WO2019242339A1 - 一种包含第三信号受体的嵌合抗原受体及其应用 - Google Patents

一种包含第三信号受体的嵌合抗原受体及其应用 Download PDF

Info

Publication number
WO2019242339A1
WO2019242339A1 PCT/CN2019/077923 CN2019077923W WO2019242339A1 WO 2019242339 A1 WO2019242339 A1 WO 2019242339A1 CN 2019077923 W CN2019077923 W CN 2019077923W WO 2019242339 A1 WO2019242339 A1 WO 2019242339A1
Authority
WO
WIPO (PCT)
Prior art keywords
car
cells
intracellular region
scfv
cd3zeta
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2019/077923
Other languages
English (en)
French (fr)
Chinese (zh)
Inventor
杨选明
傅阳心
汪鑫
叶圣勤
李民
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Longyao Bio-Tech Inc
Original Assignee
Shanghai Longyao Bio-Tech Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Longyao Bio-Tech Inc filed Critical Shanghai Longyao Bio-Tech Inc
Priority to EP19823459.3A priority Critical patent/EP3812401A4/en
Priority to KR1020217001094A priority patent/KR102853978B1/ko
Priority to JP2020571580A priority patent/JP7433656B2/ja
Priority to CN201980041282.0A priority patent/CN112673025B/zh
Priority to CN202210196195.6A priority patent/CN114516922A/zh
Publication of WO2019242339A1 publication Critical patent/WO2019242339A1/zh
Priority to US17/127,283 priority patent/US11524034B2/en
Anticipated expiration legal-status Critical
Priority to US18/047,750 priority patent/US20230134345A1/en
Ceased legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4202—Receptors, cell surface antigens or cell surface determinants
    • A61K40/4221—CD20
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70521—CD28, CD152
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/715—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons
    • C07K14/7155—Receptors; Cell surface antigens; Cell surface determinants for cytokines; for lymphokines; for interferons for interleukins [IL]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2887—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD20
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/58—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation
    • A61K2039/585—Medicinal preparations containing antigens or antibodies raising an immune response against a target which is not the antigen used for immunisation wherein the target is cancer
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2851—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2869—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against hormone receptors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2878—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/033—Fusion polypeptide containing a localisation/targetting motif containing a motif for targeting to the internal surface of the plasma membrane, e.g. containing a myristoylation motif
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
    • C12N2740/15041—Use of virus, viral particle or viral elements as a vector
    • C12N2740/15043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/10—Plasmid DNA
    • C12N2800/106—Plasmid DNA for vertebrates
    • C12N2800/107—Plasmid DNA for vertebrates for mammalian

