34results about How to "Avoid off-target effects" patented technology

The invention relates to a method for preparing a CAR-T cell by CRISPR/Cas9. The method comprises integrating a CAR molecule into a first intron of a human 19th chromosome AAVS1 site by a CRISPR/Cas9 technology, and integrating antibody molecules of various tumor surface antigens into a human T cell genome AAVS1 site by CRISPR/Cas9. The method realizes accurate integration of the CAR molecule into a human T cell genome specific safe port site, does not influence human normal gene functions, and prevents a series of clinical risks such as viral vector safety hidden trouble and genetic toxicity and immunogenicity caused by insertion of an exogenous gene into a genome.

The invention provides a nucleic acid molecule for encoding a chimeric antigen receptor targeting CD22. The chimeric antigen receptor comprises an extracellular region, a transmembrane region and a intracellular signal transduction region, the extracellular region encoded by the nucleic acid molecule comprises a CD22 binding domain, and the CD22 binding domain is amino acid sequence as shown in SEQ ID No. 3. By using flow cytometry, a degranulation analysis experiment and ELISA to detect cell factors secreted by a T cell, a fact that the CAR-T cell has a high lethal effect on B cell lymphoma cells and acute B cell lymphoma cell leukemia cells expressing CD22 and hardly has a lethal effect on cells which do not express CD22, and an off-target effect is prevented effectively is proved. The chimeric antigen receptor CD22scFv-CD8alpha-4-1BB-CD3zeta can be used for treating CD22<+>B cell hematologic neoplasms and is applicable to combined treatment with a CD19 CAR-T cell.

The present invention provides a nucleic acid molecule encoding a chimeric antigen receptor targeting CD5, the chimeric antigen receptor comprises an extracellular domain, a transmembrane domain and an intracellular signal transduction domain, and the extracellular domain encoded by the chimeric antigen receptor comprises a CD5 binding domain which is an amino acid sequence as shown in SEQ ID NO:3. The chimeric antigen receptor comprises an extracellular domain, a transmembrane domain and an intracellular signal transduction domain, and the extracellular domain encoded by the chimeric antigenreceptor comprises a CD5 binding domain which is an amino acid sequence shown in SEQ ID NO:3. The cytokines secreted by NK cells were detected by flow cytometry, degranulation assay and ELISA. The results showed that the CD5 CAR NK cells had strong cytotoxic effect on hematological tumor cells expressing CD5, but had weak cytotoxic effect on cells not expressing CD5. The cytokines secreted by CD5CAR NK cells could effectively prevent the missed target effect. A chimeric antigen receptor CD5 scFv-CD8[alpha]-4-1BB-CD 3 Xi can be used in the treatment of lymphocytic hematologic tumors with positive CD 5.

The invention relates to an early drug screening method based on fluorescence. The method comprises the following steps of: 1, providing target cells which stably express ion channel targets; 2, carrying out incubation on a voltage-sensitive dye and drugs with different concentrations together with the target cells; 3, measuring a fluorescence baseline value and a time-varying value of the incubated cells by utilizing a fluorescence measurement device; and 4, carrying out data fitting on the obtained values and determining whether the drugs have an exciting effect or an inhibiting effect on the ion channel targets. By utilizing the method, dozens of ion channel targets can be subjected to drug screening with an intermediate flux through utilizing a microplate reader with low cost; and theearly drug screening method is particularly applicable to early drug screening.

The invention provides a targeting responsive release system. The targeting responsive release system comprises a chimeric apoptotic corpuscle, the chimeric apoptotic corpuscle comprises an apoptoticcorpuscle membrane and mesoporous nanoparticles wrapped in the apoptotic corpuscle membrane, the apoptotic corpuscle membrane is an apoptotic corpuscle membrane derived from immune cells, and the mesoporous nanoparticles are loaded with active substances. The system provides an effective means for constructing a specific treatment product with a targeted delivery function and an accurate drug release function.

