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

Anti-PD1 and TGF beta bifunctional antibody and preparation method thereof, and pharmaceutical composition containing same

A bifunctional antibody, PD-1 technology, applied in the direction of drug combinations, chemical instruments and methods, botany equipment and methods, etc., can solve the problems of mediocre experimental results and high clinical toxicity, and achieve good specificity and low in vivo toxicity , the effect of improving the clinical response rate

Active Publication Date: 2021-06-22
GUANGDONG FEIPENG PHARM CO LTD
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although TGFβ antibodies have been proven to be effective in tumor therapy, the preclinical results of antibody therapy targeting TGFβ receptor 2 alone are not effective, and the phase I clinical toxicity is very high

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
  • Anti-PD1 and TGF beta bifunctional antibody and preparation method thereof, and pharmaceutical composition containing same
  • Anti-PD1 and TGF beta bifunctional antibody and preparation method thereof, and pharmaceutical composition containing same
  • Anti-PD1 and TGF beta bifunctional antibody and preparation method thereof, and pharmaceutical composition containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1. Expression of fusion protein PD-1 / TGFβRII trap

[0077] Using the extracellular domain of the TGFβRII receptor (SEQ ID NO: 1) as the immunomodulatory molecular part of the fusion protein, the PD-1-op antibody targeting PD1 (secreted anti-PD-1κ light chain polypeptide sequence and heavy chain The polypeptide sequences are shown in SEQ ID NO:2 and SEQ ID NO:3), PD-1-76-C2 antibody, PD-1-97-C2 antibody, PD-1-112-C2 antibody (the constant regions are all Shown in SEQ ID NO:73 and 74) respectively as the targeting part of the fusion protein, with (Gly 4 Ser) 4 Gly is used as a linking sequence to connect the C-terminal of the sequence shown in SEQ ID NO:3 with the N-terminal of the sequence shown in SEQ ID NO:1 to form four fusion proteins. These four proteins have basically the same experimental performance. The following is the same as PD-1 -op antibody fusion formation is represented by Anti-PD1-TGFβRII extracellular region fusion protein, indicating its expe...

Embodiment 2

[0080] Example 2. Expression of fusion protein PD-1 / TGFβRII trap

[0081] The expression plasmid was constructed to transiently transfect human embryonic kidney HEK 293 cells, and the anti-PD1-TGFβRIITrap produced by the isolated and purified cells had a band molecular weight of about 170kD on SDS-PAGE under non-reducing conditions ( figure 1 In the B panel), the molecular weight of the band on SDS-PAGE under reducing conditions is about 80kD. There was a small peak after the 190kD peak on size exclusion chromatography, which was identified by mass spectrometry as the anti-PD1-TGFBR2 trap antibody moiety cleaved at a site in the N-terminal portion of TGFβRII.

Embodiment 3

[0082] Example 3. ELISA detection of binding activity of PD1-TGFβRII trap antibody end and trap end in vitro

[0083] The antigens used for the binding detection of antibody end and trap end are self-produced PD1-His (SEQ ID NO: 12), PDL1-His (SEQ ID NO: 13), human TGFβ1 (CA59), human TGFβ2 (CJ79), human TGFβ3 (CJ44) (TGFβ protein was purchased from Novoprotein), and the detection process was as follows:

[0084] Dilute PD1-His (SEQ ID NO:12), TGFβ1, TGFβ2, TGFβ3 to 0.5 μg / ml with 1× phosphate buffered saline (PBS), coat 96-well microtiter plate with 100 μl / well, overnight at 4°C;

[0085] Wash 3 times with 250 μl 1×PBST (PBS+0.5% Tween20), add 200 μl PBS containing 2% bovine serum albumin (BSA) to block for 1 hour at room temperature;

[0086] Wash 3 times with 250 μl 1×PBST, add serially diluted PD1-TGFβRII trap, PD1 antibody as positive control, incubate at room temperature for 2 hours;

[0087] Wash 3 times with 250 μl 1×PBST, add 100 μl diluted Goat-anti-human Fc-HRP co...

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

No PUM Login to View More

Abstract

The invention relates to the technical field of antibodies, in particular to an anti-PD1 and TGF beta bifunctional antibody, a preparation method thereof and a pharmaceutical composition containing the same. The bifunctional antibody comprises a first protein functional region targeting PD-1 and a second protein functional region targeting TGF beta, the first protein functional region is a PD-1 antibody, and the second protein functional region is an extracellular structural domain of TGF beta RII; and an antibody heavy chain C end of the first protein functional area is connected with the N end of the second protein functional area.

Description

technical field [0001] The present invention relates to the technical field of antibodies, in particular to a bifunctional antibody against PD1 and TGFβ, a preparation method thereof, and a pharmaceutical composition containing the same. Background technique [0002] Traditional cancer treatment options are mainly radiotherapy and chemotherapy. Chemotherapy and radiotherapy have a strong killing effect on normal tissues while killing cancer cells due to lack of targeting. Since then, targeted therapy has been developed on the basis of chemotherapy, and targeted therapy is mainly aimed at overexpressed proteins or proto-oncogenes that are important for tumor survival and growth. Although targeted therapy is effectively reduced, there are a series of disadvantages such as drug resistance and organ toxicity. [0003] Cancer immunotherapy is an emerging concept of cancer treatment, which reorganizes the host immune response, especially the adaptive T cell response, to provide i...

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): C07K16/44C12N15/13C12P21/02A61K39/395A61P35/00
CPCC07K16/2818C07K16/22A61P35/00C07K2317/31C07K2317/92C07K2317/76A61K2039/505
Inventor 路力生张轶博霍永庭潘志福欧颖烨芦迪涂晶晶
Owner GUANGDONG FEIPENG PHARM CO LTD
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