CN113862292A - 嵌合抗原受体、组合物以及方法 - Google Patents
嵌合抗原受体、组合物以及方法 Download PDFInfo
- Publication number
- CN113862292A CN113862292A CN202111106136.7A CN202111106136A CN113862292A CN 113862292 A CN113862292 A CN 113862292A CN 202111106136 A CN202111106136 A CN 202111106136A CN 113862292 A CN113862292 A CN 113862292A
- Authority
- CN
- China
- Prior art keywords
- cells
- cell
- chimeric antigen
- polynucleotide
- icd
- 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.)
- Pending
Links
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 title claims abstract description 94
- 238000000034 method Methods 0.000 title claims abstract description 20
- 239000000203 mixture Substances 0.000 title description 13
- 210000000822 natural killer cell Anatomy 0.000 claims abstract description 63
- 239000000427 antigen Substances 0.000 claims abstract description 33
- 102000036639 antigens Human genes 0.000 claims abstract description 33
- 108091007433 antigens Proteins 0.000 claims abstract description 33
- 230000003834 intracellular effect Effects 0.000 claims abstract description 19
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 13
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 9
- 230000019491 signal transduction Effects 0.000 claims abstract description 4
- 230000011664 signaling Effects 0.000 claims description 15
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 12
- 108091033319 polynucleotide Proteins 0.000 claims description 11
- 239000002157 polynucleotide Substances 0.000 claims description 11
- 102000040430 polynucleotide Human genes 0.000 claims description 11
- 206010028980 Neoplasm Diseases 0.000 claims description 10
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 8
- 125000006850 spacer group Chemical group 0.000 claims description 8
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 claims description 5
- 101000809875 Homo sapiens TYRO protein tyrosine kinase-binding protein Proteins 0.000 claims description 5
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 claims description 5
- 102100038717 TYRO protein tyrosine kinase-binding protein Human genes 0.000 claims description 5
- 101000851370 Homo sapiens Tumor necrosis factor receptor superfamily member 9 Proteins 0.000 claims description 4
- 102100036856 Tumor necrosis factor receptor superfamily member 9 Human genes 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 229920001184 polypeptide Polymers 0.000 claims description 4
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 4
- 201000010099 disease Diseases 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 238000009169 immunotherapy Methods 0.000 claims description 3
- 108010017411 Interleukin-21 Receptors Proteins 0.000 claims description 2
- 102000004527 Interleukin-21 Receptors Human genes 0.000 claims 1
- 210000004899 c-terminal region Anatomy 0.000 claims 1
- 239000003814 drug Substances 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 48
- 102000003735 Mesothelin Human genes 0.000 description 16
- 108090000015 Mesothelin Proteins 0.000 description 16
- 102000005962 receptors Human genes 0.000 description 11
- 108020003175 receptors Proteins 0.000 description 11
- 230000003013 cytotoxicity Effects 0.000 description 9
- 231100000135 cytotoxicity Toxicity 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 8
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 231100000433 cytotoxic Toxicity 0.000 description 5
- 230000001472 cytotoxic effect Effects 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- 102100029360 Hematopoietic cell signal transducer Human genes 0.000 description 4
- 101000990188 Homo sapiens Hematopoietic cell signal transducer Proteins 0.000 description 4
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 4
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 4
- 101000589305 Homo sapiens Natural cytotoxicity triggering receptor 2 Proteins 0.000 description 4
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 4
