WO2023200266A1 - 키메릭 항원 수용체 및 hgf 중화항체의 조합 요법 - Google Patents
키메릭 항원 수용체 및 hgf 중화항체의 조합 요법 Download PDFInfo
- Publication number
- WO2023200266A1 WO2023200266A1 PCT/KR2023/004996 KR2023004996W WO2023200266A1 WO 2023200266 A1 WO2023200266 A1 WO 2023200266A1 KR 2023004996 W KR2023004996 W KR 2023004996W WO 2023200266 A1 WO2023200266 A1 WO 2023200266A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- car
- cancer
- domain
- yyb
- hgf
- Prior art date
Links
- 108010019670 Chimeric Antigen Receptors Proteins 0.000 title claims abstract description 69
- 238000002648 combination therapy Methods 0.000 title 1
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 50
- 201000011510 cancer Diseases 0.000 claims abstract description 34
- 230000003472 neutralizing effect Effects 0.000 claims abstract description 23
- 238000002560 therapeutic procedure Methods 0.000 claims abstract description 21
- 239000012642 immune effector Substances 0.000 claims abstract description 6
- 229940121354 immunomodulator Drugs 0.000 claims abstract description 6
- 210000004027 cell Anatomy 0.000 claims description 67
- 239000000427 antigen Substances 0.000 claims description 34
- 102000036639 antigens Human genes 0.000 claims description 34
- 108091007433 antigens Proteins 0.000 claims description 34
- 206010033128 Ovarian cancer Diseases 0.000 claims description 29
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 29
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 24
- -1 EGP-40 Proteins 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- 230000011664 signaling Effects 0.000 claims description 15
- 230000001086 cytosolic effect Effects 0.000 claims description 11
- 102100029215 Signaling lymphocytic activation molecule Human genes 0.000 claims description 10
- 230000000139 costimulatory effect Effects 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 102100032816 Integrin alpha-6 Human genes 0.000 claims description 9
- 102100020793 Interleukin-13 receptor subunit alpha-2 Human genes 0.000 claims description 9
- 101710112634 Interleukin-13 receptor subunit alpha-2 Proteins 0.000 claims description 9
- 102100022339 Integrin alpha-L Human genes 0.000 claims description 8
- 101000994365 Homo sapiens Integrin alpha-6 Proteins 0.000 claims description 7
- 239000003446 ligand Substances 0.000 claims description 7
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 claims description 6
- 206010009944 Colon cancer Diseases 0.000 claims description 6
- 241000701022 Cytomegalovirus Species 0.000 claims description 6
- 101000994375 Homo sapiens Integrin alpha-4 Proteins 0.000 claims description 6
- 101000935040 Homo sapiens Integrin beta-2 Proteins 0.000 claims description 6
- 101000633786 Homo sapiens SLAM family member 6 Proteins 0.000 claims description 6
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 claims description 6
- 102100032818 Integrin alpha-4 Human genes 0.000 claims description 6
- 102100025390 Integrin beta-2 Human genes 0.000 claims description 6
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 claims description 6
- 102100029197 SLAM family member 6 Human genes 0.000 claims description 6
- 101710163413 Signaling lymphocytic activation molecule Proteins 0.000 claims description 6
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 claims description 6
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 claims description 6
- 208000029742 colonic neoplasm Diseases 0.000 claims description 6
- 102100038080 B-cell receptor CD22 Human genes 0.000 claims description 5
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 claims description 5
- 102100038078 CD276 antigen Human genes 0.000 claims description 5
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 claims description 5
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 claims description 5
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 claims description 5
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 claims description 5
- 101000914496 Homo sapiens T-cell antigen CD7 Proteins 0.000 claims description 5
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 claims description 5
- 101000851370 Homo sapiens Tumor necrosis factor receptor superfamily member 9 Proteins 0.000 claims description 5
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 claims description 5
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 claims description 5
- 102100027208 T-cell antigen CD7 Human genes 0.000 claims description 5
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 claims description 5
- 102100036856 Tumor necrosis factor receptor superfamily member 9 Human genes 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 102100024263 CD160 antigen Human genes 0.000 claims description 4
- 102100038077 CD226 antigen Human genes 0.000 claims description 4
- 101000761938 Homo sapiens CD160 antigen Proteins 0.000 claims description 4
- 101000884298 Homo sapiens CD226 antigen Proteins 0.000 claims description 4
- 101001078158 Homo sapiens Integrin alpha-1 Proteins 0.000 claims description 4
- 101001035237 Homo sapiens Integrin alpha-D Proteins 0.000 claims description 4
- 101001046687 Homo sapiens Integrin alpha-E Proteins 0.000 claims description 4
- 101000935043 Homo sapiens Integrin beta-1 Proteins 0.000 claims description 4
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 claims description 4
- 101000971538 Homo sapiens Killer cell lectin-like receptor subfamily F member 1 Proteins 0.000 claims description 4
- 101000633780 Homo sapiens Signaling lymphocytic activation molecule Proteins 0.000 claims description 4
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 claims description 4
- 102100025323 Integrin alpha-1 Human genes 0.000 claims description 4
- 102100039904 Integrin alpha-D Human genes 0.000 claims description 4
- 102100022341 Integrin alpha-E Human genes 0.000 claims description 4
- 102100022338 Integrin alpha-M Human genes 0.000 claims description 4
- 102100022297 Integrin alpha-X Human genes 0.000 claims description 4
- 102100025304 Integrin beta-1 Human genes 0.000 claims description 4
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 claims description 4
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 claims description 4
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 claims description 4
- 102100021458 Killer cell lectin-like receptor subfamily F member 1 Human genes 0.000 claims description 4
- 102100038082 Natural killer cell receptor 2B4 Human genes 0.000 claims description 4
- 102000014128 RANK Ligand Human genes 0.000 claims description 4
- 108010025832 RANK Ligand Proteins 0.000 claims description 4
- 102100027744 Semaphorin-4D Human genes 0.000 claims description 4
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 claims description 4
- 102100028785 Tumor necrosis factor receptor superfamily member 14 Human genes 0.000 claims description 4
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 claims description 4
- 102100031585 ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Human genes 0.000 claims description 3
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 claims description 3
- 206010005003 Bladder cancer Diseases 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 3
- 208000026310 Breast neoplasm Diseases 0.000 claims description 3
- 102100032912 CD44 antigen Human genes 0.000 claims description 3
- ODINCKMPIJJUCX-UHFFFAOYSA-N Calcium oxide Chemical compound [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 claims description 3
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 claims description 3
- 108010009685 Cholinergic Receptors Proteins 0.000 claims description 3
- 108010066687 Epithelial Cell Adhesion Molecule Proteins 0.000 claims description 3
- 102000018651 Epithelial Cell Adhesion Molecule Human genes 0.000 claims description 3
- 102100031940 Epithelial cell adhesion molecule Human genes 0.000 claims description 3
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 claims description 3
- 102100030595 HLA class II histocompatibility antigen gamma chain Human genes 0.000 claims description 3
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 claims description 3
- 101000777636 Homo sapiens ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Proteins 0.000 claims description 3
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 claims description 3
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 claims description 3
- 101000920667 Homo sapiens Epithelial cell adhesion molecule Proteins 0.000 claims description 3
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 claims description 3
- 101001082627 Homo sapiens HLA class II histocompatibility antigen gamma chain Proteins 0.000 claims description 3
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 claims description 3
- 101001103039 Homo sapiens Inactive tyrosine-protein kinase transmembrane receptor ROR1 Proteins 0.000 claims description 3
- 101001078143 Homo sapiens Integrin alpha-IIb Proteins 0.000 claims description 3
- 101000623901 Homo sapiens Mucin-16 Proteins 0.000 claims description 3
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 claims description 3
- 101001103036 Homo sapiens Nuclear receptor ROR-alpha Proteins 0.000 claims description 3
- 101000873418 Homo sapiens P-selectin glycoprotein ligand 1 Proteins 0.000 claims description 3
- 101000610551 Homo sapiens Prominin-1 Proteins 0.000 claims description 3
- 101001136592 Homo sapiens Prostate stem cell antigen Proteins 0.000 claims description 3
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims description 3
- 101000633782 Homo sapiens SLAM family member 8 Proteins 0.000 claims description 3
- 101000874179 Homo sapiens Syndecan-1 Proteins 0.000 claims description 3
- 101000946843 Homo sapiens T-cell surface glycoprotein CD8 alpha chain Proteins 0.000 claims description 3
- 102100039615 Inactive tyrosine-protein kinase transmembrane receptor ROR1 Human genes 0.000 claims description 3
- 102100025306 Integrin alpha-IIb Human genes 0.000 claims description 3
- 208000008839 Kidney Neoplasms Diseases 0.000 claims description 3
- 206010023825 Laryngeal cancer Diseases 0.000 claims description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 3
- 102100023123 Mucin-16 Human genes 0.000 claims description 3
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 claims description 3
- 206010061306 Nasopharyngeal cancer Diseases 0.000 claims description 3
- 102000003729 Neprilysin Human genes 0.000 claims description 3
- 108090000028 Neprilysin Proteins 0.000 claims description 3
- 108010012255 Neural Cell Adhesion Molecule L1 Proteins 0.000 claims description 3
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 claims description 3
- 102100024964 Neural cell adhesion molecule L1 Human genes 0.000 claims description 3
- 102100034925 P-selectin glycoprotein ligand 1 Human genes 0.000 claims description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 3
- 208000000821 Parathyroid Neoplasms Diseases 0.000 claims description 3
- 102100040120 Prominin-1 Human genes 0.000 claims description 3
- 102100036735 Prostate stem cell antigen Human genes 0.000 claims description 3
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims description 3
- 206010038389 Renal cancer Diseases 0.000 claims description 3
- 102100029214 SLAM family member 8 Human genes 0.000 claims description 3
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- 102100035721 Syndecan-1 Human genes 0.000 claims description 3
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 claims description 3
- 208000024770 Thyroid neoplasm Diseases 0.000 claims description 3
- 208000023915 Ureteral Neoplasms Diseases 0.000 claims description 3
- 206010046392 Ureteric cancer Diseases 0.000 claims description 3
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 3
- 102000034337 acetylcholine receptors Human genes 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 3
- 201000009036 biliary tract cancer Diseases 0.000 claims description 3
- 208000020790 biliary tract neoplasm Diseases 0.000 claims description 3
- 230000001605 fetal effect Effects 0.000 claims description 3
- 229940014144 folate Drugs 0.000 claims description 3
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 3
- 235000019152 folic acid Nutrition 0.000 claims description 3
- 239000011724 folic acid Substances 0.000 claims description 3
- 206010017758 gastric cancer Diseases 0.000 claims description 3
- 208000014829 head and neck neoplasm Diseases 0.000 claims description 3
- 201000010982 kidney cancer Diseases 0.000 claims description 3
- 206010023841 laryngeal neoplasm Diseases 0.000 claims description 3
- 201000007270 liver cancer Diseases 0.000 claims description 3
- 208000014018 liver neoplasm Diseases 0.000 claims description 3
- 201000005202 lung cancer Diseases 0.000 claims description 3
- 208000020816 lung neoplasm Diseases 0.000 claims description 3
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 3
- 208000026045 malignant tumor of parathyroid gland Diseases 0.000 claims description 3
- 201000002528 pancreatic cancer Diseases 0.000 claims description 3
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 3
- 229920001481 poly(stearyl methacrylate) Polymers 0.000 claims description 3
- 201000000849 skin cancer Diseases 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 101150047061 tag-72 gene Proteins 0.000 claims description 3
- 201000002510 thyroid cancer Diseases 0.000 claims description 3
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 3
- 108010056102 CD100 antigen Proteins 0.000 claims description 2
- 108010017009 CD11b Antigen Proteins 0.000 claims description 2
- 102100027207 CD27 antigen Human genes 0.000 claims description 2
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 claims description 2
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 claims description 2
- 101150013553 CD40 gene Proteins 0.000 claims description 2
- 108010062802 CD66 antigens Proteins 0.000 claims description 2
