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CN-122011370-A - PH-sensitive polyamino acid nano micelle and preparation method and application thereof

CN122011370ACN 122011370 ACN122011370 ACN 122011370ACN-122011370-A

Abstract

The application provides a pH sensitive polyamino acid nano micelle, a preparation method and application thereof, wherein a triblock copolymer consisting of polyethylene glycol, polylysine and phenylalanine is provided, the N-end is a polyethylene glycol chain with the length of 113 repeating units, the middle section is a polylysine chain consisting of 10 lysine residues, and the C-end is a phenylalanine chain consisting of N phenylalanine residues, so as to form the polyethylene glycol-polylysine-phenylalanine triblock copolymer. The polymer has no toxicity to tumor cells, and the drug-loaded nano micelle can effectively inhibit proliferation of tumor cells. In addition, confocal laser scanning microscopy shows that the nano-micelle can be effectively endocytosed by tumor cells, and the drug can be rapidly released from the nano-micelle and passively diffused into the cell nucleus. The synthesis of the material provides a new idea for the field of drug sustained release.

Inventors

  • ZHANG HAOHAO
  • CHEN YAJUAN
  • Zhou Piqian
  • WANG MIAO
  • OUYANG YIFAN
  • WANG JIWEI

Assignees

  • 福州大学

Dates

Publication Date
20260512
Application Date
20260130

Claims (9)

  1. 1. A pH-sensitive polyamino acid nano-micelle is characterized by comprising three sections of polyethylene glycol, polylysine and polyphenylalanine, and has the specific structure that the N-end is a polyethylene glycol chain with the length of 113 repeating units and is connected with the next chain section through an amido bond, the middle section is a polylysine chain formed by 10 lysine residues, the side chain of the polylysine chain contains amino groups, the benzyloxycarbonyl group in the polylysine chain polymer of the lysine residues is removed, the pH-sensitive polyamino acid nano-micelle has cationic property, the C-end is a polyphenylalanine chain formed by N phenylalanine residues, wherein n=10-30, the main chain of the whole molecule is a polyamino acid-polyethylene glycol block copolymer formed by connecting peptide bonds, and the structure can be named as PEG 113 –(Lys) 10 –(Phe) n , namely a 'polyethylene glycol-polylysine-polyphenylalanine' triblock copolymer.
  2. 2. The pH-sensitive polyamino acid nanomicelle according to claim 1 wherein n of the triblock copolymer is 10, 20 or 30 respectively and corresponds to the formula PEG 113 –(Lys) 10 –(Phe) 10 、PEG 113 –(Lys) 10 –(Phe) 20 、PEG 113 –(Lys) 10 –(Phe) 30 .
  3. 3. The pH-sensitive polyamino acid nanomicelle of claim 1 wherein the polylysine is L-Lys (Cbz) -NCA, the polyethylene glycol is mPEG 113 -NH 2 , and the phenylalanine is L-Phe-NCA.
  4. 4. A method for preparing pH sensitive polyamino acid nano micelle, comprising the following steps: 1) Synthesis of copolymer Dissolving L-Lys (Cbz) -NCA in DMF, vacuumizing the system, replacing the system with argon for three times to keep the system in an inert gas atmosphere, dissolving mPEG 113 -NH 2 in anhydrous DMF, adding the anhydrous DMF into a flask, continuously reacting at 35 ℃ for 60: 60 h, dissolving L-Phe-NCA in DMF, slowly adding the DMF into the system for second-stage polymerization, continuously reacting again for 60: 60 h, concentrating by reduced pressure distillation after the whole process is finished, precipitating white powder solid by using diethyl ether as a poor solvent, and naming the white powder solid as P-Cbz, wherein the molar ratio of L-Lys (Cbz) -NCA to mPEG 113 -NH 2 to L-Phe-NCA is 10:1:10-30; 2) Removal of benzyloxycarbonyl groups Dissolving the P-Cbz in the step 1) in TFA, stirring for 30 min, injecting HBr/ACOH into a flask under ice bath condition, continuously stirring for 4h under a closed state, and precipitating the target product P by taking diethyl ether as a poor solvent, wherein the product is light yellow powdery solid.
  5. 5. The preparation method of the pH-sensitive polyamino acid nano micelle is characterized in that the molar ratio of L-Lys (Cbz) -NCA to mPEG 113 -NH 2 to L-Phe-NCA is 10:1:10-10, a white powder solid is obtained, the target product is named as P1-Cbz, the molar ratio of L-Lys (Cbz) -NCA to mPEG 113 -NH 2 to L-Phe-NCA is 10:1:10-20, a white powder solid is obtained, the target product is named as P12Cbz, the target product is P2, the molar ratio of L-Lys (Cbz) -NCA to mPEG 113 -NH 2 to L-Phe-NCA is 10:1:10-30, and the target product is named as P3-Cbz.
  6. 6. The method for preparing the pH-sensitive polyamino acid nano-micelle according to claim 4, wherein the chemical synthesis process of the nano-micelle is as follows: 。
  7. 7. a pH-sensitive polyamino acid nano-micelle drug-loading application, a nano-micelle according to any one of claims 1 to 3 or a nano-micelle loaded Doxorubicin (DOX) model drug obtained by the preparation method according to any one of claims 4 to 6.
  8. 8. The method according to claim 7, wherein the nanomicelle is loaded with Doxorubicin (DOX) model drug, 8mg DOX. Mu.l of HCl is dissolved in a conical flask with a certain amount of DMSO, 5. Mu.l of triethylamine is injected and stirred for 5 hours, 30mg of polyamino acid nanomicelle P is dissolved in a certain amount of DMF and transferred into a reaction bottle, 24mg of the mixture is stirred in a dark place and h mg of the mixture is slowly added into the flask with 5mL pH =7.4 of PBS solution to complete the self-assembly process of the polymer, then the mixed solution is added into a dialysis bag for dialysis for 2 days, and finally the solution is filtered by a 0.45 μm aqueous phase filter membrane to remove sediment and impurities, and freeze-dried to obtain the drug-loaded nanoparticle P-M.
  9. 9. The use according to claim 8, wherein the nanomicelle carries Doxorubicin (DOX) model drug, the chemical synthesis process of which is as follows: 。

