US20110110852A1 - Multivalent Antibodies and Uses Therefor - Google Patents
Multivalent Antibodies and Uses Therefor Download PDFInfo
- Publication number
- US20110110852A1 US20110110852A1 US12/939,117 US93911710A US2011110852A1 US 20110110852 A1 US20110110852 A1 US 20110110852A1 US 93911710 A US93911710 A US 93911710A US 2011110852 A1 US2011110852 A1 US 2011110852A1
- Authority
- US
- United States
- Prior art keywords
- antibody
- binding protein
- tnf
- antibodies
- cells
- 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.)
- Abandoned
Links
- 239000000427 antigen Substances 0.000 claims abstract description 163
- 108091007433 antigens Proteins 0.000 claims abstract description 163
- 102000036639 antigens Human genes 0.000 claims abstract description 163
- 230000027455 binding Effects 0.000 claims abstract description 106
- 230000001225 therapeutic effect Effects 0.000 claims abstract description 14
- 210000004027 cell Anatomy 0.000 claims description 154
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 152
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 146
- 229920001184 polypeptide Polymers 0.000 claims description 142
- 102000005962 receptors Human genes 0.000 claims description 73
- 108020003175 receptors Proteins 0.000 claims description 73
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 71
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 48
- 239000003446 ligand Substances 0.000 claims description 44
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 claims description 41
- 102000014914 Carrier Proteins Human genes 0.000 claims description 41
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 claims description 41
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims description 41
- 108091008324 binding proteins Proteins 0.000 claims description 41
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims description 38
- 102000004127 Cytokines Human genes 0.000 claims description 28
- 108090000695 Cytokines Proteins 0.000 claims description 28
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 26
- 102100024598 Tumor necrosis factor ligand superfamily member 10 Human genes 0.000 claims description 25
- 101150013553 CD40 gene Proteins 0.000 claims description 24
- 108700012411 TNFSF10 Proteins 0.000 claims description 24
- 150000007523 nucleic acids Chemical class 0.000 claims description 23
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 claims description 22
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 claims description 22
- 102000039446 nucleic acids Human genes 0.000 claims description 21
- 108020004707 nucleic acids Proteins 0.000 claims description 21
- 239000003814 drug Substances 0.000 claims description 20
- 108060003951 Immunoglobulin Proteins 0.000 claims description 19
- 102000018358 immunoglobulin Human genes 0.000 claims description 19
- 229940127089 cytotoxic agent Drugs 0.000 claims description 18
- 239000012634 fragment Substances 0.000 claims description 17
- 239000003795 chemical substances by application Substances 0.000 claims description 16
- 102100032937 CD40 ligand Human genes 0.000 claims description 14
- 239000002254 cytotoxic agent Substances 0.000 claims description 14
- 231100000599 cytotoxic agent Toxicity 0.000 claims description 14
- 108010029697 CD40 Ligand Proteins 0.000 claims description 12
- 239000003102 growth factor Substances 0.000 claims description 12
- 108010065805 Interleukin-12 Proteins 0.000 claims description 10
- 102000013462 Interleukin-12 Human genes 0.000 claims description 10
- 102400001320 Transforming growth factor alpha Human genes 0.000 claims description 10
- 101800004564 Transforming growth factor alpha Proteins 0.000 claims description 10
- 102100024584 Tumor necrosis factor ligand superfamily member 12 Human genes 0.000 claims description 10
- 102100031988 Tumor necrosis factor ligand superfamily member 6 Human genes 0.000 claims description 10
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 claims description 10
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 claims description 10
- 239000000556 agonist Substances 0.000 claims description 10
- 229940079593 drug Drugs 0.000 claims description 10
