38results about How to "Eliminates the possibility of self-replication" patented technology

The invention discloses a CAR-T transgene vector based on replication defective recombinant lentivirus. The CAR-T transgene vector comprises an original nuclear replicon pUCOri sequence, a resistance gene AmpR sequence containing ampicillin, a virus replicon SV40 Ori sequence, a lentivirus packaging cis element, ZsGreen1 green fluorescent protein, an IRES ribosome binding sequence, a human EF1 alpha promoter , a chimeric antigen receptor of second-generation CAR or third-generation CAR and a regulating element, wherein the original nuclear replicon pUCOri sequence is used for plasmid replication; the resistance gene AmpR sequence is used for massively proliferating target strains; the virus replicon SV40 Ori sequence is used for enhancing replication in eukaryocyte; the lentivirus packaging cis element is used for lentivirus packaging; the ZsGreen1 green fluorescent protein is used for expressing green fluorescent for eukaryocyte; the IRES ribosome binding sequence is used for jointly transcribing and expressing protein; the human EF1 alpha promoter is used for conducting eukaryotic transcription on antigen receptor genes; the chimeric antigen receptor is used for forming the second-generation CAR or the third-generation CAR integrating recognition, transfer and start; the regulating element is used for enhancing expression efficiency of transgenes and used after eWPRE-enhanced type woodchuck hepatitis b virus is transcribed. Besides, the invention further discloses a construction method and application of the vector. By means of the CAR-T transgene vector and the construction method and application of the vector, secretion of cell factors and an in vitro killing effect of CAR-T cells can be remarkably improved, and the clinical treatment effect is remarkable.

The invention discloses an anti-CD20 chimeric antigen receptor, an encoding gene, a recombinant expression vector, a construction method of the recombinant expression vector and an application. The anti-CD20 chimeric antigen receptor comprises CD8 leader chimeric receptor signal peptide, CD20 VL, Optimal Linker C, a CD20 single-chain antibody heavy chain VH, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulated factor and a TCR chimeric receptor T cell activating domain which are connected in series. Moreover, the invention discloses an encoding gene of the anti-CD20 chimeric antigen receptor, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. Secretion of cell factors and an in-vitro killing effect of CAR-T cells are obviously improved, and the CAR-T cells have an outstanding clinical treatment effect.

The invention discloses an anti-EGFRvIII chimeric antigen receptor, an encoding gene, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. The anti-EGFRvIII chimeric antigen receptor comprises CD8 leader chimeric receptor signal peptide, an EGFRvIII single-chain antibody light chain VL, Optimal Linker C, an EGFRvIII single-chain antibody heavy chain VH, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulated factor and a TCR chimeric receptor T cell activating domain which are connected in series. Moreover, the invention discloses an encoding gene of the anti-EGFRvIII chimeric antigen receptor, a recombinant expression vector, a construction method of the recombinant expression vector, and an application. Secretion of cell factors and an in-vitro killing effect of CAR-T cells are obviously improved, and the CAR-T cells have an outstanding clinical treatment effect.

The invention discloses a CD 123-targeting replication-defective recombinant lentivirus CAR-T transgenic vector. The CD 123-targeting replication-defective recombinant lentivirus CAR-T transgenic vector comprises a prokaryotic replicor pUC Ori sequence used for plasmid duplication; an amicillin resistance gene AmpR sequence used for amplification of a large number of target strains; a virus replicor SV40 Ori sequence used for enhancing the replication in eukaryocytes; a lentivirus packaging cis element used for lentivirus packaging; ZsGreen 1 green fluorescent protein used for green fluorescence expression of eukaryocytes; an IRES ribosome binding sequence used for co-transcription and co-expression of protein; a human EF1 alpha promoter used for eukaryotic transcription of genes of a chimeric antigen acceptor; the chimeric antigen acceptor used for forming second-generation CAR or third-generation CAR integrating recognition, delivery and promoting; an eWPRE element used for enhancing the expression efficiency of transgenes. In addition, the invention further discloses a construction method and applications of the vector. With the vector, the secretion of the cell factors and the in-vitro lethal effect of the CAR-T cells can be obviously improved, and the effect in treatingacute myelogenous leukemia (AML) clinically is outstanding.