- 108010004217 Natural Cytotoxicity Triggering Receptor 1 Proteins 0.000 description 4
- 102100032870 Natural cytotoxicity triggering receptor 1 Human genes 0.000 description 4
- 102100032851 Natural cytotoxicity triggering receptor 2 Human genes 0.000 description 4
- 206010033128 Ovarian cancer Diseases 0.000 description 4
- 206010061535 Ovarian neoplasm Diseases 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000002147 killing effect Effects 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 101100273212 Arabidopsis thaliana CAR7 gene Proteins 0.000 description 2
- 241000701022 Cytomegalovirus Species 0.000 description 2
- 101710112752 Cytotoxin Proteins 0.000 description 2
- 101000971538 Homo sapiens Killer cell lectin-like receptor subfamily F member 1 Proteins 0.000 description 2
- 101001023379 Homo sapiens Lysosome-associated membrane glycoprotein 1 Proteins 0.000 description 2
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 241000701806 Human papillomavirus Species 0.000 description 2
- 102100030704 Interleukin-21 Human genes 0.000 description 2
- 108020004684 Internal Ribosome Entry Sites Proteins 0.000 description 2
- 102100021458 Killer cell lectin-like receptor subfamily F member 1 Human genes 0.000 description 2
- 102100035133 Lysosome-associated membrane glycoprotein 1 Human genes 0.000 description 2
- 101100005008 Mus musculus Ca7 gene Proteins 0.000 description 2
- 230000006051 NK cell activation Effects 0.000 description 2
- 101100439689 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) chs-4 gene Proteins 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 208000009956 adenocarcinoma Diseases 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000022534 cell killing Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000016396 cytokine production Effects 0.000 description 2
- 231100000599 cytotoxic agent Toxicity 0.000 description 2
- 239000002619 cytotoxin Substances 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 208000006454 hepatitis Diseases 0.000 description 2
- 231100000283 hepatitis Toxicity 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000012212 insulator Substances 0.000 description 2
- 108010074108 interleukin-21 Proteins 0.000 description 2
- 230000036210 malignancy Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011325 microbead Substances 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000003151 transfection method Methods 0.000 description 2
- 239000003981 vehicle Substances 0.000 description 2
- 101100273214 Arabidopsis thaliana CAR9 gene Proteins 0.000 description 1
- 108010031480 Artificial Receptors Proteins 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 102100038077 CD226 antigen Human genes 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102100024644 Carbonic anhydrase 4 Human genes 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000884298 Homo sapiens CD226 antigen Proteins 0.000 description 1
- 101000760567 Homo sapiens Carbonic anhydrase 4 Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102100030699 Interleukin-21 receptor Human genes 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 206010027406 Mesothelioma Diseases 0.000 description 1
- 101100165851 Mus musculus Ca9 gene Proteins 0.000 description 1
- 108010004222 Natural Cytotoxicity Triggering Receptor 3 Proteins 0.000 description 1
- 102100032852 Natural cytotoxicity triggering receptor 3 Human genes 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 240000007019 Oxalis corniculata Species 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 108010092262 T-Cell Antigen Receptors Proteins 0.000 description 1
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 1
- 102000008579 Transposases Human genes 0.000 description 1
- 108010020764 Transposases Proteins 0.000 description 1
- 108010084455 Zeocin Proteins 0.000 description 1
- 239000003070 absorption delaying agent Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 108091005764 adaptor proteins Proteins 0.000 description 1