- 102100036922 Tumor necrosis factor ligand superfamily member 13B Human genes 0.000 claims description 9
- 102100038080 B-cell receptor CD22 Human genes 0.000 claims description 8
- 102000018697 Membrane Proteins Human genes 0.000 claims description 8
- 108010052285 Membrane Proteins Proteins 0.000 claims description 8
- 102000014128 RANK Ligand Human genes 0.000 claims description 8
- 108010025832 RANK Ligand Proteins 0.000 claims description 8
- 101000884305 Homo sapiens B-cell receptor CD22 Proteins 0.000 claims description 7
- 102100023050 Nuclear factor NF-kappa-B p105 subunit Human genes 0.000 claims description 7
- 101710097155 Tumor necrosis factor ligand superfamily member 12 Proteins 0.000 claims description 7
- 102100033725 Tumor necrosis factor receptor superfamily member 16 Human genes 0.000 claims description 7
- 230000001640 apoptogenic effect Effects 0.000 claims description 7
- 230000003053 immunization Effects 0.000 claims description 7
- 102000006495 integrins Human genes 0.000 claims description 7
- 108010044426 integrins Proteins 0.000 claims description 7
- 229940124597 therapeutic agent Drugs 0.000 claims description 7
- 239000003053 toxin Substances 0.000 claims description 7
- 231100000765 toxin Toxicity 0.000 claims description 7
- 108700012359 toxins Proteins 0.000 claims description 7
- 108010039471 Fas Ligand Protein Proteins 0.000 claims description 6
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 claims description 6
- 108010032605 Nerve Growth Factor Receptors Proteins 0.000 claims description 6
- 229960000485 methotrexate Drugs 0.000 claims description 6
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 claims description 6
- 239000013598 vector Substances 0.000 claims description 6
- 108010028006 B-Cell Activating Factor Proteins 0.000 claims description 5
- 108010017987 CD30 Ligand Proteins 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 claims description 5
- 101000679921 Homo sapiens Tumor necrosis factor receptor superfamily member 21 Proteins 0.000 claims description 5
- 102000004388 Interleukin-4 Human genes 0.000 claims description 5
- 108090000978 Interleukin-4 Proteins 0.000 claims description 5
- 108010002616 Interleukin-5 Proteins 0.000 claims description 5
- 102100039897 Interleukin-5 Human genes 0.000 claims description 5
- 108090001005 Interleukin-6 Proteins 0.000 claims description 5
- 102100037795 Interleukin-6 receptor subunit beta Human genes 0.000 claims description 5
- 102000004890 Interleukin-8 Human genes 0.000 claims description 5
- 108090001007 Interleukin-8 Proteins 0.000 claims description 5
- 108010002335 Interleukin-9 Proteins 0.000 claims description 5
- 102100026871 Interleukin-9 Human genes 0.000 claims description 5
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 claims description 5
- 229940088597 hormone Drugs 0.000 claims description 5
- 239000005556 hormone Substances 0.000 claims description 5
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 claims description 4
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 claims description 4
- 102000003951 Erythropoietin Human genes 0.000 claims description 4
- 108090000394 Erythropoietin Proteins 0.000 claims description 4
- 102000003886 Glycoproteins Human genes 0.000 claims description 4
- 108090000288 Glycoproteins Proteins 0.000 claims description 4
- 108010051696 Growth Hormone Proteins 0.000 claims description 4
- 101000878605 Homo sapiens Low affinity immunoglobulin epsilon Fc receptor Proteins 0.000 claims description 4
- 101000611023 Homo sapiens Tumor necrosis factor receptor superfamily member 6 Proteins 0.000 claims description 4
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 claims description 4
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 claims description 4
- 102100020880 Kit ligand Human genes 0.000 claims description 4
- 102100038007 Low affinity immunoglobulin epsilon Fc receptor Human genes 0.000 claims description 4
- 102100038803 Somatotropin Human genes 0.000 claims description 4
- 108010000499 Thromboplastin Proteins 0.000 claims description 4
- 102100030859 Tissue factor Human genes 0.000 claims description 4
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims description 4
