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A kind of exosome targeting lymphoma cells and its preparation method and application

A technology of lymphoma cells and exosomes, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulas, etc., can solve the problems of accurate detection and low drug targeting efficiency, and achieve High drug loading rate, prolonged circulation time in vivo, and good biocompatibility

Active Publication Date: 2021-12-10
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the Fc segment of the complete CD22mAb is easily phagocytized and cleared by mononuclear phagocytes, which leads to low drug targeting efficiency and accuracy.

Method used

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  • A kind of exosome targeting lymphoma cells and its preparation method and application
  • A kind of exosome targeting lymphoma cells and its preparation method and application
  • A kind of exosome targeting lymphoma cells and its preparation method and application

Examples

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Effect test

Embodiment 1

[0043] A CD22-F (ab') carrying doxorubicin 2 A method for preparing an exosome-targeted drug-carrying system of antibody fragments, the method comprising the following steps:

[0044] Step 1: Select 293T cells of the human kidney epithelial cell line as exosome-derived cells, and replace the culture medium of exosome-derived cells with serum-free medium 12-48 hours before extracting exosomes. After culturing for 24-48 hours, collect the cell suspension;

[0045] Step 2: Centrifuge the cell suspension (300g, 10min, 4°C), discard the cells and debris, take the supernatant and centrifuge again (2000g, 10min, 4°C), take the supernatant and centrifuge again (12000g, 30min, 4°C ), the supernatant after the third centrifugation was transferred to a 100kD ultrafiltration tube to balance, and concentrated by centrifugation (4000g, 15min, 4°C). Centrifuge the concentrate at an ultrahigh speed (100,000g, 90min, 4°C), discard the supernatant, resuspend and wash, then centrifuge at the s...

Embodiment 2

[0058] A CD22-F(ab') carrying cyclophosphamide 2 A method for preparing an exosome drug-carrying system of an antibody fragment, the method comprising the following steps:

[0059] Step 1: Select 293T cells of the human kidney epithelial cell line as exosome-derived cells, and replace the culture medium of exosome-derived cells with serum-free medium 12-48 hours before extracting exosomes. After culturing for 24-48 hours, collect the cell suspension;

[0060] Step 2: Centrifuge the cell suspension (300g, 10min, 4°C), discard the cells and debris, take the supernatant and centrifuge again (2000g, 10min, 4°C), take the supernatant and centrifuge again (12000g, 30min, 4°C ), the supernatant after the third centrifugation was transferred to a 100kD ultrafiltration tube to balance, and concentrated by centrifugation (4000g, 15min, 4°C). Centrifuge the concentrate at an ultrahigh speed (100,000g, 90min, 4°C), discard the supernatant, resuspend and wash, then centrifuge at the same...

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Abstract

The invention belongs to the field of biopharmaceutical carrier preparation and discloses a CD22-F(ab') linker 2 Exosomes of antibody fragments whose surface undergoes CD22-F(ab') 2 The antibody fragment modification can not only retain the CD22mAb binding ability, but also avoid the phagocytosis of phagocytic cells mediated by the Fc segment, thereby prolonging the in vivo circulation time of the drug-loaded particles. In addition, an exosome drug delivery system was innovatively constructed. The drug-loading system can effectively cross the blood-brain barrier, and has the advantages of good biocompatibility, precise and active targeting, and long circulation time. Anti-central nervous system lymphoma curative effect, reduce peripheral side effects, will establish a general platform for the effective delivery of various types of anti-central lymphoma chemotherapy drugs in vivo, and provide a powerful means for clinical targeted therapy of central nervous system lymphoma.

Description

technical field [0001] The invention belongs to the field of biopharmaceutical carrier preparation, and relates to an exosome targeting the central nervous system and a preparation method and application thereof. Background technique [0002] Primary central nervous system lymphoma (PCNSL) is a highly aggressive non-Hodgkin's lymphoma, and more than 90% of its subtypes are diffuse large B-cell lymphoma (DLBCL). Primary central nervous system lymphoma accounts for about 1%-5% of primary intracranial tumors and 1% of all lymphomas. Due to the existence of the blood-brain barrier, many drugs that can be applied to DLBCL are difficult to reach the intracranial lesions, the five-year survival rate of primary central nervous system lymphoma is only 20%-30%, and about 25% of PCNSL occurs primary Drug resistance, nearly 50% will eventually relapse. At present, the treatment requires super-large doses of chemotherapy drugs to ensure that the chemotherapy drugs entering the central ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/46A61K47/42A61K31/704A61K31/675A61P35/00
CPCA61K47/46A61K47/42A61K31/704A61K31/675A61P35/00
Inventor 许佩佩柳旭夏天陈兵欧阳建
Owner NANJING DRUM TOWER HOSPITAL
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