- 102100027217 CD82 antigen Human genes 0.000 claims description 2
- 101710139831 CD82 antigen Proteins 0.000 claims description 2
- 102100035793 CD83 antigen Human genes 0.000 claims description 2
- 102100037904 CD9 antigen Human genes 0.000 claims description 2
- 102100024533 Carcinoembryonic antigen-related cell adhesion molecule 1 Human genes 0.000 claims description 2
- 102100027816 Cytotoxic and regulatory T-cell molecule Human genes 0.000 claims description 2
- 102100025137 Early activation antigen CD69 Human genes 0.000 claims description 2
- 101000585551 Equus caballus Pregnancy-associated glycoprotein Proteins 0.000 claims description 2
- 102100022086 GRB2-related adapter protein 2 Human genes 0.000 claims description 2
- 102100026122 High affinity immunoglobulin gamma Fc receptor I Human genes 0.000 claims description 2
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 claims description 2
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 claims description 2
- 101000738354 Homo sapiens CD9 antigen Proteins 0.000 claims description 2
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 claims description 2
- 101000900690 Homo sapiens GRB2-related adapter protein 2 Proteins 0.000 claims description 2
- 101000913074 Homo sapiens High affinity immunoglobulin gamma Fc receptor I Proteins 0.000 claims description 2
- 101001046683 Homo sapiens Integrin alpha-L Proteins 0.000 claims description 2
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 claims description 2
- 101001046668 Homo sapiens Integrin alpha-X Proteins 0.000 claims description 2
- 101001015037 Homo sapiens Integrin beta-7 Proteins 0.000 claims description 2
- 101001043809 Homo sapiens Interleukin-7 receptor subunit alpha Proteins 0.000 claims description 2
- 101000777628 Homo sapiens Leukocyte antigen CD37 Proteins 0.000 claims description 2
- 101001047640 Homo sapiens Linker for activation of T-cells family member 1 Proteins 0.000 claims description 2
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 claims description 2
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 claims description 2
- 101001090688 Homo sapiens Lymphocyte cytosolic protein 2 Proteins 0.000 claims description 2
- 101001109503 Homo sapiens NKG2-C type II integral membrane protein Proteins 0.000 claims description 2
- 101000589305 Homo sapiens Natural cytotoxicity triggering receptor 2 Proteins 0.000 claims description 2
- 101001124867 Homo sapiens Peroxiredoxin-1 Proteins 0.000 claims description 2
- 101000692259 Homo sapiens Phosphoprotein associated with glycosphingolipid-enriched microdomains 1 Proteins 0.000 claims description 2
- 101000702132 Homo sapiens Protein spinster homolog 1 Proteins 0.000 claims description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 claims description 2
- 101000633778 Homo sapiens SLAM family member 5 Proteins 0.000 claims description 2
- 101000633784 Homo sapiens SLAM family member 7 Proteins 0.000 claims description 2
- 101000934346 Homo sapiens T-cell surface antigen CD2 Proteins 0.000 claims description 2
- 101000934341 Homo sapiens T-cell surface glycoprotein CD5 Proteins 0.000 claims description 2
- 101000596234 Homo sapiens T-cell surface protein tactile Proteins 0.000 claims description 2
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 claims description 2
- 101000795169 Homo sapiens Tumor necrosis factor receptor superfamily member 13C Proteins 0.000 claims description 2
- 101000648507 Homo sapiens Tumor necrosis factor receptor superfamily member 14 Proteins 0.000 claims description 2
- 101000801234 Homo sapiens Tumor necrosis factor receptor superfamily member 18 Proteins 0.000 claims description 2
- 101000679857 Homo sapiens Tumor necrosis factor receptor superfamily member 3 Proteins 0.000 claims description 2
- 108010041100 Integrin alpha6 Proteins 0.000 claims description 2
- 108010030465 Integrin alpha6beta1 Proteins 0.000 claims description 2
- 102100033016 Integrin beta-7 Human genes 0.000 claims description 2
- 102100021593 Interleukin-7 receptor subunit alpha Human genes 0.000 claims description 2
- 102100031586 Leukocyte antigen CD37 Human genes 0.000 claims description 2
- 102100024032 Linker for activation of T-cells family member 1 Human genes 0.000 claims description 2
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 claims description 2
- 102100034709 Lymphocyte cytosolic protein 2 Human genes 0.000 claims description 2
- 108010061593 Member 14 Tumor Necrosis Factor Receptors Proteins 0.000 claims description 2
- 108090000015 Mesothelin Proteins 0.000 claims description 2
- 102000003735 Mesothelin Human genes 0.000 claims description 2
- 101100236305 Mus musculus Ly9 gene Proteins 0.000 claims description 2
- 108091008877 NK cell receptors Proteins 0.000 claims description 2
- 102100022683 NKG2-C type II integral membrane protein Human genes 0.000 claims description 2
- 108010004217 Natural Cytotoxicity Triggering Receptor 1 Proteins 0.000 claims description 2
- 108010004222 Natural Cytotoxicity Triggering Receptor 3 Proteins 0.000 claims description 2
- 102100032870 Natural cytotoxicity triggering receptor 1 Human genes 0.000 claims description 2
- 102100032851 Natural cytotoxicity triggering receptor 2 Human genes 0.000 claims description 2
- 102100032852 Natural cytotoxicity triggering receptor 3 Human genes 0.000 claims description 2
- 101710141230 Natural killer cell receptor 2B4 Proteins 0.000 claims description 2
- 102100021462 Natural killer cells antigen CD94 Human genes 0.000 claims description 2
- 102100026066 Phosphoprotein associated with glycosphingolipid-enriched microdomains 1 Human genes 0.000 claims description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 claims description 2
- 102100029216 SLAM family member 5 Human genes 0.000 claims description 2
- 102100029198 SLAM family member 7 Human genes 0.000 claims description 2
- 108091008874 T cell receptors Proteins 0.000 claims description 2
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims description 2
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 claims description 2
- 102100025244 T-cell surface glycoprotein CD5 Human genes 0.000 claims description 2
- 102100035268 T-cell surface protein tactile Human genes 0.000 claims description 2
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 claims description 2
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 claims description 2
- 102100029690 Tumor necrosis factor receptor superfamily member 13C Human genes 0.000 claims description 2
- 102100033728 Tumor necrosis factor receptor superfamily member 18 Human genes 0.000 claims description 2
- 102100033733 Tumor necrosis factor receptor superfamily member 1B Human genes 0.000 claims description 2
- 101710187830 Tumor necrosis factor receptor superfamily member 1B Proteins 0.000 claims description 2
- 102100022156 Tumor necrosis factor receptor superfamily member 3 Human genes 0.000 claims description 2
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 2
- 101001038499 Yarrowia lipolytica (strain CLIB 122 / E 150) Lysine acetyltransferase Proteins 0.000 claims description 2
- 108010072917 class-I restricted T cell-associated molecule Proteins 0.000 claims description 2
- 102000003675 cytokine receptors Human genes 0.000 claims description 2
- 108010057085 cytokine receptors Proteins 0.000 claims description 2
- 102000006495 integrins Human genes 0.000 claims description 2
- 108010044426 integrins Proteins 0.000 claims description 2
- 230000000849 parathyroid Effects 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 210000001685 thyroid gland Anatomy 0.000 claims description 2
- 102000003298 tumor necrosis factor receptor Human genes 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 102100028976 HLA class I histocompatibility antigen, B alpha chain Human genes 0.000 claims 1
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 claims 1
- 210000000988 bone and bone Anatomy 0.000 claims 1
- 210000004556 brain Anatomy 0.000 claims 1
- 210000003128 head Anatomy 0.000 claims 1
- 208000026037 malignant tumor of neck Diseases 0.000 claims 1
- 210000003739 neck Anatomy 0.000 claims 1
- 210000002307 prostate Anatomy 0.000 claims 1
- 210000003491 skin Anatomy 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 8
- 239000007787 solid Substances 0.000 abstract description 8
- 230000001093 anti-cancer Effects 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 6
- 210000000987 immune system Anatomy 0.000 abstract description 5
- 230000001965 increasing effect Effects 0.000 abstract description 4
- 230000003993 interaction Effects 0.000 abstract description 3
- 229940124597 therapeutic agent Drugs 0.000 abstract description 2
- 230000002489 hematologic effect Effects 0.000 abstract 1
- 102000003745 Hepatocyte Growth Factor Human genes 0.000 description 35
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 35
- 238000011357 CAR T-cell therapy Methods 0.000 description 20
- 150000001413 amino acids Chemical group 0.000 description 15
- 239000002246 antineoplastic agent Substances 0.000 description 14
- 229940041181 antineoplastic drug Drugs 0.000 description 13
- 239000000126 substance Substances 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 11
- 102000003816 Interleukin-13 Human genes 0.000 description 10
- 108090000176 Interleukin-13 Proteins 0.000 description 10
- 238000012360 testing method Methods 0.000 description 9
- 238000009169 immunotherapy Methods 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 238000000684 flow cytometry Methods 0.000 description 6
- 238000010276 construction Methods 0.000 description 5
- 238000002054 transplantation Methods 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 229940024606 amino acid Drugs 0.000 description 4
- 238000002659 cell therapy Methods 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 235000002374 tyrosine Nutrition 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 206010008342 Cervix carcinoma Diseases 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 3
- 201000010881 cervical cancer Diseases 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 3
- 150000002333 glycines Chemical class 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 3
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- 206010005949 Bone cancer Diseases 0.000 description 2
- 208000018084 Bone neoplasm Diseases 0.000 description 2
- 208000003174 Brain Neoplasms Diseases 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 102220474088 Protection of telomeres protein 1_S67D_mutation Human genes 0.000 description 2
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 102000006426 human interleukin-13 receptor Human genes 0.000 description 2
- 108010044118 human interleukin-13 receptor Proteins 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 108010082117 matrigel Proteins 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000003449 preventive effect Effects 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 102200024855 rs886037778 Human genes 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 description 1
- 101000898034 Homo sapiens Hepatocyte growth factor Proteins 0.000 description 1
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 1
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 108091054437 MHC class I family Proteins 0.000 description 1
- 206010061309 Neoplasm progression Diseases 0.000 description 1
- 208000007125 Neurotoxicity Syndromes Diseases 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- 206010044221 Toxic encephalopathy Diseases 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 101150058049 car gene Proteins 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000001268 conjugating effect Effects 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 102000057308 human HGF Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000004068 intracellular signaling Effects 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000007651 self-proliferation Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000005751 tumor progression Effects 0.000 description 1
- 150000003668 tyrosines Chemical class 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
Images
Classifications
-
- 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
-
- 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
- A61K39/0005—Vertebrate antigens
- A61K39/0011—Cancer antigens
-
- 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
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- 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
-
- 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
- C07K14/70503—Immunoglobulin superfamily
- C07K14/70517—CD8
-
- 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
- C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
-
- 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
-
- 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
- C07K16/2866—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
-
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5156—Animal cells expressing foreign proteins
-
- 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
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/515—Animal cells
- A61K2039/5158—Antigen-pulsed cells, e.g. T-cells
-
- 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
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/80—Vaccine for a specifically defined cancer
- A61K2039/892—Reproductive system [uterus, ovaries, cervix, testes]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- 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
Definitions
- the present invention relates to the prevention or treatment of solid cancer using CAR-T cell therapy, a third-generation immunotherapy. Specifically, it is characterized by increasing the anticancer effect of CAR-T cell therapy by affecting the interaction between cancer cells and the immune system by combining hepatocyte growth factor (HGF) neutralizing antibodies.
