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CN-121971647-A - CAR-T cell membrane modified nano drug delivery system and preparation method and application thereof

CN121971647ACN 121971647 ACN121971647 ACN 121971647ACN-121971647-A

Abstract

The invention relates to the technical field of biology, in particular to a nano drug delivery system modified by a CAR-T cell membrane, which comprises a compound formed by self-assembly of nano particles and gelatin, an anti-tumor drug coated in a compound network, and the CAR-T cell membrane coated on the surface of the compound network, wherein the nano particles are at least one selected from artificially synthesized lithium alginate and neodymium ion doped artificially synthesized lithium alginate. The nano-carrier has the advantages of high drug loading rate, uniform particle size and long circulation time, can be used for transporting more drugs into a body, is highly targeted by modifying the CAR-T cell membrane, and can replace different CAR-T membranes or load other drugs by a modularized design.

Inventors

  • CUI JINGYUAN
  • LI YULIN
  • LI MINGPEI

Assignees

  • 上海大学温州研究院

Dates

Publication Date
20260505
Application Date
20251224

Claims (10)

  1. 1. A CAR-T cell membrane modified nanomedicine delivery system comprising: A complex formed by self-assembly of nanoparticles and gelatin; Antitumor drugs entrapped in the complex network, and A CAR-T cell membrane coated on the surface of the complex network; Wherein the nano particles are at least one selected from artificially synthesized lithium algae soil and neodymium ion doped artificially synthesized lithium algae soil.
  2. 2. The CAR-T cell membrane modified nano drug delivery system according to claim 1 is characterized in that the preparation method of neodymium ion doped artificial lithium algae soil comprises the steps of dissolving magnesium chloride hexahydrate and neodymium nitrate hexahydrate with deionized water to obtain solution A, dissolving lithium chloride in the deionized water, stirring and dissolving the solution A to form solution A-B, adding tetraethoxysilane into the solution A to dissolve the solution A, adding dilute hydrochloric acid dropwise to adjust pH to 3-4 to hydrolyze the solution A-B to obtain solution C, slowly adding the solution C dropwise into the solution A-B to form milky suspension by stirring, slowly adding sodium hydroxide solution to pH to 10.0-10.5 to supplement deionized water, performing hydrothermal reaction, centrifuging, washing, drying and grinding to obtain the nano drug delivery system.
  3. 3. The CAR-T cell membrane modified nano-drug delivery system of claim 1, wherein the anti-tumor drug comprises at least one of doxorubicin, paclitaxel, daunorubicin, and irinotecan.
  4. 4. The CAR-T cell membrane modified nano-drug delivery system of claim 1, wherein the CAR-T cell membrane comprises a CAR-T cell membrane of HER2 and/or a CAR-T cell membrane of TYRP 1.
  5. 5. A method of preparing a CAR-T cell membrane modified nanomedicine delivery system according to any one of claims 1-4, comprising the steps of: (1) Dissolving gelatin in water to prepare gelatin water solution, adding nanoparticle dispersion liquid, stirring to enable gelatin and nanoparticles to self-assemble to form complex dispersion liquid; (2) Adding the composite dispersion liquid obtained in the step (1) into acetone or water, and stirring to form nano dispersion liquid; (3) Adding glutaraldehyde solution into the nano dispersion liquid obtained in the step (2) for stirring and crosslinking; (4) Adding an anti-tumor drug solution into the crosslinked system in the step (3) for incubation, embedding the drug into a complex network, and then separating and removing free drug to obtain drug-loaded nano particles; (5) Extracting cell membrane vesicles from CAR-T cells targeted to a specific tumor antigen; (6) And (3) co-extruding the drug-loaded nano particles obtained in the step (4) and the cell membrane vesicles obtained in the step (5) to coat cell membranes on the surfaces of the drug-loaded nano particles, thereby obtaining the nano drug delivery system.
  6. 6. The method according to claim 5, wherein in the step (1), the concentration of the gelatin aqueous solution is 0.1-10 mg/mL, the stirring time is 5-60 minutes, the mass ratio of gelatin to artificially synthesized lithium algae soil is 10.0-0.3, the volume ratio of the complex dispersion liquid to acetone is 0.05-1.0, and the stirring time is 5-30 minutes.
  7. 7. The method according to claim 5, wherein in the step (3), the concentration of the glutaraldehyde solution is 0.02-2% (w/v), the mass ratio of glutaraldehyde to gelatin is 0.05-0.25, and the stirring time is 5-30 minutes.
  8. 8. The method of claim 5, wherein in step (5), cell membrane vesicles are extracted from CAR-T cells by cyclic freeze thawing in combination with differential centrifugation at a rotational speed of 5000-20000 rpm for a centrifugation time of 10-100 min.
  9. 9. A pharmaceutical composition comprising the CAR-T cell membrane-modified nano-drug delivery system of any one of claims 1-4 and a pharmaceutically acceptable carrier or excipient.
  10. 10. Use of the CAR-T cell membrane modified nano-drug delivery system of any one of claims 1-4 or the pharmaceutical composition of claim 9 in the manufacture of a medicament for the treatment of a tumor.