- 102000035181 adaptor proteins Human genes 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000000139 costimulatory effect Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 230000001024 immunotherapeutic effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000010212 intracellular staining Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- CWCMIVBLVUHDHK-ZSNHEYEWSA-N phleomycin D1 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC[C@@H](N=1)C=1SC=C(N=1)C(=O)NCCCCNC(N)=N)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C CWCMIVBLVUHDHK-ZSNHEYEWSA-N 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- 231100000588 tumorigenic Toxicity 0.000 description 1
- 230000000381 tumorigenic effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
-
- 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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4613—Natural-killer cells [NK or NK-T]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/463—Cellular immunotherapy characterised by recombinant expression
- A61K39/4631—Chimeric Antigen Receptors [CAR]
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464402—Receptors, cell surface antigens or cell surface determinants
-
- 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
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
- A61K39/464466—Adhesion molecules, e.g. NRCAM, EpCAM or cadherins
- A61K39/464468—Mesothelin [MSLN]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4747—Apoptosis related proteins
-
- 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
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
-
- 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
-
- 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
-
- 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
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour 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
- 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
- C12N5/0646—Natural killers cells [NK], NKT 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
- 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
- C12N5/0696—Artificially induced pluripotent stem cells, e.g. iPS
-
- 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
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/31—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterized by the route of administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/38—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the dose, timing or administration schedule
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
-
- 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
- C07K2319/33—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
- C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
-
- 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
Abstract
本公开描述了用于在自然杀伤(NK)细胞中表达的嵌合抗原受体,包括经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)的药物组合物,以及涉及这些嵌合抗原受体的方法。通常,所述嵌合抗原受体包括包含抗原识别区的胞外结构域、与该胞外结构域连接的跨膜结构域,以及与该跨膜结构域连接的胞内结构域。所述胞内结构域可包括激活NK细胞的信号传导肽。
Description
相关申请的交叉引用
本申请是申请号为201680008013.0的中国专利申请的分案申请。本申请要求2015年1月29日提交的美国临时专利申请62/109,281的的优先权,通过引用将其并入本文。
发明内容
在一个方面,本公开描述了用于在自然杀伤(NK)细胞中表达的嵌合抗原受体。通常,所述嵌合抗原受体包括包含抗原识别区的胞外结构域、连接至所述胞外结构域的跨膜结构域,以及连接至所述跨膜结构域的胞内结构域。所述胞内结构域可包含激活NK细胞的信号传导肽(signaling peptide)。
在一些实施方式中,所述抗原识别结构域可特异性地结合与疾病相关的抗原。
在一些实施方式中,所述抗原识别结构域可特异性地结合肿瘤抗原。
在一些实施方式中,所述胞外结构域还可包括信号肽(signal peptide)或前导序列(leader sequence)和/或间隔序列(spacer)。
在一些实施方式中,所述胞内结构域可包括诸如2B4、DAP10、DAP12、IL21R、CD137(41BB)或CD3ζ的NK细胞膜结合信号接头蛋白(NK cell membrane-bound signalingadaptor protein)的信号传导结构域(signaling domain)。
在一些实施方式中,所述跨膜结构域可包括诸如CD16、NKp44、NKp46或NKG2D的在NK细胞中表达的自然细胞毒性受体(natural cytotoxicity receptor)的跨膜区。
在另一方面,本公开描述了包括被修饰以便表达上文总结的嵌合抗原受体的任何实施方式的NK细胞(和/或iPSC)的药物组合物。
在另一方面,本公开描述了向有病况(condition)的受试者提供免疫治疗的方法。通常,该方法包括将上文总结的治疗性组合物施用于该受试者,其中所述嵌合抗原受体的抗原识别区特异性地结合与该病况相关的抗原。
以上的总结无意描述本发明的每个公开的实施方式或者每种实践。以下的说明书更具体地例举了阐释性实施方式。在该申请的若干处,通过列出实施例来提供指导,这些实施例可以以各种组合使用。每种情形下,所给出的列表仅用作代表性组,而不应解释为排他性列表。
附图简要说明