- HGF hepatocyte growth factor
- the CAR-T cell therapy of the present invention can also be effectively used to treat solid tumors, which are recognized as difficult to use as immunotherapy.
- first-generation chemical anticancer drugs to treat cancer, they attack and kill cancer cells with cytotoxic substances, but they have severe side effects because they damage not only cancer cells but also normal cells.
- the second-generation targeted anti-cancer drugs which are intended to overcome these shortcomings of the first-generation chemical anti-cancer drugs, target and attack specific substances in cancer cells, so they have fewer side effects compared to the first-generation chemical anti-cancer drugs, but they have the major disadvantage of developing resistance.
- third-generation immunotherapy drugs use our body's immune system, there are fewer toxicity problems with first-generation chemical anticancer drugs and resistance problems with second-generation targeted anticancer drugs, and there are significantly fewer side effects.
- second-generation targeted anti-cancer drugs show a high survival rate in the early stages, but their durability is low due to resistance issues, while immuno-anti-cancer drugs maintain their anti-cancer effect, so patients who respond well to immuno- anti-cancer drugs are closer to cure.
- third-generation immunotherapy drugs contribute to improving the quality of life of cancer patients as well as the long-term survival rate of cancer patients due to their excellent efficacy and fewer side effects.
- a chimeric antigen receptor is composed of an antibody fragment, a hinge region, a transmembrane domain, and an intracellular signaling domain.
- T cells expressing the chimeric antigen receptor are immunotherapy drugs designed to attack only cancer cells. It is one of the CAR-T cell therapy is a very important cell therapy product in that it significantly increases the aggressiveness of immune T cells that specifically attack cancer cells, reducing damage to normal cells and targeting only cancer cells.
- immune cell therapies such as CAR T cell therapy have shown high cure rates for some blood cancers, but have not been effective in treating solid cancers, which account for most cancers, as the human body tends to suppress strong immune responses. It is known to be because immune cells cannot function sufficiently.
- Patent Document 1 Republic of Korea Patent No. 10-0556660
- Patent Document 2 Republic of Korea Patent No. 10-2011789
- the present inventors studied a treatment method that allows CAR-T cell therapy to be effective even in solid cancers such as ovarian cancer, and as a result, a third-generation treatment with less toxicity problems of first-generation chemical anticancer drugs and less resistance problems of second-generation targeted anticancer drugs was discovered. It was confirmed that combining a CAR-T cell therapy, an immunotherapy, with a neutralizing antibody that binds to a neutralizing epitope of HGF can lead to an improvement in the inhibition or reduction of tumor progression compared to the CAR-T cell therapy alone, and the present invention Completed.
- the present invention aims to provide a CAR therapy for the treatment of solid tumors, including ovarian cancer, and provides a chimeric antigen for use in the treatment of cancer in a subject in combination with a neutralizing antibody that binds to a neutralizable epitope of HGF.
- a specific challenge is to provide a CAR therapy comprising a population of immune effector cells expressing the receptor (CAR).
- the present invention discloses the following means.
- the present invention provides a CAR therapy comprising a population of immune effector cells expressing a chimeric antigen receptor (CAR) for use in the treatment of cancer in a subject in combination with a neutralizing antibody that binds to a neutralizing epitope of HGF. begins.
- CAR chimeric antigen receptor
- CAR includes IL13R ⁇ 2 antigen binding domain; hinge area; transmembrane domain; costimulatory domain; And it may include, but is not limited to, a cytoplasmic signaling domain.
- the CAR is represented by the amino acid sequence of SEQ ID NO: 9 or SEQ ID NO: 10.
- the heavy chain of the HGF neutralizing antibody is represented by the amino acid sequence of SEQ ID NO: 13
- the light chain is represented by the amino acid sequence of SEQ ID NO: 14. Additionally, known linker sequences can be added.
- the present invention discloses a CAR therapy comprising a population of immune effector cells expressing a chimeric antigen receptor (CAR) for use in the treatment of cancer in a subject in combination with a neutralizing antibody that binds a neutralizable epitope of HGF.
- CAR chimeric antigen receptor
- the CAR includes an antigen binding domain; hinge area; transmembrane domain; costimulatory domain; And it may include, but is not limited to, a cytoplasmic signaling domain.
- the antigen binding domain of the CAR is CD19, MUC16, MUCl, CAlX, CEA, CDS, CD7, CD10, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, Cyto Megalovirus (CMV) infected cell antigen, EGP-2, EGP-40, EpCAM, erb-B2,3,4, FBP, fetal acetylcholine receptor, folate receptor-a, GD2, GD3, HER-2, hTERT, K-light chain, KDR, LeY, L1 cell adhesion molecule, MAGE-A1, mesothelin, NKG2D ligand, NY-ES0-1, carcinoembryonic antigen (h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, It may comprise an antigen binding domain that binds an antigen selected from WT-1, CD276 or
- the transmembrane domain of the CAR includes the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, and a transmembrane domain selected from CD137 or CD154.
- the costimulatory domain of the CAR includes MHC class I molecules, TNF receptor protein, immunoglobulin-like protein, cytokine receptor, integrin, signaling lymphocytic activation molecule (SLAM), activating NK cell receptor, BTLA ( B an T lymphocyte attenuator), Tolllike ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8 alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4,
- the cytoplasmic signaling domain of the CAR may include a cytoplasmic signaling domain selected from the functional signaling domain of 4-1BB, CD28, OX40, CD3 ⁇ , or a combination thereof.
- the CAR of the invention comprises an IL13R ⁇ 2 antigen binding domain; hinge area; transmembrane domain; costimulatory domain; and a cytoplasmic signaling domain.
- the CAR may be represented by the amino acid sequence of SEQ ID NO: 9 or the amino acid sequence of SEQ ID NO: 10.
- the HGF neutralizing antibody of the present invention may have the amino acid sequence of SEQ ID NO: 13 for the heavy chain and SEQ ID NO: 14 for the light chain.
- cancer includes ovarian cancer, breast cancer, stomach cancer, lung cancer, liver cancer, biliary tract cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, kidney cancer, colon cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, and head cancer. It may be cervical, skin, thyroid, parathyroid, or ureteral cancer.
- “Chimeric antigen receptor (CAR)” includes an antigen-binding domain that recognizes an antigen; A hinge region (or spacer) connecting the antigen-binding domain and the transmembrane domain; transmembrane domain; costimulatory domain; and a second-generation CAR consisting of a cytoplasmic signaling domain.
- the antigen-binding domain is the site where the main signal is transmitted and is located outside the cell membrane and recognizes cancer cells expressing a specific antigen. Therefore, in cancer treatment using CAR-T cells, the specific treatment target is determined by the antigen binding domain.
- the antigen binding to this antigen binding domain is CD19, MUC16, MUCl, CAlX, CEA, and CDS.
- the sequence of the antigen-binding domain that binds to IL13R ⁇ 2 in the CAR according to the present invention is the same as SEQ ID NO: 2, and positions 11, 64, 67, and 107 of the antigen-binding wild type IL-13 sequence that binds to IL13R ⁇ 2 are E11K, respectively. It is a mutation replaced by .R64D.S67D.R107K.
- the amino acid substituted at the position may be replaced with an amino acid with similar properties to the amino acid specified above.
- the CAR according to the present invention has three additional glycines introduced between the antigen-binding domain and the hinge region to increase the solubility of the CAR protein and thus increase the expression of the chimeric antigen receptor.
- the above three glycines (G) can be replaced with amino acids with similar properties: alanine (A), valine (V), leucine (L), or isoleucine (I).
- the co-stimulatory domain of CAR is a site where a co-stimulatory signal is transmitted, and allows CAR-T cells that recognize a specific antigen bound to the antigen-binding domain to trigger an immune response, assist self-proliferation, and increase the remaining time in the body. It plays a role in transmitting signals.
- the costimulatory domain of SEQ ID NO: 6 is used.
- the cytoplasmic signaling domain of CAR used the CD3 ⁇ signaling domain of a normal person containing additional glutamine, rather than the CD3 ⁇ signaling domain of Jurkat T cells, and the additional glutamine is represented by SEQ ID NO: In 7, it means glutamine (Q) at position 50.
- CD3 ⁇ has a total of three immunoreceptor tyrosine-based activation motif (ITAM- YxxL/Ix6-8YxxL/I) sequences, the second and third of the three YxxL/Ix6-8YxxL/I. It can be used by mutating tyrosine (Y) to phenylalanine (F) (SEQ ID NO: 8).
- ITAM- YxxL/Ix6-8YxxL/I immunoreceptor tyrosine-based activation motif
- Nucleic acid sequences of polypeptides constituting the domains disclosed herein can be obtained using recombinant methods known in the art, for example, by screening libraries from cells expressing the genes using standard techniques. , can be obtained by inducing a gene from a vector known to contain the same gene, or by isolating it directly from cells and tissues containing the same gene. Alternatively, the gene of interest can be generated synthetically rather than cloning.
- Expression vectors can be rapidly introduced into host cells by any method known in the art.
- CAR-T cells can be produced by conjugating the finally constructed CAR gene fragment to an MFG retroviral expression vector cleaved with BamH1/NotI. It is to be understood that the present invention includes any number of variants for each component of the construct.
