RM@Sun exhibited an effective anti-cell migration effect, and the drug release in the presence of hydrogen peroxide further enhanced the anti-cell migration effect, indicating that the ROS-responsive micelles (RM@Sun) carrying anti-angiogenic drugs can effectively inhibit angiogenesis and promote vascular normalization.
[0053] Example 5
[0054] The killing effect of large and small GSH-responsive micelles carrying tumor stem cell killing drugs on tumor stem cells: First, tumor stem cell spheres were obtained by suspension sphere culture. 4T1 cells in good logarithmic growth phase were digested, counted, and resuspended in serum-free DMEM / F12 medium (containing 5 μg / mL insulin, 20 ng / mL epidermal growth factor, 20 ng / mL basic fibroblast growth factor, 1X B27, and 0.4% (w / v) bovine serum albumin). Cells were seeded at a density of 40,000 cells / well in low-adsorption 6-well plates and cultured at 37°C and 5% CO2. Serum-free DMEM / F12 medium was added every 3 days to maintain the nutrients required for microsphere growth. After 7 days, the cells were passaged. Uniformly sized 4T1 tumor stem cell spheres were selected after 3 passages for tumor stem cell killing experiments.
[0055] To investigate the effect of size on tumor stem cell killing, a control material, large-sized GSH-responsive drug-loaded micelles (BGM@Sal), was prepared via reverse nanoprecipitation. The method was as follows: 5 mg of hPC-CB-CBA polymer was dissolved in 1 mL of purified water and added to 10 mL of acetone with stirring. Reverse nanoprecipitation was then used to form large-sized micelles with an opalescent appearance. Photoinitiator I2959 was then added to the micelle solution, and the mixture was crosslinked under UV light for 20 min. After irradiation, the acetone was removed by rotary evaporation to obtain the large-sized GSH-responsive micelles (BGM). The average particle size of the micelles was measured to be 152 nm using dynamic light scattering. The particle size distribution is shown in the attached figure. Figure 6 Freshly prepared GSH-responsive micelles were slowly added to a 50 mg / mL MDSO solution of salinomycin under stirring. After stirring for half an hour, the unloaded drug and organic solvent were removed by dialysis with high-purity water to obtain the control material, a large-sized GSH-responsive micelle loaded with the tumor stem cell killing drug (BGM@Sal).
[0056] Uniformly sized tumor stem cell spheres were selected and placed in low-adsorption 24-well plates. GM@Sal or BGM@Sal (Sal: 8 μg / mL) was added to serum-free DMEM / F12 medium. The size of the tumor stem cell spheres was recorded by photograph every 24 hours.
[0057] As attached Figure 7The small-sized GM@Sal, leveraging its size effect, can effectively penetrate into the interior of 3D tumor spheres, achieving highly efficient killing of tumor stem cells. GM@Sal's destructive ability against tumor spheres is far superior to that of the larger-sized BGM@Sal, indicating that this small-sized GSH-responsive micelle (GM@Sal) carrying anti-tumor stem cell drugs has therapeutic advantages in tumor stem cell therapy. In subsequent animal treatments, it can be combined with ROS-responsive micelles (RM@Sun) to normalize blood vessels, allowing for deeper penetration into the tumor and synergistic killing of tumor stem cells.
[0058] Example 6
[0059] Therapeutic effects of combined tumor angiogenesis and tumor stem cell killing therapy in 4T1 tumor-bearing mice
[0060] Female BALB / c mice were subcutaneously injected with 100 μL of a suspension of 4T1 cells in the logarithmic growth phase (1 × 10⁻⁶ cells). 6 (Cells / Mouse). When the mouse tumor grew to 80 mm 3 Mice were randomly assigned to six groups (n=6): PBS, RM-Sun, BGM-Sal, GM-Sal, RM-Sun+BGM-Sal, and RM-Sun+GM-Sal (15 mg Sun equiv. / kg; 5 mg Sal equiv. / kg). Treatment was administered via tail vein on days 0, 2, 4, 6, and 8. On day 24 of treatment, tumor-bearing mice were sacrificed, and tumors were collected, photographed, weighed, and analyzed.
[0061] As attached Figure 8 As shown, the tumor volume and weight after treatment with vascular normalization plus small-sized stem cell nanomedicine (RM-Sun+GM-Sal) were significantly smaller than those after treatment with small-sized stem cell nanomedicine alone (GM-Sal), indicating that tumor vascular normalization effectively improved the therapeutic effect of small-sized anti-tumor stem cell nanomedicine. However, the tumor volume and weight after treatment with vascular normalization plus large-sized stem cell nanomedicine (RM-Sun+BGM-Sal) were similar to those after treatment with large-sized stem cell nanomedicine (BGM-Sal), with no significant difference, indicating that tumor vascular normalization did not improve the therapeutic effect of large-sized anti-tumor stem cell nanomedicine. These results suggest that in vascular normalization therapy, the size effect of the nanocarrier influences the combined therapeutic effect. Only small-sized nanomedicines can benefit from the increased tumor accumulation and penetration brought about by vascular normalization, thereby increasing the drug concentration near tumor stem cells at the tumor center. The application of small-sized drug-loaded micelles enabled the combined application of vascular normalization and tumor stem cell killing therapy to exhibit a significant tumor-suppressive effect in tumor-bearing mice.
Claims
1. A ROS / GSH-responsive zwitterionic drug delivery system, characterized in that, The drug delivery system acylates hyperbranched zwitterionic polycarbonate and acrylate oxaloyl chloride to obtain ROS-responsive hyperbranched zwitterionic polycarbonate, and performs Michael addition reaction of acrylate functional hyperbranched zwitterionic polycarbonate with dimercapto compounds and cystamine bisacrylamide to obtain GSH-responsive hyperbranched zwitterionic polycarbonate. ROS-responsive and GSH-responsive hyperbranched zwitterionic polycarbonate were self-assembled and photocrosslinked in aqueous solution to obtain ROS-responsive and GSH-responsive zwitterionic micelles. By loading anti-angiogenic drugs onto ROS-responsive zwitterions and anti-tumor stem cell drugs onto GSH-responsive zwitterions, a ROS / GSH-responsive zwitterion drug delivery system for combined therapy of vascular normalization and tumor stem cell killing is obtained. The ROS-responsive hyperbranched zwitterionic polycarbonate is: ； The GSH-responsive hyperbranched zwitterionic polycarbonate is: 。 2. The zwitterionic micelle drug delivery system according to claim 1, characterized in that, The anti-angiogenic drug is selected from tyrosine kinase inhibitors of the tyrosine kinase class, and the anti-tumor stem cell drug is salinomycin, thioridazine, all-trans retinoic acid, or curcumin.
3. The zwitterionic micelle drug delivery system according to claim 1, characterized in that, The ROS / GSH-responsive zwitterionic micelles are small micelles ranging from 5 to 25 nm in size.
4. A method for preparing the ROS / GSH-responsive hyperbranched zwitterionic drug delivery system according to claim 1, characterized in that, Includes the following steps: (1) ROS-responsive hyperbranched zwitterionic polycarbonate was obtained by acylation reaction of hyperbranched zwitterionic polycarbonate and acrylate oxaloyl chloride; GSH-responsive hyperbranched zwitterionic polycarbonate was obtained by Michael addition reaction of acrylate functional hyperbranched zwitterionic polycarbonate with dimercapto compound and cystamine bisacrylamide in sequence. (2) The ROS-responsive hyperbranched zwitterionic polycarbonate and the GSH-responsive hyperbranched zwitterionic polycarbonate are obtained by self-assembly and photocrosslinking in aqueous solution, respectively. (3) Loading anti-angiogenic drugs onto ROS-responsive zwitterions and anti-tumor stem cell drugs onto GSH-responsive zwitterions yields the ROS / GSH-responsive zwitterion drug delivery system.
5. The manufacturing method according to claim 4, characterized in that, The reaction formula for the synthesis of ROS-responsive hyperbranched zwitterionic polycarbonate is as follows: 。 6. The manufacturing method according to claim 4, characterized in that, The reaction formula for the synthesis of GSH-responsive hyperbranched zwitterionic polycarbonate is as follows: 。 7. The use of the ROS / GSH-responsive zwitterionic drug delivery system of claim 1 in the preparation of a drug for combined therapy that promotes vascular normalization and tumor stem cell killing.