Definitions

  • the invention relates to the technical field of cellular immunotherapy, in particular to a chimeric antigen receptor containing a third signal receptor and application thereof.
  • chimeric antigen receptor T cells were first proposed by Gross, Waks, and Eshhar in 1989. They expressed antibodies that recognize TNP on T cells, achieving the activation and effects of antigen-specific, non-MHC-restricted T cells. Enhancement and put forward the concept of the application of CAR-T technology in tumor treatment. According to this principle, embedding tumor-specific antibodies into T cells will give T cells new tumor-killing capabilities. Later, CAR-T technology was introduced into anti-tumor clinical trials. However, early CAR-T cells contained only the first signal because of their intracellular signalling domain, and the selected tumor type was solid tumor. The final clinical results were not Too ideal.
  • the structure of CAR consists of an extracellular antigen recognition domain, an extracellular hinge region, a transmembrane region, and an intracellular signaling domain.
  • the extracellular antigen recognition domain is usually composed of a single chain antibody, which specifically recognizes the surface molecules of tumor cell membranes, and may also be a ligand or receptor for certain tumor-specific antigens.
  • the extracellular hinge region is a space structure that separates the antigen recognition domain from the transmembrane region. The purpose is to provide a suitable spatial location so that the extracellular antigen recognition domain can maintain the correct structure before and after recognition of the antigen and conduct intracellularly. signal.
  • the transmembrane region is a domain that ensures the localization of CAR molecules on the membrane surface.
  • the intracellular signaling domain is a key part of mediating CAR signaling, usually one or several first signals (recognition of TCR and MHC-I-peptide complexes), and second signals (co-stimulatory receptors and co-stimulatory receptors). Body recognition).
  • the first generation CAR contains only the first signal
  • the second generation CAR has a first signal and a second signal
  • the third generation CAR has a first signal and two second signal domains.
  • CAR-T for B cell surface targeting molecules CD19 and CD20 prepared from patient's own blood cells has been relatively mature in the treatment of B cell leukemia, but although the response rate is high, there are a large number of recurrences. Tumor treatment efficiency is relatively low, which is related to the immunosuppressive microenvironment in solid tumors.
  • tumor microenvironment In solid tumors, there are a variety of immune cells, tumor cells and stromal cells, which together constitute the tumor microenvironment.
  • the tumor microenvironment is usually immunosuppressive. It can inhibit endogenous anti-tumor T cell responses or adoptive T cells (such as CAR-T) at multiple levels. For example, T cells are depleted and tumor-killing cells are lost. Function, eventually T cells are cleared. How to enhance the activation capacity of CAR-T in solid tumors so that it can fight against immunosuppression in the tumor microenvironment is an important idea and direction for the expansion of CAR-T to solid tumors.
  • CAR-T domains use novel regulatory molecules such as IL-12, 4 -1BBL, etc. In addition to affecting CAR-T, these molecules will also have non-specific activation of other non-CAR-T cells, potentially causing immune side effects.
  • the purpose of the present invention is to overcome the defects in the prior art, provide a chimeric antigen receptor of a third signal receptor and its application, and provide a CAR constructed from a recombinant expression vector such as the chimeric antigen receptor.
  • -T cells in which the activation of T cells is subjected to a first signal (recognition of TCR and MHC-I-peptide complex), a second signal (recognition of co-stimulatory receptors and co-stimulatory ligands), and 3
  • the regulation of signals cytokine receptor and cytokine recognition
  • these three synergistically achieve a large number of T cells to expand, exert effector functions, and clear infections or tumors.
  • the present invention adopts the following technical solutions:
  • a first object of the present invention is to provide a chimeric antigen receptor including a third signal receptor, and the structure of the chimeric antigen receptor is scFv (X)-(Y) CD3zeta-MN;
  • X includes a tumor-targeting antibody or a ligand and a receptor capable of specifically binding to the tumor
  • Y is an intracellular region of a co-stimulatory receptor selected from ICOS, CD28, CD27, HVEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, CD226
  • M is the intracellular region of the gamma chain family cytokine receptor, the cytokine receptor is selected from IL2Ra, IL2Rb, IL4Ra, IL7Ra, IL9Ra, IL15Ra, IL21Ra
  • N is the intracellular region of IL2Rg.
  • the technical measures taken by the present invention also include:
  • the X is selected from the group consisting of an anti-CD19 antibody, an anti-CD20 antibody, an EGFR antibody, an HER2 antibody, an EGFRVIII antibody, an anti-PSMA antibody, an anti-BCMA antibody, an anti-CD22 antibody, and an anti-CD30 antibody. It can be understood that X may also be another protein capable of specifically binding to a tumor.
  • the X is an anti-CD20 antibody
  • the Y is a 4-1BB intracellular region
  • the M is selected from the group consisting of an IL2Rb intracellular region, an IL4Ra intracellular region, an IL7Ra intracellular region, an IL9Ra intracellular region, and an IL21Ra cell.
