Preparation method of nano delivery system for ROS (reactive oxygen species) response drug-loading dual-targeting treatment of diabetic nephropathy

Abstract

The invention discloses a preparation method of a nano delivery system for ROS-responsive drug-loading dual-targeting treatment of diabetic nephropathy, and belongs to the technical field of nano drug loading and biological medicine. The preparation method comprises the following steps: preparing a metal complex TCPP-Fe < 3 + >-Ce < 3 + > by taking FeCl3, Ce (NO3) 3.6 H2O and tetra (4-carboxyphenyl) porphyrin (TCPP) as raw materials, activating carboxyl on the metal complex, carrying out grafting reaction on the carboxyl and amino, and linking a diselenide bond capable of being subjected to ROS response to obtain a nano-carrier TCPP-Fe < 3 + >-Ce < 3 + > (at) Se-Se, combining the amino on the TCPP-Fe < 3 + >-Ce < 3 + > (at) Se-Se with a specific peptide ligand of a targeted kidney tissue, and further loading a medicine, so that the targeted kidney tissue is prepared. The nano delivery system for targeted treatment of diabetic nephropathy through active oxygen response drug loading is obtained.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and particularly relates to a preparation method of a ROS-responsive drug-loaded dual-targeting nanodelivery system for the treatment of diabetic kidney disease. Background Art

[0002] Diabetic kidney disease (DKD) is one of the main microvascular complications of diabetes and an important cause of chronic kidney disease and end-stage kidney disease. The progression of DKD is closely related to the overproduction of mitochondrial reactive oxygen species (ROS), mitochondrial DNA mutations, mitochondrial abnormalities, excessive mitochondrial fission caused by disorders of mitochondrial respiratory chain complexes, and mitochondrial dyskinesia. It has been confirmed that regulating mitochondrial homeostasis in DKD can alleviate lipid accumulation.

[0003] Since the 1990s, nanoparticles have benefited various diseases such as cancer and tumors as drug delivery carriers, but the research on nanodrugs for DKD is still very limited. This is because the glomerular filtration barrier (GFB) is the primary obstacle for nanodrugs to passively target the kidney. In addition to designing the physicochemical properties of nanoparticles to passively target the kidney, active targeting and specific recognition of the sites of kidney disease can also be achieved through the functionalization of targeting ligands to improve drug delivery for treating specific kidney diseases. In addition, the selection of nanodrug carriers and the bioavailability of the nanosystem are also major problems to be solved currently. The choice of nanocarrier will affect the drug release and the degradation cycle of the system. Therefore, there is an urgent need to develop a more precise, safer, and more efficient treatment strategy for DKD.

[0004] To effectively treat DKD, preparing a drug delivery system with dual targeting of renal tubules and glomeruli, good biocompatibility, sustained-release performance, and degradability, and intervening in key targets with key drugs can provide a certain basis for the further development of drugs for diabetic kidney disease. Summary of the Invention

[0005] The object of the present invention is to provide a preparation method of a ROS-responsive drug-loaded dual-targeting nanodelivery system for the treatment of diabetic kidney disease. Using FeCl 3 , Ce(NO 3 ) 3 ·6H 2 O and tetrakis(4-carboxyphenyl)porphyrin (TCPP) as raw materials to prepare the metal complex TCPP-Fe 3+ -Ce 3+ , by activating the carboxyl group on the metal complex and grafting it with an amino group and linking a ROS-responsive diselenide bond to obtain the nanocarrier TCPP-Fe 3+ -Ce 3+ @Se-Se, using TCPP-Fe3+ -Ce 3+ The amino group on @Se-Se binds to the specific peptide ligand targeting the renal tissue, and then further loads the drug to obtain a reactive oxygen species-responsive drug-loaded targeted nanodelivery system for the treatment of diabetic nephropathy.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The technical solution of the present invention: provides a preparation method of a ROS-responsive drug-loaded dual-targeted nanodelivery system for the treatment of diabetic nephropathy, comprising the following steps:

[0008] Step 1: Preparation of the nanocarrier

[0009] Dissolve ferric chloride and cerium nitrate in 5 mL of an organic solvent, add TCPP and stir for 1 - 3 h (rotation speed 600 - 1500 rpm) and sonicate for 5 - 30 min. Subsequently, add polyvinylpyrrolidone PVP and glacial acetic acid, stir for 1 - 3 h (rotation speed 600 - 1500 rpm) and sonicate for 5 - 30 min to form a uniformly dispersed system. Add the above mixture to a hydrothermal reaction device and heat it in a water bath at 50 - 100 °C for 10 - 48 h. After the reaction, collect the sample, centrifuge for 5 - 30 min (10000 - 15000 rpm, 0 - 4 °C), take the supernatant and dialyze (MWCO: 3500 Da) for 12 - 48 h to obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ 。