- the CAR according to the present invention may be represented by the amino acid sequence of SEQ ID NO: 9 or SEQ ID NO: 10, and was produced by reflecting the content disclosed in International Patent Publication WO 2017/023138, the entire contents of which are incorporated by reference.
- the HGF neutralizing antibody of the present invention exhibits the activity of neutralizing HGF by binding to a neutralizable epitope, and includes an antibody that specifically binds to HGF or an antigen-binding fragment thereof.
- the antibody or antigen-binding fragment thereof that specifically binds to HGF of the present invention has a V H region represented by the amino acid sequence of SEQ ID NO: 11 and a V L region represented by the amino acid sequence of SEQ ID NO: 12, and has four frameworks There is a framework region (FR) and three antigen binding regions (complementarity determining region (CDR)) (SEQ ID NOs: 15 to 20).
- FR framework region
- CDR complementarity determining region
- the heavy chain of the HGF neutralizing antibody used in combination with the CAR therapy of the present invention can be represented by the amino acid sequence of SEQ ID NO: 13, and the light chain can be represented by the amino acid sequence of SEQ ID NO: 14, and were manufactured according to the contents disclosed in Korean Patent No. 556660. , this entire text is incorporated by reference.
- Cancers that can be prevented or treated by the CAR therapy of the present invention include various cancers known in the art, such as ovarian cancer, breast cancer, stomach cancer, lung cancer, liver cancer, biliary tract cancer, bronchial cancer, nasopharyngeal cancer, It includes, but is not limited to, laryngeal cancer, pancreatic cancer, bladder cancer, kidney cancer, colon cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, skin cancer, thyroid cancer, parathyroid cancer, or ureteral cancer.
- cancer that can be prevented or treated by the CAR therapy of the present invention is cancer that expresses IL13Ra2 and secretes HGF, and more specifically, ovarian cancer that expresses IL13Ra2 and secretes HGF.
- the CAR-T cell therapy of the present invention goes through several steps before being injected into a cancer patient. After extracting T cells from the patient's blood through a process of separating and collecting white blood cells, the CAR-designed gene is injected into the T cells using an expression vector, the CAR-T cells are proliferated, and then injected into the patient.
- the HGF neutralizing antibody of the present invention is administered simultaneously or sequentially with the injection of the CAR-T cell therapy agent, for example, in any order.
- the combination is administered at predetermined treatment intervals.
- the HGF neutralizing antibody is administered within 2 weeks before/after administration of the CAR-T cell therapy or simultaneously with the CAR-T cell therapy.
- the appropriate dosage of the CAR-T cell therapy and HGF neutralizing antibody of the present invention depends on factors such as administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity. There are many, and a normally skilled physician can easily determine and prescribe effective dosages for the desired treatment or prevention. According to a specific embodiment of the present invention, a single dose of the CAR-T cell therapy of the present invention is 1 x 10 7 to 10 8 cells/kg, and a single dose of the HGF neutralizing antibody is 10 to 30 mg/kg. am.
- the CAR therapy agent of the present invention exhibits a preventive or therapeutic effect on solid cancer, including ovarian cancer. More specifically, by combining HGF neutralizing antibodies, the anti-cancer effect of CAR-T cell therapy is increased by increasing the therapeutic effect in the interaction between cancer cells and the immune system, making it useful as a preventive or therapeutic agent for solid cancers, including ovarian cancer. You can. In particular, there is a special effect in that CAR-T cell therapy, a third-generation immunotherapy that has fewer toxicity problems of first-generation chemical anticancer drugs and less resistance problems of second-generation targeted anticancer drugs, can be used for solid cancer.
- Figure 1 shows the results of flow cytometry analysis showing the expression rate of IL13Ra2 (human interleukin 13 receptor alpha 2) in an ovarian cancer cell line (A2780).
- IL13Ra2 human interleukin 13 receptor alpha 2
- FIG. 2 shows the results of hepatocyte growth factor (HGF) ELISA analysis in an ovarian cancer cell line (A2780).
- HGF hepatocyte growth factor
- Figure 3 is a diagram showing the structures of YYB-103 and YYB-103-1XX.
- Figure 4 shows the results of flow cytometry analysis confirming the IL13 expression rate of YYB-103 CAR-T and YYB-103-1XX CAR-T cells.
- Figure 5 shows the results of observing survival by single administration of CAR-T or YYB-101 and combined administration of CAR-T and YYB-101.
- Figure 6 shows the results of observing survival when CAR-T was administered alone at a high concentration.
- Figure 7 shows the sequences of YYB-101, YYB-103 and YYB-103-1XX of the present application.
- Example 1 Confirmation of IL13Ra2 (human interleukin 13 receptor alpha 2) expression in ovarian cancer cell line (A2780)
- IL13Ra2 monoclonal antibody
- FITC-conjugated anti-human IL13Ra2 monoclonal antibody R&D, Cat. No., FAB614F
- the cells were washed once, and the expression rate of IL13Ra2 was checked in the ovarian cancer cell line (A2780).
- an isotype control sample R&D, Cat. No., IC108F was included.
- Table 1 and Figure 1 show the results of confirming the IL13Ra2 expression rate using the ovarian cancer cell line (A2780) according to the experimental method.
- the expression rate of IL13Ra2 in the ovarian cancer cell line (A2780) was 69.1%, and in the case of the isotype control used as a control, it was confirmed to be 0.9%.
- Ovarian cancer cell line (A2780) IL13Ra2 expression rate Unstained 0.7% Isotype control 0.9% Anti-IL13Ra2 69.1%
- HGF Hepatocyte growth factor
- HGF hepatocyte growth factor
- RPMI medium containing 2% fetal bovine serum (FBS, Gibco, 10082-147) was used.
- FBS fetal bovine serum
- A2780 was cultured for 2 days at 32°C and 6% CO 2 conditions. After 2 days, the culture medium was centrifuged at 1,500 rpm for 5 minutes, and the supernatant was transferred to a new 1.5 mL tube.
- human HGF Quantikine ELISA kit R&D system, DHG00B
- RPMI a culture medium for ovarian cancer cell line (A2780) was included as a control for the ELISA analysis results.
- the ELISA analysis results are shown in Figure 2.
- the culture medium of the ovarian cancer cell line (A2780) used as a control the amount of HGF was confirmed at a concentration of 4 ng/mL, and in the case of the ovarian cancer cell line (A2780), the amount of HGF was confirmed at a concentration of 164 ng/mL, which is approximately 41 times higher. It has been done.
- an ovarian cancer cell line (A2780) expressing IL13Ra2 and secreting HGF was selected for the combination test of YYB-103 CAR-T cells and YYB-101.
- YYB-103 a second-generation (IL13.E11K.R64D.S67D.R107K.TNFRSF9.CD3 ⁇ ) chimeric antigen receptor that specifically binds to IL13Ra2, has three additional glycines (G) between the antigen-binding domain and the hinge region. It is introduced, and the antigen-binding domain binds to IL13R ⁇ 2, but as shown in SEQ ID NO: 2, positions 11, 64, 67, and 107 of the antigen-binding wild type IL-13 sequence that binds to IL13R ⁇ 2 are lysine (K), respectively. Aspartic acid (D), aspartic acid (D) and lysine (K) were substituted (SEQ ID NO: 9). YYB-103 expressing T cells were produced according to the disclosure in International Patent Publication WO2017/023138 (FIG. 3), the entire text of which is incorporated by reference.
- G glycines
- CD3 ⁇ a component of T cells, has a total of three immunoreceptor tyrosine-based activation motif (ITAM-YxxL/Ix6-8YxxL/I) sequences, and YYB-103-1XX is the sequence of YYB-103.
- ITAM-YxxL/Ix6-8YxxL/I immunoreceptor tyrosine-based activation motif
- YYB-103-1XX is the sequence of YYB-103.
- the second and third tyrosines (Y) were mutated to phenylalanine (F) (SEQ ID NO: 10).
- YYB-103-1XX was produced according to the contents disclosed in International Publication Patent WO 2019/133969 (FIG. 3)
- YYB-103-1XX expressing T cells were produced according to the contents disclosed in International Publication Patent WO2017/023138, the full text of which is referenced. is integrated into
- YYB-103 CAR-T and YYB-103-1XX CAR-T (donor number, YY93) 1 x 10 6 cells cultured in T cell culture medium were centrifuged. Afterwards, the supernatant was removed, and YYB-103 CAR-T and YYB-103-1XX CAR-T cells were washed twice using PBS containing 2% bovine serum albumin. After washing was completed, the expression of transduced T cells was checked through flow cytometry for surface IL13 expression of YYB-103 CAR-T and YYB-103-1XX CAR-T through IL13. The results are shown in Table 2 and Figure 4. shown in As a control for the flow cytometry results, samples that were not transduced with YYB-103 CAR or YYB-103-1XX CAR virus (Untransduced T cells) were included.
- Flow cytometric analysis was performed to confirm the expression of YYB-103 CAR-T or YYB-103-1XX CAR-T cells (donor number, YY93), and results showed that there was no transduction in YYB-103 CAR or YYB-103-1XX CAR.
- the expression of IL13 in untransduced T cells was less than 0.3% on DAY 11, indicating no IL13 expression, whereas the expression rate of IL13 in YYB-103 CAR-T or YYB-103-1XX CAR-T cells was 0.3% or less on DAY 11.
- -T showed 60.6%, YYB-103-1XX CAR-T showed 54.9%.
- Example 4 Production of a neutralizing antibody (YYB-101) that binds to a neutralizing epitope of HGF (hepatocyte growth factor)
- YYB-101 (SEQ ID NO: 13 and SEQ ID NO: 14), an HGF-neutralizing antibody that interferes with the binding between HGF and its receptor, was produced according to the disclosure in Republic of Korea Patent No. 556660, the entire contents of which are incorporated by reference.
- An ovarian cancer cell line (A2780) expressing IL13R ⁇ 2 and secreting HGF was cultured in a CO 2 incubator (Thermo, 371) at 32°C and 6% CO 2 using DMEM medium containing 10% FBS and 1% antibiotics.
- Ovarian cancer cell line (A2780) tumor transplantation and population construction
- Engraftment was induced by direct transplantation into the ovaries of NSGA mice using 8 ul of 1x10 5 A2780 cells and 2 ul of matrigel. Three days after engraftment, the weight of the test animals was measured and the tumors were distributed evenly using in vivo luciferase imaging.
- the test group consists of one control group (Vehicle) and five test substance administration groups (Untransduced T cells, YYB-103 CAR-T, YYB-103-1XX CAR-T, YYB-101 and YYB-103-1XX CAR-T and (YYB-101 combined use), and 3 animals were distributed to each group.
- each untransduced T cell and CAR-T cell were prepared after being washed twice with PBS and diluted with PBS.
