Polypeptide and application thereof in preparation of immunomodulatory drugs

A drug and composition technology, applied in the field of polypeptide and its application in the preparation of immunomodulatory drugs, can solve the problems of scarcity of polypeptide immunomodulatory drugs and low safety of immunomodulatory drugs, and achieve small molecular weight, easy preparation, and stable good sex effect

Active Publication Date: 2021-10-15
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to provide a novel polypeptide PEP20 encoded by the host gene of long-chain non-coding RNA LINC01871, which can specifically target CD8 in an inflammatory environment + T cells, suppress CD8 + The differentiation of T cells into Tc17 cells can be applied to the preparation of immunomodulatory drugs to solve the problems of low safety of existing immunomodulatory drugs and scarcity of polypeptide immunomodulatory drugs

Method used

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  • Polypeptide and application thereof in preparation of immunomodulatory drugs
  • Polypeptide and application thereof in preparation of immunomodulatory drugs
  • Polypeptide and application thereof in preparation of immunomodulatory drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1, PEP20 sequence analysis and in vitro synthesis

[0088] 1. PEP20 sequence analysis

[0089] The sequence of the host gene of LINC 01871 is as follows (SEQ ID NO: 1):

[0090]

[0091]

[0092]

[0093] In the above sequence, the underlined part in bold is the nucleotide sequence of PEP20. The sequence translated into amino acids is: MVEEIQASLMWQQAREREGE (SEQ ID NO: 2).

[0094] 2. In vitro synthesis of PEP20

[0095] The conventional solid-phase peptide synthesis method is used to synthesize the peptide according to the amino acid sequence of SEQ ID NO: 2, and the correctness and purity of the obtained peptide are analyzed by mass spectrometry and HPLC detection. The result is as figure 1 As shown, the mass spectrometry analysis confirmed that the amino acid was correct, and its molecular weight was 2420.69 ( figure 1 A) of A); The purity of the obtained polypeptide detected by HPLC is 98.65% ( figure 1 B). Dissolve in HBSS before use.

Embodiment 2

[0096] Example 2, PEP20 inhibits Tc17 cell differentiation

[0097] The PEP20 obtained by the solid-phase polypeptide synthesis method in Example 1 was tested for its effect on the differentiation of Tc17 cells.

[0098] Obtain mouse spleen cells, and use immunomagnetic beads to isolate mouse naive CD8 + T cells were cultured in RPMI-1640 medium at 37°C, and anti-CD3 (5 μg / ml), anti-CD28 (2 μg / ml), TFG-β (10 ng / ml), IL -23 (20 ng / ml), anti-IL-4 (5 μg / ml) and anti-IFN-γ (10 μg / ml) were cultured for 3 days to obtain Tc17 cells.

[0099] Tc17 cells were cultured in RPMI-1640 medium at 37°C and divided into three culture groups: Untreated group, Scramble group and PEP20 group, in which PEP20 group was added with 10 μM concentration of PEP20, and Scramble group was added with 10 μM concentration of Scramble Peptides (randomly scrambled sequences with the same 20 amino acids as PEP20), the Untreated group did not receive any treatment. The effect of PEP20 on Tc17 differentiation ...

Embodiment 3

[0101] Example 3, PEP20 and PEP20-PEG can effectively alleviate the signs of psoriasis in mice induced by IMQ

[0102] SPF level mice were randomly divided into 6 groups, including Vehicle group (i.e. solvent group), 3 positive control groups, PEP20 group and PEP20-PEG group, and IMQ was used to smear the inside and outside of mouse ear skin, 30mg / mouse, total After smearing for 7 days, an IMQ-induced psoriasis model in mice was established, and solvent, ainti-IL17, ainti-IL23p19, ainti-TNFα, PEP20 and PEP20-PEG were respectively performed on the second day after IMQ administration (a small amount of dissolution first In HBSS, then dilute with normal saline to use the concentration of 80μg / 200μl) tail vein administration, the dose is 80μg / 20g mice, once a day, wherein antibody drugs ainti-IL17, ainti-IL23p19, ainti-TNFα are used as positive control drugs . Ear skin thickness was measured regularly every day from the day of induction. The mice were treated on the 7th day of I...

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Abstract

The invention provides a polypeptide and application thereof in preparation of an immunomodulatory drug. The functional polypeptide is a novel polypeptide PEP20 which is coded by a host gene of a long-chain non-coding RNA LINC01871; the polypeptide can specifically target CD8 < + > T cells in an inflammatory environment, inhibit differentiation of the CD8 < + > T cells to Tc17 cells, and inhibit over-differentiation and over-proliferation of the Tc17 cells and over-expression of IL-17A. The invention has an immune regulation function and can be used for preventing or treating autoimmune diseases, such as psoriasis, rheumatoid arthritis and multiple sclerosis. The PEP20 disclosed by the invention is small in molecular weight and can easily enter cells to play a role; the invention can be synthesized by a chemical method, is easy to prepare in a large scale, has good stability, and is lower in price and safer compared with the existing antibody for treating autoimmune inflammatory diseases such as psoriasis.

Description

technical field [0001] The present invention relates to the field of biomedicine, more specifically, the present invention relates to a polypeptide and its application in the preparation of immunomodulatory drugs. Background technique [0002] Since its discovery in 1993, the pro-inflammatory cytokine IL-17A has been the focus of research on autoimmune inflammatory diseases. At first it was found that it was caused by CD4 + A specific subset of T cells (so-called Th17 cells) is produced, however, other immune cell subsets are also known to synthesize and express IL-17, including CD8 + T cells. CD8 expressing IL-17 + The presence of T cells (also known as Tc17 cells) has been described in several human inflammatory diseases. [0003] An earlier study reported that CD8 + Presence of IL-17 mRNA in T cell clones. Later assays using flow cytometry revealed that skin plaques from patients with psoriasis contained increased amounts of IL-17's CD8 + T cells, which were not ob...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/12A61K38/10A61P37/02A61P17/06A61P17/00A61P11/06A61P1/00A61P19/02A61P29/00A61P25/00
CPCC07K7/08A61P37/02A61P17/06A61P17/00A61P11/06A61P1/00A61P19/02A61P29/00A61P25/00A61K38/00
Inventor 王宏林孙洋楼方舟孙力波
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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