The invention discloses a nucleic acid molecule for coding a chimeric antigen receptor targeting CD123. The chimeric antigen receptor comprises an extracellular region, a transmembrane region and an intracellular signal transduction region; the extracellular region coded by the chimeric antigen receptor comprises a CD123 binding structural domain; the CD123 binding structural domain is an anti-CD123 single-chain antibody variable region fragment; and the anti-CD123 single-chain antibody variable region fragment is an amino acid sequence shown as SEQ ID NO.13 or a sequence with 90%-99% identity with the amino acid sequence shown as SEQ ID NO.13, an amino acid sequence shown as SEQ ID NO.14 or a sequence with 90%-99% identity with the amino acid sequence shown as SEQ ID NO.14, or an amino acid sequence shown as SEQ ID NO.15 or a sequence with 90%-99% identity with the amino acid sequence shown as SEQ ID NO.15. According to the nucleic acid molecule for coding the chimeric antigen receptor targeting the CD123, the chimeric antigen receptor can be used for treating CD123+ and/or CD33+ hematologic tumors, so that the off-target effect is effectively prevented, the treatment effect is better, and escape is not prone to occurring.

The invention discloses a nucleic acid molecule encoding a chimeric antigen receptor targeting CD22 and CD19. The chimeric antigen receptor comprises an extracellular region, a transmembrane region and an intracellular signal transduction region, wherein the extracellular region encoded by the chimeric antigen receptor comprises a CD22 and CD19 binding domain, and the CD22 and CD19 binding domainis composed of an antibody single-chain variable region fragment of CD22 and an antibody single-chain variable region fragment of CD19; and the antibody single-chain variable region fragment of CD22 and the antibody single-chain variable region fragment of CD19 are arranged in the sequence of an amino acid sequence shown in SEQ ID No. 9, an amino acid sequence shown in SEQ ID No. 10, an amino acidsequence shown in SEQ ID No. 11 or an amino acid sequence shown in SEQ ID No. 12. The chimeric antigen receptor disclosed by the invention can be used for treating CD19 <+> and CD22 <+> B cell bloodtumors and combined treatment with CD19 CAR-T cells or CD22 CAR-T cells.

The invention discloses an encoding chimeric antigen receptor nucleic acid molecule. The chimeric antigen receptor comprises an extracellular region, a transmembrane domain and an intracellular signaltransduction domain, wherein the encoded extracellular region comprises an FLT3 binding structural domain, and the FLT3 binding structural domain is an FLT3 ligand or an amino acid sequence with the90 to 99% identity as the FLT3 ligand. A flow cytometry, a degranulation analysis experiment and cytokines secreted by ELISA detection T cells prove that T cells modified by the chimeric antigen receptor have a strong killing effect of leukemia cells for expressing FLT3, especially have a specific killing effect on AML cells carrying FLT3 mutant and can effectively prevent an off-target effect. The chimeric antigen receptor FLT3L-CD8 alpha-4-1BB-CD3 zeta disclosed by the invention can be applied to treating FLT3+leukemia, especially leukemia carrying the FLT3 mutant.

The present invention relates to an RNA interference-inducing nucleic acid and the use thereof, and more particularly to an RNA interference-inducing nucleic acid comprising at least one single strand of double strands, the at least one single strand comprising a modification substituted to a spacer, which is unable to form a base pair, in the 5′ end or the 3′ end region. The RNA interference-inducing nucleic acid of the present invention is a novel modified form of nucleotide provided to prevent off-target effects, offering a method to selectively repress target gene expression. In addition, according to the present invention, the RNA interference-inducing nucleic acid provides novel modified forms with target selectivity and specificity as a method to block the off-target effects while silencing the target gene expression, whereas the usage of conventional RNA interference-inducing nucleic acids cause inaccuracy and adverse effects through off-targets, thereby the RNA interference-inducing nucleic acid was offered to solve the problem, wherein it will be widely used as a method for repressing gene expression in research and for gene therapy without concerning the off-target effects.