- T cells not transduced with CAR virus (Untransduced T cells), YYB-103 CAR-T, and YYB-103-1XX CAR-T cells were administered intravenously as a single dose at a concentration of 1.5 x 10 7 cells.
- YYB-103-1XX CAR-T and YYB-101 combination group YYB-101 is administered simultaneously with YYB-103-1XX CAR-T (1st treatment) and administered one additional time (2nd treatment). The administration was repeated intravenously twice a week (20 mpk).
- Ovarian cancer cell line (A2780) was cultured in a CO 2 incubator (Thermo, 371) at 32°C and 6% CO 2 using DMEM medium containing 10% FBS and 1% antibiotics.
- Ovarian cancer cell line (A2780) tumor transplantation and population construction
- Engraftment was induced by directly transplanting 8 ul of 1x10 5 A2780 cells and 2 ul of matrigel into the ovaries of NSGA mice. Three days after engraftment, it was confirmed that the tumor size was evenly distributed by measuring the weight of the test animals and using in vivo luciferase imaging.
- the test group consisted of one control group (Vehicle) and one test substance administration group (YYB-103 CAR-T), with 5 animals distributed to each group.
- CAR-T cell therapy CAR-T cells were prepared after being washed twice with PBS and diluted with PBS.
- YYB-103 CAR-T cells were administered intravenously as a single dose at a concentration of 5 x 10 7 cells.
- the CAR therapy according to the present invention exhibits an anticancer effect on solid cancers, including ovarian cancer, and can be used as a medicine such as an anticancer agent.
Abstract
본 발명은 HGF의 중화가능 에피토프에 결합하는 중화 항체와 조합하여 대상체에서 암의 치료에 사용하기 위한 키메릭 항원 수용체(CAR)를 발현하는 면역 이펙터 세포의 집단을 포함하는 CAR 요법제에 관한 것으로서, HGF 중화항체를 조합함으로써 암세포와 면역계의 상호 작용에 있어서 치료효과를 상승시켜 CAR-T 세포치료제의 항암 효과를 증가시켜 혈액암 뿐만 아니라 고형암 치료에 효과적으로 사용될 수 있다.
Description
본 발명은 3세대 면역항암제인 CAR-T 세포치료제의 고형암 예방 또는 치료에 관한 것이다. 구체적으로, 간세포성장인자 (Hepatocyte growth factor, HGF) 중화항체를 조합하여 암세포와 면역계의 상호 작용에 영향을 줌으로써 CAR-T 세포치료제의 항암 효과를 증가시키는 것을 특징으로 한다.
본 발명의 CAR-T 세포치료제는 면역항암제가 활용되기 어렵다고 인식되어 있는 고형암 치료에도 효과적으로 사용될 수 있다.
수십 년 동안 암을 치료하는 방법들은 꾸준히 변화하고 발전해왔다. 1800년대에서부터 1900년대까지는 외과적인 수술 (Surgery), 화학요법 (Chemotherapy), 그리고 방사선 요법 (Radiation therapy)과 같은 방법들이 주로 이뤄졌지만, 이들에 대한 한계점들이 드러나기 시작하여, 최근 면역세포 요법으로 체내의 면역세포를 꺼내서, 강화시키거나 유전공학적으로 변형시켜 다시 넣어주는 세포치료 방식이 개발되고 있다.
보다 구체적으로, 암을 치료하기 위한 1세대 화학항암제의 경우 세포독성물질로 암세포를 공격해 사멸시키는 방법인데, 암세포 뿐만 아니라 정상세포도 같이 손상을 주기 때문에 부작용이 심한 문제가 있다. 이러한 1세대 화학항암제의 단점을 극복하기 위한 2세대 표적항암제의 경우 암세포의 특정 물질을 목표로 공격하기 때문에 1세대 화학항암제에 비해 부작용은 적지만 내성이 생긴다는 큰 단점을 지니고 있다. 반면, 3세대 면역항암제는 우리 몸의 면역체계를 이용하기 때문에 1세대 화학항암제의 독성 문제와 2세대 표적항암제의 내성 문제가 적고 부작용도 현저히 적다. 또한, 2세대 표적항암제는 초기에 높은 생존율을 보이지만, 내성문제로 지속성이 떨어지는 반면 면역항암제는 항암효과가 유지돼 면역항암제에 대한 반응이 좋은 환자는 완치에 가까워진다. 이처럼 3세대 면역항암제는 탁월한 효능과 적은 부작용으로 인해 암환자의 장기 생존율은 물론 암환자의 삶의 질을 높이는데 기여한다.
키메릭 항원 수용체(chimeric antigen receptor, CAR)는 항체의 단편, 힌지 영역, 막통과 도메인 및 세포 내 신호전달 도메인으로 구성되는데, 상기 키메릭 항원 수용체가 발현된 T 세포는 암세포만 공격하게 설계된 면역항암제 중 하나이다. CAR-T 세포치료제는 암세포를 특이적으로 공격하는 면역 T 세포의 공격성을 크게 높였기 때문에 정상세포의 손상은 줄이고 암세포만을 표적 삼아 공격한다는 점에서 매우 중요한 세포치료제이다.
현재까지 CAR T 세포치료제와 같은 면역세포치료가 일부 혈액암에서 높은 완치율을 보였지만, 암 대부분을 차지하는 고형암에서는 제대로 치료효과를 발휘하지 못하였는데, 이는 인체가 강한 면역반응을 억제하는 경향이 있어 투여된 면역세포가 충분히 활동할 수 없기 때문으로 알려져 있다.
[선행기술문헌]
[특허문헌]
(특허문헌 1) 대한민국 등록특허 제10-0556660호
(특허문헌 2) 대한민국 등록특허 제10-2011789호
이에, 본 발명자들은 CAR-T 세포치료제를 난소암 등 고형암에서도 치료 효과가 발휘될 수 있도록 하는 치료 방법을 연구한 결과, 1세대 화학항암제의 독성 문제와 2세대 표적항암제의 내성 문제가 적은 3세대 면역항암제인 CAR-T 세포치료제에 HGF의 중화가능 에피토프에 결합하는 중화항체를 조합할 경우 CAR-T 세포치료제 단독에 비해 종양 진행의 억제 또는 감소의 개선을 초래할 수 있다는 것을 확인하고, 본 발명을 완성하였다.
따라서, 본 발명은 난소암을 포함한 고형암 치료를 위한 CAR 요법제를 제공하는 것을 해결과제로 하며, HGF의 중화가능 에피토프에 결합하는 중화 항체와 조합하여 대상체에서 암의 치료에 사용하기 위한 키메릭 항원 수용체(CAR)를 발현하는 면역 이펙터 세포의 집단을 포함하는 CAR 요법제를 제공하는 것을 구체적인 해결과제로 한다.
상기 과제를 해결하기 위하여, 본 발명에서는 하기와 같은 수단을 개시한다.
본 발명은 HGF의 중화가능 에피토프에 결합하는 중화 항체와 조합하여 대상체에서 암의 치료에 사용하기 위한 키메릭 항원 수용체(chimeric antigen receptor, CAR)를 발현하는 면역 이펙터 세포의 집단을 포함하는 CAR 요법제를 개시한다.
본 발명에서 CAR는 IL13Rα2 항원 결합 도메인; 힌지 영역; 막관통 도메인; 보조 자극 도메인; 및 세포질 신호 전달 도메인을 포함할 수 있으며, 이에 한정되는 것은 아니다.
상기 CAR는 서열번호 9 또는 서열번호 10의 아미노산 서열로 표시된다.
상기 HGF 중화 항체의 중쇄는 서열번호 13, 경쇄는 서열번호 14의 아미노산 서열로 표시된다. 또한, 공지의 링커 서열이 추가될 수 있다.
본 발명은 HGF의 중화가능 에피토프에 결합하는 중화 항체와 조합하여 대상체에서 암의 치료에 사용하기 위한 키메릭 항원 수용체(CAR)를 발현하는 면역 이펙터 세포의 집단을 포함하는 CAR 요법제를 개시한다.
본 발명에서, 상기 CAR는 항원 결합 도메인; 힌지 영역; 막관통 도메인; 보조 자극 도메인; 및 세포질 신호 전달 도메인을 포함할 수 있으며, 이에 한정되는 것은 아니다.
상기 CAR의 항원 결합 도메인은 CD19, MUC16, MUCl, CAlX, CEA, CDS, CD7, CD10, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, 사이토메갈로바이러스(CMV) 감염된 세포 항원, EGP-2, EGP-40, EpCAM, erb-B2,3,4, FBP, 태아 아세틸콜린 수용체, 폴레이트 수용체-a, GD2, GD3, HER-2, hTERT, K-경쇄, KDR, LeY, L1 세포 부착 분자, MAGE-A1, 메소텔린, NKG2D 리간드, NY-ES0-1, 암태아 항원(h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, WT-1, CD276 또는 IL13Ra2로부터 선택되는 항원과 결합하는 항원 결합 도메인을 포함할 수 있다.
상기 CAR의 막관통 도메인은 T-세포 수용체의 알파, 베타 또는 제타 쇄, CD28, CD3엡실론, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 또는 CD154로부터 선택되는 막관통 도메인을 포함할 수 있다.
상기 CAR의 보조 자극 도메인은 MHC 클래스 I 분자, TNF 수용체 단백질, 이뮤노글로불린-유사 단백질, 시토카인 수용체, 인테그린, 신호전달 림프구성 활성화 분자 (signaling lymphocytic activation molecule, SLAM), 활성화 NK 세포 수용체, BTLA(B an T lymphocyte attenuator), 톨-유사 리간드 수용체(Tolllike ligand receptor), OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8알파, CD8베타, IL2R 베타, IL2R 감마, IL7R 알파, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, 또는 CD83과 특이적으로 결합하는 리간드로부터 선택되는 보조 자극 도메인을 포함할 수 있다.
상기 CAR의 세포질 신호 전달 도메인은 4-1BB, CD28, OX40, CD3ζ의 기능적 신호 전달 도메인, 또는 이들의 조합으로부터 선택되는 세포질 신호 전달 도메인을 포함할 수 있다.
구체적인 양태에서, 본 발명의 CAR는 IL13Rα2 항원 결합 도메인; 힌지 영역; 막관통 도메인; 보조 자극 도메인; 및 세포질 신호 전달 도메인을 포함할 수 있다. 이때, 상기 CAR는 서열번호 9의 아미노산 서열 또는 서열번호 10의 아미노산 서열로 표시될 수 있다.
또한, 본 발명의 HGF 중화항체는 중쇄가 서열번호 13, 경쇄가 서열번호 14의 아미노산 서열로 표시될 수 있다.
본 발명에서 암은 난소암, 유방암, 위암, 폐암, 간암, 담도암, 기관지암, 비인두암, 후두암, 췌장암, 방광암, 신장암, 대장암, 결장암, 자궁경부암, 뇌암, 전립선암, 골암, 두경부암, 피부암, 갑상선암, 부갑상선암 또는 요관암일 수 있다.
