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CN-121971662-A - Drug-loaded vesicle for treating thyroid cancer and preparation method and application thereof

CN121971662ACN 121971662 ACN121971662 ACN 121971662ACN-121971662-A

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a medicine-carrying vesicle for treating thyroid cancer, and a preparation method and application thereof. The drug-loaded vesicle comprises a vesicle, wherein radioactive iodine is loaded in a cavity of the vesicle, and the membrane surface of the vesicle is modified with a) a first single-chain antibody fragment capable of specifically binding to thyroglobulin, b) a second single-chain antibody fragment capable of specifically binding to thyroid stimulating hormone receptor, and c) CD47 protein. After intravenous injection, the drug-loaded vesicle can be enriched in the thyroid cancer focus area of RAIR-DTC patients, and can continuously release radioactive iodine while inhibiting cancer cell proliferation, thereby playing a long-term treatment role.

Inventors

  • MING JIE
  • Li Jiexiao
  • ZHANG JIANING
  • TANG KE
  • TANG ZIMEI

Assignees

  • 华中科技大学同济医学院附属协和医院

Dates

Publication Date
20260505
Application Date
20260403

Claims (10)

  1. 1. A drug-loaded vesicle, comprising: a vesicle loaded with radioactive iodine in its cavity, the membrane surface of said vesicle being modified with: a) A first single chain antibody fragment capable of specifically binding thyroglobulin; b) A second single chain antibody fragment capable of specifically binding to thyroid stimulating hormone receptor; c) CD47 protein; The amino acid sequence of the first single-chain antibody fragment is shown as SEQ ID NO. 1, and the amino acid sequence of the second single-chain antibody fragment is shown as SEQ ID NO. 2.
  2. 2. The drug-loaded vesicle according to claim 1, wherein the membrane surface of the vesicle is modified with a bispecific chimeric antigen receptor comprising the first single chain antibody fragment and a second single chain antibody fragment, wherein the first single chain antibody fragment and the second single chain antibody fragment are linked by a flexible linker peptide.
  3. 3. The drug-loaded vesicle of claim 2, wherein the flexible connecting peptide is (Gly 4 Ser） n , wherein n is not less than 3.
  4. 4. The drug-loaded vesicle of claim 2, wherein the bispecific chimeric antigen receptor has a molecular weight of 60-70 kDa.
  5. 5. The drug-loaded vesicle of claim 2, wherein the amino acid sequence of the bispecific chimeric antigen receptor is shown in SEQ ID NO. 3.
  6. 6. The drug-loaded vesicle of claim 2, wherein the particle size of the vesicle is 100-300 nm.
  7. 7. A method for preparing a drug-loaded vesicle, which is characterized by comprising the following steps: Constructing nucleotide sequences for encoding a first single-chain antibody fragment, a second single-chain antibody fragment and CD47 protein, and introducing the nucleotide sequences into a carrier plasmid, wherein the amino acid sequence of the first single-chain antibody fragment is shown as SEQ ID NO. 1, and the amino acid sequence of the second single-chain antibody fragment is shown as SEQ ID NO. 2; packaging the vector plasmid into a lentivirus, and transferring into a host cell, and screening to obtain an engineering cell co-expressing the first single-chain antibody fragment, the second single-chain antibody fragment and the CD47 protein; Inducing the engineered cell to secrete vesicles; radioiodine is encapsulated into the chamber of the vesicle using electroporation.
  8. 8. The method of claim 7, wherein the engineered cell secretion vesicles are induced by starvation and UV irradiation.
  9. 9. The method of claim 7, wherein the electroporation parameters are 400-600V and 400-600 μF.
  10. 10. Use of a drug-loaded vesicle according to any one of claims 1 to 6 for the preparation of a medicament for the treatment of thyroid cancer.

