Checkpoint regulator antagonists

An antibody and antigen technology, applied in the field of bispecific and trispecific checkpoint regulator antagonists, checkpoint regulator antagonists, can solve the problem that the host cannot eliminate cancer cells, etc.

Pending Publication Date: 2019-07-09
GENSUN BIOPHARMA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Host inability to eliminate cancer cells remains a major concern

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0347] Example 1: Production of Monoclonal Antibodies

[0348] Monoclonal antibodies (mAbs) of the present application are generated and screened using techniques well known in the art, see eg Harlow and Lane (1988) Antibodies, A Laboratory Manual, (Antibodies, A Laboratory Manual) Cold Spring Harbor Publishing, New York. Antigen-specific hybridoma monoclonal antibodies are cloned, sequenced and engineered using techniques well known in the art, see e.g. Lo.B.K.C Methods in Molecular Biology TM , Volume 248 2004 Antibody Engineering (Methods in Molecular Biology TM . Volume 248 2004. Antibody Engineering).

Embodiment 2

[0349] Example 2: TIGIT Binder Screening Assay

[0350] Coat 1 μg / ml TIGIT-His protein overnight at 4°C and block with 1% BSA in PBS for 1 hour at room temperature, then incubate with 50 μl hybridoma supernatant for 1 hour at room temperature, pass through anti-mouse IgG HRP Detect mouse IgG for 30 minutes, then add TMB substrate and pass through the addition of 2N H 2 SO 4 Stop the reaction. Wells with an OD of at least 5 times background were selected as positive binders.

Embodiment 3

[0351] Example 3: TIGIT Blocker Screening Assay

[0352] 3 x 10 at 4°C 4TIGIT+CHO-K1 cells were incubated in 50 μl hybridoma supernatant for 20 minutes, and then human PVR human Fc tag fusion protein was added to a final concentration of 0.6 μg / ml; after incubation at 4°C for 30 minutes, the cells were incubated with FACS buffer ( 0.5% BSA, 2mM EMTA in PBS) to wash the cells, and then incubate the cells with 1 μg / ml PE-labeled anti-human Fc antibody at 4°C for 20 minutes. Cells were then washed with FACS buffer, then resuspended in 7-amino-actinomycin D (7AAD) solution, and analyzed with the iQue intellicyt system. Wells that completely blocked TIGIT and PVR (Fc tag) binding were selected as blocking agents. In the first screen, 18 blockers were identified from 65 binders. In the second screen, 17 blockers were identified from 30 binders. figure 1 Exemplary results of competition assays of various mouse anti-TIGIT monoclonal antibodies are shown.

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Abstract

Checkpoint regulator antagonists that bind specifically to TIGIT, PD-1 and / or PD-L1 are disclosed. Also disclosed are methods of making and using the checkpoint regulator inhibitors, including monospecific, bispecific and trispecific checkpoint regulator antagonists thereof.

Description

[0001] This application claims priority to US Patent Application Serial No. 15 / 858,963, filed December 29, 2017. The entire content of the above application is incorporated herein by reference. technical field [0002] This application relates generally to cancer therapy, and in particular, to checkpoint regulator antagonists, including anti-T cell Ig and ITIM domain (TIGIT) inhibitors, PD-1 inhibitors, PD-L1 inhibitors, including their bispecific and trispecific checkpoint regulator antagonists. Background technique [0003] The inability of the host to eliminate cancer cells remains a major problem. Although a growing number of therapeutic monoclonal antibodies have been approved for the treatment of various cancers, the emergence of resistance to these antibodies is often observed given the many different molecular pathways by which cancers grow and progress to metastasis. Although the immune system is the primary mechanism for cancer prevention, cancer cells can resist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N15/13C12P21/02A61K39/395A61P35/00
CPCC07K16/18A61P35/00C07K2317/76C07K2317/92A61K2039/505A61K39/0011C07K16/2803C07K16/2818A61K2039/507C07K2317/31C07K2317/33C07K2317/74A61K38/1774A61K39/39558
Inventor 盛泽琪刘波盛亦清
Owner GENSUN BIOPHARMA INC
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