본 발명에 따른 “키메릭 항원 수용체(chimeric antigen receptor, CAR)”는 항원을 인식하는 항원 결합 도메인; 항원 결합 도메인과 막통과 도메인을 연결하는 힌지 영역(또는 스페이서); 막통과 도메인; 보조 자극 도메인; 및 세포질 신호 도메인으로 구성된 2세대 CAR이다.
본 발명의 CAR 구조에서 항원 결합 도메인은 주신호가 전달되는 부위로 세포막 외부에 있으며 특정 항원을 발현하는 암세포를 인식하는 부분이다. 따라서, CAR-T 세포를 이용한 암 치료에 있어서, 구체적인 치료대상은 항원 결합 도메인에 의하여 결정되는데, 본 발명에서, 이와 같은 항원 결합 도메인에 결합하는 항원은 CD19, MUC16, MUCl, CAlX, CEA, CDS, CD7, CD10, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, 사이토메갈로바이러스(CMV) 감염된 세포 항원, EGP-2, EGP-40, EpCAM, erb-B2,3,4, FBP, 태아 아세틸콜린 수용체, 폴레이트 수용체-a, GD2, GD3, HER-2, hTERT, K-경쇄, KDR, LeY, L1 세포 부착 분자, MAGE-A1, 메소텔린, NKG2D 리간드, NY-ES0-1, 암태아 항원(h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, WT-1, CD276 또는 IL13Ra2가 포함될 수 있으며, 이에 제한되지 않고, 바람직하게는 IL13Rα2에 특이적으로 결합할 수 있는 키메릭 항원 수용체를 개시한다.
본 발명에 따른 CAR에서 IL13Rα2와 결합하는 항원 결합 도메인의 서열은 서열번호 2와 같은데, IL13Rα2와 결합하는 항원 결합 wild type IL-13 서열의 11번, 64번, 67번 및 107번 위치가 각각 E11K.R64D.S67D.R107K로 치환된 돌연변이다. 해당 위치에서 치환되는 아미노산은 상기 특정된 아미노산과 유사한 성질의 아미노산으로 대체될 수 있다.
또한, 본 발명에 따른 CAR는 CAR 단백질의 용해도를 높여 키메릭 항원 수용체의 발현을 증가시키기 위해서 항원 결합 도메인과 힌지 영역 사이에 3개의 글리신을 추가적으로 도입한 것이다. 위 3개의 글리신(G)은 이와 유사한 성질의 아미노산인 알라닌(A), 발린(V), 류신(L) 또는 이소류신(I)으로 치환될 수 있다.
본 발명에 따른 CAR의 보조 자극 도메인은 보조 자극 신호가 전달되는 부위로서 항원 결합 도메인과 결합한 특정 항원을 인식한 CAR-T 세포가 면역반응을 일으키며 자가 증식을 돕고, 체내에 잔존하는 시간을 늘리도록 신호를 전달하는 역할을 한다. 본 발명에서는 서열번호 6의 보조 자극 도메인을 사용한다.
본 발명에 따른 CAR의 세포질 신호 전달 도메인은 Jurkat T 세포의 CD3ζ 신호전달 도메인이 아닌, 추가의 글루타민이 포함된 정상 인간(normal person)의 CD3ζ 신호전달 도메인을 사용하였고, 상기 추가의 글루타민은 서열번호 7에 있어서 50번 위치의 글루타민(Q)를 의미한다. 또한, CD3ζ는 총 3개의 면역 수용체 티로신 기반 활성화 모티프 (Immunoreceptor tyrosine-based activation motif, ITAM- YxxL/Ix6-8YxxL/I) 서열을 지니고 있는데, 3개의 YxxL/Ix6-8YxxL/I 중 2번째와 3번째의 tyrosine (Y)을 phenylalanine (F)으로 돌연변이(Mutation) 하여 사용될 수 있다 (서열번호 8). 이와 같은 돌연변이는 WO 2019/133969에 개시된 내용으로 제작되었고, 이 전문은 참고로 통합된다.
본 명세서에서 개시하는 도메인을 구성하는 폴리펩타이드의 핵산 서열은 당해 기술분야에 공지된 재조합 방법을 이용하여 수득될 수 있으며, 예를 들어 표준 기법을 이용하여 상기 유전자를 발현하는 세포로부터 라이브러리를 스크리닝하거나, 상기 동일한 유전자를 포함하도록 공지된 벡터로부터 유전자를 유도하거나, 상기 동일한 유전자를 포함하는 세포 및 조직으로부터 직접 단리함으로써 수득될 수 있다. 대안적으로는, 상기 관심이 있는 유전자는 클로닝이 아닌 합성에 의해 생성될 수 있다.
세포 내로 유전자를 도입하고 발현하는 방법은 당해 기술분야에 공지되어 있다. 발현 벡터는 당해 기술분야에 공지된 임의의 방법에 의해 숙주 세포 내로 신속하게 도입될 수 있다. 예를 들면, 본 발명에서는 최종적으로 제작된 CAR 유전자 단편을 BamH1/NotI로 절단된 MFG 레트로바이러스 발현 벡터에 접합시켜 CAR-T 세포를 제조할 수 있다. 본 발명은 구조체의 성분 각각에 대한 다수의 임의의 변이체를 포함하는 것으로 이해되어야 한다.
또한, 본 발명에 따른 CAR는 서열번호 9 또는 서열번호 10의 아미노산 서열로 표시될 수 있으며, 국제공개특허 WO 2017/023138에 개시된 내용을 반영하여 제작되었으며, 이 전문은 참조로 통합된다.
본 발명의 HGF 중화항체는 중화가능 에피토프에 결합하여 HGF를 중화시킬 수 있는 활성을 나타내는 것으로, HGF에 특이적으로 결합하는 항체 또는 그의 항원 결합 단편을 포함한다.
본 발명의 HGF에 특이적으로 결합하는 항체 또는 그의 항원 결합 단편은 서열번호 11의 아미노산 서열로 표시되는 VH 영역 및 서열번호 12의 아미노산 서열로 표시되는 VL 영역을 갖는 것으로, 4개의 프레임워크 영역(framework region; FR)과 3개의 항원결합부위(complementarity determining region; CDR)가 존재(서열번호 15 내지 20)한다.
구체적으로, 본 발명의 CAR 요법제와 조합하여 사용되는 HGF 중화항체의 중쇄는 서열번호 13, 경쇄는 서열번호 14의 아미노산 서열로 표시될 수 있으며, 대한민국 등록특허 제556660호에 개시된 내용으로 제작되었으며, 이 전문은 참조로 통합된다.
본 발명의 CAR 요법제에 의해 예방 또는 치료될 수 있는 암은 당업계에 공지된 다양한 암을 포함하며, 예를 들어 난소암, 유방암, 위암, 폐암, 간암, 담도암, 기관지암, 비인두암, 후두암, 췌장암, 방광암, 신장암, 대장암, 결장암, 자궁경부암, 뇌암, 전립선암, 골암, 두경부암, 피부암, 갑상선암, 부갑상선암 또는 요관암을 포함하나, 이에 한정되는 것은 아니다.
구체적으로, 본 발명의 CAR 요법제에 의해 예방 또는 치료될 수 있는 암은 IL13Ra2 를 발현하면서 HGF를 분비하는 암이고, 보다 구체적으로 IL13Ra2를 발현하면서 HGF를 분비하는 난소암이다.
본 발명의 CAR-T 세포치료제는 암 환자에게 주입하기까지는 여러 단계를 거친다. 환자의 혈액에서 백혈구성분 분리채집 과정을 거쳐 T 세포를 추출한 뒤, 발현 벡터를 이용하여 CAR로 디자인된 유전자를 T 세포에 주입하고 이 CAR-T 세포를 증식시킨 후, 이를 환자에게 주입하게 된다.
또한, 본 발명의 HGF 중화항체는 CAR-T 세포치료제 주입과 동시에 또는 순차적으로,예를 들어 임의의 순서로 투여된다. 일 실시형태에서, 조합물은 소정의 치료 간격에서 투여된다. 본 발명의 구체적인 구현예에 따르면, HGF 중화항체의 투여는 CAR-T 세포치료제의 투여 전/후 2주 이내 또는 CAR-T 세포치료제와 동시에 투여된다.
본 발명의 CAR-T 세포치료제 및 HGF 중화항체의 적합한 투여량은 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 구체적인 구현예에 따르면, 본 발명의 CAR-T 세포치료제의 1회 투여량은 1 x 107~108 cells/ kg 이고, HGF 중화항체의 1회 투여량은 10~30 mg/kg 이다.
본 발명에 CAR 요법제는 난소암을 포함하는 고형암의 예방 또는 치료 효과를 나타낸다. 보다 특별하게는, HGF 중화항체를 조합함으로써 암세포와 면역계의 상호 작용에 있어서 치료 효과 상승을 통해 CAR-T 세포치료제의 항암 효과를 증가시켜 난소암을 포함하는 고형암의 예방 또는 치료제로서 유용하게 활용될 수 있다. 특히, 1세대 화학항암제의 독성 문제와 2세대 표적항암제의 내성 문제가 적은 3세대 면역항암제인 CAR-T 세포치료제를 고형암에 활용할 수 있다는 특별한 효과가 있다.
도 1은 난소암 세포주 (A2780)에서 IL13Ra2 (human interleukin 13 receptor alpha 2)발현률을 나타내는 유세포 분석 결과이다.
도 2는 난소암 세포주 (A2780)에서 간세포 성장인자 (Hepatocyte growth factor, HGF) ELISA 분석 결과이다.
도 3은 YYB-103 및 YYB-103-1XX의 구조를 나타낸 도면이다.
도 4는 YYB-103 CAR-T 및 YYB-103-1XX CAR-T 세포의 IL13 발현률을 확인한 유세포 분석 결과이다.
도 5는 CAR-T 또는 YYB-101 단독투여 및 CAR-T와 YYB-101 병용투여에 의한 생존을 관찰한 결과이다.
도 6은 고농도로 CAR-T 단독 투여한 경우 생존을 관찰한 결과이다.
도 7은 본 출원의 YYB-101, YYB-103 및 YYB-103-1XX의 서열에 대한 것이다.
이하, 본 발명을 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
실시예 1. 난소암 세포주 (A2780)에서 IL13Ra2 (human interleukin 13 receptor alpha 2) 발현 확인
실험 방법
유세포 분석을 위해, FITC-conjugated anti-human IL13Ra2 단일클론 항체 (R&D, Cat. No., FAB614F)를 첨가하기 전 세포를 2% bovine serum albumin을 함유한 PBS에 1회 세척하였다. 세척 후 빛이 차단된 상태에서 4℃, 30 분간 각각의 항체와 반응한 후, 세포를 1회 세정하고, 난소암 세포주 (A2780)에서 IL13Ra2 발현률을 체크하였다. 대조군으로 isotype control 시료 (R&D, Cat. No., IC108F)를 포함시켰다.