The invention provides a bispecific chimeric antigen receptor (CAR) targeting ROR1 and CD33 and application thereof, the bispecific chimeric antigen receptor (CAR) has the following structure: ROR1 scFv-CD33 scFv-H-TM-C-CD3 zeta, in which '-' is a connecting peptide or a peptide bond; h is a hinge area; tM is a transmembrane domain; c is a costimulatory signal molecule; cD3zeta is an intracellular signal transduction sequence. The chimeric antigen receptor can inhibit tumor growth in vivo and in vitro, prolong the life cycle, promote cytokine secretion and play a synergistic anti-tumor role.

The invention provides a bispecific chimeric antigen receptor (CAR) targeting ROR1 and CD7 and application thereof, the bispecific chimeric antigen receptor (CAR) has the following structure: ROR1 scFv-CD7 scFv-H-TM-C-CD3 zeta, in which "-" is a connecting peptide or a peptide bond; H is a hinge area; TM is a transmembrane domain; C is a costimulatory signal molecule; and CD3zeta is an intracellular signal transduction sequence. The chimeric antigen receptor can inhibit tumor growth in vivo and in vitro, prolong the life cycle, promote cytokine secretion and play a synergistic anti-tumor role.

The invention discloses a nucleic acid molecule for coding a chimeric antigen receptor targeting BCMA, the chimeric antigen receptor comprises an extracellular region, a transmembrane region and an intracellular signal transduction region, the extracellular region coded by the chimeric antigen receptor comprises a BCMA binding structural domain, the BCMA binding structural domain is a single-chain antibody fragment specifically binding to BCMA, and the intracellular signal transduction region is a single-chain antibody fragment specifically binding to BCMA. The single-chain antibody fragment comprises a light chain variable region amino acid sequence as shown in SEQ ID No. 1 and a heavy chain variable region amino acid sequence as shown in SEQ ID No. 2. The T cell modified by the chimeric antigen receptor has a very strong killing effect on multiple myeloma cells expressing BCMA, almost has no killing effect on cells not expressing BCMA, and effectively prevents an off-target effect. The chimeric antigen receptor disclosed by the invention can be used for treatment of BCMA + multiple myeloma cell hematologic tumors and combined treatment of CAR-T cells such as CD38, CD138 and CD19.

The present invention provides short activating RNA molecules which up-regulate albumin production. The present invention also provides methods of up-regulating albumin production, such methods involving the use of short activating RNA molecules capable of increasing the expression of albumin. The present invention also provides the use of the short activating RNA molecules in therapy, such as treating or preventing a hyperproliferative disorder and/or a disorder characterised by hypoalbuminemia.

The invention discloses an HLA-G specific chimeric antigen receptor, nucleic acid encoding the HLA-G specific chimeric antigen receptor, an expression plastid of the HLA-G specific chimeric antigen receptor, a cell expressing the HLA-G specific chimeric antigen receptor, an application of the cell and a composition, The present invention relates to the HLA-G specific chimeric antigen receptor, an isolated nucleic acid, the expression plastid of the HLA-G specific chimeric antigen receptor, the cell expressing the HLA-G specific chimeric antigen receptor, a pharmaceutical composition for treatment of cancer, and the application of the cell expressing the HLA-G specific chimeric antigen receptor. The HLA-G specific chimeric antigen receptor and a human leukocyte antigen G are specifically bound. The cell expressing the HLA-G specific chimeric antigen receptor is obtained by transducing the HLA-G specific chimeric antigen receptor into an immune cell. The pharmaceutical composition for treatment of cancer comprises the cell expressing the HLA-G specific chimeric antigen receptor and a pharmaceutically acceptable carrier. Therefore, the cell expressing the HLA-G specific chimeric antigen receptor can be used for inducing tumor cell death of mammals.