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Application of tdgf-1 truncated small molecule polypeptide in anti-hepatic fibrosis

A TDGF-1, anti-hepatic fibrosis technology, applied in the field of biomedicine, can solve problems such as functional limitation

Active Publication Date: 2020-06-02
JILIN UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, as a carcinoembryonic antigen, the function of the CFC domain plays a major role, while the amino acid sequence of the EGF domain has a relatively clear mutation sequence relative to the EGF molecule, causing its functional limitation
[0006] In the prior art, there is no report on the correlation between TDGF-1 protein and its protein domain and liver fibrosis

Method used

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  • Application of tdgf-1 truncated small molecule polypeptide in anti-hepatic fibrosis
  • Application of tdgf-1 truncated small molecule polypeptide in anti-hepatic fibrosis
  • Application of tdgf-1 truncated small molecule polypeptide in anti-hepatic fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 TDGF-1 protein and its truncated polypeptide synthesis

[0064]The full-length protein of human TDGF-1 (sequence shown in SEQ ID NO.3) was obtained by prokaryotic expression in Escherichia coli, and the TDGF-1 truncated polypeptide was synthesized in cooperation with Jikai Gene (Shanghai): EGF- Like domain short peptide (sequence shown in SEQ ID NO.1) and CFC domain short peptide (sequence shown in SEQ ID NO.2).

[0065] SEQ ID NO. 1: PPMGIQHSKELNRTCCLNGGTCML;

[0066] SEQ ID NO. 2: PSFYGRNCEHDVRKENCGSVPHDTWLPKKCSLCK.

[0067] SEQ ID NO.3:

[0068] MDCRKMARFSYSVIWIMAISKVFELGLVAGLGHQEFARPSRGYLAFRDDSIWPQEEPAIRPRSSQRVPPMGIQHSKELNRTCCLNGGTCMLGSFCACPPSFYGRNCEHDVRKENCGSVPHDTWLPKKCSLCKCWHGQLRCFPQAFLPGCDGLVMDEHLVASRTPELPPSARTTTFMLVGICLSIQSYY

[0069] Protein and peptide electrophoresis figure 1 shown.

Embodiment 2

[0070] Example 2 In vitro cell experiments to study the function of TDGF-1 protein and its truncated polypeptide

[0071] 1. Immunofluorescence experiment

[0072] Method: Divide 1×10 5 LX2 (human hepatic stellate cell line) cells were planted in a 6-well plate at 37°C, 5% CO 2 Cultivate in a sterile environment for 24 hours until completely adhered to the wall, remove the culture supernatant, replace with DMEM medium containing 3% fetal bovine serum to continue the culture, add PBS and 50ng Cripto-1 whole protein to different cell supernatants (R&D Company), 50ng EGF-like domain short peptide, 50ng CFC domain short peptide, incubated for 48 hours. Remove the supernatant, wash the cells three times with PBS, add pre-cooled 4% paraformaldehyde to each well to fix for 5 minutes, wash with PBS three times, add PBS containing 0.5% TritonX-100, 1ml per well, incubate at room temperature for 10 minutes; Aspirate 0.5% TritonX-100 PBS, add 1ml 5% BSA (diluted in PBS) to each well, ...

Embodiment 3

[0085] Example 3 In vivo experiments to study the function of TDGF-1 protein and its truncated polypeptide

[0086] 1. Construction of mouse model of liver fibrosis

[0087] Healthy C57 male mice, weighing between 18-22g, were adaptively fed for 3-5 days. Create a mouse model of liver fibrosis by subcutaneous injection of carbon tetrachloride or intraperitoneal injection (15 μl carbon tetrachloride / 85 μl olive oil), inject twice a week, the total volume of each injection is 100 μl, and sacrifice after continuous injection for 8 weeks From mice, the liver and peripheral blood were collected for the analysis of liver fibrosis evaluation indicators (source of model establishment scheme: J Hepatol.2017Oct; 67(4):770-779).

[0088] 2. Detection

[0089] From the 4th week of liver fibrosis mouse modeling, different time points (4-10 weeks) were used to intervene with small molecules of TDGF-1 truncated polypeptide to mice with liver fibrosis, intraperitoneally injected (10ng / g bod...

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Abstract

The invention discloses an application of TDGF-1 (teratocarcinoma-derived growth factor-1) truncated micro-molecule polypeptide in anti-hepatic fibrosis. An experiment proves that the TDGF-1 truncatedmicro-molecule polypeptide has the capabilities of blocking a TGF(transforming growth factor)-[beta] signal pathway and lowering [alpha]-SMA (smooth muscle action) expression in a hepatic stellate cell and is an ideal polypeptide medicine for anti-fibrosis treatment. Because the TDGF-1 truncated micro-molecule polypeptide disclosed by the invention lacks a tumor-prompted structural domain, and therefore, the TDGF-1 truncated micro-molecule polypeptide is safe in the anti-fibrosis application, and also has a good market application prospect.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of TDGF-1 truncated small molecular polypeptide in anti-hepatic fibrosis. Background technique [0002] Liver fibrosis is a pathological change common to almost all chronic liver diseases progressing to the end stage, and its essence is the fibrous tissue scar after liver inflammation and necrosis. Chronic liver diseases that lead to liver fibrosis include alcoholic liver disease, nonalcoholic steatohepatitis (NASH), and chronic hepatitis caused by viral infection. If the disease continues to progress, liver fibrosis will develop into cirrhosis and liver cancer. Therefore, timely and active intervention in patients with liver fibrosis is very important to reduce the incidence of end-stage liver disease. [0003] The essence of liver fibrosis is a scar reaction, which can be caused by various liver injuries, including poisoning, metabolism and virus attack. Underlying th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P1/16
CPCA61K38/00A61P1/16C07K14/47
Inventor 石莹张语桐
Owner JILIN UNIV FIRST HOSPITAL
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