图1.(A)示例性自然细胞毒性受体以及这些受体与它们配体的结合对NK细胞脱颗粒和极化以及靶细胞杀伤的影响。(B)自然细胞毒性受体的代表性实例和它们相应的信号接头。(C)用在iPSC衍生的NK细胞中的第三代嵌合T细胞抗原受体构建体。
图2.(A)NK激活CAR(NK-activating CAR)的一般化示意图。(B)新型嵌合抗原受体构建体的示意图。将嵌合抗原受体克隆进pkt2载体,该载体含有与SB100X转座酶一起使用的IR/DR、mCAGs启动子、嵌合抗原受体(CAR)序列、内部核糖体进入位点(IRES),以及GFP:Zeo选择标记。嵌合抗原受体片段从UniProt获得,并通过gBlock合成和传统限制酶克隆(IDT)进行组装。
图3.在NK92和iPS细胞中嵌合抗原受体的表面表达。采用SB100X使用SleepingBeaut转座子系统转染NK92细胞或iPS细胞。然后使用博莱霉素(Zeocin)筛选细胞,并进行流式细胞分析以评估各种嵌合抗原受体的细胞表面表达。采用生物素偶联的识别小鼠IgGF(ab’)2片段的多克隆山羊抗小鼠抗体来评估表达(Jackson ImmunoResearchLaboratories,Inc.,West Grove,PA,cat#115-065-072)。采用与荧光染料偶联的链霉亲和素来检测所结合的抗体。
图4.NK92细胞中CD107A的释放和IFN-γ的产生。NK92细胞脱颗粒和细胞因子生成采用流式细胞分析评价。将NK92细胞与间皮素(mesothelin)阴性(MA148)、间皮素阳性(A1847)卵巢癌靶细胞,或者与间皮素/Fc嵌合蛋白偶联或未偶联的蛋白A微珠以1:1混合。将细胞针对CD107a染色,针对IFN-γ的产生进行细胞内染色。
图5.采用NK92细胞的Cr-51释放测定。将NK92、NK92/28/41BB/CD3或NK92/CAR4细胞与K562、K562间皮素+、MA148或A1847细胞以标明的比例一起温育4小时。然后检测Cr-51的释放以评估细胞杀伤。本实验依照Woll等,2009,Blood113(24):6094-6101进行,除了用iPSC衍生的NK细胞替换hESC衍生的NK细胞。
图6.示例性的其他NK激活嵌合抗原受体。
图7.比较第三代T细胞CAR和图2中反映的示例性NK CAR构建体的示意图。
图8.示例性NK CAR构建体的示意图。
图9.示例性NK CAR构建体的示意图。
图10.显示NK CAR对K562细胞的细胞毒性的数据。
图11.显示NK CAR对两种卵巢癌细胞系的细胞毒性的数据。
图12.显示通过诱导多能干细胞表达示例性NK CAR的数据。
图13.显示通过iPSC衍生的NK细胞表达而进行的示例性CAR的表面表达的数据。
图14.示例性的一般化的NK CAR载体构建体。
图15.示例性的一般化的NK CAR载体构建体。串联cHS4绝缘子可抑制CAR载体沉默,由此改善该CAR在NK细胞和iPSC中的表达。
具体实施方式
本公开描述了经设计特异性地并入NK细胞激活结构域的嵌合抗原受体。嵌合抗原受体可并入细胞内区和/或跨膜区,包括例如与来自例如2B4(CD244)、CD137(41BB)、IL21、DAP10、DAP12和/或CD3ζ的共刺激或信号传导结构域相连接的来自CD16、NKp44、NKp46和/或NKG2D的细胞内区和/或跨膜区。
嵌合抗原受体(CAR)为工程化人工受体,其可为表达该CAR的免疫细胞提供工程化特异性。通常,可从患有特定形式癌症的受试者收集免疫细胞群。所收集的免疫细胞可经修饰以表达与肿瘤细胞表达的抗原特异性地结合的嵌合抗原受体,然后回输进该受试者。表达嵌合抗原受体的经修饰的免疫细胞能够更好地识别并杀灭表达被该嵌合抗原受体特异性地识别的抗原的肿瘤细胞。
嵌合抗原受体已被设计来激活T细胞,用于处理难治性ALL(靶向CD19)、胰腺癌(靶向间皮素),以及其它恶性肿瘤。存在几种嵌合抗原受体构建体,但多数是设计来激活T细胞。
相对而言,本文描述的嵌合抗原受体经设计为在诱导性多功能干细胞(iPSC)中表达,这些细胞然后分化为NK细胞。它们也可直接表达进外周血(PB)-NK细胞、NK-92细胞,或另外的合适NK细胞系中。NK-92细胞或其它NK细胞系已经用在临床研究中用于抗癌治疗。表达嵌合抗原受体的NK细胞接着可用作免疫治疗用于处理多种癌症。本文描述的嵌合抗原受体可包括可与各种靶向抗体的抗原识别部分一起使用的信号传导结构域。具体地,本公开描述了反映靶向间皮素的嵌合抗原受体用于治疗卵巢癌的示例性实施方式。但是,由于间皮素在很多腺癌上表达,所描述的实施方式可具有更广泛的用途。此外,所描述的间皮素靶向结构域仅仅是示例性的;其它的单链可变片段(scFV)可被工程化进该NK特异性的嵌合抗原受体(NK-CAR)信号构建体中以靶向基本上任何恶性肿瘤。
本文描述的NK细胞嵌合抗原受体的一个特征为可绕过自然细胞毒性受体(natural cytotoxicity receptor)启动信号转导所需的接头分子/辅助蛋白(adaptormolecules/accessory proteins)。另外一种方式或除此之外,包括有通常与接头分子/辅助蛋白结合的跨膜结构域使得辅助蛋白也能进行结合,让信号转导更可能启动。设计为包括例如CD3ζ的NK细胞嵌合抗原受体使得能够绕过其它的自然细胞毒性受体。并入跨膜结构域和其它的细胞内结构域可使得这些NK细胞嵌合抗原受体能够与接头蛋白结合并通过激活多种途径而提供比单独CD3ζ更佳的信号传导。
尽管一些T细胞嵌合抗原受体构建体由于共享的信号传导结构域而可在一定程度上激活NK细胞,但是本文描述的嵌合抗原受体是特异性地设计为激活NK细胞。经设计为特异性地激活NK细胞的嵌合抗原受体可在NK细胞免疫疗法(例如,细胞介导的难治性肿瘤杀灭)中改善NK功能和受体用途。
嵌合抗原受体典型地包括胞外结构域、跨膜结构域以及胞内结构域。所述胞内结构域通常驻留在细胞的细胞质中。一旦抗原被胞外结构域所识别,该胞内结构域就将激活信号传递至NK细胞,这诱导NK细胞破坏所靶向的肿瘤细胞。示例性信号传导胞内结构域包括例如膜结合信号接头蛋白(包括例如,2B4(CD244)、CD137(41BB)、IL21、DAP10、DAP12和/或CD3ζ)的信号传导结构域,或其部分,包括例如免疫受体酪氨酸活化基序(ITAM)、YxxM基序、TxYxxV/I基序、FcRγ、NKp80(通过非典型hemi-ITAM信号传导)和/或DNAM等。
跨膜结构域横跨质膜,将胞内结构域连接至胞外结构域。示例性跨膜结构域例如包括自然细胞毒性受体(NCR)(例如包括CD16、NKp44、NKp46、NKG2D、NKp30、NKp80和/或DNAM-1)的细胞内和/或跨膜结构域,或其包括例如一个或更多个带电荷氨基酸的部分。在一些实施方式中,该带电荷的氨基酸可以是赖氨酸和/或精氨酸残基。在一些情况下,跨膜区可以来自跨膜蛋白,意味着其天然具有细胞外C末端,而不是细胞外N末端。这种情况下,可反转跨膜区的方向,例如在图6中以“Rev TM”标明的,以便嵌合抗原受体在NK细胞膜中正确定向。
胞外结构域通常包括信号肽和抗原识别区。在很多实施方式中,胞外结构域还可包括间隔序列(spacer)。信号肽将新生多肽引导进内质网,以便其可被正确糖基化并锚定进质膜。一般来讲,可使用任何真核细胞信号肽,只要其引导蛋白至内质网。一个示例性信号肽包括CD8α前导序列,但其它的信号肽序列也可能是适合的。当间隔序列存在时,其将抗原识别结构域连接至跨膜结构域。该间隔序列通常提供柔韧性,从而抗原识别区可自由定向至不同方向,由此使得抗原识别区能够结合抗原靶。一个示例性间隔序列包括CD8α铰链序列,但其它的Ig铰链区也可能是适合的。抗原识别区可包括能够特异性地结合指定靶的任何肽序列。如本文所用,“特异性地结合”及其变型指对特定靶具有任何程度的差异化的或非一般的亲和力。因此,抗原识别区可包括特异性地结合特定抗原(例如,肿瘤抗原、病毒抗原、经修饰的自身抗原等)的抗体片段,例如scFv或Fab。在一些实施方式中,scFv可来自单克隆抗体。嵌合抗原受体可设计为包括可特异性地结合任何指定靶的抗原识别区。因此,尽管图2、图6、图7和图8显示了设计为特异性地结合间皮素的实施方式,但是NK激活嵌合抗原受体可被设计为特异性地结合并由此靶向与准备作为NK细胞介导杀伤标靶的细胞(例如致瘤细胞或病毒感染细胞)相关的任何抗原。例如,NK细胞和/或CAR已显示出针对各种各样的实体瘤和病毒感染细胞的活性,包括但不限于,HIV(人免疫缺陷病毒)、乙型肝炎、丙型肝炎、CMV(巨细胞病毒)、EBV(Epstein-Barr病毒)、HPV(人乳头瘤病毒),等等。
于是,例如,为了更好地介导针对包括表达间皮素的细胞的肿瘤(例如,卵巢癌、胰腺癌、肺癌、结肠腺癌、间皮瘤,以及其它表达间皮素的腺癌)的NK细胞毒性,可以设计并在NK细胞中表达例如图2、图6、图7和图8中显示的嵌合抗原受体。所展示的嵌合抗原受体构建体含有NK细胞特异性跨膜结构域和激活结构域(activating domain),并能够在NK细胞肿瘤系NK92中表达。跨膜区和细胞内区取自CD16、NKp44、NKp46和/或NKG2D,同时以试图最大化激活NK细胞的方式组合2B4、DAP10、DAP12和/或CD3ζ的激活结构域。图3显示NK92和iPS细胞表达图2中显示的嵌合抗原受体。
为了评估嵌合抗原受体的功能,针对抗原包覆微珠和间皮素表达细胞系测试了表达该嵌合抗原受体的NK细胞。图4显示,当表达嵌合抗原受体的NK细胞与间皮素阳性靶混合时,图2的嵌合抗原受体增强了NK细胞的脱颗粒和细胞因子生成。
