WO2023000170A1 - Anticorps cd147 et cellules cd147-car-t - Google Patents

Anticorps cd147 et cellules cd147-car-t Download PDF

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WO2023000170A1
WO2023000170A1 PCT/CN2021/107395 CN2021107395W WO2023000170A1 WO 2023000170 A1 WO2023000170 A1 WO 2023000170A1 CN 2021107395 W CN2021107395 W CN 2021107395W WO 2023000170 A1 WO2023000170 A1 WO 2023000170A1
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car
cells
seq
amino acid
antibody
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Xiang Chen
Mingzhu YIN
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Central South University
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    • C07K2317/622—Single chain antibody (scFv)
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    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
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    • C12N2510/00—Genetically modified cells

Definitions

  • the present invention relates to CD147-specific antibodies and anti-CD147-CAR-T Cells, which are useful in the field of adoptive immunity gene therapy for tumors.
  • Immunotherapy is emerging as a highly promising approach for the treatment of cancer.
  • T cells or T lymphocytes the armed forces of our immune system, constantly look for foreign antigens and discriminate abnormal (cancer or infected cells) from normal cells.
  • Genetically modifying T cells with CAR (Chimeric antigen receptor) constructs is the most common approach to design tumor-specific T cells.
  • CAR-T cells targeting tumor-associated antigens TAA
  • TAA tumor-associated antigens
  • ACT adoptive cell transfer
  • the advantage of CAR-T technology compared with chemotherapy or antibody is that reprogrammed engineered T cells can proliferate and persist in the patient ( “aliving drug” ) [1, 2] .
  • CARs usually consist of a monoclonal antibody-derived single-chain variable fragment (scFv) at the N-terminal part, hinge, transmembrane domain, and a number of intracellular co-activation domains: (i) CD28, (ii) 4-1BB, CD27 or other co-stimulatory domains, in tandem with a activation CD3-zeta domain. ( Figure 1) [2, 3] .
  • the evolution of CARs went from first generation (with no costimulatory domains) to second generation (with one co-stimulation domain) to third generation CAR (with several costimulatory domains) .
  • Generating CARs with multiple costimulatory domains have led to increased cytolytic CAR-T cell activity, improved persistence of CAR-T cells leading to its augmented antitumor activity.
  • CD147 protein (also known by other names EMMPRIN, Basigin, M6, or Neurothelin) is a member of immunoglobulin superfamily and plays important role in development, cancer and neurological diseases (6) .
  • the CD147 gene or BSG (for Basigin) is located on chromosome 19p13.3 and encodes 27-29 kDa protein that can have higher molecular weight (35-69 kDa) depending on the level of post-translational glycosylation (6) . It has several isoforms which are generated by alternative splicing; for example: isoform 1, Long isoform (Basigin-2) (P35613-1) and Short isoform 2 or Basigin-1 (P35613-2) with missing 24-139 amino-acids.
  • CD147 The Long 42 kDa isoform is shown below with 1-21 aa-signaling peptide; 138-323-extracellular domain; 324-344 transmembrane domain; 345-385-cytoplasmic domain.
  • CD147 was shown to be overexpressed in several types of cancer and induced angiogenesis and metastasis (6) .
  • CD147 enhances the invasion and survival of cancer cells through either induction of MMPs, Ras, Raf, ERK1/2 and/or PI3K, AKT, vascular endothelial growth factor (VEGF) , integrin, cyclophilin or other cross-linked signal transduction pathways.
  • the CD147 protein is involved in angiogenesis, proliferation, invasion, survival, glucose metabolism, and drug resistance pathways.
  • CD147 The protein portion of CD147 is 28 kDa, but its high glycosylation increases its molecular weight to about 43-66 kDa. There are three Asn glycosylation sites in the extracellular region. The glycan portion differs and the glycosylation difference is responsible for various CD147 molecular weights depending on the protein source. CD147 has two Ig domains in the extracellular region. There are four known isoforms of Cd147 protein with longest isoform-1 is shown below with 1-21 signaling peptide; 138-323 extracellular domain (underlined) , 324-344 aa-transmembrane domain; and 345-385 aa-cytoplasmic domain ( Figure 2) .
