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TIGIT immunoadhesin and preparation method and use thereof

A technology of immunoadhesin and fragments, which is applied in the preparation of TIGIT-Ig immunoadhesin and the treatment of immune-related diseases to achieve the effect of preventing degradation

Inactive Publication Date: 2019-01-15
PHARCHOICE THERAPEUTICS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether soluble fragments of the TIGIT protein can play a therapeutic role in immune-related diseases is unknown and needs further study

Method used

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  • TIGIT immunoadhesin and preparation method and use thereof
  • TIGIT immunoadhesin and preparation method and use thereof
  • TIGIT immunoadhesin and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1, Construction and expression of TIGIT immunoadhesin

[0067] Such as figure 1 As shown, TIGIT immunoadhesin is a homodimer with antibody IgG Fc, which is composed of TIGIT protein analog and human IgG antibody Fc region. The specific construction and expression steps are as follows:

[0068] (1) Whole gene synthesis of human TIGIT protein extracellular segment (amino acid sequence and nucleotide sequence as shown in SEQ ID NO: 1-2), mouse TIGIT protein extracellular segment (amino acid sequence and nucleotide sequence as shown in SEQ ID NO : 7-8), to obtain human and mouse TIGIT protein analogues respectively. The Fc fragment of human immunoglobulin IgG (nucleotide sequence and amino acid sequence are shown in SEQ ID NO: 13-14) is synthesized from the whole gene.

[0069] (2) Using the extracellular segment of the TIGIT protein obtained in the above step (1) and the obtained Fc segment as templates, the above segments were fused and loaded into expression v...

Embodiment 2

[0073] Embodiment 2.Biacore analysis

[0074] Human TIGIT immunoadhesin and chimeric TIGIT immunoadhesin were respectively coated on CM5M5 chips (GE Company), and after capturing the detected antibodies, the affinity of each immunoadhesin was detected with Biacore T100 (GE Healthcare). The detection affinity values ​​are shown in Table 1.

[0075] Table 1. Biacore analysis results

[0076]

Embodiment 3

[0077] Example 3: Therapeutic effect of TIGIT immunoadhesin on lupus mouse model

[0078] As a representative immune system disease, lupus nephritis has an incidence of about 50 / 100,000, accounting for about 0.7‰ of the population in my country. More than 90% of lupus nephritis occurs in women, mainly young and middle-aged women. It is generally believed that people under the age of 30 have a high rate of kidney involvement, and about 70% of patients have different degrees of clinical manifestations of kidney damage, with varying degrees of proteinuria, Microscopic hematuria is common, often accompanied by cast urine and renal dysfunction, which seriously affects the normal life of patients.

[0079] The lupus mouse model is mostly produced by crossing NZB female mice with NZW male mice. The first-generation (NZB×NZW) F1 hybrid can produce typical lupus symptoms including lupus nephritis. It is currently recognized as an animal model for lupus nephritis research. one. The es...

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Abstract

The invention relates to the technical field of biomedical engineering, and provides a TIGIT immunoadhesin and a preparation method and use thereof. The TIGIT immunoadhesin is a homologous dimer withan IgGFc antibody, and the monomer structure general formula is Z1-Z2-Z3 or Z1-Z3, wherein Z1 is a TIGIT protein analogue, and derived from a full-length protein or fragment of natural TIGIT, Z2 is apeptide junction with 10 to 25 amino acids, at least 50% of which are glycine residues; Z3 is a Fc region of a human IgG antibody. The TIGIT immunoadhesin is simple in construction and expression process, can be constructed either as a human TIGIT immunoadhesin or as a chimeric TIGIT immunoadhesin. Experiments show that the human TIGIT immunoadhesin and the chimeric TIGIT immunoadhesin both have good therapeutic effects on immune-related diseases, can effectively treat immune-related diseases by used in combination with other anti-immune-related diseases drugs, and have broad clinical application prospects.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a novel TIGIT-Ig immunoadhesin and its preparation method and application, especially the application for treating immune-related diseases. Background technique [0002] There are many important membrane molecules on the surface of T cells, which play an important role in the activation, proliferation, differentiation and effector function of T cells. According to different functions, it can be mainly divided into the following categories: (1) TCR-CD3 complex, which enables T cells to recognize the antigen peptide-MHC molecular complex on the antigen-presenting cell and transmit activation signals to the cell; (2) ) CD4 molecule and CD8 molecule: respectively assist TCR of CD4+ and CD8+ T cells to recognize antigens and participate in the transduction of T cell activation signals; (3) co-stimulatory molecules: such as CD28, CTLA-4, ICOS and PD-1, etc., for T cells...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62A61K38/17A61P37/02
CPCA61K38/00A61P37/02C07K14/47C07K2319/30
Inventor 胡适傅文燕丁敏
Owner PHARCHOICE THERAPEUTICS INC
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