图5显示,与第三代T细胞特异嵌合抗原受体(NK92/28/41BB/CD3ζ)或非转染NK92细胞相比,表达如本文描述的NK特异性嵌合抗原受体的NK92细胞改善了对间皮素阳性靶细胞的体外杀伤。
图10显示了示例性NK CAR对K562细胞(左上插图)和作为该CAR的标靶的表达间皮素的K562细胞(右上插图)的细胞毒性。这些结果显示了以间皮素特异性方式明显改善了杀伤,以CAR7和CAR9最为显著。下方的插图为对以溶解单位表示的结果的总结(Bryant等,1992,J Immunol Methods 146(1):91-103)。CAR7和CAR9显示了比以前的研究中使用的第三代T细胞CAR(NK92 meso 3rd)明显更高的细胞毒性。按之前描述的Cr-51释放测定法测量细胞毒性(Knorr等,2013,Stem Cells Transl Med 2(4):274-283;Woll等,2009,Blood113(24):3094-6101;Woll等,2005,J Immunol 175(8):5095-5103)。图11显示了采用两种卵巢癌细胞系meso-高(A1497)和meso-低(MA148)的类似结果。具有不同的基于NK细胞的抗meso CAR的NK92以meso特异性的方式杀伤。底部插图也反映了以溶解单位表示的总结。此外,当以如图10和图11中的标靶刺激时,NK CAR介导了增强的CD107a和/或IFN-γ表达(数据未示出)。
在一些实施方式中,如本文描述的NK特异的嵌合抗原受体可在iPSC中表达,这些iPSC可随后分化成NK细胞。可以按Knorr等,2013,Stem Cells Transl Med.2(4):274-283或Ni等,2014,Stem Cells 32(4):1021-1031中所描述分化iPSC。图12显示通过诱导多能干细胞(iPSCs)表达示例性NK CAR,如通过CD45+CD56+细胞的产生所显示的(首行,第5插图)。图13显示通过iPSC衍生的NK细胞进行的CAR表面表达。图13的底部三行显示,与不表达CAR的PB-NK细胞和iPSC-NK细胞(未修饰的对照细胞,第4列,第4和第5行)相比,仅在iPSC-CAR4v2(第4列,底部三行)表面上的CAR表达。图12和13中的其它插图显示iPSC-CAR4v2上的其它典型NK细胞表面抗原/受体与存在于未修饰对照细胞上的那些相类似。这些结果表明,iPSC衍生的NK细胞可展示出与图10和图11中的靶向间皮素的CAR表达NK细胞(mesothelin-targeted-CAR-expressing NK cells)相同的靶特异性细胞毒性。
可以采用常规转染方法将编码NK CAR构建体的多核苷酸引入NK细胞或iPSC。由此,尽管本文是在采用Sleeping Beauty转座子系统将编码CAR的多核苷酸转染进细胞的示例性实施方式情形下描述,但可以采用任何适合的转染方法修饰NK细胞(和/或iPSC)。图14显示了可以用于修饰NK细胞(和/或iPSCs)以表达嵌合抗原受体的示例性载体构建体(vector construct)。图15显示了备选的示例性载体的构建,该载体还包括串联cHS4绝缘子(Aker等,2007,Hum Gene Ther 18(4):333-343),其可抑制CAR载体沉默,由此改善该CAR在NK细胞和iPSC中的表达。
本文描述的经修饰以表达嵌合抗原受体的NK细胞和/或iPSs可以与“载体(carrier)”一起配制成药物组合物,用于递送至受试者,该受试者具有至少部分地以细胞可以是NK细胞毒性的标靶为特征的病况。如本文所用,“载体”包括任何溶剂、分散介质、媒介、包衣、稀释剂、抗菌剂和/或抗真菌剂、等渗剂、吸收延缓剂、缓冲剂、载体溶液、混悬剂、胶体等。将这些介质和/或试剂用于药学上有活性的物质在本领域内是公知的。除了任何常规媒介或试剂与该活性成分不相容之外,考虑将该活性成分用在治疗性组合物中。补充性活性成分也可掺入到该组合物中。
“药学上可接受的”指并非生物学上或其它方面不利的物质,即,该物质可以与经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)一起施用于个体,而不引起任何不希望的生物效果或不与其所在的药物组合物中的任何其它组分以有害方式相互作用。
该药物组合物可以配制为适合于优选施用途径的各种形式。由此,组合物可以通过已知途径施用,例如包括胃肠外(例如,皮内、经皮、皮下、肌内、静脉内、腹膜内等等)或局部施用(例如,气管内、肺内等等)。组合物也可通过持续或延缓释放施用。
制剂可以方便地以单位剂型呈现,并且可以通过药学领域公知的方法制备。以药学上可接受的载体制备组合物的方法包括让经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)与构成一种或更多种辅助成分的载体结合的步骤。一般来讲,可以通过让NK细胞(和/或iPSC)均匀地和/或密切地与例如液体载体结合来制备制剂。
包括经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)的药物组合物可以以适合的形式提供,包括但不限于溶液、混悬液、乳剂、喷雾剂、气溶胶,或任何形式的混合物。该组合物可以在制剂中与任何药学上可接受的赋形剂、载体或媒介一起递送。
施用于受试者的经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)的量可随各种因素变动,包括但不限于,受试者的体重、身体状况,和/或年龄,是否施用一种或更多种嵌合抗原受体,和/或施用途径。因此,包括在给定单位剂型中的NK细胞(和/或iPSC)的绝对量可大范围变动,并取决于一些因素,例如受试者的物种、年龄、体重和身体状况,以及施用方法。相应地,普遍性地阐述对每个和/或所有可能的应用有效的经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)的量是不切实际的。然而,本领域普通技术人员通过适当考虑这些因素可容易地确定合适的量。
在一些实施方式中,该方法可包括施用足够数量的经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC),以向受试者提供例如约105个细胞/kg至约1010个细胞/kg的剂量,尽管在一些实施方式中可以通过施用超出该范围的剂量的NK细胞(和/或iPSC)来实施该方法。在这些实施方式的一些中,该方法包括施用足够数量的经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC),以向受试者提供例如约107个细胞/kg至约108个细胞/kg的剂量,例如,约1×107个细胞/kg至约8×107个细胞/kg的剂量。
另外一种方式,可以采用在治疗过程开始前获得的实际体重来计算该剂量。对于以此方式计算的剂量,在治疗过程开始之前采用Dubois法计算身体表面积(m2):m2=(重量kg0.425×身高cm0.725)×0.007184。
在一些实施方式中,包括经修饰以表达嵌合抗原受体的NK细胞(和/或iPSC)的药物组合物可以例如以每周单剂量至多剂量进行施用,尽管在一些实施方式中药物组合物可通过以超出该范围的频率施用来实施该方法。在某些实施方式中,该药物组合物可以每月施用约1次至每周施用约5次。
通常,以病况的症状或临床迹象任何程度上的有效减少、限制其进展、缓解或消退的量和给药方案向受试者施用药物组合物。如本文所用,“缓解”指特定病况特征性的症状或临床征象在程度、严重性、频率,和/或可能性方面的任何减轻。“症状”指疾病或患者病况的任何主观证据(subjective evidence)。“征象”或“临床征象”指非患者本人发现的与具体状况相关的客观身体检查结果。
在前面的描述中,为了清楚起见,可以孤立地描述特定实施方式。除非另有明确指明特定实施方式的特征与另一实施方式的特征不相容,否则某些实施方式可包括本文结合一个或更多个实施方式描述的相容特征的组合。
对于本文描述的包括分开步骤的任何方法,这些方法可以以任何可行的顺序进行。视情况,两个或更多个步骤的任意组合可以同时进行。
本发明通过以上描述的示例性实施方式来阐释。应当理解,特定的实例、物质、量、以及操作应根据本文描述的发明的范围和精神在广义上解释。
如本文所用,术语“和/或”指所列出的要素的一个或全部或所列出的要素的任意两个或更多个的组合;当出现在说明书和权利要求书中时,术语“包含”及其变型不具有限制性含义;除非另有指出,“a”、“an”、“the”以及“至少一个”可互换使用,指一个或多于一个;通过端点对数值范围的叙述包括归纳在该范围内的所有数字(例如,1至5包括1、1.5、2、2.75、3、3.80、4、5等等)。
将本文提及的所有专利、专利申请和出版物以及以电子方式提供的材料(例如,包括例如GenBank和RefSeq中的核苷酸序列提交,例如SwissProt、PIR、PRF、PDB中氨基酸序列提交,以及GenBank和RefSeq中来自注释编码区域的翻译)的完整公开通过引用以其全文并入。如果本申请的公开与通过引用并入本文的任何文献的公开之间存在任何不一致,则适用本申请的公开。仅出于清楚理解的目的给出了前文的详细说明和实施例。不应从中理解不必要的限制。本发明不限于所示和所描述的精确细节,因为本领域技术人员显而易见的变化将被包括在由权利要求书所定义的发明内。
