CN-121975072-A - Method for preparing biomacromolecule drug delivery carrier based on simple and efficient interface crosslinking system

Abstract

The invention discloses a method for preparing a biomacromolecule drug delivery carrier based on a simple and efficient interface crosslinking system. According to the invention, the amphiphilic polymer monomer containing the epoxy group is prepared through RAFT polymerization, so that the amphiphilic polymer monomer can exist on an oil-water interface, and meanwhile, the three-arm cross-linking agent which has environmental sensitivity and is subjected to amino end capping is also prepared, and the amino group on the cross-linking agent and the epoxy group on the amphiphilic monomer are subjected to cross-linking reaction on the oil-water interface to form the drug delivery carrier. The carrier can encapsulate biological macromolecules, the tumor is precisely targeted through a high-permeability long-retention effect, glutathione rich in the tumor microenvironment breaks disulfide bonds in the carrier, and the encapsulated medicines are released, so that the purpose of treating the tumor is achieved. Therefore, the invention provides a simple and convenient drug delivery carrier for efficiently encapsulating biological macromolecules and having high stability, the prepared carrier can be efficiently absorbed by tumor cells so as to regulate and control apoptosis of the tumor cells, and the safety is high.

Inventors

• CHENG DU
• LI HAOWEN
• ZHANG ZHEN

Assignees

• 中山大学

Dates

Publication Date

20260505

Application Date

20260407

Claims (10)

1. 1. A method for preparing a biological macromolecule drug delivery carrier based on a simple and efficient interface crosslinking system is characterized in that an amphiphilic monomer containing an epoxy group and a three-arm crosslinking agent with an amino group are subjected to interface polymerization reaction at an oil-water interface to prepare the biological macromolecule drug delivery carrier; the structural formula of the amphiphilic monomer containing the epoxy group is shown as follows: , wherein m=0.5 to 4, n=0 to 2, p=0.5 to 10; The structural formula of the three-arm cross-linking agent with amino is shown as follows: 。
2. 2. The method of claim 1, wherein the molar ratio of amphiphilic monomer to amino-bearing three-arm crosslinker in the biomacromolecule drug delivery vehicle is 8:1-1:8.
3. 3. The method according to claim 1, wherein the preparation method of the amphiphilic monomer containing epoxy groups is characterized in that a RAFT reagent 2- [ dodecylthio (thiocarbonyl) thio ] -2-methylpropanoic acid is taken as a chain transfer agent and gradually polymerized with ethylene oxide-2-methyl methacrylate, lauryl methacrylate and methacryloyl ethyl sulfobetaine to prepare the amphiphilic monomer consisting of a hydrophilic block, a hydrophobic block and a block carrying epoxy groups.
4. 4. The method according to claim 1, wherein the preparation method of the three-arm crosslinking agent with amino groups is as follows: s1, reacting acrylonitrile with ethylenediamine to obtain an intermediate product No. 1; s2, reacting borane dimethyl sulfide with the intermediate product 1 in the step S1 to obtain an intermediate product 2; s3, reacting the intermediate product No. 2 in the step S2 with (2- ((2-aminoethyl) disulfide alkyl) ethyl) carbamic acid tert-butyl ester to obtain an intermediate product No. 3; And S4, reacting the intermediate product No. 3 in the step S3 with trifluoroacetic acid to obtain the three-arm crosslinking agent with amino.
5. 5. The method according to claim 2, wherein the molar ratio of 2- [ dodecylthio (thiocarbonyl) thio ] -2-methylpropanoic acid, methyl oxiran-2-ylmethacrylate, lauryl methacrylate and methacryloyl ethyl sulfobetaine is 1:3-5:3-5:5-10.
6. 6. The method according to claim 4, wherein the molar ratio of acrylonitrile to ethylenediamine in the step S1 is 4-50:1.
7. 7. The method according to claim 4, wherein the molar ratio of the intermediate product No. 1 to the borane dimethyl sulfide in the step S2 is 1:150-300.
8. 8. The method according to claim 4, wherein the molar ratio of the intermediate product No. 2 to the tert-butyl (2- ((2-aminoethyl) disulfanyl) ethyl) carbamate in step S3 is 1:3-4.
9. 9. The drug-loaded nanoparticle is characterized by comprising the biomacromolecule drug delivery carrier prepared by the method of any one of claims 1-8 and a loaded macromolecular drug, wherein the macromolecular drug is selected from RNA, DNA, plasmid or protein.
10. 10. Use of the drug-loaded nanoparticle of claim 9 for the preparation of a medicament for the treatment of cancer.