[0010] Step 2: Preparation of the diselenide bond-modified nanocarrier

[0011] Take the above dialyzed product, add carbodiimide (EDC), adjust the pH to 4 - 6 with NaOH, activate for 0.5 - 2 h (600 - 1500 rpm), and then add a crosslinking stabilizer and stir for 2 - 24 h (600 - 1500 rpm). Obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ @Se-Se;

[0012] Step 3: Loading of the specific ligand

[0013] Add the specific ligand to the TCPP-Fe 3+ -Ce 3+ @Se-Se solution, add EDC and a crosslinking stabilizer, stir for 2 - 24 h (600 - 1500 rpm), and dialyze (MWCO: 3500 Da) for 12 - 48 h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep..

[0014] Step 4: Drug Loading

[0015] Weigh the drug and dissolve it in an organic solvent, vortex at high speed for 30 - 300 s, and add it to the TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. solution, and stir for 2 - 12 h. Perform dialysis for 10 - 24 h (3500 Da) to obtain the final product TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep.@Dr.

[0016] The organic solvents in Step 1 and Step 4 are one or more of the organic solvents that retain their original water solubility, such as dimethylformamide (DMF), dihydrolevoglucosenone, ethyl acetate, methyl ethyl ketone (MEK), etc.;

[0017] In Step 1, the molecular weight of PVP is 10000 - 50000 Mw, and the mass ratio of PVP to glacial acetic acid is 0 - 2:1.

[0018] The crosslinking stabilizers in Step 2 and Step 3 are one or more of the carbodiimide crosslinking stabilizers such as N-hydroxysuccinimide (NHS) and selenocystamine dihydrochloride, etc.

[0019] The specific ligands in Step 3 are one or more of the polypeptides such as Cyclo(RGDfC), PKNGSDP, and ELRGD(R / M)AX(W / L) that can dual-target glomerular podocytes and renal tubular interstitial endothelial cells.

[0020] The drugs in Step 4 are one or more of the powerful antioxidants and anti-inflammatory agents such as celastrol, vitamin C, vitamin E, tea polyphenols, anthocyanins, coenzyme Q10, sulforaphane, lipoic acid, etc., or other drugs that can be used for the treatment of diabetic nephropathy.

[0021] The beneficial technical effects of the present invention are as follows:

[0022] In the present invention, we are committed to designing a drug delivery platform for the treatment of diabetic nephropathy with a nano-polymer having dual-targeting characteristics of the glomerulus and renal tubules and ROS responsiveness. By utilizing the high specificity of the specific peptide ligand and the complementary target protein of the target cell and the ROS responsiveness of the nano-carrier, the bioavailability and effectiveness of the drug treatment for diabetic nephropathy are improved.

[0023] In the present invention, we form a ROS-responsive substrate by coordinating organic polymer TCPP with metal particles and linking disulfide bonds, which endows the system with higher cycle stability, drug accumulation ability and oxidative stress responsiveness while ensuring grafting with polypeptides, providing a favorable guarantee for the treatment of diabetic nephropathy. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 TEM images of TCPP-Fe 3+ -Ce 3+ in Examples 1, 2, 3 and 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d).

[0025] Figure 2 XRD patterns of TCPP-Fe 3+ -Ce 3+ in Examples 1, 2, 3 and 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d).

[0026] Figure 3 Particle size diagrams of TCPP-Fe 3+ -Ce 3+ in Examples 1, 2, 3 and 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d). DETAILED DESCRIPTION OF THE INVENTION

[0027] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be construed as a limitation on the present invention, but rather as a more detailed description of certain aspects, features and embodiments of the present invention. It should be understood that the terms used in the present invention are only for describing particular embodiments and are not intended to limit the present invention.

[0028] In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0029] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention.

[0030] Regarding the terms "comprising", "including", "having", "containing", etc. used in this text, they are all open-ended terms, meaning including but not limited to.