除非另有说明,在说明书和权利要求书中使用的表示组分数量、分子量等的所有数字应理解为在所有情况下都被术语“约”修饰。因此,除非有相反说明,在说明书和权利要求书中列出的数值参数为近似值,它们可以随本发明所寻求获得的期望性能而变化。最起码,并且并非试图将等同原则的应用限于权利要求的范围,每一数值参数应至少按照所报告的有效数以及应用普通舍入法来理解。
尽管陈述本发明的广泛范围的数值范围和参数是近似值,但是具体实施例中列出的数值为尽可能精确地报告。然而,所有数值固有地包含由其各自测量中发现的标准偏差所必然产生的范围。
除非另有说明,所有的标题都是为了方便阅读者,而不应用于限制该标题后文本的含义。
Claims (13)
1.编码嵌合抗原受体(CAR)中包含的多肽的多核苷酸,其中所述多肽按从N末端至C末端方向包括:
(a)跨膜结构域,其包含与天然具有胞外C末端的天然NKG2D相比处于反转的方向的NKG2D的跨膜区;以及
(b)胞内结构域,其包含激活自然杀伤细胞的信号传导肽,其中所述胞内结构域包含下述项的至少一部分:
(i)2B4(CD244)的细胞内结构域(ICD);
(ii)41BB(CD137)ICD;
(iii)DAP12 ICD;
(iv)2B4 ICD和41BB ICD;
(v)IL21R ICD;或
(vi)41BB ICD和2B4 ICD。
2.如权利要求1所述的多核苷酸,所述多肽进一步包含胞外结构域,所述胞外结构域在N末端包含抗原识别区。
3.如权利要求2所述的多核苷酸,其中所述抗原识别区特异性地结合与疾病相关的抗原。
4.如权利要求2所述的多核苷酸,其中所述抗原识别区特异性地结合肿瘤抗原。
5.如权利要求1-4中任一项所述的多核苷酸,其中所述胞外结构域还包含信号肽或前导序列。
6.如权利要求1-5中任一项所述的多核苷酸,其中所述胞外结构域还包含间隔序列。
7.如权利要求1-6中任一项所述的多核苷酸,其中所述胞内结构域还包含CD3ζ的信号传导结构域的至少一部分。
8.包含如权利要求1-7任一项所述的多核苷酸的诱导多能干细胞(iPSC)或NK细胞。
9.生产如权利要求8所述的iPSC或NK细胞的方法,包括向iPSC或NK细胞引入如权利要求1-7任一项所述的多核苷酸。
10.如权利要求9所述的方法,进一步包括使iPSC分化为包含所述CAR的NK细胞。
11.包含如权利要求8所述的iPSC或NK细胞的药物组合物。
12.如权利要求11所述的药物组合物,其中所述NK细胞来自于iPSC。
13.权利要求11的药物组合物在制备用于向有病况的受试者提供免疫治疗的药物中的用途,其中所述嵌合抗原受体包含特异性地结合与所述病况相关的抗原的抗原识别区。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201562109281P | 2015-01-29 | 2015-01-29 | |
US62/109,281 | 2015-01-29 | ||
CN201680008013.0A CN107428843B (zh) | 2015-01-29 | 2016-01-28 | 嵌合抗原受体、组合物以及方法 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201680008013.0A Division CN107428843B (zh) | 2015-01-29 | 2016-01-28 | 嵌合抗原受体、组合物以及方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113862292A true CN113862292A (zh) | 2021-12-31 |
Family
ID=55358136
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201680008013.0A Active CN107428843B (zh) | 2015-01-29 | 2016-01-28 | 嵌合抗原受体、组合物以及方法 |
CN202111106136.7A Pending CN113862292A (zh) | 2015-01-29 | 2016-01-28 | 嵌合抗原受体、组合物以及方法 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201680008013.0A Active CN107428843B (zh) | 2015-01-29 | 2016-01-28 | 嵌合抗原受体、组合物以及方法 |
Country Status (8)
Country | Link |
---|---|
US (2) | US10640570B2 (zh) |
EP (2) | EP3250587B1 (zh) |
JP (3) | JP6849600B6 (zh) |
CN (2) | CN107428843B (zh) |
AU (3) | AU2016211438C1 (zh) |
CA (1) | CA2975384A1 (zh) |
ES (1) | ES2789330T3 (zh) |
WO (1) | WO2016123333A1 (zh) |
Families Citing this family (30)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2948544A4 (en) | 2013-01-28 | 2016-08-03 | St Jude Childrens Res Hospital | CHIMERIC RECEPTOR WITH NKG2D SPECIFICITY FOR CELL THERAPY AGAINST CANCER AND INFECTION DISEASES |
US10428305B2 (en) | 2014-05-15 | 2019-10-01 | National University Of Singapore | Modified natural killer cells that express IL15 and uses thereof |
US11478555B2 (en) | 2015-08-17 | 2022-10-25 | Seoul National University R&Db Foundation | Chimeric antigen receptor to which anti-cotinine antibody is linked, and use thereof |
CA3035660A1 (en) | 2016-09-06 | 2018-03-15 | The Children's Medical Center Corporation | Immune cells derived from induced pluripotent stem cell |
JP2019532640A (ja) * | 2016-09-29 | 2019-11-14 | ナントクエスト インコーポレイテッド | 免疫原性が低下したhlaクラスi欠損nk−92細胞 |
CA3061898A1 (en) * | 2016-12-28 | 2019-10-29 | Green Cross Lab Cell Corporation | Chimeric antigen receptor and natural killer cells expressing same |
CN110475857B (zh) * | 2017-01-05 | 2023-07-18 | 韩国生命工学研究院 | 表达抗-可替宁嵌合抗原受体的天然杀伤细胞 |
CA3056591A1 (en) | 2017-03-27 | 2018-10-04 | National University Of Singapore | Stimulatory cell lines for ex vivo expansion and activation of natural killer cells |
CA3056439A1 (en) | 2017-03-27 | 2018-10-04 | National University Of Singapore | Truncated nkg2d chimeric receptors and uses thereof in natural killer cell immunotherapy |
US20200283501A1 (en) * | 2017-10-26 | 2020-09-10 | Regents Of The University Of Minnesota | Recombinant immune cells, methods of making, and methods of use |
CN107759701B (zh) * | 2017-10-27 | 2021-07-02 | 杭州优善生物科技有限公司 | 嵌合抗原受体、其修饰的NK细胞、编码DNA、mRNA、表达载体、制备方法和应用 |
