Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Antibody targeting PSMA protein, chimeric antigen receptor, drug and application

A chimeric antigen receptor and targeting technology, which can be used in anti-tumor drugs, drug combinations, antibody medical components, etc. The effect of enhancing curative effect and good killing effect

Pending Publication Date: 2022-02-22
中国人民解放军海军军医大学第一附属医院
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Prostate-specific membrane antigen (PSMA) is highly expressed in prostate cancer and is an ideal target for immunotherapy of solid tumors. CAR-T cells targeting PSMA have great specificity and target potential in tumor immunotherapy. Junghans R et al. (2016) and Zuccolotto G (2014) proved the effectiveness and safety of CAR-T cells in preclinical research and clinical trials, but their The application is still limited, so further research and exploration are needed

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antibody targeting PSMA protein, chimeric antigen receptor, drug and application
  • Antibody targeting PSMA protein, chimeric antigen receptor, drug and application
  • Antibody targeting PSMA protein, chimeric antigen receptor, drug and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, antibody screening

[0055] 1. Phage library screening

[0056] The fully human antibody library was panned using 293T-PSMA cells. First, use 293T cells to perform negative panning on the established human tumor-specific antibody library for prostate cancer. The specific steps are as follows:

[0057] Incubate at 4°C for 2 hours, and the input amount of phage is 10 12 Phage, after centrifugation, take the supernatant, then add the supernatant to the 293T-PSMA overexpression cell line, and incubate at 4°C for 2 hours. After centrifugation, use PBS to wash away the phages not bound to the cell surface, then use 0.1M HCl-Glycine to elute the phages bound to the cells, then use Tris-HCl to neutralize the eluate, and take a part of the phage infection logarithm Escherichia coli TG1 in the growth phase was used to prepare phage for the next round of panning. Gradually increase the screening intensity of each round, and stop panning when the enrichment degree...

Embodiment 2

[0096] Embodiment 2, construction contains the plasmid vector expressing chimeric antigen receptor

[0097] An expression cassette expressing a chimeric antigen receptor targeting prostate-specific membrane antigen (such as Image 6 Shown): PSMA-CAR includes: CD8α signal peptide, PSMA single-chain antibody heavy chain variable region, Linker, PSMA single-chain antibody light chain, CD8 hinge region, CD8α transmembrane domain, intracellular co-stimulation of 4-1BB The element and the intracellular domain of CD3ζ, the above sequences are connected sequentially, and the Kozac sequence and the corresponding enzyme cutting site are introduced at the front end. Using XbaI and SalI double digestion, the expression cassette was transferred to the lentiviral packaging shuttle plasmid (Shanghai Bangyao Biotechnology Co., Ltd.), and two chimeric antigen receptor expression vectors pELPS-PSMA-BBz-HTS0101 and pELPS- PSMA-BBz-HTS0103. Wherein, the sequence of each element of the expressio...

Embodiment 3

[0120] Embodiment 3, construct the virus that expresses chimeric antigen receptor

[0121] Use Escherichia coli to amplify the above pELPS-PSMA-BBz-HTS0101 and pELPS-PSMA-BBz-HTS0103 as the two target plasmids and lentiviral packaging helper plasmids pMD2.G and psPAX2, extract the plasmids and perform agarose gel electrophoresis and sequencing Verify the correctness of the plasmid. Select the 293T cells in good condition and the first generation as the lentivirus packaging cells, and use the transfection reagent PEI to transfect the 293T cells with the above three plasmids. The transfection is completed in a 10cm culture dish with a total system of 10mL, and the cell transfection mixture in each dish should be prepared into a 1mL system with serum-free DMEM, so that the psPAX2 plasmid: pMD2.G plasmid: pELPS-PSMA-BBz-HTS0101 or pELPS-PSMA -BBz-HTS0103 plasmid: PEI=5μg: 3μg: 5μg: 50μl, mix the transfection mixture at room temperature, let it stand for 20min, then slowly add it ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Titeraaaaaaaaaa
Titeraaaaaaaaaa
Login to View More

Abstract

The invention discloses a PSMA-targeting antibody, a chimeric antigen receptor, a drug and application of the PSMA-targeting antibody and the chimeric antigen receptor. A heavy chain variable region of the antibody comprises an amino acid sequence as shown in SEQ ID NO.1 or 5; and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO.3 or 7. According to the invention, human PSMA protein is taken as an antigen, a phage library display technology is combined, and two new scFv antibodies are obtained through panning. Experiments prove that the two antibodies both have the capability of combining the PSMA protein, and the CAR-T prepared from the antibodies has a specific killing effect on prostate cancer target cells with positive PSMA protein. CAR-T cells of targeted PSMA protein prepared by using the two scFv antibodies have a good killing effect on human prostate cancer cells, and if methods such as androgen deprivation therapy are combined for synergistic effect, the curative effect on castration-resistant prostate cancer can be greatly enhanced.

Description

technical field [0001] The invention relates to the technical field of cellular immunotherapy, in particular to an antibody targeting PSMA protein, a chimeric antigen receptor, a drug and its application. Background technique [0002] Prostate cancer is one of the most common tumors of the male genitourinary system. The global incidence rate accounts for the second male malignant tumor, and the global mortality rate accounts for the fifth malignant tumor. It is the cancer with the highest incidence rate in the United States and the second leading cause of death. cancer. Data show that in the past 10 years, my country's prostate cancer has shown a high incidence trend, and large cities have become "hardest hit areas". Based on the characteristics of an aging population base and age structure, along with the increase in the incidence of prostate cancer, the incidence of prostate cancer in China The rate of increase is worrying. [0003] Traditional prostate cancer treatments ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/30C12N15/13C07K19/00C12N15/62C12N15/867C12N5/10A61K39/00A61P35/00A61P13/08
CPCC07K16/3069C07K14/7051C07K14/70517C07K14/70578C12N15/86C12N5/0636A61K39/001195A61P35/00A61P13/08C07K2317/622C07K2319/02C07K2319/03C07K2319/33C12N2740/15043C12N2800/107C12N2510/00A61K2039/5156A61K2039/884
Inventor 任善成张娜杜冰刘明耀肖雨田
Owner 中国人民解放军海军军医大学第一附属医院
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com