40 results about "HBx" patented technology

The present invention relates to therapeutic protocols and pharmaceutical compositions designed to target HBx mediated activation of Src kinase, members of the Src tyrosine kinase family and components of the Src kinase family signal transduction pathways for the treatment of HBV infection and related disorders and diseases, such as HCC. The invention further relates to pharmaceutical compositions for the treatment of HBV infection targeted to HBx and its essential activities required to sustain HBV replication. The invention is based, in part, on the Applicants' discovery that activation of Src kinase signaling cascades play a fundamental role in mammalian hepadnavirus replication. Applicants have demonstrated that HBx mediates activation of the Src family of kinases and that this activation is a critical function provided by HBx for mammalian hepadnavirus replication.

The invention belongs to the field of gene therapy and aims at providing a novel targeted gene vaccine for treating HBV (Hepatitis B Virus)-relevant liver cancer and application of the novel targeted gene vaccine in preparation of a targeted gene immunotherapeutic drug for the HBV-relevant liver cancer. The main active component of the liver caner-resistant vaccine disclosed by the invention is a recombinant vector of a gene coded with an HBx protein and a gene for coding a human IL-12 protein; and the recombinant vector can be used for simultaneously expressing the HBx protein and the human IL-12 protein in a eukaryotic cell. Provided by the experiment, a recombinant adenovirus vaccine disclosed by the invention can be used for selectively killing HBx-electropositive liver cancer cell and expresses a favorable liver cancer-resisting effect. The recombinant adenovirus vaccine provides a new selection for targeted gene immunization therapy of the HBV-relevant liver cancer and has a favorable application prospect.

The invention discloses a Stat3 target CRISPR / Cas carrier, a Stat3-HBx dual-target CRISPR / Cas carrier, and two repeated-target CRISPR / Cas carriers targeting HBx, and particularly relates to an Stat3 target single-function plasmid, an Stat3-HBx dual-target CRISPR / Cas carrier, two CRISPR / Cas carriers repeatedly targeting the HBx, and a construction method of the carriers. Through tests, the inventorproves that the constructed carriers have a specific target treatment effect on HBV-related liver cancer cells, and the silent gene carrier, the carrier construction method, double-silence strategiesof the carriers for Stat3 and the HBx and application of the carriers to preparation of HBV chronic infection and anti-liver cancer related drugs are within the scope of right protection correspondingly.

The invention relates to an siRNA (small interfering RNA) directed at HBx and a liposome for transferring the siRNA. The liposome contains siRNA plasmids, cationic lipid 98N12-5, cholesterol and PEGylated phospholipid. The liposome not only has good in vitro-in vivo stability, can well reach the target part, but also acquires high transfection efficiency, thus being in favor of playing a role in conveying siRNA into cells to inhibit hepatitis B virus gene HBx expression. In addition, the liposome also has low toxicity, thus being beneficial to its application in-vivo study.

The invention belongs to the field of a biological technology, and particularly relates to a purification method of an HBx protein core segment HBx delta N delta C in HBV and an HBx delta N delta C monoclonal antibody. A formula and purification process of various buffered solutions in an original purification method are changed, so that HBx delta N delta C samples being active, stable, uniform, high in purity and solitary are obtained, and standard large-scale production and preparation are convenient. The obtained HBx delta N delta C are used for further preparing a mouse sourced HBx delta Ndelta C monoclonal antibody and hybridoma strains which can stably express the mouse sourced HBx delta N delta C monoclonal antibody. The HBx delta Ndelta C and the HBx delta N delta C monoclonal antibody can be used as a scientific research reagent, the mouse sourced HBx delta N delta C monoclonal antibody can also be used as a detection reagent of hepatitis b virus HBV infection diseases, and the mouse sourced HBx delta N delta C monoclonal antibody is humanized, so that humanized HBx delta N delta C monoclonal antibody can be obtained and can be used as a scientific research reagent, a detection reagent for HBV infection diseases and an auxiliary treatment preparation.

Provided herein are, inter alia, antibodies, antigen-binding antibody fragments, cells, polynucleotides, compositions, kits, and methods relating to the detection of HBV protein X (HBx), e.g., in vitro and in vivo. Included are antibodies and fragments thereof that bind HBx, as well as kits, cells, and compositions comprising such antibodies and fragments.

The invention belongs to the technical field of biology, and particularly relates to a dissolving and purifying method of an HBx protein inclusion body derived from Hepatitis B Virus HBV. The preparation method comprises the following steps: expressing HBx by using a recombinant expression system; collecting cells containing the HBx protein which has been subjected to recombinant expression, breaking the cells by using a lysis buffer solution under a cooling condition, and collecting a part containing an HBx protein inclusion body; adding a dissolution buffer solution into the obtained part containing the HBx protein inclusion body for resuspension and denaturation; performing renaturationon on the product, removing nucleic acid, performing purification, and obtaining HBx protein; whereinthe steps are completed under the environmental condition of less than or equal to 10 DEG C. By using the method provided by the invention, a stable and uniform monomer HBx sample can be obtained, andthe concentration can reach 15mg / ml. The protein sample can be used as a scientific research reagent, and a monoclonal antibody of HBx can be prepared by using the protein sample.