WO2019094360A1 (en) * | 2017-11-07 | 2019-05-16 | The Board Of Regents Of The University Of Texas System | Targeting lilrb4 with car-t or car-nk cells in the treatment of cancer |
US11649294B2 (en) | 2017-11-14 | 2023-05-16 | GC Cell Corporation | Anti-HER2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same |
CN110028589B (zh) * | 2018-02-07 | 2023-07-21 | 阿思科力(苏州)生物科技有限公司 | 嵌合抗原受体、表达该嵌合抗原受体的nkg2d car-nk细胞及其制备方法和应用 |
CN110903399B (zh) * | 2018-09-17 | 2022-02-01 | 台湾中国医药大学附设医院 | 嵌合抗原受体、其核酸、表达质体、细胞、用途及组合物 |
CN109265561B (zh) * | 2018-09-25 | 2021-05-25 | 山东兴瑞生物科技有限公司 | 抗EGFRvⅢ安全型嵌合抗原受体、其制备方法、利用其修饰的NK细胞及应用 |
SG11202104287RA (en) * | 2018-11-06 | 2021-05-28 | Univ Washington | Chimeric antigen receptor memory-like (carml) nk cells and methods of making and using same |
CN109762844A (zh) * | 2019-01-31 | 2019-05-17 | 北京呈诺医学科技有限公司 | 一种靶标间皮素的car-nk细胞制备方法 |
WO2020180882A1 (en) | 2019-03-05 | 2020-09-10 | Nkarta, Inc. | Cd19-directed chimeric antigen receptors and uses thereof in immunotherapy |
AU2020244775A1 (en) * | 2019-03-22 | 2021-09-30 | The Regents Of The University Of California | Switchable chimeric antigen receptor-engineered human natural killer cells |
CN112300288B (zh) * | 2019-07-29 | 2022-08-02 | 济南赛尔生物科技股份有限公司 | 一种cik细胞的嵌合抗原受体car及其应用 |
GB201913697D0 (en) * | 2019-09-23 | 2019-11-06 | King S College London | DAP10/DAP12 fusion polypeptides |
JP2022550899A (ja) * | 2019-10-07 | 2022-12-05 | フェイト セラピューティクス,インコーポレイテッド | 免疫のための増強されたキメラ抗原受容体エフェクター細胞操作およびその使用 |
JP2022552314A (ja) * | 2019-10-17 | 2022-12-15 | フェイト セラピューティクス,インコーポレイテッド | 免疫のための増強されたキメラ抗原受容体エフェクター細胞操作およびその使用 |
WO2021201679A1 (en) | 2020-04-01 | 2021-10-07 | Kiadis Pharma Intellectual Property B.V. | Compositions and methods targeting coronaviruses |
WO2021201677A1 (en) | 2020-04-01 | 2021-10-07 | Kiadis Pharma Intellectual Property B.V. | Compositions and methods targeting influenza |
EP4284394A1 (en) * | 2021-01-26 | 2023-12-06 | Cytocares (Shanghai) Inc. | Chimeric antigen receptor (car) constructs and nk cells expressing car constructs |
CN113896800A (zh) * | 2021-09-30 | 2022-01-07 | 中国人民解放军陆军军医大学 | 靶向叶酸受体α嵌合抗原受体、其制备方法及其应用 |
WO2023215748A2 (en) * | 2022-05-03 | 2023-11-09 | H. Lee Moffitt Cancer Center And Research Institute Inc. | Chimeric antigen receptor (car) constructs with nk receptor signaling domain |
WO2024064824A2 (en) | 2022-09-21 | 2024-03-28 | Yale University | Compositions and methods for identification of membrane targets for enhancement of nk cell therapy |
Family Cites Families (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6444789B1 (en) | 1995-05-03 | 2002-09-03 | Applied Research Systems Ars Holding N.V. | CD16-II variants |
US7829084B2 (en) | 2001-01-17 | 2010-11-09 | Trubion Pharmaceuticals, Inc. | Binding constructs and methods for use thereof |
US20050113564A1 (en) * | 2003-11-05 | 2005-05-26 | St. Jude Children's Research Hospital | Chimeric receptors with 4-1BB stimulatory signaling domain |
JP2008005722A (ja) * | 2006-06-27 | 2008-01-17 | Institute Of Physical & Chemical Research | ウイルス感染症の予防・治療 |
WO2010040091A1 (en) | 2008-10-03 | 2010-04-08 | Arizona Board of Regents, a body corporate acting for and on behalf of Arizona State University | Novel dna nanostructures that promote cell-cell interaction and use thereof |
CN106220739A (zh) * | 2010-12-09 | 2016-12-14 | 宾夕法尼亚大学董事会 | 嵌合抗原受体‑修饰的t细胞治疗癌症的用途 |
AU2013222284A1 (en) * | 2012-02-22 | 2014-08-07 | The Trustees Of The University Of Pennsylvania | Use of the CD2 signaling domain in second-generation chimeric antigen receptors |
UY35468A (es) | 2013-03-16 | 2014-10-31 | Novartis Ag | Tratamiento de cáncer utilizando un receptor quimérico de antígeno anti-cd19 |
HUE048312T2 (hu) | 2013-05-03 | 2020-07-28 | Ohio State Innovation Foundation | CS-1 specifikus kiméra antigén receptor módosított immuneffektor sejtek |
WO2015142661A1 (en) * | 2014-03-15 | 2015-09-24 | Novartis Ag | Regulatable chimeric antigen receptor |
US10301370B2 (en) * | 2014-05-02 | 2019-05-28 | The Trustees Of The University Of Pennsylvania | Compositions and methods of chimeric autoantibody receptor T cells |