Description

CAR-T cell membrane modified nano drug delivery system and preparation method and application thereof Technical Field The invention relates to the technical field of biology, in particular to a CAR-T cell membrane modified nano drug delivery system, a preparation method and application thereof. Background Cancer has become a global major public health problem in which solid tumors are a treatment bottleneck due to the presence of dense extracellular matrix and abnormal tumor microenvironment, which makes it difficult for chemotherapeutic drugs to effectively penetrate and enrich. Although chemotherapeutic drugs such as Doxorubicin (DOX) are widely applied to clinic, the problems of poor targeting property, strong toxic and side effects, easiness in generating multi-drug resistance and the like exist, and development of a novel drug delivery system is needed to solve the defects. The nanotechnology provides a new direction for tumor treatment, and nano carriers such as mesoporous silica, micelle and the like are used for drug delivery, but most carriers have fixed sizes, so that the requirements of long circulation in blood circulation and deep penetration of tumor tissues are difficult to meet at the same time. The nano-carrier with the changeable size can realize tumor enrichment through a large size, and deep penetration through a small size, so that the treatment effect is remarkably improved. However, the existing nano-carriers have single or double stimulus responsiveness, and are difficult to accurately match with complex tumor microenvironment, so that the controllability of drug release is insufficient. The high specific recognition capability of the CAR-T cell therapy to specific tumor antigens is of great interest, and the nano-particles modified by the CAR-T cell membrane can realize active targeting by means of chimeric antigen receptors on the surface of the cell membrane, avoid immune system clearance and prolong blood circulation time. Gelatin is used as a natural biological material, has good biocompatibility and enzyme responsiveness, can be degraded by matrix metalloproteinase (MMP-2/MMP-9) over-expressed by tumors to realize tumor microenvironment response, and Laponite is used as artificial synthetic lithium algae soil, has the advantages of high purity, uniform structure, high stability and the like compared with natural lithium algae soil, has outstanding electrochemical performance, and can enhance ionic conductivity and the like. Combining the two to construct a size-convertible carrier and combining the targeting advantage of the CAR-T cell membrane is expected to solve the bottleneck of the existing drug delivery system. Based on the above, the invention provides a CAR-T cell membrane modified nano drug delivery system, and a preparation method and application thereof. Disclosure of Invention In order to achieve the aim, the invention provides a CAR-T cell membrane modified nano drug delivery system, a preparation method and application thereof, and the invention utilizes strong electrostatic interaction of doxorubicin, laponite and gelatin to construct a novel nano delivery system with proper size, and solves the problems of low drug encapsulation rate, poor colloid stability nano particles and poor targeting of the existing lithium algae soil nano carrier by modifying nano particles through the CAR-T cell membrane. The high targeting of the tumor part is realized through the CAR-T cell membrane, and the rapid release of the medicine is realized by utilizing the tumor acidity and the microenvironment stimulation of the over-expression of the matrix metalloproteinase, and meanwhile, the small-size Laponite nano unit with the size of 20-30 nanometers is cracked to further penetrate into the deep tumor. In one aspect, the invention provides a CAR-T cell membrane modified nanomedicine delivery system comprising: A complex formed by self-assembly of nanoparticles and gelatin; Antitumor drugs entrapped in the complex network, and A CAR-T cell membrane coated on the surface of the complex network; Wherein the nano particles are at least one selected from artificially synthesized lithium algae soil and neodymium ion doped artificially synthesized lithium algae soil. The preparation method of the neodymium ion doped artificial synthetic lithium algae soil comprises the steps of dissolving magnesium chloride hexahydrate and neodymium nitrate hexahydrate with deionized water to obtain solution A, dissolving lithium chloride in the deionized water, stirring and dissolving the solution A, pouring the solution A into the solution A to form mixed solution A-B, dissolving tetraethoxysilane in the deionized water, adding dilute hydrochloric acid dropwise to adjust pH to 3-4, hydrolyzing to obtain solution C, slowly adding the solution C dropwise into the mixed solution A-B, stirring to form milky suspension, slowly adding sodium hydroxide solution to pH to 10.0-10.5, supplementing the deionized wa