A hepatitis B virus antisense RNA expression vector targeted toward liver cancer is a gene therapeutic matter for hepatitis B virus related liver cell cancer. The present invention adopts EB virus expressing plasmid pEBAF as vector and positive liver cancer tissue expressing vector controlled by AFP promotor controlling and specifically expressed in AFP only to construct three kinds of HBV antisence RNA expression vectors pEBAF-as-preS2, pEBAF-as-preS1 and pEBAF-as-X, which contain HBV preS2, preS1 and X segment genes separately. Antisense RNA expression vector is prepared through amplifying a destination gene, designing, a primer, and a PCR amplification of HBpreS2, HBpreS1 and HBX genes. The antisense RNA expression vector is used for treating hepatitis B virus related liver cell cancer and hepatitis B virus infection.

The present invention relates to therapeutic protocols and pharmaceutical compositions designed to target HBx mediated activation of Src kinase, members of the Src tyrosine kinase family and components of the Src kinase family signal transduction pathways for the treatment of HBV infection and related disorders and diseases, such as HCC. The invention further relates to pharmaceutical compositions for the treatment of HBV infection targeted to HBx and its essential activities required to sustain HBV replication. The invention is based, in part, on the Applicants' discovery that activation of Src kinase signaling cascades play a fundamental role in mammalian hepadnavirus replication. Applicants have demonstrated that HBx mediates activation of the Src family of kinases and that this activation is a critical function provided by HBx for mammalian hepadnavirus replication.

The invention discloses a Stat3-targeting CRISPR / Cas carrier, a Stat3-HBx dual-targeting CRISPR / Cas carrier and two repeat-targeting HBx-targeting CRISPR / Cas carriers, in particular to a Stat3-targeting single function Plasmids, a Stat3-HBx dual-targeting CRISPR / Cas vector, two repeating HBx-targeting CRISPR / Cas vectors, and methods for constructing the above-mentioned vectors. The inventors have proved through experiments that the vector constructed in this invention has a specific targeted therapeutic effect on HBV-related liver cancer cells. The present invention correspondingly protects the above-mentioned silencing gene vector, the construction method of the vector, the dual silencing strategy of the vector for Stat3 and HBx, and The application of the carrier in the preparation of HBV chronic infection and anti-liver cancer related drugs.

The invention discloses a method for evaluating therapeutic effects of lapatinib on liver cancer comprising: obtaining a liver cancer biopsy from a patient; determining level of a biomarker in the liver cancer biopsy obtained from the patient ex vivo; comparing the determined level of the biomarker in the liver cancer biopsy obtained from the patient to a reference level of the biomarker; and predicting therapeutic effect of lapatinib on liver cancer according to the comparison between the determined level and the reference level of the biomarker; wherein the reference level of the biomarker is level of the biomarker in a liver biopsy obtained from a normal, non-cancerous subject; wherein the biomarker is HBx or ErbB3.

The invention belongs to the field of biotechnology and genetic engineering, and relates to a method for evaluating an X protein (HBx protein) binding molecule of human hepatitis C virus. The method provided by the invention comprises the following steps: expressing a secreting type fusion protein containing the HBx protein and luciferase NanoLuciferase (Nluc) in a cell; and quantifying the levelof the secreted HBx protein by detecting the activity of luciferase of the fusion protein. The method can be used for screening and identifying the HBx protein binding molecule, provides a support forresearching an HBx protein targeted drug, and has good application prospect.

The invention discloses siRNA (small interfering RNA) for treating HBV (hepatitis B virus), which has a sequence shown in SEQ ID NO.3. The siRNA can be artificially synthesized or synthesized through in vivo transcription. After the siRNA transfects HepG2.2.15 cell for 24 hours and 48 hours, the siRNA can reduce the expression of HBx gene, the content of HBsAg and HBeAg in the cell and supernate is also obviously reduced, and the effect of 3p-siRNA is more obvious than that of chemically synthesized siRNA. The siRNA can be applied to medicaments for resisting the HBV.

Provided herein are, inter alia, antibodies, antigen-binding antibody fragments, cells, polynucleotides, compositions, kits, and methods relating to the detection of HBV protein X (HBx), e.g., in vitro and in vivo. Included are antibodies and fragments thereof that bind HBx, as well as kits, cells, and compositions comprising such antibodies and fragments.

The invention provides a group of deletion mutants of hepatitis B HBx gene, wherein SEQ ID NO.1 is a mutant in which 264 bases are deleted between HBV nucleotide positions 1577-1840 in HBV gene; SEQ ID NO.2 is HBx A mutant in which 19 bases are deleted between nucleotide positions 382-400 in the gene; SEQ ID NO.3 is a mutant in which 15 bases are deleted in nucleotide positions 431-445 in the HBx gene. This group of deletion mutants all have obvious tumorigenic effects, and can be used for gene detection of canceration in hepatitis B virus carriers and chronic hepatitis B patients, as well as preparation of drugs for preventing liver cancer and treating HBV chronic infection.

The invention discloses siRNA for treating HBV, which has a sequence shown in SEQ ID NO.1 or SEQ ID NO.2 or SEQ ID NO.3. The siRNA can be artificially synthesized or synthesized through in vivo transcription. After the siRNA transfects HepG2.2.15 cell for 24 hours and 48 hours, the siRNA can reduce the expression of HBx gene, the content of HBsAg and HBeAg in the cell and supernate is also obviously reduced, and the effect of 3p-siRNA is more obvious than that of chemically synthesized siRNA. The siRNA can be applied to medicaments for resisting the HBV.