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims description 4
- 102100040403 Tumor necrosis factor receptor superfamily member 6 Human genes 0.000 claims description 4
- 239000003242 anti bacterial agent Substances 0.000 claims description 4
- 229940105423 erythropoietin Drugs 0.000 claims description 4
- 239000000122 growth hormone Substances 0.000 claims description 4
- 238000002649 immunization Methods 0.000 claims description 4
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 claims description 4
- 108010082808 4-1BB Ligand Proteins 0.000 claims description 3
- 102100022464 5'-nucleotidase Human genes 0.000 claims description 3
- 102100025218 B-cell differentiation antigen CD72 Human genes 0.000 claims description 3
- 102100029945 Beta-galactoside alpha-2,6-sialyltransferase 1 Human genes 0.000 claims description 3
- 102100024217 CAMPATH-1 antigen Human genes 0.000 claims description 3
- 108010046080 CD27 Ligand Proteins 0.000 claims description 3
- 108010065524 CD52 Antigen Proteins 0.000 claims description 3
- 102100027221 CD81 antigen Human genes 0.000 claims description 3
- 102100035793 CD83 antigen Human genes 0.000 claims description 3
- 102100032768 Complement receptor type 2 Human genes 0.000 claims description 3
- 101150064015 FAS gene Proteins 0.000 claims description 3
- 102100030595 HLA class II histocompatibility antigen gamma chain Human genes 0.000 claims description 3
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 claims description 3
- 101000934359 Homo sapiens B-cell differentiation antigen CD72 Proteins 0.000 claims description 3
- 101000863864 Homo sapiens Beta-galactoside alpha-2,6-sialyltransferase 1 Proteins 0.000 claims description 3
- 101000914479 Homo sapiens CD81 antigen Proteins 0.000 claims description 3
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 claims description 3
- 101001082627 Homo sapiens HLA class II histocompatibility antigen gamma chain Proteins 0.000 claims description 3
- 101000777628 Homo sapiens Leukocyte antigen CD37 Proteins 0.000 claims description 3
- 101000884271 Homo sapiens Signal transducer CD24 Proteins 0.000 claims description 3
- 101000851376 Homo sapiens Tumor necrosis factor receptor superfamily member 8 Proteins 0.000 claims description 3
- 102000000589 Interleukin-1 Human genes 0.000 claims description 3
- 108010002352 Interleukin-1 Proteins 0.000 claims description 3
- 102000003814 Interleukin-10 Human genes 0.000 claims description 3
- 108090000174 Interleukin-10 Proteins 0.000 claims description 3
- 108090000177 Interleukin-11 Proteins 0.000 claims description 3
- 108010002350 Interleukin-2 Proteins 0.000 claims description 3
- 102100026879 Interleukin-2 receptor subunit beta Human genes 0.000 claims description 3
- 102100039064 Interleukin-3 Human genes 0.000 claims description 3
- 102000004889 Interleukin-6 Human genes 0.000 claims description 3
- 108010002586 Interleukin-7 Proteins 0.000 claims description 3
- 102100021592 Interleukin-7 Human genes 0.000 claims description 3
- 102100031586 Leukocyte antigen CD37 Human genes 0.000 claims description 3
- 102100026238 Lymphotoxin-alpha Human genes 0.000 claims description 3
- 102000015731 Peptide Hormones Human genes 0.000 claims description 3
- 108010038988 Peptide Hormones Proteins 0.000 claims description 3
- 108091000080 Phosphotransferase Proteins 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 3
- 102100038081 Signal transducer CD24 Human genes 0.000 claims description 3
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 claims description 3
- 102100036857 Tumor necrosis factor receptor superfamily member 8 Human genes 0.000 claims description 3
- 229940100198 alkylating agent Drugs 0.000 claims description 3
- 239000002168 alkylating agent Substances 0.000 claims description 3
- 230000000340 anti-metabolite Effects 0.000 claims description 3
- 229940100197 antimetabolite Drugs 0.000 claims description 3
- 239000002256 antimetabolite Substances 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 239000000813 peptide hormone Substances 0.000 claims description 3
- 102000020233 phosphotransferase Human genes 0.000 claims description 3