[0031] Example 1

[0032] Step 1: Preparation of nanocarriers

[0033] Dissolve 7.66 mg of ferric chloride and 20.52 mg of cerium nitrate in 5 mL of DMF organic solvent, add 5 mg of TCPP, stir for 1 h (rotation speed 1000 rpm), and ultrasonically treat for 5 min. Add the above mixture to a hydrothermal reaction device and heat it in a water bath at 65 °C for 10 h. After the reaction, collect the sample, centrifuge for 10 min (15000 rpm, 0 °C), take the supernatant and dialyze (MWCO: 3500 Da) for 12 h to obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ .

[0034] Step 2: Preparation of diselenide bond-modified nanocarriers

[0035] Take the above dialyzed product, add 9.66 mg of carbodiimide (EDC), adjust the pH to 4.5 with NaOH, activate for 2 h (1000 rpm), then add 58.01 mg of NHS and 1 mg of selenocystamine dihydrochloride and stir for 12 h (1000 rpm). Obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ @Se-Se;

[0036] Step 3: Loading of specific ligands

[0037] In the TCPP-Fe 3+ -Ce 3+ @Se-Se solution, add 2.15 mg of the specific ligand polypeptide cyclo, add 1.2 mg of EDC and 7.1 mg of NHS, stir for 8 h (1000 rpm), and dialyze (MWCO: 3500 Da) for 48 h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep..

[0038] Step 4: Loading of drugs

[0039] Weigh 3 mg of celastrol, dissolve it in DMF organic solvent, vortex at high speed for 30 s, add it to the TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. solution, and stir for 8 h. Dialyze for 12 h (3500 Da) to obtain the final product TCPP-Fe 3+ -Ce 3+@Se-Se@Pep.@Dr.

[0040] Example 2

[0041] Step 1: Preparation of nanocarriers

[0042] Dissolve 7.66 mg of ferric chloride and 20.52 mg of cerium nitrate in 5 mL of DMF organic solvent, add 5 mg of TCPP and stir for 1 h (rotation speed 1000 rpm) and sonicate for 5 min. Subsequently, add 300 mg of polyvinylpyrrolidone PVP (MW: 10000) and 200 μL of glacial acetic acid, stir for 1 h (rotation speed 600 rpm) and sonicate for 5 min to form a uniformly dispersed system. Add the above mixture to a hydrothermal reaction device and heat it in a water bath at 65 °C for 10 h. After the reaction, collect the sample, centrifuge for 10 min (15000 rpm, 0 °C), take the supernatant and dialyze (MWCO: 3500 Da) for 12 h to obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ .

[0043] Step 2: Preparation of diselenide bond-modified nanocarriers

[0044] Take the above dialyzed product, add 9.66 mg of carbodiimide (EDC), adjust the pH to 4.5 with NaOH, activate for 2 h (1000 rpm), and then add 58.01 mg of NHS and 1 mg of selenocystamine dihydrochloride and stir for 12 h (1000 rpm). Obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ @Se-Se;

[0045] Step 3: Loading of specific ligands

[0046] In the TCPP-Fe 3+ -Ce 3+ @Se-Se solution, add 2.15 mg of the specific ligand polypeptide cyclo, add 1.2 mg of EDC and 7.1 mg of NHS, stir for 8 h (1000 rpm), and dialyze (MWCO: 3500 Da) for 48 h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep..

[0047] Step 4: Loading of drugs

[0048] Weigh 3 mg of vitamin E and dissolve it in DMF organic solvent, vortex at high speed for 30 s, and add it to the TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. solution, and stir for 8 h. Dialyze for 12 h (3500 Da) to obtain the final productTCPP-Fe 3+ -Ce 3+ @Se-Se@ Pep.@Dr.。

[0049] Example 3

[0050] Step 1: Preparation of nanocarriers

[0051] Dissolve 7.66 mg of ferric chloride and 20.52 mg of cerium nitrate in 5 mL of DMF organic solvent, add 5 mg of TCPP and stir for 1 h (rotation speed 1000 rpm) and sonicate for 5 min. Subsequently, add 300 mg of polyvinylpyrrolidone PVP (MW: 40000) and 200 μL of glacial acetic acid, stir for 1 h (rotation speed 600) and sonicate for 5 min to form a homogeneous dispersion system. Add the above mixture to a hydrothermal reaction device and heat it in a water bath at 65 °C for 10 h. After the reaction is completed, collect the sample, centrifuge for 10 min (15000 rpm, 0 °C), take the supernatant and dialyze (MWCO: 3500 Da) for 12 h to obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ .

[0052] Step 2: Preparation of diselenide bond-modified nanocarriers

[0053] Take the above dialysis product, add 9.66 mg of carbodiimide (EDC), adjust the pH to 4.5 with NaOH, activate for 2 h (1000 rpm), and then add 58.01 mg of NHS and 1 mg of selenocystamine dihydrochloride and stir for 12 h (1000 rpm). Obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ @Se-Se;

[0054] Step 3: Loading of specific ligands

[0055] In the TCPP-Fe 3+ -Ce 3+ @Se-Se solution, add 2.15 mg of the specific ligand polypeptide cyclo, add 1.2 mg of EDC and 7.1 mg of NHS, stir for 8 h (1000 rpm), and dialyze (MWCO: 3500 Da) for 48 h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep..

[0056] Step 4: Loading of drugs

[0057] Weigh 3 mg of tea polyphenols and dissolve them in DMF organic solvent, vortex at high speed for 30 s, and add them to TCPP-Fe 3+ -Ce 3+In the @Se-Se@Pep. solution, stir for 8 h. Perform dialysis for 12 h (3500 Da) to obtain the final product TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep.@Dr.

[0058] Example 4

[0059] Step 1: Preparation of nanocarriers

[0060] Dissolve 7.66 mg of ferric chloride and 20.52 mg of cerium nitrate in 5 mL of DMF organic solvent, add 5 mg of TCPP and stir for 1 h (rotation speed 1000 rpm) and sonicate for 5 min. Subsequently, add 300 mg of polyvinylpyrrolidone PVP (MW: 50000) and 200 μL of glacial acetic acid, stir for 1 h (rotation speed 600) and sonicate for 5 min to form a uniformly dispersed system. Add the above mixture to a hydrothermal reaction device and heat it in a water bath at 65 °C for 10 h. After the reaction is completed, collect the sample, centrifuge for 10 min (15000 rpm, 0 °C), take the supernatant and dialyze (MWCO: 3500 Da) for 12 h to obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ 。

[0061] Step 2: Preparation of nanocarriers modified with diselenide bonds

[0062] Take the above dialyzed product, add 9.66 mg of carbodiimide (EDC), adjust the pH to 4.5 with NaOH, activate for 2 h (1000 rpm), and then add 58.01 mg of NHS and 1 mg of selenocystamine dihydrochloride and stir for 12 h (1000 rpm). Obtain the product, denoted as TCPP-Fe 3+ -Ce 3+ @Se-Se;

[0063] Step 3: Loading of specific ligands

[0064] In the TCPP-Fe 3+ -Ce 3+ @Se-Se solution, add 2.15 mg of the specific ligand polypeptide cyclo, add 1.2 mg of EDC and 7.1 mg of NHS, stir for 8 h (1000 rpm), and dialyze (MWCO: 3500 Da) for 48 h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep.

[0065] Step 4: Loading of drugs

[0066] Weigh 3 mg of celastrol and dissolve it in DMF organic solvent. Vortex at high speed for 30 s, and then add it to the TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. solution, and stir for 8 h. Perform dialysis for 12 h (3500 Da) to obtain the final product TCPP-Fe 3+ -Ce 3+ @Se-Se@ Pep.@Dr.。

[0067] Figure 1 which are the transmission electron microscopy images of TCPP-Fe 3+ -Ce 3+ in Example 1, Example 2, Example 3, and Example 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d). As Figure 1 shown, PVP was not added in Example 1, PVP (Mw 10000) was added in Example 2, PVP (Mw 40000) was added in Example 3, and PVP (Mw 50000) was added in Example 4. Different morphologies of the nanocarrier TCPP-Fe 3+ -Ce 3+ were synthesized due to the different addition amounts and molecular weights of polyvinylpyrrolidone PVP.

[0068] Figure 2 which are the XRD patterns of TCPP-Fe 3+ -Ce 3+ in Example 1, Example 2, Example 3, and Example 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d). The XRD patterns show that the diffraction peaks of TCPP appear in the nanocarriers synthesized in all four examples, indicating successful competitive coordination of metal particles.

[0069] Figure 3 which are the particle size diagrams of TCPP-Fe 3+ -Ce 3+ in Example 1, Example 2, Example 3, and Example 4. Example 1 (a), Example 2 (b), Example 3 (c), Example 4 (d). PVP was not added in Example 1, PVP (Mw 10000) was added in Example 2, PVP (Mw 40000) was added in Example 3, and PVP (Mw 50000) was added in Example 4. Nanocarriers with different particle sizes of TCPP-Fe 3+ -Ce 3+ were synthesized due to the different addition amounts and molecular weights of polyvinylpyrrolidone PVP.

