Use of epirubicin hydrochloride in the preparation of an inhibitor capable of blocking the interaction between p97 and npl4

A technology of epirubicin hydrochloride and application, applied in the biological field, can solve the problems such as no report of epirubicin hydrochloride, no p97 effect and the like

Active Publication Date: 2019-03-12
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] But the role of p97 in virus infection immunity has not been seen in the prior art
At the same time, in the prior art, the effect of epirubicin hydrochloride (CAS No. 56390-09-1) on virus infection immunity has never been reported.

Method used

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  • Use of epirubicin hydrochloride in the preparation of an inhibitor capable of blocking the interaction between p97 and npl4
  • Use of epirubicin hydrochloride in the preparation of an inhibitor capable of blocking the interaction between p97 and npl4
  • Use of epirubicin hydrochloride in the preparation of an inhibitor capable of blocking the interaction between p97 and npl4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 p97 negatively regulates type I interferon signaling pathway depends on its ATPase activity

[0087] 1. Experimental purpose: to determine whether p97 is involved in the type I interferon response and the signal pathway of antiviral immunity.

[0088] 2. Experimental method:

[0089] (1) Cell infection experiment: human HEK293, HEK293T, mouse MEF cells were purchased from the cell bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. The cell culture method is as described in Basic Experiment Method 10. Sendai virus (SeV) and GFP-Vesicular stomatitis virus with green fluorescent vesicular stomatitis virus (GFP-VSV) were gifted by Professor Shu Hongbing of Wuhan University. SeV and VSV-GFP are both commercially available. The VSV-GFP modification method is the same Construction and expression of high titer pseudotyped GFP-retroviruses by cotransfection of VSV-G gene Journal of Shanghai Jiaotong University (Medical Science)" 2007-07. poly...

Embodiment 2

[0109] Example 2 Npl4 cooperates with p97 to regulate antiviral immune response

[0110] 1. Experimental purpose: p97 is combined with different cofactors to participate in the regulation of different signal pathways. The purpose of this example is to determine the cofactors that p97 binds in the regulation of the antiviral immune response.

[0111] 2. Experimental method:

[0112] (1) siRNA knocks out p97, Npl4, Ufd1, p47, FAF1 in the cell: The above siRNA is synthesized by Shanghai Jima Pharmaceutical Technology Co., Ltd. The sequence of p97siRNA is shown in Table 6, and Npl4, Ufd1, p47, FAF1 are shown in Table 7. .

[0113] Table 7 siRNA sequence

[0114]

[0115] (2) The activity of the reporter gene and the detection of the transcription level of the gene are the same as in Example 1.

[0116] (3) Construction of Npl4 vector overexpressed in cells

[0117] Using the method and system in Example 1 to extract total RNA from cells, and reverse transcribed into cDNA, use Takara high-fid...

Embodiment 3

[0135] Example 3 Structural analysis of p97-Npl4 complex

[0136] 1. Experimental purpose: through the method of structural biology, the p97 and Npl4 protein assembly complex obtained by in vitro purification is used to analyze the structure and study the protein interaction mode.

[0137] 2. Experimental method:

[0138] (1) Construction of expression vector for complex structure analysis:

[0139] HT- stands for pET28a-HisTEV vector, which is a vector constructed by the laboratory and modified from the commercial vector pET28a (Merck). For plasmid information, see (Shi, Z., et al. Structure of the MST4 in complex with MO25 provides insights into its activation mechanism. Structure 21, 449-461, 2013) Drosophila HT-Ter94-N (20-186), Drosophila HT- Npl4-UBD(1-77) uses Drosophila Ter94 full-length, reverse-transcribed cDNA as a template, and is used in Takara high-fidelity DNA polymerase PrimeSTAR The corresponding primers are used for PCR amplification under the action of, PCR primer...

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Abstract

The invention relates to the technical field of biology, and more specifically relates to a purpose of epirubicin hydrochloride in preparation of an inhibitor for blocking interaction p97 and Npl4. The function of epirubicin hydrochloride which can effectively block the interaction of p97 and Npl4 is firstly found, a RIG-I mediated IFNbeta signal pathway is promoted so as to increase the anti-virus infection capability, so that the treatment effect for virus infection is enhanced, and the epirubicin hydrochloride has clinical and market value.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to the use of epirubicin hydrochloride in preparing inhibitors capable of blocking the interaction between p97 and Npl4. Background technique [0002] Viral infection can cause a strong antiviral immune response of the body. Innate immunity is the body's first barrier against viral infections. It recognizes receptors in the body through pathogens, including toll-like receptors (TLR) and RIG-I-like receptors ( RLRs) can recognize viral nucleic acids. The activated receptor can initiate the signal transduction pathway of downstream transcription factor NF-κB and interferon regulatory factor IRF to produce a series of pro-inflammatory cytokines and type I interferons (type I interferons, type I IFN). The release of type I interferon can promote the transcription of downstream genes to effectively inhibit further virus replication. At the same time, the secretion of type I interferon c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/704A61P31/12A61P31/14
Inventor 周兆才焦石
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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