Description

Method for preparing biomacromolecule drug delivery carrier based on simple and efficient interface crosslinking system Technical Field The invention belongs to the technical field of biological medicine, and particularly relates to a method for preparing a biological macromolecule drug delivery carrier based on a simple and efficient interface crosslinking system. Background Biological macromolecular drugs play an important role in the treatment of a variety of important diseases, including cancer, cardiovascular and cerebrovascular diseases, neurodegenerative diseases, immune diseases, and the like. Compared with small molecular medicines, the biological macromolecular medicines mainly comprise polypeptides, proteins, RNA, DNA and the like, have the advantages of lower toxicity, obvious curative effect, weaker immunogenicity and the like, and can generate treatment effects on various complex diseases. While the importance of biomacromolecule drugs is becoming increasingly prominent, the development and use thereof still face a number of challenges. For example, the drugs often have the problems of poor in vitro stability, difficult separation and purification, complex structure, difficult crossing of various physiological barriers in human bodies and the like, and the full play of the drug effect is limited. Therefore, in order to improve the bioavailability of the biomacromolecule drug, the bioactivity of the biomacromolecule drug is kept to the maximum extent, so that the biomacromolecule drug can effectively cross the physiological barrier and exert the therapeutic effect, and the development of an efficient drug delivery system is particularly critical. The method is not only an important way for realizing the high-efficiency utilization of the biomacromolecule drug, but also a key ring for promoting the clinical transformation and application of the biomacromolecule drug. Delivery systems for biological macromolecular drugs can be largely divided into two major classes, viral and non-viral vectors. Common non-viral vectors include liposomes, polymeric nanoparticles, nanovesicles, and the like. The virus vector has the problems of low drug carrying efficiency, strong potential immunogenicity, complex purification process, high biosafety risk and the like although the application is earlier. In contrast, non-viral vectors do not typically cause genetic mutations or other deleterious biological interactions and are therefore considered more promising delivery systems, as well as the focus of current research. The liposome is a micro-nano vesicle formed by phospholipid bilayer, has good biocompatibility and degradability, can flexibly design the structure, can encapsulate hydrophilic and hydrophobic drugs at the same time, and becomes one of biological macromolecule delivery carriers with wider clinical application. However, liposomes still face multiple challenges such as complex manufacturing processes, limited drug encapsulation efficiency, low in vivo delivery efficiency, and inadequate stability. Therefore, development of new preparation strategies and carrier systems to improve delivery efficiency and clinical applicability of biomacromolecule drugs has become an urgent need to promote their wide application. The polymer nano particles are used as an important biomacromolecule drug delivery system, have wide application prospects in the field of biological medicine in recent years, and particularly have the advantage of being difficult to replace in treating complex diseases such as tumors. The biological macromolecule drug delivery system has the characteristics of controllable structure, easy functional modification on the surface, flexible and various drug delivery modes, good biocompatibility and the like, can remarkably improve the stability of the biological macromolecule drug, enhance the targeting property and the bioavailability, is beneficial to realizing the precise delivery of the macromolecule drug and the cooperative treatment of other drug molecules, and has unique comprehensive advantages in the biological macromolecule drug delivery system. The preparation method of the polymer nano-particles comprises a double-emulsification method, a microfluidic method, an emulsion/interface polymerization method and the like. (1) Double emulsion method Double emulsion is a common technique for preparing polymer nanoparticles, and mainly comprises two types, namely water-in-oil-in-water (W/O/W) and oil-in-water-in-oil (O/W/O). Taking water-in-oil-in-water (W/O/W) as an example, the method comprises the steps of dispersing an inner water phase (containing hydrophilic drugs or molecules to be encapsulated) in an oil phase to form colostrum (W/O) and dispersing the colostrum in an outer water phase to form emulsion drops with multiple structures of water, oil and water. The organic solvent in the oil phase is then removed, causing the polymer dissolved in the oil phase to precipitate and solidify,