[0070] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for preparing a ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy, characterized in that: The following steps are involved: Preparation of metal complex TCPP-Fe from FeCl3, Ce(NO3)3·6H2O and tetrakis(4-carboxyphenyl)porphyrin (TCPP) 3 + -Ce 3+ The nanocarrier TCPP-Fe was obtained by activating the carboxyl group on the metal complex and grafting it with the amino group and linking it with a ROS-responsive diselenide bond. 3+ -Ce 3+ @Se-Se, using TCPP-Fe 3+ -Ce 3+ The amino group on @Se-Se is combined with a specific peptide ligand targeting renal tissue, and then further loaded with drugs to obtain the diabetic nephropathy nanodelivery system.

2. The method for preparing a ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 1, characterized in that: The mass ratio of tetrakis(4-carboxyphenyl)porphyrin (TCPP), FeCl3 and Ce(NO3)3·6H2O is 1-5:0.1-3:0.1-3.

3. The method for preparing a ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 1, characterized in that: The specific steps of the preparation method of the nano-delivery system include: Step 1: Preparation of nanocarriers Dissolve ferric chloride and cerium nitrate in 5 mL of organic solvent, add TCPP, stir for 1-3 hours (speed 600-1500 rpm) and ultrasonicate for 5-30 minutes. Then add polyvinyl pyrrolidone PVP and glacial acetic acid, stir for 1-3 hours (speed 600-1500 rpm) and ultrasonicate for 5-30 minutes to form a uniform dispersion system. Add the above mixture to the hydrothermal reaction equipment, heat in a 50-100°C water bath for 10-48 hours. After the reaction is completed, collect the sample, centrifuge for 5-30 minutes (10000-15000 rpm, 0-4°C), take the supernatant and dialyze (MWCO: 3500Da) for 12-48 hours to obtain the product, recorded as TCPP-Fe 3+ -Ce 3+ . Step 2: Preparation of diselenide bond-modified nanocarriers Take the above dialyzed product, add carbodiimide (EDC), adjust the pH to 4-6 with NaOH, activate for 0.5-2h (600-1500rpm), then add the crosslinking stabilizer and stir for 2-24h (600-1500rpm). The product is recorded as TCPP-Fe 3+ -Ce 3+ @Se-Se; Step 3: Loading of specific ligand In TCPP-Fe 3+ -Ce 3+ Add specific ligands to the Se-Se solution, add EDC and cross-linking stabilizer, stir for 2-24h (600-1500rpm), and dialyze (MWCO: 3500Da) for 12-48h to obtain TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. Step 4: Drug loading Weigh the drug and dissolve it in an organic solvent, vortex at high speed for 30-300s, and add it to TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep. solution, stirred for 2-12h. Dialyzed for 10-24h (3500Da) to obtain the final product TCPP-Fe 3+ -Ce 3+ @Se-Se@Pep.@Dr.

4. The method for preparing the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 3, characterized in that: The organic solvent in step 1 and step 4 is one or more of dimethylformamide (DMF), dihydrolevoglucosenone, ethyl acetate, butanone (MEK) and other organic solvents that retain their original water solubility.

5. The method for preparing the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 3, characterized in that: In the step 1, the molecular weight of PVP is 10000-50000Mw, and the mass ratio of PVP to glacial acetic acid is 0-2:

1.

6. The method for preparing the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 3, characterized in that: The cross-linking stabilizer in step 2 and step 3 is one or more carbodiimide cross-linking stabilizers such as N-hydroxysuccinimide NHS and selenocystamine dihydrochloride.

7. The method for preparing the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 3, characterized in that: The specific ligand in step 3 is one or more of the polypeptides Cyclo (RGDfC), PKNGSDP and ELRGD (R / M) AX (W / L) that can dual-target to glomerular podocytes and tubular interstitial endothelial cells.

8. The method for preparing the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to claim 3, characterized in that: The drugs in step 4 are one or more potent antioxidants and anti-inflammatory agents including tripterygium wilfordii, vitamin C, vitamin E, tea polyphenols, anthocyanins, coenzyme Q10, sulforaphane, lipoic acid, etc., or other drugs that can be used for the treatment of diabetic nephropathy.

9. A preparation method and application of the ROS-responsive drug-loaded dual-targeted nano-delivery system for treating diabetic nephropathy according to any one of claims 1 to 8.

10. A method for preparing a diselenide bond-bridged active oxygen scavenging drug-loaded targeted nano-delivery platform as claimed in claim 1 and its application in biomedicine.

Images