실험 결과
실험방법에 따라 난소암 세포주 (A2780)을 이용하여 IL13Ra2 발현률을 확인한 결과를 표 1 및 도 1에 나타내었다. 난소암 세포주 (A2780)의 IL13Ra2의 발현률은 69.1% 이고, 대조군으로 사용한 isotype control의 경우 0.9%로 확인되었다.
난소암 세포주(A2780) | IL13Ra2 발현률 |
Unstained | 0.7% |
Isotype control | 0.9% |
Anti-IL13Ra2 | 69.1% |
실시예 2.
난소암 세포주 (A2780)에서 간세포 성장인자 (Hepatocyte growth factor, HGF) 분석
실험 방법
난소암 세포주 (A2780)에서 간세포 성장인자 (Hepatocyte growth factor, HGF)가 분비되는 양을 확인하기 위해서 2% Fetal bovine serum (FBS, Gibco, 10082-147)가 포함된 RPMI 배지를 이용하여 난소암 세포주 (A2780)을 32℃, 6% CO2 조건에서 2일간 배양하였다. 2일후 배양 배지를 1,500 rpm 조건으로 5분간 원심분리 한 후 상층액을 새로운 1.5 mL tube에 옮겼다. 난소암 세포주 (A2780)에서 분비되는 HGF의 양을 확인하기 위해서 human HGF Quantikine ELISA kit (R&D system, DHG00B)을 이용하였다. ELISA 분석 결과에 대한 대조군으로 난소암 세포주 (A2780)의 배양액인 RPMI을 포함시켰다.
실험 결과
ELISA 분석 결과를 도 2에 나타내었다. 대조군으로 사용한 난소암 세포주 (A2780)의 배양액인 RPMI의 경우 4 ng/mL의 농도로 HGF 양이 확인되었으며, 난소암 세포주 (A2780)의 경우 164 ng/mL 농도로 약 41배 높은 HGF 양이 확인되었다.
실시예 1 및 실시예 2의 결과를 종합하여, IL13Ra2 발현 및 HGF를 분비하는 난소암 세포주 (A2780)가 YYB-103 CAR-T 세포와 YYB-101의 병용 시험을 위해 선정되었다.
실시예 3. YYB-103 그리고 YYB-103-1XX 유전자 발현 T세포의 제작
YYB-103 발현 T 세포의 제작
IL13Ra2에 특이적으로 결합하는 2세대 (IL13.E11K.R64D.S67D.R107K.TNFRSF9.CD3ζ) 키메릭 항원 수용체인 YYB-103은 항원 결합 도메인과 힌지 영역 사이에 추가로 3개의 글리신(G)이 도입되어 있으며, 항원 결합 도메인은 IL13Rα2와 결합하되, 서열번호 2와 같이 IL13Rα2와 결합하는 항원 결합 wild type IL-13 서열의 11번, 64번, 67번 및 107번 위치가 각각 리신(K), 아스파르트산(D), 아스파르트산(D) 및 리신(K)으로 치환되었다(서열번호 9). YYB-103 발현 T 세포는 국제공개특허 WO2017/023138에 개시된 내용으로 제작되었으며(도 3), 이 전문은 참조로 통합된다.
YYB-103-1XX 발현 T 세포의 제작
T 세포의 구성요소인 CD3ζ 는 총 3개의 면역 수용체 티로신 기반 활성화 모티프 (Immunoreceptor tyrosine-based activation motif, ITAM- YxxL/Ix6-8YxxL/I) 서열을 지니고 있는데, YYB-103-1XX는 YYB-103의 3개의 YxxL/Ix6-8YxxL/I 중 2번째와 3번째의 tyrosine (Y)을 phenylalanine (F)로 돌연변이(Mutation) 하였다(서열번호 10). YYB-103-1XX는 국제공개특허 WO 2019/133969에 개시된 내용으로 제작되었고(도 3), YYB-103-1XX 발현 T 세포는 국제공개특허 WO2017/023138에 개시된 내용으로 제작되었으며, 이 전문들은 참고로 통합된다.
실험방법
T 세포 배양 배지에 배양 중인 YYB-103 CAR-T 및 YYB-103-1XX CAR-T (공여자 번호, YY93) 1 x 106 세포를 원심분리 하였다. 그 후 상층액을 제거하고, 2% bovine serum albumin을 함유한 PBS을 이용하여 YYB-103 CAR-T 및 YYB-103-1XX CAR-T 세포를 2회 세척하였다. 세척이 완료된 후 형질 도입된 T 세포의 발현은 IL13을 통해서 YYB-103 CAR-T 및 YYB-103-1XX CAR-T의 surface IL13 발현을 유세포 분석을 통해 체크하였으며, 그 결과는 표 2 및 도 4에 나타내었다. 유세포 분석 결과에 대한 대조군으로 YYB-103 CAR 또는 YYB-103-1XX CAR 바이러스에 형질도입 되지 않은 (Untransduced T 세포) 시료를 포함시켰다.
공여자 번호 (YY93) | CAR-T 발현 (%, IL13) |
Untransduced T 세포 | 0.3 |
YYB-103 CAR-T | 60.6 |
YYB-103-1XX CAR-T | 54.9 |
실험 결과
YYB-103 CAR-T 또는 YYB-103-1XX CAR-T 세포 (공여자 번호, YY93)의 발현을 확인하기 위해서 유세포 분석을 진행한 결과, YYB-103 CAR 또는 YYB-103-1XX CAR에 형질도입 되지 않은 untransduced T 세포의 IL13 발현은 DAY 11에서 0.3% 이하로 IL13이 발현하지 않았는데 반해, YYB-103 CAR-T 또는 YYB-103-1XX CAR-T 세포의 IL13의 발현률은 DAY 11에서 YYB-103 CAR-T는 60.6%, YYB-103-1XX CAR-T는 54.9%을 보였다.
실시예 4. HGF(hepatocyte growth factor; 간세포성장인자)의 중화가능 에피토프에 결합하는 중화 항체(YYB-101)의 제작
HGF와 그 수용체간의 결합을 방해하는 HGF 중화 항체인 YYB-101(서열번호 13 및 서열번호 14)은 대한민국 등록특허 제556660호에 개시된 내용으로 제작되었으며, 이 전문은 참조로 통합된다.
실험예 1. CAR-T 또는 YYB-101 단독투여 및 CAR-T와 YYB-101 병용 투여
난소암 세포주 (A2780) 준비
IL13Rα2 발현 및 HGF 분비하는 난소암 세포주 (A2780)를 10% FBS와 1% antibiotics가 포함된 DMEM 배지를 사용하여 CO2 incubator (Thermo, 371) 내에서 32℃, 6% CO2 조건에서 배양하였다.
난소암 세포주 (A2780) 종양이식 및 군구성
1x105 A2780 세포 8 ul와 matrigel 2 ul를 이용하여 NSGA 마우스의 난소에 직접 이식하여 생착을 유도하였다. 생착 3일 후, 시험동물의 체중 측정 및 in vivo luciferase 이미징을 이용하여 종양의 크기가 고르게 분포되도록 분배하였다. 시험군은 1개의 대조군 (Vehicle)과 5개의 시험물질의 투여군 (Untransduced T 세포, YYB-103 CAR-T, YYB-103-1XX CAR-T, YYB-101 및 YYB-103-1XX CAR-T와 YYB-101 병용)으로 구성하였으며, 각 군당 3마리씩 분배하였다.
시험물질 조제
CAR-T 세포치료제에 대하여 PBS을 이용하여 2회 세척 후 각각의 Untransduced T 세포 및 CAR-T 세포를 PBS로 희석한 후 준비하였다.
실험 방법
종양이식 후 3일에 CAR 바이러스에 형질도입 되지 않은 T 세포 (Untransduced T 세포), YYB-103 CAR-T 및 YYB-103-1XX CAR-T 세포를 1.5 x 107 세포 농도로 정맥으로 단회 투여하였다. 또한, YYB-103-1XX CAR-T와 YYB-101 병용군의 경우 YYB-101은 YYB-103-1XX CAR-T와 동시에 투여(1차)를 시작하여 추가(2차)로 1회 더 투여하여 총 주 2회 (20 mpk) 정맥으로 반복 투여하였다.
실험 결과
Untransduced T 세포군과 YYB-101만을 처리한 군의 경우 종양 생착 후 20~30일 안에 모든 개체가 사망하였다. 반면, YYB-103 CAR-T 및 YYB-103-1XX CAR-T의 경우 마지막 개체가 50일 이후 사망함에 따라서 Untransduced T 세포 군보다 약 20일 생존이 길어질 수 있음을 확인하였다. YYB-103-1XX CAR-T와 YYB-101을 병용한 경우에는 마지막 개체의 사망일 기준으로 YYB-103-1XX CAR-T 단독군에 비해서 17일정도 더 생존하였다.
그 결과는 도 5에 나타난 바와 같이, YYB-103-1XX CAR-T 세포치료제와 HGF을 특이적으로 인식하는 항체인 YYB-101의 병용하였을 때 CAR-T 세포치료제 단독 투여군에 비해 항암 효과가 증가하는 것을 확인할 수 있었다.
실험예 2. 고용량 CAR-T 단독 투여
난소암 세포주 (A2780) 준비
난소암 세포주 (A2780)를 10% FBS와 1% antibiotics가 포함된 DMEM 배지를 사용하여 CO2 incubator (Thermo, 371) 내에서 32℃6% CO2 조건에서 배양하였다.
난소암 세포주 (A2780) 종양이식 및 군구성
NSGA 마우스에 1x105 A2780 세포 8 ul와 matrigel 2 ul를 이용하여 NSGA 마우스의 난소에 직접 이식하여 생착을 유도하였다. 생착 3일 후, 시험동물의 체중 측정 및 in vivo luciferase 이미징을 이용하여 종양의 크기가 고르게 분배되었음을 확인하였다. 시험군은 1개의 대조군 (Vehicle)과 1개의 시험물질의 투여군 (YYB-103 CAR-T)으로 구성하였으며, 각 군당 5마리씩 분배하였다.
시험물질 조제
CAR-T 세포치료제에 대하여 PBS을 이용하여 2회 세척 후 CAR-T 세포를 PBS로 희석한 후 준비하였다.
실험 방법
종양이식 후 3일에, YYB-103 CAR-T 세포를 5 x 107 세포 농도로 정맥으로 단회 투여하였다.
실험 결과
YYB-103 CAR-T 세포를 고농도(5 x 107 세포)로 정맥으로 단회 투여한 경우, 도 6에 나타난 바와 같이 마지막 개체가 약 70일에 사망함을 확인할 수 있었다. 즉, 5 x 107 CAR-T 세포 투여에 의한 마우스의 중간 생존이 상기 실험예 1의 1.5 x 107 YYB-103-1XX CAR-T와 YYB-101의 병용과 유사한 결과를 나타냄을 알 수 있었다. 이와 같은 결과는, YYB-103-1XX CAR-T와 YYB-101의 병용을 통해 저용량의 CAR-T로도 효과적으로 암세포를 사멸시킬 수 있기 때문에, 면역 효과 세포 관련 신경 독성 증후군(ICANS) 및 사이토카인 방출 증후군(CRS)와 같은 CAR-T 주입에 따른 부작용을 최소화 할 수 있다는 것을 의미한다.