The invention discloses a CD22-taregted replication-defective recombinant lentivirus CAR-T transgenic vector. The vector comprises a prokaryotic replicon pUC Ori sequence for plasmid replication, an ampicillin-containing resistance gene AmpR sequence for mass amplification of a target strain, a virus replicon SV400ri sequence for enhancing replication within eukaryotic cells, a lentivirus packaging cis element for lentivirus packaging, ZsGreen1 green fluorescence protein for expression of green fluorescence of the eukaryotic cells, an IRES ribosome binding sequence for common transcriptional expression of the protein, a human EF1[alpha] promoter for eukaryotic transcription of a chimeric antigen receptor gene, a chimeric antigen receptor for constituting the second or the third generation of CAR integrating identification, transferring and promoting, and an eWPRE element for enhancing a transgenic expression efficiency. In addition, the invention also discloses a construction method and an application of the vector. The vector disclosed by the invention can significantly improve secretion of cell factors and an in vitro killing effect of CAR-T cells; and the vector is obvious in effect on the clinical treatment of precursor B-cell acute lymphocytic leukemia.

The invention discloses an OCTS (One CAR (Chimeric Antigen Receptor) with two ScFvs (Single-chain variable Fragments)) technique based CAR-T (Chimeric Antigen Receptor-T cell immunotherapy) therapeutic vector for glioblastoma. The OCTS technique based CAR-T therapeutic vector comprises a lentiviral skeleton plasmid, a human EF1 [alpha] promoter (SEQ ID NO. 14), an OCTS chimeric receptor structural domain and a PDL1 single-chain antibody, wherein the OCTS chimeric receptor structural domain comprises a CD8 leader chimeric receptor signal peptide (SEQ ID NO. 15), a PDL1 single-chain antibody light chain VL (SEQ ID NO. 16), a PDL1 single-chain antibody heavy chain VH (SEQ ID NO. 17), an EGFRvIII single-chain antibody light chain VL (SEQ ID NO. 18), an EGFRvIII single-chain antibody heavy chain VH (SEQ ID NO. 19), an antibody Inner-Linker (SEQ ID NO. 20), a single-chain antibody Inter-Linker (SEQ ID NO. 21), a CD8 Hinge chimeric receptor linker (SEQ ID NO. 22), a CD8 Transmembrane chimeric receptor transmembrane domain (SEQ ID NO. 23), a TCR (T Cell Receptor) chimeric receptor T cell activation domain (SEQ ID NO. 26) and a chimeric receptor co-stimulator domain. In addition, the invention also discloses a construction method of the vector and application of the vector to the preparation of a medicine for treating the glioblastoma.

The invention discloses an anti-CD138 chimeric antigen receptor, a coding gene, a recombinant expression vector and a construction method and an application of the recombinant expression vector. The anti-CD138 chimeric antigen receptor comprises a CD8leader chimeric receptor signal peptide, a light chain VH of the CD138 single-chain antibody, an Optimal Linker C, a heavy chain VL of a CD138 single-chain antibody, a CD8Hinge chimeric receptor hinge, a CD8Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulation factor and a TCR chimeric receptor T-cell activation domain, which are serially connected in sequence. In addition, the invention discloses the coding gene of the anti-CD138 chimeric antigen receptor, the recombinant expression vector and the construction method and the application of the recombinant expression vector. According to the anti-CD138 chimeric antigen receptor, the secretion of cell factors and the in vitro killing effect of CAR-T (Chimeric Antigen Receptor T-Cell Immunotherapy) cells can be remarkably improved, and the clinical treatment effect is outstanding.

The invention discloses a marker used for in-vivo tracing and manual removal of CAR-T cells. The marker comprises a trace-linker 1 with a sequence as shown in SEQ ID No. 18, a trace-linker 2 with a sequence as shown in SEQ ID No. 19 and a trace-linker 3 with a sequence as shown in SEQ ID No. 19. The invention also discloses a CAR-T therapy vector including the marker and a construction method thereof. Moreover, the invention further discloses application of the marker to preparation of the CAR-T therapy vector and drugs used for treating triple-negative breast cancer. The marker provided by the invention can realize controllable in-vivo tracing, in-vitro separation and manual removal of CAR-T cells without influence on the tumor killing effect of the CAR-T cells, so the security of CAR-T cell therapy is greatly improved, and a powerful pool can be provided for deeper analysis of the process of CAR-T cell therapy. The vector provided by the invention can express a targeting chimeric antigen receptor of CD117 in human T lymphocytes, guides and activates killing effect of T lymphocytes on CD117 positive cells, and is applicable to treatment of triple-negative breast cancer in clinical practice.