MA41433A (fr) * | 2015-01-26 | 2017-12-05 | Baylor College Medicine | Cellules immunitaires universelles pour l'immunothérapie anticancéreuse |
-
2016
- 2016-01-28 JP JP2017540182A patent/JP6849600B6/ja active Active
- 2016-01-28 AU AU2016211438A patent/AU2016211438C1/en active Active
- 2016-01-28 WO PCT/US2016/015351 patent/WO2016123333A1/en active Application Filing
- 2016-01-28 EP EP16704526.9A patent/EP3250587B1/en active Active
- 2016-01-28 CA CA2975384A patent/CA2975384A1/en active Pending
- 2016-01-28 CN CN201680008013.0A patent/CN107428843B/zh active Active
- 2016-01-28 EP EP20157464.7A patent/EP3674315A1/en active Pending
- 2016-01-28 US US15/546,177 patent/US10640570B2/en active Active
- 2016-01-28 CN CN202111106136.7A patent/CN113862292A/zh active Pending
- 2016-01-28 ES ES16704526T patent/ES2789330T3/es active Active
-
2020
- 2020-04-01 US US16/837,661 patent/US20200291125A1/en active Pending
- 2020-11-18 AU AU2020270507A patent/AU2020270507B2/en active Active
-
2021
- 2021-03-04 JP JP2021034578A patent/JP7227290B2/ja active Active
-
2023
- 2023-02-09 JP JP2023018349A patent/JP2023053110A/ja active Pending
- 2023-12-20 AU AU2023285795A patent/AU2023285795A1/en active Pending
Also Published As
Publication number | Publication date |
---|---|
JP2021087452A (ja) | 2021-06-10 |
JP6849600B6 (ja) | 2021-06-30 |
CA2975384A1 (en) | 2016-08-04 |
AU2016211438A1 (en) | 2017-08-10 |
AU2020270507B2 (en) | 2023-09-21 |
AU2016211438B2 (en) | 2020-08-27 |
EP3674315A1 (en) | 2020-07-01 |
JP2023053110A (ja) | 2023-04-12 |
AU2023285795A1 (en) | 2024-01-25 |
AU2016211438C1 (en) | 2021-03-04 |
CN107428843A (zh) | 2017-12-01 |
EP3250587A1 (en) | 2017-12-06 |
US20200291125A1 (en) | 2020-09-17 |
JP6849600B2 (ja) | 2021-03-24 |
AU2020270507A1 (en) | 2020-12-17 |
EP3250587B1 (en) | 2020-03-25 |
CN107428843B (zh) | 2021-10-15 |
JP2018505174A (ja) | 2018-02-22 |
ES2789330T3 (es) | 2020-10-26 |
US20180002438A1 (en) | 2018-01-04 |
JP7227290B2 (ja) | 2023-02-21 |
US10640570B2 (en) | 2020-05-05 |
WO2016123333A1 (en) | 2016-08-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107428843B (zh) | 嵌合抗原受体、组合物以及方法 | |
JP6968389B2 (ja) | Bcmaに結合するキメラ抗原受容体(car)及びその応用 | |
JP7048494B2 (ja) | 修飾t細胞に対する条件的活性型キメラ抗原受容体 | |
CN109415409B (zh) | Flag标记的cd19-car-t细胞 | |
CN113286879A (zh) | 用于细胞疗法之多样化抗原结合域、新颖平台及其他增强子 | |
EP3599251B1 (en) | Construction of chimeric antigen receptor targeting cd20 antigen and activity identification of engineered t cells thereof | |
JP2021510063A (ja) | Nyeso tcr | |
CA2969384A1 (en) | Inhibitory chimeric antigen receptor (icar or n-car) expressing non-t cell transduction domain | |
CN110818802A (zh) | 一种嵌合t细胞受体star及其应用 | |
CN105407902A (zh) | 溶瘤病毒 | |
AU2019336031A1 (en) | Chimeric antigen receptor for solid cancer and T cells expressing chimeric antigen receptor | |
JP2021509290A (ja) | 双方向活性化共刺激分子受容体及びその用途 | |
TW202140532A (zh) | 一種辨識afp的t細胞受體及其編碼序列 | |
JP2022516710A (ja) | Car t細胞の方法及び構築物 | |
JP2022514815A (ja) | CDR1領域に突然変異したヒト化CD19 scFvを有するCAR-T細胞 | |
CN112041432A (zh) | Foxp3靶向剂组合物以及用于过继细胞疗法的使用方法 | |
WO2023125813A1 (zh) | 抗间皮素纳米抗体嵌合抗原受体及其应用 | |
WO2023241522A1 (zh) | 靶向kras g12v突变多肽的t细胞受体及其用途 | |
WO2023226921A1 (zh) | 靶向bcma-cd19的双特异性嵌合抗原受体及其应用 | |
WO2022166904A1 (zh) | 一种识别hpv的t细胞受体 | |
WO2023125766A1 (zh) | 靶向cs1的嵌合抗原受体、靶向bcma/cs1的双特异性嵌合抗原受体及其应用 | |
WO2023088359A1 (zh) | 靶向bcma的嵌合抗原受体及其应用 | |
WO2023010068A2 (en) | Multiprotein-engineered cells secreting a multispecific antibody | |
WO2023139253A1 (en) | Antigen recognizing construct that binds specific peptide with determinable affinity and t cell receptor having antigenic specificity for kk-lc-1 as well as corresponding nucleic acid sequence, vector, host cell, pharmaceutical composition and kit | |
KR20240034205A (ko) | 항EGFRviii 항체, 폴리펩타이드, 상기 폴리펩타이드를 발현하는 세포, 상기 세포를 포함하는 의약 조성물, 상기 세포의 제조 방법 및 상기 폴리펩타이드를 코딩하는 염기서열을 포함하는 폴리뉴클레오티드 또는 벡터 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 40064903 Country of ref document: HK |