본 발명에 따른 CAR 요법제는 난소암을 포함하는 고형암에 대한 항암 효과를 나타내므로, 항암제와 같은 의약품으로 이용 가능하다.
Claims (11)
- HGF의 중화가능 에피토프에 결합하는 중화 항체와 조합하여 대상체에서 암의 치료에 사용하기 위한 키메릭 항원 수용체(CAR)를 발현하는 면역 이펙터 세포의 집단을 포함하는 CAR 요법제.
- 제 1항에 있어서, 상기 CAR가 항원 결합 도메인; 힌지 영역; 막관통 도메인; 보조 자극 도메인; 및 세포질 신호 전달 도메인을 포함하는 CAR 요법제.
- 제 2항에 있어서, 상기 CAR의 항원 결합 도메인이 CD19, MUC16, MUCl, CAlX, CEA, CDS, CD7, CD10, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, 사이토메갈로바이러스(CMV) 감염된 세포 항원, EGP-2, EGP-40, EpCAM, erb-B2,3,4, FBP, 태아 아세틸콜린 수용체, 폴레이트 수용체-a, GD2, GD3, HER-2, hTERT, K-경쇄, KDR, LeY, L1 세포 부착 분자, MAGE-A1, 메소텔린, NKG2D 리간드, NY-ES0-1, 암태아 항원(h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, WT-1, CD276 또는 IL13Ra2로부터 선택되는 항원과 결합하는 항원 결합 도메인을 포함하는 CAR 요법제.
- 제 2항에 있어서, 상기 CAR의 막관통 도메인이 T-세포 수용체의 알파, 베타 또는 제타 쇄, CD28, CD3엡실론, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 또는 CD154로부터 선택되는 막관통 도메인을 포함하는 CAR 요법제.
- 제 2항에 있어서, 상기 CAR의 보조 자극 도메인이 MHC 클래스 I 분자, TNF 수용체 단백질, 이뮤노글로불린-유사 단백질, 시토카인 수용체, 인테그린, 신호전달 림프구성 활성화 분자 (signaling lymphocytic activation molecule, SLAM), 활성화 NK 세포 수용체, BTLA(B an T lymphocyte attenuator), 톨-유사 리간드 수용체(Tolllike ligand receptor), OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8알파, CD8베타, IL2R 베타, IL2R 감마, IL7R 알파, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, 또는 CD83과 특이적으로 결합하는 리간드로부터 선택되는 보조 자극 도메인을 포함하는 CAR 요법제.
- 제 2항에 있어서, 상기 CAR의 세포질 신호 전달 도메인이 4-1BB, CD28, OX40, CD3ζ의 기능적 신호 전달 도메인, 또는 이들의 조합으로부터 선택되는 세포질 신호 전달 도메인을 포함하는 CAR 요법제.
- 제 2항에 있어서, 상기 CAR가 IL13Rα2 항원 결합 도메인; 힌지 영역; 막관통 도메인; 보조 자극 도메인; 및 세포질 신호 전달 도메인을 포함하는 CAR 요법제.
- 제 7항에 있어서, 상기 CAR가 서열번호 9의 아미노산 서열로 표시되는 CAR 요법제.
- 제 7항에 있어서, 상기 CAR가 서열번호 10의 아미노산 서열로 표시되는 CAR 요법제.
- 제 1항에 있어서, 상기 HGF 중화항체의 중쇄가 서열번호 13, 경쇄가 서열번호 14의 아미노산 서열로 표시되는 CAR 요법제.
- 제 1항 내지 제10항 중 어느 한 항에 있어서, 상기 암이 난소암, 유방암, 위암, 폐암, 간암, 담도암, 기관지암, 비인두암, 후두암, 췌장암, 방광암, 신장암, 대장암, 결장암, 자궁경부암, 뇌암, 전립선암, 골암, 두경부암, 피부암, 갑상선암, 부갑상선암 또는 요관암인 CAR 요법제.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2022-0046299 | 2022-04-14 | ||
KR1020220046299A KR20230148433A (ko) | 2022-04-14 | 2022-04-14 | 키메릭 항원 수용체 및 hgf 중화항체의 조합 요법 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2023200266A1 true WO2023200266A1 (ko) | 2023-10-19 |
Family
ID=88330022
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2023/004996 WO2023200266A1 (ko) | 2022-04-14 | 2023-04-13 | 키메릭 항원 수용체 및 hgf 중화항체의 조합 요법 |
Country Status (2)
Country | Link |
---|---|
KR (1) | KR20230148433A (ko) |
WO (1) | WO2023200266A1 (ko) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100556660B1 (ko) * | 2003-11-11 | 2006-03-10 | 국립암센터 | Hgf의 중화가능 에피토프 및 이에 결합하는 중화 항체 |
KR20170142995A (ko) * | 2015-08-05 | 2017-12-28 | 주식회사 유영제약 | 키메라 항원 수용체 및 키메라 항원 수용체가 발현된 t 세포 |
-
2022
- 2022-04-14 KR KR1020220046299A patent/KR20230148433A/ko unknown
-
2023
- 2023-04-13 WO PCT/KR2023/004996 patent/WO2023200266A1/ko unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100556660B1 (ko) * | 2003-11-11 | 2006-03-10 | 국립암센터 | Hgf의 중화가능 에피토프 및 이에 결합하는 중화 항체 |
KR20170142995A (ko) * | 2015-08-05 | 2017-12-28 | 주식회사 유영제약 | 키메라 항원 수용체 및 키메라 항원 수용체가 발현된 t 세포 |
Non-Patent Citations (4)
Title |
---|
KIM HYORI, HONG SUNG HEE, KIM JUNG YONG, KIM IN-CHULL, PARK YOUNG-WHAN, LEE SONG-JAE, SONG SEONG-WON, PARK GUNWOO, KIM TAE MIN, KI: "Preclinical development of a humanized neutralizing antibody targeting HGF", EXPERIMENTAL & MOLECULAR MEDICINE, vol. 49, no. 3, pages e309 - e309, XP093098245, DOI: 10.1038/emm.2017.21 * |
KIM KIWAN, GWAK HO-SHIN, HAN NAYOUNG, HONG EUN KYUNG, CHOI BEOM K., LEE SANGEUN, CHOI SOYOUNG, PARK JU-HWANG, SEOK JI-HYE, JEON YE: "Chimeric Antigen Receptor T Cells With Modified Interleukin-13 Preferentially Recognize IL13Rα2 and Suppress Malignant Glioma: A Preclinical Study", FRONTIERS IN IMMUNOLOGY, vol. 12, XP093098252, DOI: 10.3389/fimmu.2021.715000 * |
MANISH CHARAN; PIYUSH DRAVID; MAREN CAM; ANTHONY AUDINO; AMY C. GROSS; MICHAEL A. ARNOLD; RYAN D. ROBERTS; TIMOTHY P. CRIPE; ALEXA: "GD2‐directed CAR‐T cells in combination with HGF‐targeted neutralizing antibody (AMG102) prevent primary tumor growth and metastasis in Ewing sarcoma", INTERNATIONAL JOURNAL OF CANCER, JOHN WILEY & SONS, INC., US, vol. 146, no. 11, 6 November 2019 (2019-11-06), US , pages 3184 - 3195, XP071290684, ISSN: 0020-7136, DOI: 10.1002/ijc.32743 * |
SEO YOON-SEOK: "CellabMed announces ‘HGF antibody + IL13Rα2 CAR-T’ ASGCT", BIOSPECTATOR, 19 May 2022 (2022-05-19), XP093098256, Retrieved from the Internet <URL:http://biospectator.com/view/news_view.php?varAtcId=16319> * |
Also Published As
Publication number | Publication date |
---|---|
KR20230148433A (ko) | 2023-10-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20230357717A1 (en) | Cell secreted minibodies and uses thereof | |
US20190321404A1 (en) | Novel chimeric antigen receptor and use thereof | |
EP3114147B1 (en) | Chimeric antigen receptor | |
US20180327470A1 (en) | Chimeric antigen receptor-modified immune effector cell carrying pd-l1 blocking agent | |
JP2021500861A (ja) | 共刺激のための新規のプラットフォーム、新規のcar設計、および養子細胞療法のための他の増強 | |
WO2022037520A9 (en) | Lymphocytes-antigen presenting cells co-stimulators and uses thereof | |
JP2018504094A (ja) | Fc受容体様5を標的とするキメラ抗原受容体およびその使用 | |
US20170267737A1 (en) | Glypican-3-specific T-cell receptors and their uses for immunotherapy of hepatocellular carcinoma | |
WO2021232200A1 (en) | Il-12 armored immune cell therapy and uses thereof | |
JP2022512450A (ja) | Gpc3を標的とする免疫エフェクター細胞およびその応用 | |
AU2020272074A1 (en) | Compositions and Methods Comprising a High Affinity Chimeric Antigen Receptor (CAR) with Cross-Reactivity to Clinically-Relevant EGFR Mutated Proteins | |
CN110461363A (zh) | 靶向cd37的嵌合抗原受体 | |
WO2023200266A1 (ko) | 키메릭 항원 수용체 및 hgf 중화항체의 조합 요법 | |
WO2023200267A1 (ko) | 키메릭 항원 수용체 및 hgf 결합 억제 물질의 조합 요법 | |
WO2021244654A1 (en) | Activation induced cytokine production in immune cells | |
EP4323387A2 (en) | Improved chimeric antigen receptors and uses thereof | |
US20210161958A1 (en) | Compositions and methods for treating melanoma with a chimeric antigen receptor | |
WO2022215919A1 (ko) | Cd47에 특이적으로 결합하는 키메릭 항원 수용체 및 이의 용도 | |
WO2023191526A1 (ko) | Cd30 유래의 세포내 신호전달 도메인을 포함하는 키메라 항원 수용체, 이를 발현하는 면역세포 및 이들의 용도 | |
CA3068636A1 (en) | Compositions and methods for adoptive cell therapy for cancer | |
US20230277589A1 (en) | Bcma-targeted car-t cell therapy for multiple myeloma | |
US20240156962A1 (en) | Bcma-targeted car-t cell therapy of multiple myeloma | |
WO2023006120A1 (zh) | 通用型t细胞及其应用 | |
WO2023182861A1 (ko) | 항-hla-g 키메라 항원 수용체 및 이의 용도 | |
WO2023077343A1 (en) | Bcma-targeted car-t cell therapy for multiple myeloma |
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: 23788615 Country of ref document: EP Kind code of ref document: A1 |