Description

PH-sensitive polyamino acid nano micelle and preparation method and application thereof Technical Field The application belongs to the technical field of polymer chemistry and pharmacy, and particularly relates to a pH-sensitive polyamino acid nano micelle and a preparation method and application thereof. Background Chemotherapy is one of the main methods for clinically treating malignant tumors at present, but conventional chemotherapeutics have poor selectivity, and can inhibit tumor growth, but damage normal tissues at the same time. The advent of nano-delivery systems effectively avoided a series of problems with traditional drugs. Nano-delivery systems rely on the small size effect and surface effect of nanoparticles to enhance drug absorption and to achieve control of drug release. The nano carrier can also change the distribution of the medicine in the body, enhance the concentration of the medicine in a focus area, and realize the targeted release of the medicine through the enhanced permeability and retention effect (EPR effect) of tumor blood vessels, thereby reducing toxic and side effects. The morphology of the nano-micelles is generally spherical, cubic or spindle-shaped, and can be better captured by cell membranes by changing the specific surface area. The nano micelle has controllable size and good dispersibility, is beneficial to circulating in blood without precipitation, and can ensure the integrity of the drug when reaching specific tissues. The polymer nano micelle is a functional system with a specific structure, and is formed by self-assembling amphiphilic block polymers in aqueous solution or co-solution. The main driving forces for self-assembly include hydrogen bonding, coordination bonding, van der Waals forces, electrical forces, solvent effects, hydrophilic and hydrophobic effects, and the like. When the concentration of the polymer in the aqueous solution is low, it is generally uniformly dispersed in the solution. When the polymer concentration is higher than the Critical Micelle Concentration (CMC), the hydrophobic segment bends to form a core to reduce the contact area with the water phase, while the hydrophilic segment covers the outside of the core to form a micelle shell. Typically, the nanomicelles are spherical core-shell structures ranging in size from 10 nm to 200 nm. The core may support the hydrophobic chemotherapeutic agent and promote its circulation in the body by forming chemical bonds or electrostatic interactions. Hydrophilic shells can prevent aggregation between different cores, and some shells can be modified by active ligands to guide medicines to be transported to lesion sites, so that toxic and side effects are reduced. Although the traditional drug-loaded nano-micelle can realize a certain drug loading, the nano-micelle lacks response to tumor environment, so that the problems of slow drug release and the like are caused. Disclosure of Invention The present application has been made in view of the above-described problems, and an object of the present application is to provide a pH-sensitive polyamino acid nanomicelle which is internalized by tumor cells in response to a minute change in tumor environment by using a polylysine moiety as a hydrophilic segment, and which has amphiphilic properties, thereby realizing targeted drug release. The first aspect of the application provides a pH-sensitive polyamino acid nano micelle, which is a triblock copolymer consisting of three sections of polyethylene glycol (PEG), polylysine (PL-lysine) and polyphenylalanine (Phe), and has the specific structure that the N-end is a polyethylene glycol (PEG) chain with the length of 113 repeating units and is connected with the next chain section through an amide bond, the middle section is a polylysine (Poly-L-lysine) chain consisting of 10 lysine residues, the side chain of the polylysine (Poly-L-lysine) chain contains amino (-NH 2), the phenyloxycarbonyl group in the polylysine (Poly-L-lysine) chain of the lysine residues is removed, the triblock copolymer has cationic property, the C-end is a polyphenylalanine (Polyphenylalanine) chain consisting of N phenylalanine residues, wherein n=10-30, and the main chain of the whole molecule is connected by peptide bonds (-CO-NH) -, thus the structure can be named as PEG 113-(Lys)10-(Phe)n. In any embodiment, n of the triblock copolymer is 10, 20 or 30, respectively, corresponding to the formula PEG113-(Lys)10-(Phe)10、PEG113-(Lys)10-(Phe)20、PEG113-(Lys)10-(Phe)30. In any embodiment, the polylysine is L-Lys (Cbz) -NCA, the polyethylene glycol is mPEG 113-NH2, and the polyphenylalanine is L-Phe-NCA. The second aspect of the application provides a method for preparing pH-sensitive polyamino acid nano-micelles, comprising the following steps: 1) Synthesis of copolymer Dissolving L-Lys (Cbz) -NCA in DMF, vacuumizing the system, replacing the system with argon for three times to keep the system in an inert gas atmosphere, dissolving mP