- 239000012217 radiopharmaceutical Substances 0.000 claims description 3
- 229940121896 radiopharmaceutical Drugs 0.000 claims description 3
- 230000002799 radiopharmaceutical effect Effects 0.000 claims description 3
- 108090001117 Insulin-Like Growth Factor II Proteins 0.000 claims description 2
- 102000048143 Insulin-Like Growth Factor II Human genes 0.000 claims description 2
- 108010074328 Interferon-gamma Proteins 0.000 claims description 2
- 108010002386 Interleukin-3 Proteins 0.000 claims description 2
- 102000008072 Lymphokines Human genes 0.000 claims description 2
- 108010074338 Lymphokines Proteins 0.000 claims description 2
- 102000013967 Monokines Human genes 0.000 claims description 2
- 108010050619 Monokines Proteins 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 108090000176 Interleukin-13 Proteins 0.000 claims 35
- 102000003816 Interleukin-13 Human genes 0.000 claims 35
- 102100040247 Tumor necrosis factor Human genes 0.000 claims 27
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims 21
- 101000756808 Homo sapiens Repulsive guidance molecule A Proteins 0.000 claims 14
- 102100022813 Repulsive guidance molecule A Human genes 0.000 claims 14
- 239000008194 pharmaceutical composition Substances 0.000 claims 11
- 239000012216 imaging agent Substances 0.000 claims 9
- 102100031585 ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Human genes 0.000 claims 8
- 102100036302 C-C chemokine receptor type 6 Human genes 0.000 claims 8
- 102100039498 Cytotoxic T-lymphocyte protein 4 Human genes 0.000 claims 8
- 101000777636 Homo sapiens ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Proteins 0.000 claims 8
- 101000716068 Homo sapiens C-C chemokine receptor type 6 Proteins 0.000 claims 8
- 102100036305 C-C chemokine receptor type 8 Human genes 0.000 claims 7
- 101000716063 Homo sapiens C-C chemokine receptor type 8 Proteins 0.000 claims 7
- 101001040964 Homo sapiens Interleukin-36 receptor antagonist protein Proteins 0.000 claims 7
- 102000003810 Interleukin-18 Human genes 0.000 claims 7
- 108090000171 Interleukin-18 Proteins 0.000 claims 7
- 102100021150 Interleukin-36 receptor antagonist protein Human genes 0.000 claims 7
- 102100031789 Myeloid-derived growth factor Human genes 0.000 claims 7
- 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 claims 6
- 102100022718 Atypical chemokine receptor 2 Human genes 0.000 claims 6
- 102100036846 C-C motif chemokine 21 Human genes 0.000 claims 6
- 102100036849 C-C motif chemokine 24 Human genes 0.000 claims 6
- 102100021936 C-C motif chemokine 27 Human genes 0.000 claims 6
- 102100025618 C-X-C chemokine receptor type 6 Human genes 0.000 claims 6
- 108010021064 CTLA-4 Antigen Proteins 0.000 claims 6
- 229940045513 CTLA4 antagonist Drugs 0.000 claims 6
- 102100035294 Chemokine XC receptor 1 Human genes 0.000 claims 6
- 101000678892 Homo sapiens Atypical chemokine receptor 2 Proteins 0.000 claims 6
- 101000713085 Homo sapiens C-C motif chemokine 21 Proteins 0.000 claims 6
- 101000804783 Homo sapiens Chemokine XC receptor 1 Proteins 0.000 claims 6
- 101001128431 Homo sapiens Myeloid-derived growth factor Proteins 0.000 claims 6
- 102100021943 C-C motif chemokine 2 Human genes 0.000 claims 5
- 102100028990 C-X-C chemokine receptor type 3 Human genes 0.000 claims 5
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 claims 5
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 claims 5
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 claims 5
- 101000916050 Homo sapiens C-X-C chemokine receptor type 3 Proteins 0.000 claims 5
- 101000998140 Homo sapiens Interleukin-36 alpha Proteins 0.000 claims 5
- 101000998126 Homo sapiens Interleukin-36 beta Proteins 0.000 claims 5
- 101000998122 Homo sapiens Interleukin-37 Proteins 0.000 claims 5
- 102100024167 C-C chemokine receptor type 3 Human genes 0.000 claims 4
- 102100037853 C-C chemokine receptor type 4 Human genes 0.000 claims 4
- 102100023702 C-C motif chemokine 13 Human genes 0.000 claims 4
- 102100023700 C-C motif chemokine 16 Human genes 0.000 claims 4
- 102100023698 C-C motif chemokine 17 Human genes 0.000 claims 4
- 102100023701 C-C motif chemokine 18 Human genes 0.000 claims 4
- 102100036848 C-C motif chemokine 20 Human genes 0.000 claims 4