  • CD147 The sequence of CD147 (Isoform 1 (identifier: P35613-1) is shown in Figure 2: 385 amino-acids, MW 42 kDa.
  • Transmembrane domain includes amino acids 324 to 344.
  • Extracellular domain includes 138-323 amino acid.
  • FIG. 1 The structures of CAR. On left, the structure of first generation (no costimulatory domains) , on the middle panel-second generation (one co-stimulatory domain CD28 or 4-BB) and on the right panel -third generation of CAR (two or several co-stimulation domains) are shown.
  • the Figure is from Golubovskaya, Wu, Cancers, 2016 [5] .
  • FIG. 1 The structure of CD147 CAR construct.
  • the second-generation CAR was used with either 41BB co-stimulatory domains.
  • FIGS 4A-4B Real-time RTCA assay demonstrated high killing activity of CD147-CAR cells (PMC909) .
  • Figure 4A SKOV-3 ovarian cancer target cell line.
  • Figure 4B A375 melanoma cancer target cell line. (#23) .
  • Figures 7A-7B In vivo efficacy study of CD147-CAR-T cells using SKOV-3 xenograft tumor model.
  • Figure 7A demonstrates xenograft tumor volume after administration of CAR-T cells. Significant tumor inhibition with elimination of tumors was caused by CD147-CAR-T cells (bottom curve) , but not by Mock CAR-T cells. *p ⁇ 0.0001, Student’s t-test, CAR-T cells versus Mock CAR-T cells.
  • a “chimeric antigen receptor (CAR)" is a receptor protein that has been engineered to give T cells the new ability to target a specific protein.
  • the receptor is chimeric because they combine both antigen-binding and T-cell activating functions into a single receptor.
  • CAR is a fused protein comprising an extracellular domain capable of binding to an antigen, a transmembrane domain, and at least one intracellular domain.
  • the "chimeric antigen receptor (CAR) " is sometimes called a "chimeric receptor” , a "T-body” , or a “chimeric immune receptor (CIR) .
  • the "extracellular domain capable of binding to an antigen” means any oligopeptide or polypeptide that can bind to a certain antigen.
  • the "intracellular domain” means any oligopeptide or polypeptide known to function as a domain that transmits a signal to cause activation or inhibition of a biological process in a cell.
  • a “domain” means one region in a polypeptide which is folded into a particular structure independently of other regions.
  • humanized antibodies are antibodies derived from non-human species whose protein sequences have been modified to increase their similarity to antibody variants produced naturally in humans. For example, after a mouse antibody is developed, the DNA coding for that antibody can be sequenced. The DNA sequence corresponding to the antibody CDRs can then be determined. The CDR sequences can be inserted into a construct containing the DNA for a human antibody variant to prepare humanized antibodies.
  • a “single chain variable fragment (scFv)" means a single chain polypeptide derived from an antibody which retains the ability to bind to an antigen.
  • An example of the scFv includes an antibody polypeptide which is formed by a recombinant DNA technique and in which Fv regions of immunoglobulin heavy chain (H chain) and light chain (L chain) fragments are linked via a spacer sequence.
  • H chain immunoglobulin heavy chain
  • L chain light chain
  • tumor antigen means a biological molecule having antigenicity, expression of which causes cancer.
  • the inventors have generated mouse anti-human monoclonal antibody specifically targeting CD147 antigen.
  • the inventors have produced CD147-CAR-T cells to target cancer cells overexpressing CD147 tumor antigen.
  • the CD147-CAR-T cells of the present invention have high cytotoxic activity against several cancer cell lines
  • the present invention is directed to a mouse monoclonal anti-human CD147 antibody or an antigen-binding fragment (e.g., Fab, (Fab) 2 , scFv) comprising V H having the amino acid of SEQ ID NO: 4 and V L having the amino acid of SEQ ID NO: 5.
