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Anti-tigit antibodies and uses thereof

An antibody and antigen technology, applied in the direction of antibodies, antibody medical components, anti-tumor drugs, etc., can solve the problems of insufficient research and no development of antibodies.

Pending Publication Date: 2020-12-29
YUHAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] So far, several anti-TIGIT antibodies have been reported (such as US 9,713,641B, US 2016 / 0176963A, US 9,499,596B), but their specific mechanisms have been understudied, and none have been developed with efficacy that can actually be used as a therapeutic agent Antibodies

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  • Anti-tigit antibodies and uses thereof
  • Anti-tigit antibodies and uses thereof
  • Anti-tigit antibodies and uses thereof

Examples

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Effect test

Embodiment 1

[0133] Example 1: Screening of anti-TIGIT antibodies

[0134] 1.1 Screening of anti-TIGIT human antibody scFv and Fab clones

[0135] Antibodies that specifically bind to TIGIT were selected by a phage display screening method using scFv and Fab human antibody libraries. A scFv library was generated referring to the description in "Construction of a Large Synthetic Human scFv Library with SixDiversified CDRs and High Functional Diversity (Yang HY et al. Molecules and Cells 27, 225-235)", and a Fab library was generated referring to the description in Korean Patent No. 1694832 . Phage display screens were performed up to a total of the fourth round, and as the number of rounds increased, the number of antigens decreased and the number of washes increased. The first and third phage display screens used the human TIGIT-ECD-Fc antigen and the second and fourth phage display screens used the mouse TIGIT-ECD-Fc antigen for an antigen crossing approach. 20 μg of TIGIT antigen dilu...

Embodiment 2

[0142] Example 2. Optimization of anti-TIGIT antibodies

[0143] 2.1 Optimization of F04 and S64 antibodies

[0144] Through the above antibody screening process, two types of clones F04 and S64 were finally screened as anti-TIGIT human antibodies. To improve the stability of these antibodies, sub-libraries were constructed based on the amino acid sequences of the F04 and S64 antibodies, and screened at high temperature using prolonged washing methods to improve stability. The sub-library of the F04 antibody was a library obtained by simultaneously shuffling CDRH1 and CDRH2, which was prepared by overlapping one species. Three types of sub-libraries were generated by the overlap PCR method, including the library obtained by simultaneously shuffling CDRH1 and CDRH2, the library obtained by simultaneously shuffling CDRL1, CDRL2, and CDRL3, and the library obtained by shuffling all but Libraries obtained by shuffling CDRs other than CDRH3. CDR regions with amino acid sequence ...

Embodiment 3

[0166] Example 3. Assays to identify binding of anti-TIGIT antibodies to TIGIT on the cell surface

[0167] In order to identify the binding ability of the three types of anti-TIGIT antibodies produced in Example 2 to TIGIT expressed on the cell surface, a CHO cell line overexpressing TIGIT (hereinafter referred to as CHO-TIGIT cell line) was treated with an anti-TIGIT antibody , and then detect the anti-TIGIT antibody bound to TIGIT on the cell surface using fluorescence flow cytometry.

[0168] Specifically, the CHO-TIGIT cell line was cultured at 5% using a chemical composition medium (CD FortiCHO chemically defined medium + 8 mM L-glutamine + 20 μg / mL blasticidin + 1% anticoagulant) CO 2 Incubate in an incubator at 37°C for 48 to 72 hours. The cultured CHO-TIGIT cell line was harvested by centrifugation, diluted in FACS solution (PBS+5% FBS), and diluted at 1×10 5 The density of cells / well was assigned in 96-well round bottom plates (Corning). Then, in order to complet...

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Abstract

Disclosed are a novel antibody specifically binding to the tumor-immunosuppressant, TIGIT (T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif [ITIM] domain) or an antigen-bindingfragment thereof, a nucleic acid encoding the antibody or the antigen-binding fragment thereof, a vector and a host cell including the nucleic acid, a method for producing the antibody or the antigen-binding fragment thereof, a pharmaceutical composition containing the antibody or the antigen-binding fragment thereof as an active ingredient, and uses of the pharmaceutical composition. The antibody specifically binding to TIGIT or the antigen-binding fragment thereof and the pharmaceutical composition containing the same as an active ingredient are preferably used for the treatment of cancer or tumors.

Description

technical field [0001] The present invention relates to novel antibodies or antigen-binding fragments thereof, encoding said antibodies or Nucleic acids of antigen-binding fragments thereof, vectors and host cells comprising the nucleic acids, methods for producing the antibodies or antigen-binding fragments thereof, pharmaceutical compositions containing the antibodies or antigen-binding fragments thereof as active ingredients, and the The purposes of above-mentioned pharmaceutical composition. [0002] The antibody or antigen-binding fragment thereof that specifically binds TIGIT and the pharmaceutical composition containing the antibody or antigen-binding fragment thereof as an active ingredient are preferably used for treating cancer or tumor, but the present invention is not limited thereto. Background technique [0003] The function of the human immune system is to protect the body by attacking pathogens or viruses (antigens) that enter from the outside, as well as ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28A61K47/68A61K39/00
CPCC07K16/2803C07K2317/622C07K2317/55C07K2317/21C07K2317/33C07K2317/565C07K2317/94C07K2317/92C07K2317/73C07K2317/76A61K2039/505A61K2039/545A61K2039/507C07K2317/52C07K16/2827A61P35/00Y02A50/30A61K47/6803
Inventor 李光薰李准珩李娜来郑恩晶朴永奉张乐豪李垠政金基鸿崔诚玹崔秉贤朴珠英宋武泳李种瑞金奎兑高凤国
Owner YUHAN
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