The invention discloses a CAR-T vector capable of targeting Her2 and blocking PD-L1 to reduce tumor immune escape. The CAR-T vector comprises a human EF1 alpha promoter for eukaryotic transcription ofa chimeric antigen receptor gene, wherein the human EF1 alpha promoter is shown in the formula of SEQ ID NO. 1, a second-generation CAR or third-generation CAR chimeric antigen receptor combining recognition, delivery and starting, wherein the chimeric antigen receptor comprises a Her2 single-chain antibody light chain VL and a Her2 single-chain antibody heavy chain VH, an IRES ribosome binding sequence shown in the formula of SEQ ID NO. 10, PD-L1 scFv and a gene transfer vector. The invention discloses a construction method of the vector and a use of the vector in preparation of drugs for reducing tumor immune escape. Through a Her2 CAR-T technology and PD-L1 scFv expressed in the T lymphocytes, the PD-L1 on the surface of the tumor cells is blocked, the interaction between PD-1 and PD-L1 is inhibited, tumor cell immune escape is inhibited and the T lymphocyte effects of killing tumor cells are improved so that the CAR-T immunotherapy effects of resisting brain glioma are improved.

The invention discloses a CAR-T vector capable of targeting Her2 and disturbing PD-L1 to reduce tumor immune escape. The CAR-T vector comprises a human EF1 alpha promoter for eukaryotic transcriptionof a chimeric antigen receptor gene, wherein the human EF1 alpha promoter is shown in the formula of SEQ ID NO. 1, PD-1 shRNA in the 3'UTR, wherein the sequence of the PD-1 shRNA is shown in the formula of SEQ ID NO. 25, a gene delivery vector, and a second-generation CAR or third-generation CAR chimeric antigen receptor combining recognition, delivery and starting, wherein the chimeric antigen receptor comprises a Her2 single-chain antibody light chain VL and a Her2 single-chain antibody heavy chain VH. The invention also discloses a construction method of the vector and a use of the vector in preparation of drugs for disturbing PD-L1 to reduce tumor immune escape. Through a Her2 CAR-T technology and interference of PD-1 expression in T lymphocytes, the tumor immune escape is inhibited and the T lymphocyte effects of killing tumor cells are improved so that the CAR-T immunotherapy effects of resisting brain glioma are improved.

The invention discloses an OCTS technology-based prostatic cancer CAR-T therapy vector which comprises a lentivirus framework plasmid, a human EP1alpha promoter (SEQ ID NO.14), an OCTS chimeric receptor structure domain and a PDL1 single-chain antibody, wherein the OCTS chimeric receptor structure domain comprises A CD8 leader chimeric receptor signal peptide (SEQ ID NO.15), a PSMA single-chain antibody light chain VL (SEQ ID NO.16), a PSMA single-chain antibody heavy chain VH (SEQ ID NO.17), a PDL1 single-chain antibody light chain VL (SEQ ID NO.18), a PDL1 single-chain antibody heavy chain VH (SEQ ID NO.19), an antibody Inner-Linker (SEQ ID NO.20), a single-chain antibody Inter-Linker (SEQ ID NO.21), a CD8 Hinge chimeric receptor hinge (SED ID NO.22), a CD8 Transmembrane chimeric receptor transmembrane domain (SEQ ID NO.23), a TCR chimeric receptor T cell activation domain (SEQ ID NO.26) and a chimeric receptor co-stimulus factor domain. In addition, the invention further discloses a construction method of the vector and an application of the vector in preparation of drugs for treating a prostatic cancer.

The invention discloses a CAR-T carrier targeted at EGFRvIII to interfere PD-1 and reduce tumor immune escape, the CAR-T carrier comprises a human DF1 alpha promoter, PD-1shRNA in 3'UTR, a gene transfer carrier and a second generation CAR or third generation CAR, wherein the human DF1 alpha promoter is used for transcribing CAR genes and shown as SEQ ID NO. 1; and the sequence of the PD-1shRNA isshown as SEQ ID NO. 25; the second generation CAR or third generation CAR is used for integrating recognition, transmitting and starting and comprises an EGFRvIII single-chain antibody light chain VLand an EGFRvIII single-chain antibody heavy chain VH. Besides, the invention discloses a construction method of the carrier and application of the carrier in preparation of drugs for interfering the PD-1 to reduce tumor immune escape. According to the CAR-T carrier targeted at the EGFRvIII to interfere the PD-1 and reduce tumor immune escape and the construction method and the application, the CAR-T technology aiming at the SGFRvIII is used, tumour cell immune escape is inhibited by interfering the expression of the PD-1 in the T lymphocyte, killing to the tumour cells by the T lymphocyte is enhanced, and accordingly, the glioma resistance effect of the CAR-T immunotherapy is improved.