  • Fab fragment of CD147 antibody
  • scFv antigen-binding fragment
  • the present invention is directed to a mouse monoclonal anti-human CD147 antibody or an antigen-binding fragment (e.g., Fab, (Fab) 2 , scFv) comprising V H having the amino acid of SEQ ID NO: 9 and V L having the amino acid of SEQ ID NO: 10.
  • Fab fragment of CD147 antibody
  • scFv antigen-binding fragment
  • the present invention is directed to a humanized anti-human CD147 antibody or an antigen-binding fragment (e.g., Fab, (Fab) 2 , scFv) comprising V H having the amino acid of SEQ ID NO: 25 and V L having the amino acid of SEQ ID NO: 28 or 29.
  • a humanized anti-human CD147 antibody or an antigen-binding fragment e.g., Fab, (Fab) 2 , scFv
  • V H having the amino acid of SEQ ID NO: 25
  • V L having the amino acid of SEQ ID NO: 28 or 29.
  • the anti-human CD147 antibody is a single-chain variable fragment (scFv) .
  • ScFv can be VH -linker-VL or VL-linker-VH.
  • the scFv has the SEQ ID NO: 3 or 8.
  • the present invention is also directed to a chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) against CD147 (the present invention) , (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.
  • the CD147 CAR structure is shown in FIG. 3.
  • the co-stimulatory domain is selected from the group consisting of CD28, 4-1BB, GITR, ICOS-1, CD27, OX-40 and DAP10.
  • a preferred the co-stimulatory domain is CD28.
  • a preferred activating domain is CD3 zeta (CD3 Z or CD3 ⁇ ) .
  • the transmembrane domain may be derived from a natural polypeptide, or may be artificially designed.
  • the transmembrane domain derived from a natural polypeptide can be obtained from any membrane-binding or transmembrane protein.
  • a transmembrane domain of a T cell receptor ⁇ or ⁇ chain, a CD3 zeta chain, CD28, CD3 ⁇ ., CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, ICOS, CD154, or a GITR can be used.
  • the artificially designed transmembrane domain is a polypeptide mainly comprising hydrophobic residues such as leucine and valine.
  • a triplet of phenylalanine, tryptophan and valine is found at each end of the synthetic transmembrane domain.
  • a short oligopeptide linker or a polypeptide linker for example, a linker having a length of 2 to 10 amino acids can be arranged between the transmembrane domain and the intracellular domain.
  • a linker sequence having a glycine-serine continuous sequence can be used.
  • the present invention provides a nucleic acid encoding the CD147-CAR.
  • the nucleic acid encoding the CAR can be prepared from an amino acid sequence of the specified CAR by a conventional method.
  • a base sequence encoding an amino acid sequence can be obtained from NCBI RefSeq IDs or accession numbers of GenBank for an amino acid sequence of each domain, and the nucleic acid of the present invention can be prepared using a standard molecular biological and/or chemical procedure. For example, based on the base sequence, a nucleic acid can be synthesized, and the nucleic acid of the present invention can be prepared by combining DNA fragments which are obtained from a cDNA library using a polymerase chain reaction (PCR) .
  • PCR polymerase chain reaction
  • a nucleic acid encoding the CAR of the present invention can be inserted into a vector, and the vector can be introduced into a cell.
  • a virus vector such as a retrovirus vector (including an oncoretrovirus vector, a lentivirus vector, and a pseudo type vector) , an adenovirus vector, an adeno-associated virus (AAV) vector, a simian virus vector, a vaccinia virus vector or a sendai virus vector, an Epstein-Barr virus (EBV) vector, and a HSV vector can be used.
  • a virus vector lacking the replicating ability so as not to self-replicate in an infected cell is preferably used.
  • a suitable packaging cell based on a LTR sequence and a packaging signal sequence possessed by the vector can be selected for preparing a retrovirus particle using the packaging cell.
  • the packaging cell include PG13 (ATCC CRL-10686) , PA317 (ATCC CRL-9078) , GP+E-86 and GP+envAm-12, and Psi-Crip.
  • a retrovirus particle can also be prepared using a 293 cell or a 293T cell having high transfection efficiency.
