Human interferon alpha receptor binding related site mutant and application thereof

Abstract

The invention belongs to the technical field of medicine and bioengineering, and relates to application of a human interferon alpha receptor binding related site mutant in preparation of an anti-hepatitis B virus preparation. The interferon alpha plays an antiviral role through a receptor compound combined by two subunits of I-type interferon receptors IFNAR1 and IFNAR2. According to the invention, mutation is carried out on a site where IFN-alpha2 is combined with IFNAR1, and prokaryotic purification expression is carried out in vitro; the recognition in an HBV infection model shows that thehuman interferon alpha receptor binding related site mutant IFN-alpha2-EIFK has stronger anti-hepatitis B virus activity than IFN-alpha2, and has no cytotoxic effect under the antiviral concentration.The IFN-alpha2 receptor disclosed by the invention is combined with a related site mutant, namely IFN-alpha2-EIFK, so that a novel anti-hepatitis B virus medicine can be further prepared.

Description

technical field [0001] The invention belongs to the technical field of medicine and bioengineering, and relates to a human alpha interferon receptor binding-related site mutant and its application, in particular to an alpha interferon mutant prepared by transforming human alpha interferon and purification and its use in the preparation of anti-alpha interferon Use in hepatitis B virus preparations for reducing or eliminating viral surface antigen (HBsAg) and DNA in hepatitis B virus-infected liver cells. Background technique [0002] Hepatitis B virus (HBV) is an important pathogen that seriously endangers human health. According to relevant statistics, there are about 240 million HBV carriers in the world, of which nearly 80 million are chronic HBV-infected in my country; although there is currently a hepatitis B vaccine that can prevent HBV infection, there are still hundreds of thousands of new chronic hepatitis B infections every year At the same time, hundreds of thousa...

AI Technical Summary

Problems solved by technology

There are more than a dozen kinds of IFNs that have been identified. According to the different receptors they bind, IFNs are roughly divided into two types: I and II: Type I interferons mainly include IFN-α and IFN-β (IFN-λ is generally classified as III type II interferon), and type II interferon is mainly IFN-γ. Among them, with the emergence and development of genetic engineering technology, recombinant human IFN-α was cloned and produced in the last century for the treatment of viral hepatitis and other diseases; Compared with another large class of drugs used to treat chronic hepatitis B—nucleoside (acid) analogues, studies have shown that IFN-α and its PEGylated product (PEG-IFN-α) not only have direct antiviral effects, but also have immune Therefore, it has the advantages of relatively higher HBsAg negative conversion and higher sustained response rate; however, clinical statistics show that the efficiency of IFN-α in the treatment of chronic hepatitis B is still low, such as long-acting interferon for 48 months of HBe Only about 30% of antigen (HBeAg)-positive patients can turn HBeAg negative, while the HBsAg negative rate is even lower than 5%. Therefore, it is a consensus in the industry that it is urgent to optimize the anti-HBV efficiency and response rate of interferon

[0004] studies have shown that interferon initiates the transduction of the downstream JAK-STAT signaling pathway by specifically binding to IFNAR on the cell surface, thereby inducing the transcription of interferon-stimulated genes (ISGs) It is known that including IFN-α2, 13 subtypes of human IFN-α have been identified successively, and their coding genes are all located on human chromosome 9. There are many similar structural domains, but about 30% of the sequences are non-conserved; some reports indicate that although different subtypes of IFN-α play a role by binding to the two subunits IFNAR1 and IFNAR2 of type I interferon receptors, but Due to the different binding affinities to the two receptor subunits, there are differences in the way and degree of activation of downstream classical or bypass signaling pathways by each subtype of IFN-α; at the same time, the sensitivity of different viruses and different cells to IFN subtypes also exists. Different; through the study of interferon and its receptor affinity, it was found that interferon usually has a high binding ability to IFNAR2, but a low binding ability to IFNAR1, and the amino acid position in the interferon amino acid sequence related to interferon receptor binding point has been basically parsed

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