Description

Drug-loaded vesicle for treating thyroid cancer and preparation method and application thereof Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to a medicine-carrying vesicle for treating thyroid cancer, and a preparation method and application thereof. Background Radioiodine (RAI) treatment is the standard therapy for Differentiated Thyroid Cancer (DTC), and its core mechanism relies on sodium-iodine symporter (NIS) to actively transport radioiodine into thyroid cells, achieving tumor-targeted killing. However, about 15% -20% of DTC patients develop radioiodinated differentiated thyroid cancer (RAIR-DTC) after long-term treatment, which is mainly characterized by a loss of NIS expression or functional inactivation. In such thyroid cancer patients, radioiodine cannot actively transport into thyroid cells, and traditional radioiodine therapy fails completely, resulting in continued tumor progression. Disclosure of Invention In order to solve the problem that the traditional radioiodine treatment fails in RAIR-DTC patients, the invention provides a drug-loaded vesicle for treating thyroid cancer, and a preparation method and application thereof, and the drug-loaded vesicle can be enriched in thyroid cancer focal areas of RAIR-DTC patients after intravenous injection, and can continuously release radioiodine while inhibiting proliferation of cancer cells, thereby playing a long-term treatment role. The technical scheme provided by the invention is as follows: In a first aspect, the present invention provides a drug-loaded vesicle comprising: a vesicle loaded with radioactive iodine in its cavity, the membrane surface of said vesicle being modified with: a) A first single chain antibody fragment capable of specifically binding thyroglobulin; b) A second single chain antibody fragment capable of specifically binding to thyroid stimulating hormone receptor; c) CD47 protein; The amino acid sequence of the first single-chain antibody fragment is shown as SEQ ID NO. 1, and the amino acid sequence of the second single-chain antibody fragment is shown as SEQ ID NO. 2. In combination with the first aspect of the invention, in some embodiments, the membrane surface of the vesicle is modified with a bispecific chimeric antigen receptor comprising the first single chain antibody fragment and a second single chain antibody fragment, the first single chain antibody fragment and the second single chain antibody fragment being linked by a flexible linker peptide. In some embodiments, in combination with the first aspect of the invention, the flexible connecting peptide is (Gly 4Ser）n, wherein n≥3). In combination with the first aspect of the present invention, in some embodiments, the bispecific chimeric antigen receptor has a molecular weight of 60 to 70 kDa. In combination with the first aspect of the invention, in some embodiments the amino acid sequence of the bispecific chimeric antigen receptor is as shown in SEQ ID NO. 3. In combination with the first aspect of the invention, in some embodiments, the particle size of the vesicle is 100-300 nm. In a second aspect, the present invention provides a method for preparing a drug-loaded vesicle, comprising the steps of: Constructing nucleotide sequences for encoding a first single-chain antibody fragment, a second single-chain antibody fragment and CD47 protein, and introducing the nucleotide sequences into a carrier plasmid, wherein the amino acid sequence of the first single-chain antibody fragment is shown as SEQ ID NO. 1, and the amino acid sequence of the second single-chain antibody fragment is shown as SEQ ID NO. 2; packaging the vector plasmid into a lentivirus, and transferring into a host cell, and screening to obtain an engineering cell co-expressing the first single-chain antibody fragment, the second single-chain antibody fragment and the CD47 protein; Inducing the engineered cell to secrete vesicles; radioiodine is encapsulated into the chamber of the vesicle using electroporation. In combination with the second aspect of the invention, in some embodiments, the engineered cells are induced to secrete vesicles by starvation and UV irradiation. In some embodiments, in combination with the second aspect of the present invention, the electroporation parameter is 400-600V and the capacitance is 400-600 muF. In a third aspect, the invention provides the use of a drug-loaded vesicle as described above in the manufacture of a medicament for thyroid cancer. Compared with the prior art, the invention has at least the following beneficial effects: The invention provides a medicine carrying vesicle independent of NIS, which realizes synchronous activation of thyroglobulin (Tg) and Thyroid Stimulating Hormone Receptor (TSHR) double target recognition and macrophage 'do not eat me' signals by biomimetically modifying vesicle surface ligand, and remarkably enhances enrichment capacity and retention time of the medic