- 102100021933 C-C motif chemokine 25 Human genes 0.000 claims 4
- 102100021935 C-C motif chemokine 26 Human genes 0.000 claims 4
- 102100032367 C-C motif chemokine 5 Human genes 0.000 claims 4
- 102100031658 C-X-C chemokine receptor type 5 Human genes 0.000 claims 4
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 claims 4
- 102100039398 C-X-C motif chemokine 2 Human genes 0.000 claims 4
- 102100036150 C-X-C motif chemokine 5 Human genes 0.000 claims 4
- 102100036153 C-X-C motif chemokine 6 Human genes 0.000 claims 4
- 102100036170 C-X-C motif chemokine 9 Human genes 0.000 claims 4
- 102100032912 CD44 antigen Human genes 0.000 claims 4
- 102100022480 Cadherin-20 Human genes 0.000 claims 4
- 102100026098 Claudin-7 Human genes 0.000 claims 4
- 102100032887 Clusterin Human genes 0.000 claims 4
- 108090000197 Clusterin Proteins 0.000 claims 4
- 102100036329 Cyclin-dependent kinase 3 Human genes 0.000 claims 4
- 102100035298 Cytokine SCM-1 beta Human genes 0.000 claims 4
- 102100028417 Fibroblast growth factor 12 Human genes 0.000 claims 4
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 claims 4
- 102100028072 Fibroblast growth factor 4 Human genes 0.000 claims 4
- 102100028075 Fibroblast growth factor 6 Human genes 0.000 claims 4
- 102100028071 Fibroblast growth factor 7 Human genes 0.000 claims 4
- 102000003817 Fos-related antigen 1 Human genes 0.000 claims 4
- 108090000123 Fos-related antigen 1 Proteins 0.000 claims 4
- 102100020997 Fractalkine Human genes 0.000 claims 4
- 102000004878 Gelsolin Human genes 0.000 claims 4
- 108090001064 Gelsolin Proteins 0.000 claims 4
- 102100039619 Granulocyte colony-stimulating factor Human genes 0.000 claims 4
- 102100034221 Growth-regulated alpha protein Human genes 0.000 claims 4
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 claims 4
- 101000777558 Homo sapiens C-C chemokine receptor type 10 Proteins 0.000 claims 4
- 101000716070 Homo sapiens C-C chemokine receptor type 9 Proteins 0.000 claims 4
- 101000978375 Homo sapiens C-C motif chemokine 16 Proteins 0.000 claims 4
- 101000978362 Homo sapiens C-C motif chemokine 17 Proteins 0.000 claims 4
- 101000978371 Homo sapiens C-C motif chemokine 18 Proteins 0.000 claims 4
- 101000897480 Homo sapiens C-C motif chemokine 2 Proteins 0.000 claims 4
- 101000713099 Homo sapiens C-C motif chemokine 20 Proteins 0.000 claims 4
- 101000713078 Homo sapiens C-C motif chemokine 24 Proteins 0.000 claims 4
- 101000897486 Homo sapiens C-C motif chemokine 25 Proteins 0.000 claims 4
- 101000897494 Homo sapiens C-C motif chemokine 27 Proteins 0.000 claims 4
- 101000922405 Homo sapiens C-X-C chemokine receptor type 5 Proteins 0.000 claims 4
- 101000856683 Homo sapiens C-X-C chemokine receptor type 6 Proteins 0.000 claims 4
- 101000889128 Homo sapiens C-X-C motif chemokine 2 Proteins 0.000 claims 4
- 101000947186 Homo sapiens C-X-C motif chemokine 5 Proteins 0.000 claims 4
- 101000746022 Homo sapiens CX3C chemokine receptor 1 Proteins 0.000 claims 4
- 101000899459 Homo sapiens Cadherin-20 Proteins 0.000 claims 4
- 101000804771 Homo sapiens Cytokine SCM-1 beta Proteins 0.000 claims 4
- 101000917234 Homo sapiens Fibroblast growth factor 12 Proteins 0.000 claims 4
- 101000854520 Homo sapiens Fractalkine Proteins 0.000 claims 4
- 101001069921 Homo sapiens Growth-regulated alpha protein Proteins 0.000 claims 4
- 101000843810 Homo sapiens Hydroxycarboxylic acid receptor 1 Proteins 0.000 claims 4
- 101001017968 Homo sapiens Leukotriene B4 receptor 1 Proteins 0.000 claims 4
- 101000947178 Homo sapiens Platelet basic protein Proteins 0.000 claims 4
- 101000582950 Homo sapiens Platelet factor 4 Proteins 0.000 claims 4
- 101000830596 Homo sapiens Tumor necrosis factor ligand superfamily member 15 Proteins 0.000 claims 4
- 102100030642 Hydroxycarboxylic acid receptor 1 Human genes 0.000 claims 4
- 108090000172 Interleukin-15 Proteins 0.000 claims 4
- 102000003812 Interleukin-15 Human genes 0.000 claims 4
- 108010065637 Interleukin-23 Proteins 0.000 claims 4
- 102100033502 Interleukin-37 Human genes 0.000 claims 4
- 102100033420 Keratin, type I cytoskeletal 19 Human genes 0.000 claims 4
- 102000016267 Leptin Human genes 0.000 claims 4