  • One of the inner zones are included in the X and the Y.
  • the scFv (X)-(Y) CD3zeta is scFv-antihCD20-20BBZ, the sequence of which is shown in SEQ ID No. 1; the sequence of the IL7Ra intracellular region is shown in SEQ ID No. 2; the IL2Rb The sequence of the intracellular region is shown in SEQ ID No. 3; the sequence of the intracellular region of IL4Ra is shown in SEQ ID No. 4; the sequence of the intracellular region of IL9Ra is shown in SEQ ID No. 5; The sequence of the region is shown in SEQ ID No. 6; the sequence of the IL2Rg intracellular region is shown in SEQ ID No. 7.
  • the extracellular hinge region of the chimeric antigen receptor is selected from a region of CD8a or IgG; the transmembrane region of the chimeric antigen receptor is selected from one of CD8a, CD28, CD137, or CD3.
  • a second object of the present invention is to provide a recombinant expression vector of any of the aforementioned chimeric antigen receptors.
  • a third object of the present invention is to provide a CAR-T cell constructed from a recombinant expression vector of any of the aforementioned chimeric antigen receptors.
  • a fourth object of the present invention is to provide a method for preparing the above CAR-T cells, which includes the following steps:
  • Step 1 Construction of lentiviral vector and virus production
  • ScFv (X)-(Y) CD3zeta, M and N form a fusion protein
  • lentiviral vectors are added at both ends, and co-transfection with the lentiviral packaging plasmid to obtain scFv (X)-(Y) CD3zeta-MN virus;
  • Step 2 Preparation of scFv (X)-(Y) CD3zeta-MN CAR-T cells;
  • the isolated and purified human PBMCs were cultured and infected with the scFv (X)-(Y) CD3zeta-MN virus obtained in step 1.
  • Cell expansion was performed under appropriate conditions to prepare scFv (X)-(Y) CD3zeta-MN.
  • CAR-T cells
  • X is a tumor-targeting antibody or other protein
  • Y is an intracellular region of a co-stimulatory receptor selected from ICOS, CD28, CD27, HVEM, LIGHT, CD40L, 4-1BB, OX40 , DR3, GITR, CD30, TIM1, SLAM, CD2, CD226
  • M is the intracellular region of the gamma chain family cytokine receptor selected from IL2Ra, IL2Rb, IL4Ra, IL7Ra, IL9Ra, IL15Ra, IL21Ra
  • N is the intracellular region of IL2Rg.
  • the technical measures adopted by the present invention further include:
  • the specific steps of constructing the lentiviral vector and virus production include: scFv (X)-(Y) CD3zeta, M and N are formed by an overlap PCR to form a fusion protein, and restriction sites are added at both ends to clone the lentiviral vector. ; The endotoxin-free clone was cloned correctly and transfected with the lentiviral packaging plasmid. The supernatant was collected at a predetermined time, and the virus was concentrated by centrifugation and centrifugation to obtain scFv (X)-(Y) CD3zeta-MN virus.
  • scFv (X)-(Y) CD3zeta M and N are formed by an overlap PCR to form a fusion protein, and EcoRI and BamHI restriction sites are added at both ends.
  • Clone the pCDH-MSCVEF vector clone the correctly cloned endotoxin-free clone and transfect 293X with the lentiviral packaging plasmid, collect the supernatant after 48 and 72 hours, and centrifuge at 25,000 RPM for 0.4 hours after 0.45 uM filtration to concentrate the virus to obtain scFv ( X)-(Y) CD3zeta-MN virus.
  • the specific steps of preparing the scFv (X)-(Y) CD3zeta-MN CAR-T cells include: after separating and purifying human PBMC, inoculating it into a culture plate with appropriate stimulation conditions, and culturing for a predetermined time, The scFv (X)-(Y) CD3zeta-MN virus produced in step 1 was infected, and the cells were expanded according to appropriate stimulation conditions. After 2 rounds of stimulation and expansion, the cells obtained were scFv (X)-(Y) CD3zeta -MN CAR-T cells.
  • the stimulating conditions of cultured, isolated and purified human PBMC are anti-hCD3 and anti-hCD28, and the stimulating conditions of expanded cells are stimulated with artificial antigen presenting cells or anti-hCD3 / 28 every 6 days.
  • the X is selected from an anti-CD19 antibody, an anti-CD20 antibody, an EGFR antibody, a HER2 antibody, and an EGFRVIII antibody.
  • the X is an anti-CD20 antibody
  • the Y is 4-1BB
  • the M is selected from one of IL2Rb, IL4Ra, IL7Ra, IL9Ra, and IL21Ra.
  • the scFv (X)-(Y) CD3zeta is scFv-antihCD20-20BBZ, the sequence of which is shown in SEQ ID No. 1; the sequence of the IL7Ra intracellular region is shown in SEQ ID No. 2; the IL2Rb The sequence of the intracellular region is shown in SEQ ID No. 3; the sequence of the intracellular region of IL4Ra is shown in SEQ ID No. 4; the sequence of the intracellular region of IL9Ra is shown in SEQ ID No. 5; The sequence of the region is shown in SEQ ID No. 6; the sequence of the IL2Rg intracellular region is shown in SEQ ID No. 7.
  • the lentiviral packaging plasmid in step 1 includes VSV-g, pMD, Gag / Pol, RSV-REV, and a Beckman ultracentrifuge and a SW28 rotor are used for centrifugation.
  • a fifth object of the present invention is to provide a preparation containing the above CAR-T cells or a CAR-T cell prepared by the above-mentioned preparation method; further, the preparation further includes a pharmaceutically acceptable diluent or excipient. Agent.
  • a sixth object of the present invention is to provide an application of the above-mentioned chimeric antigen receptor, the above-mentioned CAR-T cell, or the CAR-T cell prepared by the above-mentioned preparation method in preparing a medicine for treating or preventing tumor.
  • the tumor is a solid tumor
  • examples of the solid tumor include, but are not limited to, lymphoma, kidney tumor, neuroblastoma, germ cell tumor, osteosarcoma, chondrosarcoma, soft tissue sarcoma, liver tumor, thymoma, Pulmonary blastoma, pancreatoblastoma, hemangioma, etc.
  • the present invention has the following beneficial effects:
  • the CAR-T cells according to the present invention significantly improve the tumor killing ability and expansion ability, and significantly improve the killing ability of solid tumors / metastatic tumors.
  • the CAR-T cells according to the invention include a third signal receptor (IL2Ra , IL2Rb, IL4Ra, IL7Ra, IL9Ra, IL15Ra, IL21Ra, etc.), which are not conventionally used ligands or secretory factors, such as the third signal receptor IL7Ra, which is mainly expressed in memory CD4 and CD8 T cells. Long-term survival and the formation of memory T cells have an important role. Integrating the third signal receptor signal into CAR-T has the potential to enhance the effect, and only affects CAR-T cells, reducing the immune side effects risks of.
  • the present invention improves the activation capacity and survival capacity of CAR-T cells in tumors compared with the currently used CAR-T technology, and the activation capacity and expansion capacity are significantly improved. Enhancement, thereby increasing the efficacy of CAR-T cells to have more excellent anti-tumor efficacy.
  • FIG. 1 is a schematic diagram illustrating a molecular structure of a chimeric antigen receptor (CAR) containing a third signal receptor in each embodiment of the present invention
  • FIG. 2 is a schematic diagram of a virus titer measured after 293 cells were infected by BBZIL2RbIL2Rg virus according to an embodiment of the present invention
  • FIG. 3 is a schematic diagram of a virus titer measured after 293 cells were infected by BBZIL4RaIL2Rg virus according to an embodiment of the present invention
  • FIG. 4 is a schematic diagram of a virus titer measured after 293 cells were infected by BBZIL7RaIL2Rg virus according to an embodiment of the present invention
  • FIG. 5 is a schematic diagram of a virus titer measured after BBZIL9RaIL2Rg virus infects 293 cells according to an embodiment of the present invention
  • FIG. 6 is a schematic diagram of a virus titer measured after 293 cells were infected by BBZIL21RaIL2Rg virus according to an embodiment of the present invention
  • FIG. 7 is a schematic diagram of the results of phenotypic analysis of BBZ CAR-T cells and BBZIL2RbIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 8 is a schematic diagram of the results of phenotypic analysis of BBZ CAR-T cells and BBZIL4RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 9 is a schematic diagram of the results of phenotypic analysis of BBZ CAR-T cells and BBZIL7RaIL2Rg CAR-T cells in an embodiment of the present invention.
  • FIG. 10 is a schematic diagram of the results of phenotypic analysis of BBZ CAR-T cells and BBZIL9RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 11 is a schematic diagram of the results of phenotypic analysis of BBZ CAR-T cells and BBZIL21RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 12 is a schematic diagram of the expansion capacity of BBZ CAR-T cells and BBZIL7RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 13 is a schematic diagram of tumor killing ability of BBZ CAR-T cells and BBZIL7RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 14 is a schematic diagram of the anti-tumor ability of BBZ CAR-T cells and BBZIL7RaIL2Rg CAR-T cells in an embodiment of the present invention
  • FIG. 15 is a schematic diagram of the in vivo viability of BBZ CAR-T cells and BBZIL7RaIL2Rg CAR-T cells in an embodiment of the present invention.
  • the present invention provides a chimeric antigen receptor including a third signal receptor.
  • the structure of the chimeric antigen receptor is scFv (X)-(Y) CD3zeta-MN; wherein X is a tumor-targeting antibody or other protein ; Y is the intracellular region of a co-stimulatory receptor selected from the group consisting of ICOS, CD28, CD27, HVEM, LIGHT, CD40L, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2, CD226; M is the intracellular region of the gamma chain family cytokine receptor selected from IL2Ra, IL2Rb, IL4Ra, IL7Ra, IL9Ra, IL15Ra, IL21Ra; N is the IL2Rg intracellular region.
  • the invention also relates to a CAR-T cell constructed from any of the above-mentioned chimeric antigen receptor recombinant expression vectors and a preparation method thereof, a preparation containing the CAR-T cell, and an application of the CAR-T cell.
  • the chimeric antigen receptor (CAR) molecules containing a third signal receptor are BBZIL2RbIL2Rg, BBZIL4RaIL2Rg, BBZIL7RaIL2Rg, BBZIL9RaIL2Rg, and BBZIL21RaIL2Rg, and their structures are shown in FIG. 1.
  • the preparation of the 20BBZIL2RbIL2Rg CAR-T cells described in this example includes the following steps:
  • ScFv-antihCD20-20BBZ (SEQ ID No. 1), IL2Rb intracellular region (SEQ ID No. 3) and IL2Rg intracellular region (SEQ ID No. 7) were formed by overlapping PCR to form a fusion protein, and EcoRI and BamHI were added at both ends.
  • the pCDH-MSCVEF vector was cloned at the restriction site. Endotoxin-free clones that were sequenced correctly were cloned with lentiviral packaging plasmids (VSV-g, pMD, Gag / Pol, RSV-REV) to transfect 293X. Supernatants were collected after 48 and 72 hours.
  • the Beckman ultracentrifuge and SW28 rotor were centrifuged at 25,000 RPM for 2 hours to concentrate the virus, namely pCDH-MSCVEF-20BBZIL2RbIL2Rg virus (referred to as 20BBZIL2RbIL2Rg virus) for subsequent CAR-T cell production.
