Compound having activity for inhibiting tgf- belta receptor activation, method for screening same, and composition for prevention or treatment of hepatitis c virus-induced diseases
一种丙型肝炎病毒、TGF-的技术,应用在化合物的筛选、药物组合、化学仪器和方法等方向,能够解决没有完成开发出病毒性疾病的物质等问题
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0108] (Example 1) Confirmation of TGF-β2-like antigen activity of NS3 protease
[0109] First, in order to investigate whether HCV-derived NS3 protease is involved in TGF-β signal transduction, the TGF-β2-like antigen of purified recombinant NS3 protein was studied by ELISA using TGF-β2Emax(R) ImmunoAssay System (Promega) active. That is, the TGF-β coating antibody attached to the kit was diluted to 1 / 1000 with carbonic acid buffer (pH 9.2), added to each 100 μl / well of a 96-well ELISA plate (manufactured by NUNC), and added at 4 After standing overnight at ℃, the plate was coated. After washing the coated plate with 0.05% Tween20-containing phosphate buffer solution (hereinafter, also referred to as "PBST"), add the TGF-β blocking solution attached to the kit at 270 μl / well each, and let it stand at 37°C 35 minutes for sealing. Next, the blocking-treated plate was washed with PBST, the prepared TGF-β2 standard solution and the above-mentioned purified recombinant NS3 prot...
Embodiment 2
[0111] (Example 2) Study on activation of TGF-β signal triggered by NS3 protease
[0112] Next, Reporter Assay was used to study whether HCV-derived NS3 protease activates TGF-β signaling in cells. That is, the TGF-β2-like activity of purified recombinant NS3 protein was investigated using a cell line into which DNA of nine transcription factors Smad, which is important for the transcription of TGF-β target genes, was inserted upstream of the luciferase gene A PG1 reporter plasmid (manufactured by Promega) obtained by combining the sequence (CAGA) was introduced into mink lung epithelial cells (CCL64 cells) and constructed (hereinafter also referred to as "×9CAGA / CCL64 cells"). Specifically, Dulbecco's Modified Eagle Medium (DMEM, manufactured by Invitrogen Corporation) supplemented with 10% fetal bovine serum (manufactured by EQUITECH-BIO Corporation) and 1% preservative (penicillin-streptomycin-glutamine solution, manufactured by Invitrogen Corporation) system) (hereinafter...
Embodiment 3
[0114] (Example 3) Study on the promotion of collagen production by NS3 protease in hepatic stellate cells
[0115]It is known that TGF-β acts on stellate cells of the liver, promotes abnormal production of collagen by the stellate cells, and causes liver fibrosis. Therefore, the relationship between NS3 protease and promotion of collagen production was next investigated. That is, Wistar rats (raised under SPF, male, 15 weeks old) were laparotomed under pentobarbital anesthesia, and a catheter was inserted into the portal vein, followed by washing solution for blood removal, 0.06% pronase solution (Carbiochem Co., Ltd.) and 0.03% collagenase (Wako Pure Chemical Industries, Ltd.) solutions were perfused in this order. Thereafter, the liver was removed, and in a solution obtained by adding 1 ml of 2 mg / ml DNaseI (manufactured by Roche Diagnostics) to a buffer for isolating hepatic stellate cells containing 0.057% pronase and 0.057% collagenase, the mixture was incubated for 30 ...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap