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

Anti-stat3 bispecific antibody having cell-penetrating ability, and pharmaceutical composition containing same

A technology of transcription activator and bispecific antibody, applied in the field of anti-signal transduction and transcription activator 3 bispecific antibody with intracellular penetration ability and pharmaceutical composition containing it

Pending Publication Date: 2020-09-22
罗菲佰欧株式会社
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] However, ordinary antibodies can only target extracellular molecules, and many important targets for treating diseases as well as for diagnosing diseases reside inside cells

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-stat3 bispecific antibody having cell-penetrating ability, and pharmaceutical composition containing same
  • Anti-stat3 bispecific antibody having cell-penetrating ability, and pharmaceutical composition containing same
  • Anti-stat3 bispecific antibody having cell-penetrating ability, and pharmaceutical composition containing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Example 1. Discovery of Phosphorylated STAT3 Peptide-Specific Antibody Sequences and Screening of Candidates Using Phage Display (Phagy Display)

[0121] In order to prepare an antibody that specifically binds to phosphorylated STAT3 phosphorylated at the 705th tyrosine residue (pY705STAT3), the present inventors first tried to screen antibodies that bind to the phosphorylated STAT3 by using phage display (Phagy Display).

[0122] To this end, first, a human scFv library provided by the Candidate Antibody Discovery Support Team of the Osong Advanced Comprehensive Drug Development Support Center was used, and pY705 STAT3 peptide was used as an antigen for phage display. Specifically, the antibody-phage library is mixed with antigens immobilized on a solid surface to induce antibody-antigen binding (Binding), and unbound phages are removed (Washing). Then, after using alkaline / acidic pH solution or competitor peptide to elute the phage bound to the antigen, infect Escheri...

Embodiment 2

[0126] Example 2. Preparation of Phosphorylated STAT3-specific Recombinant Antibody

[0127] The present inventors prepared recombinant diabodies specific to phosphorylated STAT3 (pY705 STAT3) using the three antibody candidate sequences finally screened in Example 1 above.

[0128] Such as Figure 1a As shown, the diabody prepared in the present invention has the following structure. The antigen-binding site at one end binds to phosphorylated STAT3, and on the other hand, the antigen-binding site at the other end binds to DNA in the nucleus. More specifically, as Figure 1b As shown, the diabody prepared in the present invention is a scFv (STAT3 scFv) that binds to phosphorylated STAT3 and a scFv (3E10 scFv) that penetrates the cell and binds to DNA in the nucleus through a linker in the form of a swivel (linker) Linked structures. The above-mentioned 3E10 scFv uses the DNA-binding domain of an autoantibody found in lupus patients, and further uses the first part of the 'he...

Embodiment 3

[0131] Example 3. Verification of the cell penetration ability of the pSTAT3 antibody

[0132] In order to verify whether the pSTAT3 antibody of the present invention actually has the function of penetrating cells, cell penetrating ability analysis was carried out. To this end, pSTAT3 diabody containing 3E10 scFv (pSTAT3-C4) or pSTAT3 antibody containing 3E10 scFv but the 31st asparagine was replaced by asparagine (D31N) was treated with DLD colorectal cancer cell line (STAT3-D31N ), the intracellular STAT3 protein (STAT3 staining) was labeled by immunocytochemical staining using each of the above pSTAT3 antibodies, and at the same time, the nuclei were stained by propidium iodide (PI) staining (PI staining fornucleus) and compared.

[0133] Such as image 3 As shown, as a result, when the pSTAT3-D31N antibody was treated, the STAT3 staining result was observed to be exactly the same as the propidium iodide staining result for the cell nucleus. From this, it was confirmed tha...

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 present invention relates to an anti-STAT3 bispecific antibody having a cell-penetrating ability, and a pharmaceutical composition containing same and, more specifically, to an anti-STAT3 bispecific antibody comprising a site specifically binding to STAT3 and a DNA binding site enabling cell-penetration, and a pharmaceutical composition containing same as an active ingredient. It has been ascertained that a STAT3 / DNA bispecific antibody according to the present invention comprises a DNA binding site, which allows penetration into cells and specifically binds to intranuclear DNA, so as to be capable of overcoming limitations of a conventional antibody drug, which only targets proteins existing outside the cells, and binds specifically only to STAT3 in the active phosphorylated form so as to be capable of inhibiting the transcription factor activity thereof, thereby being effectively usable in the development of: therapeutic agents, having no side effects, for associated diseases caused by the activation of STAT3, for example, therapeutic agents for various carcinomas associated with the activation of STAT3; drugs capable of inhibiting cancer metastasis induced by the activationof STAT3; and therapeutic agents, having no side effects, for inflammatory diseases caused by the activation of STAT3, including rheumatoid arthritis.

Description

technical field [0001] The present invention relates to an anti-STAT3 (signal transducer and activator of transcription 3) bispecific antibody with intracellular penetration ability and a pharmaceutical composition comprising the same. An anti-STAT3 bispecific antibody with a DNA (deoxyribose nucleic acid) binding site capable of penetrating cells and a pharmaceutical composition comprising the same as an active ingredient. Background technique [0002] Proteins are made using the genetic information of deoxyribonucleic acid through the processes of transcription and translation. The produced protein is a structural component of cells and also performs functions such as cytoskeleton formation, but many proteins are active, and the regulation of protein activity plays a very important role in intracellular signal transduction for regulating intracellular functions. [0003] Regulation of protein-based intracellular signal transduction can be achieved by on / off of protein act...

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
Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P29/00A61P19/02C07K16/18C07K16/44G01N33/68A61K39/00
CPCA61P19/02A61P29/00C07K16/18C07K16/44C07K16/468A61K2039/507C07K2317/31C07K2317/77C07K2317/30C07K2317/76G01N33/6872C07K2317/565C07K2317/622C07K2317/92G01N33/68
Inventor 咸圣镐洪镇泰
Owner 罗菲佰欧株式会社
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