  • Many kinds of retrovirus vectors produced based on retroviruses and packaging cells that can be used for packaging of the retrovirus vectors are widely commercially available from many companies.
  • a CAR-T cell binds to a specific antigen via the CAR, thereby a signal is transmitted into the cell, and as a result, the cell is activated.
  • the activation of the cell expressing the CAR is varied depending on the kind of a host cell and an intracellular domain of the CAR, and can be confirmed based on, for example, release of a cytokine, improvement of a cell proliferation rate, change in a cell surface molecule, or the like as an index.
  • release of a cytotoxic cytokine (atumor necrosis factor, lymphotoxin, etc. ) from the activated cell causes destruction of a target cell expressing an antigen.
  • release of a cytokine or change in a cell surface molecule stimulates other immune cells, for example, a B cell, a dendritic cell, a NK cell, and a macrophage.
  • the cell expressing the CAR can be used as a therapeutic agent for a disease.
  • the therapeutic agent comprises the cell expressing the CAR as an active ingredient, and it may further comprise a suitable excipient.
  • CD147-ScFv-41BB-CD3-CAR-T CD147-CAR-T cells against cancer cells overexpressing CD147 protein.
  • CD147-CAR-T cells express higher cytotoxic activity against CD147-positive cancer cells than against non-transduced T cells and Mock-CAR-T cells.
  • CD147 monoclonal antibody or CD147-ScFv of the present invention over other known CD147 antibodies is that the present antibody has high binding activity to CD147 antigen, and it is highly specific against CD147-positive cancer cells.
  • the CD147 antibody is highly potent as a therapeutic agent in many clinical applications.
  • the present monoclonal mouse anti-human CD147 antibody detects CD147 in CD147-positive cancer cells.
  • the present CD147 antibody can be used for immunotherapy applications: toxin/drug-conjugated antibody, monoclonal therapeutic antibody, humanization of CD147 antibody, CAR-T cell immunotherapy
  • CD147-CAR-T cells using the present CD147 antibody can be effectively used to target CD147 antigen in CD147-positive cell lines: lymphoma, or leukemia.
  • CD147-CAR-T can be used in combination with different chemotherapy: checkpoint inhibitors; targeted therapies, small molecule inhibitors, antibodies
  • CD147 antibody can be modified with site-directed mutagenesis for affinity tuning; it can be used for humanization and for complete human antibody generation.
  • CD147-CAR-T cells can be used clinically for CD147-positive cells.
  • CD28, 4-1BB and others Modifications of co-activation domains: CD28, 4-1BB and others can be used to increase its efficacy.
  • Tag-conjugated CD147 scFv can be used for CAR generation
  • Third generation CAR-T or other co-activation signaling domains can be used for the same CD147-scFv inside CAR.
  • the present invention provides T cells, NK cells, macrophages, or hematopoietic cells, modified to express CD147-CAR.
  • Humanized CD147 can be used for generation of CD147-CAR.
  • CD47 with other CAR targeting other tumor antigens or tumor microenvironment (VEGFR-1-3) , PDL-1, CD80 or bi-specific scFv-CAR can be used to enhance activity of monotherapy CD147-CAR.
  • VEGFR-1-3 tumor antigens or tumor microenvironment
  • PDL-1, CD80 or bi-specific scFv-CAR can be used to enhance activity of monotherapy CD147-CAR.
  • Bi-specific antibodies with CD147 and CD3 or other antigens can be generated for therapy.
  • the CD147-CAR can be used for generating other types of cells such as CAR-natural killer cells, CD147-CAR-macrophages, and other cells.
  • Monoclonal antibodies were produced by the hybridoma method. Briefly, Balb/c mice were immunized with extracellular CD147 antigen which was expressed in E. coli and the antibody titers were subsequently evaluated by ELISA using the antigen. Once an acceptable titer was obtained, splenocytes from the treated mice producing the best response were hybridized with myeloma cells. Supernatants from ten hybridoma clones were evaluated by Western blotting against SDS-PAGE-separated proteins. From these screenings, several clones were chosen for antibody production, purification, and evaluation. Clones yielding the best responses were sub-cultured in vitro yielding a hybridoma with single antibody specificity.
  • the lentivirus was produced by the standard procedure using 293 cells as described in (5) . Expression of CAR was first confirmed by transducing 293 cells with lentivirus and performing FACS.
  • the PBMC were washed once with CAR-T media (AIM V-AlbuMAX (BSA) (Life Technologies) , with 5%AB serum and 1.25 ug/mL amphotericin B (Gemini Bioproducts, Woodland, CA) , 100 U/mL penicillin, and 100 ug/mL streptomycin) and were frozen at -80°C.
  • CAR-T media AIM V-AlbuMAX (BSA) (Life Technologies) , with 5%AB serum and 1.25 ug/mL amphotericin B (Gemini Bioproducts, Woodland, CA) , 100 U/mL penicillin, and 100 ug/mL streptomycin
  • PBMC cells were wash once in CAR-T medium, without huIL-2, before resuspending to a final concentration of 5x10 5 cells/mL in CAR-T medium with 300bU/mL huIL2 (from a 1000x stock; Invitrogen) .
  • PBMC were activated with CD3-CD28 beads which were mixed at a 1: 1 bead-to-cell ratio and incubated at 37°C in the presence of CO 2 for 24 hr before viral transduction.
  • PBMC cells were incubated for 24 hr at 37°C, 5%CO 2 .
  • 5x10 6 lentivirus, and 2 ⁇ L/mL of media of Transplus (Alstem, Richmond, CA) (afinal dilution of 1: 500) were added to 1x10 6 of T cells.
  • Cells were incubated for an additional 24 hours before repeating addition of virus.
  • Cells were then grown in the continued presence of 300 U/ml of IL-2. Fresh medium with IL-2 was used for a period of 12-14 days.
  • FACS buffer Phosphate-buffered saline (PBS) plus 0.1%sodium azide and 0.4%BSA
  • Fc receptors were blocked with normal goat Ig (LifeTechnologies) .
  • 100 ⁇ l of 1: 1000 diluted normal goat lgG was added to each tube and incubated in ice for 10 min. Then 1.0 ml FACS buffer was added to each tube, mixed well and spin down at 300g for 5 min.
  • Biotin-labeled polyclonal goat anti-mouse-F (ab) 2 antibodies were used to detect CD147 scFv; biotin-labeled normal polyclonal goat IgG antibodies (Life Technologies) used as an isotype control. (1: 200 dilution, reaction volume of 100 ⁇ l) . Cells were incubated at 4°C for 25 minutes and washed once with FACS buffer. The cells were stained with phycoerythrin (PE) -labeled streptavidin (BD Pharmingen, San Diego, CA) and allophycocyanin (APC) -labeled CD3 antibody (eBiocience, San Diego, CA) .
  • PE phycoerythrin
  • streptavidin BD Pharmingen, San Diego, CA
  • APC allophycocyanin
  • the cytotoxicity was performed using ACEA machine according to manufacturer’s protocol as described (7) .
  • Adherent target cells were seeded into 96-well E-plates (Acea Biosciences, San Diego, CA, USA) at 1 ⁇ 10 4 cells per well and monitored in culture overnight with the impedance-based real-time cell analysis (RTCA) xCELLigence system (Acea Biosciences) . The next day, the medium was removed and re-placed with AIM V-AlbuMAX medium containing 10%FBS ⁇ 1 ⁇ 105 effector cells (CAR-T cells or non-transduced T cells) in triplicate. The cells in the E-plates were monitored for another 24–48 h with the RTCA system, and impedance was plotted over time. Cytotoxicity was calculated as (impedance of target cells without effector cells-impedance of target cells with effector cells) ⁇ 100/impedance of target cells without effector cells.
  • RTCA real-time cell analysis
  • CD147 antibodies clone 2D3A9 (supernatant #23) and clone 2D8C11 (supernatant #26) .
  • the structure of CD147 scFv is: VH-linker-VL.
  • the mouse monoclonal CD147 antibody (#23, clone 2D3A9) is IgG1 type.
  • the nucleotide sequence of scFv is provided below. The bold highlights the nucleotide sequence of V H ; the underlined highlights the nucleotide sequence of V L , in between in italics font is the nucleotide sequence encoding a linker.
  • CD147 scFv #23 amino acid sequence (SEQ ID NO: 3) , VH in bold; VL underlined
  • linker amino sequence is 3xG4S (SEQ ID NO: 6) :
  • nucleotide sequence of scFv of antibody #26 is provided below.
  • the bold highlights the nucleotide sequence of V H ;
  • the underlined highlights the nucleotide sequence of V L , in between in italics font is the nucleotide sequence encoding a linker.
  • Nucleotide sequence CD147 scFv #26 (SEQ ID NO: 7) .
  • CD147 scFv #26 (SEQ ID NO: 8) , VH is shown in bold font; VL underlined
  • the linker amino sequence is 3xG4S (SEQ ID NO: 6) .
  • Example 10A Mouse CD147-CAR Sequences (PMC909, Antibody #23)
  • CD147-CAR (PMC909) construct The scheme of CD147-CAR (PMC909) construct is shown on Figure 3.
  • Lentiviral vector Lentiviral vector with MNDU3 promoter was used for cloning of scFv CAR sequences.
  • the following nucleotide sequence shows CD147 ScFv-CD8 hinge-TM28-41BB-CD3 zeta of the present invention.
  • the structure includes Human CD8 signaling peptide, CD147 scFv (V H -Linker 3x (G4S) -V L ) , CD8 hinge, CD28 transmembrane, 41BB costimulatory and CD3 zeta activation domains ( Figure 3) .
  • VH is shown in bold, VL underlined
  • Example 10B Mouse CD147-CAR Sequences (PMC896, Antibody #26)
  • Mouse CD147 CAR construct (PMC896) was designed the same as Example 10A except with scFv of antibody #26 (2D8C11) .
  • the antibody CD147, clone 2D3A9 detected extracellular CD147 protein by ELISA (OD 450 nm reading with CD147 protein was 2.23, while with negative unrelated control protein was 0.081) .
  • the two other antibodies CD147, clone #2D8C11 had binding by ELISA with CD147 antigen of 1.8, and with negative control antigen of 0.168; and clone 2D8A9 (supernatant #25) had binding by ELISA with CD147 antigen of 2.19, and with negative antigen of 0.14.
  • the antibody detects extracellular domain, IgG1 type.
  • the affinity with KD of binding of antibody #23 was detected by Biacore equal to 2 nM.
  • Example 14 The CD147-CAR-T cells express CD147 ScFv
  • CD147 antibody and generated scFv of CD147 antibody that is shown in Example 9.
  • the CD147 scFv sequence was inserted with 41BB and CD3 zeta domains inside CAR and lentiviral CAR were transduced into T cells.
  • the CD147-CAR cells (PMC909, Example 10A) were effectively expanded in vitro (not shown) .
  • Mock control CAR-T cells with no scFv and with 3xTF tag were used as a negative control in cytotoxicity and cytokine assay.
  • the CD147-CAR-positive cells were detected by FACS with mouse anti-FAB antibody (99.3%positive) , which detects extracellular ScFv CAR domain.
  • Mock control CAR-T cells (with TF tag and without scFv) were detected with TF tag antibody.
  • Example 15 Mouse CD147-CAR-T cells (PMC909) expressed high cytotoxic activity against CD147-positive cells
  • the RTCA cytotoxicity assay was performed using human ovarian SKOV-3 cancer cells (Figure 4A) and human melanoma A375 cancer cells (Figure 4B) as target cells with mouse CD147-41BB-CD3-CAR-T cells PMC909. CD147-CAR-T cells (PMC909) had high killing activity with both target cell lines ( Figures 4A and 4B) .
  • Example 16 Mouse CD147-CAR-T cells (PMC896) expressed low cytotoxic activity against CD147-positive cells
  • CAR-T cells (PMC896) with scFv of clone #26 that had high FACS staining with CD147 antibody did not have high RTCA activity with SKOV-3 cells ( Figure 5) and HT29, HCT116 target cells (not shown) .
  • CD147h1 hVH1-linker-hVL1
  • Example 18 Mouse CD147-CAR (PMC909) secreted high level of IFN-gamma against CD147-positive cancer cells.
  • FIG. 7A The in vivo efficacy of mouse CD147-CAR-T cells (PMC909) is shown on Figure 7A.
  • 4x10 6 SKOV-3 cells were injected subcutaneously into mouse leg flank, and when tumors reached palpable tumor volume, 1x10 7 CAR-T cells were injected intravenously into tail vein.
  • CD147-CAR-T cells completely blocked SKOV-3 xenograft tumor volume compared with Mock CAR-T cells (Figure 7A) .
  • Figure 7A shows average tumor volume of untreated, mock CAR-T-cells-treated, and CD147 CAR-T cells-treated (from top to bottom) .
  • There was no visible toxicity after CAR-T cell injection as no mouse body weight loss was observed by CD147-CAR-T cells ( Figure 7B) .
  • T cells expressing chimeric antigen receptors can cause anaphylaxis in humans. Cancer Immunol Res 1, 26-31.

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Abstract

L'invention concerne un anticorps monoclonal anti-CD147 humain de souris ou un fragment variable à chaîne unique (scFv), comprenant VH ayant l'acide aminé de SEQ ID NO : 4 et VL ayant l'acide aminé de SEQ ID NO : 5, ou VH ayant l'acide aminé de SEQ ID NO : 9 et VL ayant l'acide aminé de SEQ ID NO : 10. L'invention concerne également un anticorps anti-CD147 humain humanisé ou un fragment variable à chaîne unique (scFv), comprenant VH ayant l'acide aminé de SEQ ID NO : 25 et VL ayant l'acide aminé de SEQ ID NO : 28 ou 29. L'invention concerne en outre une protéine de fusion d'un récepteur antigénique chimérique comprenant de l'extrémité N-terminale à l'extrémité C-terminale : (i) un fragment variable de chaîne unique (scFv) selon la présente invention, (ii) un domaine transmembranaire, (iii) au moins un domaine costimulateur, et (iv) un domaine d'activation.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010036460A2 (fr) * 2008-09-29 2010-04-01 Centocor Ortho Biotech Inc. Anticorps anti-cd147, procédés, et utilisations
CN104086654A (zh) * 2014-07-04 2014-10-08 中国人民解放军第四军医大学 人源化修饰型抗CD147嵌合抗体HcHAb18及其应用
CN110746509A (zh) * 2019-10-10 2020-02-04 中国人民解放军第四军医大学 一种抗人cd147 car-t细胞、制备方法和应用
US20200362033A1 (en) * 2017-07-27 2020-11-19 Daiichi Sankyo Company, Limited Anti-cd147 antibody

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010036460A2 (fr) * 2008-09-29 2010-04-01 Centocor Ortho Biotech Inc. Anticorps anti-cd147, procédés, et utilisations
CN104086654A (zh) * 2014-07-04 2014-10-08 中国人民解放军第四军医大学 人源化修饰型抗CD147嵌合抗体HcHAb18及其应用
US20200362033A1 (en) * 2017-07-27 2020-11-19 Daiichi Sankyo Company, Limited Anti-cd147 antibody
CN110746509A (zh) * 2019-10-10 2020-02-04 中国人民解放军第四军医大学 一种抗人cd147 car-t细胞、制备方法和应用

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Title
LIU, YAN; CHEN, XIUXIU; WANG, YUXIAO; DING, LI; FU, KAIFEI: "Screening and biological characteristics identification of human anti-CD147 single chain antibody fragments", IMMUNOLOGICAL JOURNAL, DI-3 JUNYI DAXUE, CHINA, vol. 34, no. 1, 1 January 2018 (2018-01-01), China , pages 59 - 64, XP009542746, ISSN: 1000-8861, DOI: 10.13431/j.cnki.immunol.j.20180009 *

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