  • pCDH-MSCVEF-20BBZ virus (20BBZ virus for short) was produced, and the obtained 20BBZIL2RbIL2Rg virus was used to infect 293 cells, and the virus titer was measured, as shown in FIG. 2.
  • the preparation of the 20BBZIL4RaIL2Rg CAR-T cells described in this example includes the following steps:
  • ScFv-antihCD20-20BBZ (SEQ ID No. 1), IL4Ra intracellular region (SEQ ID No. 4) and IL2Rg intracellular region (SEQ ID No. 7) were formed by an overlap PCR to form a fusion protein, and EcoRI and BamHI were added at both ends.
  • the pCDH-MSCVEF vector was cloned at the restriction site. Endotoxin-free clones that were sequenced correctly were cloned with lentiviral packaging plasmids (VSV-g, pMD, Gag / Pol, RSV-REV) to transfect 293X. Supernatants were collected after 48 and 72 hours.
  • the Beckman ultracentrifuge and SW28 rotor were centrifuged at 25,000 RPM for 2 hours to concentrate the virus, namely pCDH-MSCVEF-20BBZIL4RaIL2Rg virus (referred to as 20BBZIL4RaIL2Rg virus) for subsequent CAR-T cell production.
  • pCDH-MSCVEF-20BBZ virus 20BBZ virus for short
  • 20BBZIL4RaIL2Rg virus was used to infect 293 cells, and the virus titer was measured, as shown in FIG. 3.
  • the preparation of the 20BBZIL7RaIL2Rg CAR-T cells described in this example includes the following steps:
  • ScFv-antihCD20-20BBZ (SEQ ID No. 1), IL7Ra intracellular region (SEQ ID No. 2) and IL2Rg intracellular region (SEQ ID No. 7) were formed by an overlap PCR to form a fusion protein, and EcoRI and BamHI were added at both ends.
  • the pCDH-MSCVEF vector was cloned at the restriction site. Endotoxin-free clones that were sequenced correctly were cloned with lentiviral packaging plasmids (VSV-g, pMD, Gag / Pol, RSV-REV) to transfect 293X. Supernatants were collected after 48 and 72 hours.
  • the Beckman ultracentrifuge and SW28 rotor were centrifuged at 25,000 RPM for 2 hours to concentrate the virus, namely pCDH-MSCVEF-20BBZIL7RaIL2Rg virus (referred to as 20BBZIL7RaIL2Rg virus) for subsequent CAR-T cell production.
  • the control pCDH-MSCVEF-20BBZ virus (20BBZ virus for short) was produced, and the obtained 20BBZIL7RaIL2Rg virus was used to infect 293 cells, and the virus titer was measured, as shown in FIG. 4.
  • the preparation of the 20BBZIL9RaIL2Rg CAR-T cells described in this example includes the following steps:
  • ScFv-antihCD20-20BBZ (SEQ ID No. 1), IL9Ra intracellular region (SEQ ID No. 5) and IL2Rg intracellular region (SEQ ID No. 7) were formed by overlapping PCR to form a fusion protein, and EcoRI and BamHI were added at both ends.
  • the pCDH-MSCVEF vector was cloned at the restriction site. Endotoxin-free clones that were sequenced correctly were cloned with lentiviral packaging plasmids (VSV-g, pMD, Gag / Pol, RSV-REV) to transfect 293X. Supernatants were collected after 48 and 72 hours.
  • the Beckman ultracentrifuge and SW28 rotor were centrifuged at 25,000 RPM for 2 hours to concentrate the virus, namely pCDH-MSCVEF-20BBZIL9RaIL2Rg virus (referred to as 20BBZIL9RaIL2Rg virus) for subsequent CAR-T cell production.
  • pCDH-MSCVEF-20BBZ virus 20BBZ virus for short
  • 20BBZIL9RaIL2Rg virus was used to infect 293 cells, and the virus titer was measured, as shown in FIG. 5.
  • the preparation of the 20BBZIL21RaIL2Rg CAR-T cells described in this example includes the following steps:
  • ScFv-antihCD20-20BBZ (SEQ ID No. 1), IL21Ra intracellular region (SEQ ID No. 6) and IL2Rg intracellular region (SEQ ID No. 7) were formed by overlapping PCR to form a fusion protein, and EcoRI and BamHI were added at both ends.
  • the pCDH-MSCVEF vector was cloned at the restriction site. Endotoxin-free clones that were sequenced correctly were cloned with lentiviral packaging plasmids (VSV-g, pMD, Gag / Pol, RSV-REV) to transfect 293X. Supernatants were collected after 48 and 72 hours.
  • the Beckman ultracentrifuge and SW28 rotor were centrifuged at 25,000 RPM for 2 hours to concentrate the virus, namely pCDH-MSCVEF-20BBZIL21RaIL2Rg virus (referred to as 20BBZIL21RaIL2Rg virus) for subsequent CAR-T cell production.
  • pCDH-MSCVEF-20BBZ virus 20BBZ virus for short
  • 20BBZIL21RaIL2Rg virus was used to infect 293 cells, and the virus titer was measured, as shown in FIG. 6.
  • the 20BBZ CAR-T cells and 20BBZIL7RaIL2Rg CAR-T cells prepared in step 2 in Example 3 were continuously cultured for 14 days, and stimulated with artificial antigen presenting cells every 6 days. The cell count was shown in FIG. 12. It can be seen from the figure that 20BBZIL7RaIL2Rg CAR-T cells have stronger proliferation ability than 20BBZCAR-T cells.
  • 20BBZ CAR-T cells and 20BBZIL7RaIL2Rg CAR-T cells prepared in step 2 in Example 3 were seeded into a 96-well plate, and Raji tumors were added according to CAR-T: tumor cell ratios 1: 1, 1: 2, and 1: 4. Cells were compared for survival ratio of tumor cells after 24 and 48 hours. The results are shown in FIG. 13. It can be seen from the figure that 20BBZIL7RaIL2Rg CAR-T cells have similar tumor-killing ability compared to 20BBZ CAR-T cells.
  • the present invention enables CAR-T cells to activate, survive, and expand CAR
  • the enhancement ability is significantly enhanced, and it has more excellent anti-tumor efficacy.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biotechnology (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • General Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Virology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
PCT/CN2019/077923 2018-06-20 2019-03-13 一种包含第三信号受体的嵌合抗原受体及其应用 Ceased WO2019242339A1 (zh)

Priority Applications (7)

Application Number Priority Date Filing Date Title
EP19823459.3A EP3812401A4 (en) 2018-06-20 2019-03-13 CHIMERIC ANTIGEN RECEPTOR WITH A THIRD SIGNAL RECEPTOR AND USE THEREOF
KR1020217001094A KR102853978B1 (ko) 2018-06-20 2019-03-13 제3 신호 수용체를 포함하는 키메릭 항원 수용체 및 이의 용도
JP2020571580A JP7433656B2 (ja) 2018-06-20 2019-03-13 第３シグナル受容体を含むキメラ抗原受容体およびその使用
CN201980041282.0A CN112673025B (zh) 2018-06-20 2019-03-13 一种包含第三信号受体的嵌合抗原受体及其应用
CN202210196195.6A CN114516922A (zh) 2018-06-20 2019-03-13 一种包含第三信号受体的嵌合抗原受体及其应用
US17/127,283 US11524034B2 (en) 2018-06-20 2020-12-18 Chimeric antigen receptor comprising third signal receptor and use thereof
US18/047,750 US20230134345A1 (en) 2018-06-20 2022-10-19 Chimeric antigen receptor comprising third signal receptor and use thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810636414.1A CN110615843B (zh) 2018-06-20 2018-06-20 一种包含第三信号受体的嵌合抗原受体及其应用
CN201810636414.1 2018-06-20

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US17/127,283 Continuation US11524034B2 (en) 2018-06-20 2020-12-18 Chimeric antigen receptor comprising third signal receptor and use thereof

Publications (1)

Publication Number Publication Date
WO2019242339A1 true WO2019242339A1 (zh) 2019-12-26

Family

ID=68920806

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2019/077923 Ceased WO2019242339A1 (zh) 2018-06-20 2019-03-13 一种包含第三信号受体的嵌合抗原受体及其应用

Country Status (6)

Country Link
US (1) US11524034B2 (https=)
EP (1) EP3812401A4 (https=)
JP (1) JP7433656B2 (https=)
KR (1) KR102853978B1 (https=)
CN (3) CN110615843B (https=)
WO (1) WO2019242339A1 (https=)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021207150A1 (en) * 2020-04-06 2021-10-14 Celledit Llc Genetically modified immune cells expressing a chimeric antigen receptor and having reduced proinflammatory cytokine signaling
EP4083073A4 (en) * 2020-02-28 2023-07-12 Nanjing Bioheng Biotech Co., Ltd Novel chimeric antigen receptor and use thereof
WO2025040094A1 (en) * 2023-08-21 2025-02-27 Hangzhou Hervor Therapeutics Co., Ltd Synthetic cytokine signaling system, preparation method, and uses thereof

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110615843B (zh) * 2018-06-20 2023-05-09 上海隆耀生物科技有限公司 一种包含第三信号受体的嵌合抗原受体及其应用
CN111518219B (zh) * 2020-05-08 2021-06-22 浙江大学 嵌合性抗原受体及表达其的巨噬细胞、调节巨噬细胞极化的方法和应用
CN112062864A (zh) * 2020-09-18 2020-12-11 樊克兴 靶向bcma肿瘤抗原受体修饰t细胞的制备方法和应用
CN114853900A (zh) * 2021-02-03 2022-08-05 南京北恒生物科技有限公司 新型嵌合抗原受体及其用途
WO2023274355A1 (zh) * 2021-07-02 2023-01-05 上海交通大学 改造的间充质干细胞和免疫效应细胞联合治疗肿瘤
EP4402160A4 (en) * 2021-09-17 2025-11-19 Univ Pennsylvania CHIMERIC ANTIGENIC RECEPTORS COMPRISING AN INTERLEUKIN-9 RECEPTOR SIGNALING DOMAIN
AU2022348588A1 (en) 2021-09-17 2024-04-04 Parker Institute For Cancer Immunotherapy Switch receptors using il-9 signaling domains
KR20250112314A (ko) * 2022-01-10 2025-07-23 페노믹 에이아이 항-콜라겐 삼중 나선 반복 함유 1(cthrc1) 항체 및 이를 사용하는 방법
CN120225568A (zh) * 2022-12-27 2025-06-27 广州瑞风生物科技有限公司 一种融合蛋白及其应用
CN117402231B (zh) * 2023-12-14 2024-04-09 南方医科大学南方医院 一种自发传递il-21信号的受体及其应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103582699A (zh) * 2011-04-08 2014-02-12 贝勒医学院 使用嵌合细胞因子受体逆转肿瘤微环境的影响
CN105777911A (zh) * 2016-04-12 2016-07-20 上海优卡迪生物医药科技有限公司 抗bcma嵌合抗原受体、编码基因、重组表达载体及其构建方法和应用
WO2017173256A1 (en) * 2016-04-01 2017-10-05 Kite Pharma, Inc. Chimeric antigen and t cell receptors and methods of use

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MX370148B (es) * 2012-10-02 2019-12-03 Memorial Sloan Kettering Cancer Center Composiciones y su uso para inmunoterapia.
CN106279434B (zh) * 2014-02-24 2020-07-03 西比曼生物科技（上海）有限公司 工程化cd20靶向性的nkt细胞及其制备方法和应用
WO2016127257A1 (en) * 2015-02-12 2016-08-18 University Health Network Chimeric antigen receptors
WO2016134284A1 (en) 2015-02-19 2016-08-25 University Of Florida Research Foundation, Inc. Chimeric antigen receptors and uses thereof
IL315940A (en) * 2015-07-28 2024-11-01 Univ Pennsylvania Altered monocytes/macrophages expressing chimeric antigen receptors and their uses
GB201514875D0 (en) * 2015-08-20 2015-10-07 Autolus Ltd Receptor
GB201518816D0 (en) 2015-10-23 2015-12-09 Autolus Ltd Receptor
CN105949317B (zh) * 2016-04-12 2019-07-02 上海优卡迪生物医药科技有限公司 抗cd20嵌合抗原受体、编码基因、重组表达载体及其构建方法和应用
WO2017184901A1 (en) * 2016-04-20 2017-10-26 Fred Hutchinson Cancer Research Center Immunomodulatory il2r fusion proteins and uses thereof
CN106397608A (zh) * 2016-10-19 2017-02-15 中国医学科学院血液病医院（血液学研究所） Cd20特异性嵌合抗原受体及其应用
MA46862A (fr) 2016-11-17 2019-09-25 Bluebird Bio Inc Convertisseur de signaux tgf béta
WO2018103734A1 (zh) * 2016-12-09 2018-06-14 北京中捭生物科技有限公司 嵌合抗原受体及其用途和制备方法
CN107058234B (zh) * 2017-05-12 2019-09-13 常州市第一人民医院 Car.il-33-t及其制备与应用
CN107245106B (zh) * 2017-07-18 2020-06-12 深圳市免疫基因治疗研究院 一种基于cd10的嵌合抗原受体及其应用
CN107400168B (zh) * 2017-07-18 2020-02-07 深圳市免疫基因治疗研究院 一种基于cd117的嵌合抗原受体及其应用
CN107384870A (zh) * 2017-07-31 2017-11-24 时力生物科技（北京）有限公司 一种靶向pd‑l1嵌合抗原受体修饰的t淋巴细胞及其制备方法和应用
CN110615843B (zh) * 2018-06-20 2023-05-09 上海隆耀生物科技有限公司 一种包含第三信号受体的嵌合抗原受体及其应用

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103582699A (zh) * 2011-04-08 2014-02-12 贝勒医学院 使用嵌合细胞因子受体逆转肿瘤微环境的影响
WO2017173256A1 (en) * 2016-04-01 2017-10-05 Kite Pharma, Inc. Chimeric antigen and t cell receptors and methods of use
CN105777911A (zh) * 2016-04-12 2016-07-20 上海优卡迪生物医药科技有限公司 抗bcma嵌合抗原受体、编码基因、重组表达载体及其构建方法和应用

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3812401A4 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4083073A4 (en) * 2020-02-28 2023-07-12 Nanjing Bioheng Biotech Co., Ltd Novel chimeric antigen receptor and use thereof
US20240277843A1 (en) * 2020-02-28 2024-08-22 Nanjing Bioheng Biotech Co., Ltd Novel chimeric antigen receptor and use thereof
WO2021207150A1 (en) * 2020-04-06 2021-10-14 Celledit Llc Genetically modified immune cells expressing a chimeric antigen receptor and having reduced proinflammatory cytokine signaling
WO2025040094A1 (en) * 2023-08-21 2025-02-27 Hangzhou Hervor Therapeutics Co., Ltd Synthetic cytokine signaling system, preparation method, and uses thereof
WO2025039159A1 (en) * 2023-08-21 2025-02-27 Hangzhou Hervor Therapeutics Co., Ltd Synthetic cytokine signaling system, preparation method, and uses thereof

Also Published As

Publication number Publication date
CN114516922A (zh) 2022-05-20
CN110615843A (zh) 2019-12-27
CN112673025A (zh) 2021-04-16
KR102853978B1 (ko) 2025-09-04
EP3812401A1 (en) 2021-04-28
EP3812401A4 (en) 2022-04-20
JP7433656B2 (ja) 2024-02-20
CN112673025B (zh) 2022-04-01
KR20210023991A (ko) 2021-03-04
US11524034B2 (en) 2022-12-13
CN110615843B (zh) 2023-05-09
US20210169933A1 (en) 2021-06-10
JP2021529176A (ja) 2021-10-28

Similar Documents

Publication Publication Date Title
US11524034B2 (en) Chimeric antigen receptor comprising third signal receptor and use thereof
CN113087806B (zh) 靶向多种肿瘤的新型car-t细胞及其制备和方法
KR20200023165A (ko) 단일 도메인 항체에 기반한 bcma 키메라 항원 수용체 및 응용
WO2022007650A1 (zh) 靶向bcma和cd19的嵌合抗原受体car或car构建体及其应用
US20230134345A1 (en) Chimeric antigen receptor comprising third signal receptor and use thereof
CN110204619B (zh) 包含FcγRⅠ的嵌合抗原受体及其应用
CN109321530B (zh) 一种安全型嵌合抗原受体t细胞及其用途
CN116041546A (zh) 嵌合抗原受体、分离的核酸、重组载体、car-t细胞及其应用
Perera Molligoda Arachchige NK cell-based therapies for HIV infection: Investigating current advances and future possibilities
US11590168B2 (en) Chimeric antigen receptor comprising co-stimulatory receptor and application thereof
JP2023522112A (ja) 汎用型ヒト化ｃａｒ１９－ｄｎｔ細胞を製造する技術およびその使用
US20210108175A1 (en) Universal car-t cell and preparation method and use thereof
HK40066387A (zh) 一种包含第三信号受体的嵌合抗原受体及其应用
HK40042771B (en) Chimeric antigen receptor comprising third signal receptor and use thereof
HK40042771A (en) Chimeric antigen receptor comprising third signal receptor and use thereof
US20230277666A1 (en) Chimeric antigen receptor comprising co-stimulatory receptor and application thereof
HK40074947A (en) Chimeric antigen receptor comprising co-stimulatory receptor and application thereof
HK40042772A (en) Chimeric antigen receptor comprising co-stimulatory receptor and application thereof
HK40042772B (en) Chimeric antigen receptor comprising co-stimulatory receptor and application thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19823459

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2020571580

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 20217001094

Country of ref document: KR

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2019823459

Country of ref document: EP

Effective date: 20210120