- 108010092277 Leptin Proteins 0.000 claims 4
- 102100033374 Leukotriene B4 receptor 1 Human genes 0.000 claims 4
- 102100035304 Lymphotactin Human genes 0.000 claims 4
- 102100028123 Macrophage colony-stimulating factor 1 Human genes 0.000 claims 4
- 102100037273 Mammaglobin-A Human genes 0.000 claims 4
- 102100037267 Mammaglobin-B Human genes 0.000 claims 4
- 102100030335 Midkine Human genes 0.000 claims 4
- 101100027996 Mus musculus Omg gene Proteins 0.000 claims 4
- 102100036154 Platelet basic protein Human genes 0.000 claims 4
- 102100030304 Platelet factor 4 Human genes 0.000 claims 4
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 claims 4
- 108010005173 SERPIN-B5 Proteins 0.000 claims 4
- 102100030333 Serpin B5 Human genes 0.000 claims 4
- 102000012883 Tumor Necrosis Factor Ligand Superfamily Member 14 Human genes 0.000 claims 4
- 108010065158 Tumor Necrosis Factor Ligand Superfamily Member 14 Proteins 0.000 claims 4
- 102100024587 Tumor necrosis factor ligand superfamily member 15 Human genes 0.000 claims 4
- 102100026890 Tumor necrosis factor ligand superfamily member 4 Human genes 0.000 claims 4
- 102100032101 Tumor necrosis factor ligand superfamily member 9 Human genes 0.000 claims 4
- 102100038234 Vascular endothelial growth factor D Human genes 0.000 claims 4
- 102100033177 Vascular endothelial growth factor receptor 2 Human genes 0.000 claims 4
- 239000004037 angiogenesis inhibitor Substances 0.000 claims 4
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 claims 4
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 claims 4
- 229940039781 leptin Drugs 0.000 claims 4
- NRYBAZVQPHGZNS-ZSOCWYAHSA-N leptin Chemical compound O=C([C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(C)C)CCSC)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CS)C(O)=O NRYBAZVQPHGZNS-ZSOCWYAHSA-N 0.000 claims 4
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 claims 4
- 102100034065 Atypical chemokine receptor 4 Human genes 0.000 claims 3
- 102100023995 Beta-nerve growth factor Human genes 0.000 claims 3
- 102100031172 C-C chemokine receptor type 1 Human genes 0.000 claims 3
- 102100035875 C-C chemokine receptor type 5 Human genes 0.000 claims 3
- 102100036301 C-C chemokine receptor type 7 Human genes 0.000 claims 3
- 102100036842 C-C motif chemokine 19 Human genes 0.000 claims 3
- 102100036850 C-C motif chemokine 23 Human genes 0.000 claims 3
- 102100032366 C-C motif chemokine 7 Human genes 0.000 claims 3
- 102100025279 C-X-C motif chemokine 11 Human genes 0.000 claims 3
- 102100036189 C-X-C motif chemokine 3 Human genes 0.000 claims 3
- 102100025221 CD70 antigen Human genes 0.000 claims 3
- 102100039196 CX3C chemokine receptor 1 Human genes 0.000 claims 3
- 108010016777 Cyclin-Dependent Kinase Inhibitor p27 Proteins 0.000 claims 3
- 102000000577 Cyclin-Dependent Kinase Inhibitor p27 Human genes 0.000 claims 3
- 102100024458 Cyclin-dependent kinase inhibitor 2A Human genes 0.000 claims 3
- 102100023688 Eotaxin Human genes 0.000 claims 3
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 claims 3
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims 3
- 102100040895 Growth/differentiation factor 10 Human genes 0.000 claims 3
- 101000798902 Homo sapiens Atypical chemokine receptor 4 Proteins 0.000 claims 3
- 101001111439 Homo sapiens Beta-nerve growth factor Proteins 0.000 claims 3
- 101000713106 Homo sapiens C-C motif chemokine 19 Proteins 0.000 claims 3
- 101000713081 Homo sapiens C-C motif chemokine 23 Proteins 0.000 claims 3
- 101000858060 Homo sapiens C-X-C motif chemokine 11 Proteins 0.000 claims 3
- 101000947193 Homo sapiens C-X-C motif chemokine 3 Proteins 0.000 claims 3
- 101000899410 Homo sapiens Cadherin-19 Proteins 0.000 claims 3
- 101000746367 Homo sapiens Granulocyte colony-stimulating factor Proteins 0.000 claims 3
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 claims 3
- 101001076292 Homo sapiens Insulin-like growth factor II Proteins 0.000 claims 3
- 101000853002 Homo sapiens Interleukin-25 Proteins 0.000 claims 3
- 101000613343 Homo sapiens Polycomb group RING finger protein 2 Proteins 0.000 claims 3
- 101000617130 Homo sapiens Stromal cell-derived factor 1 Proteins 0.000 claims 3
- 101000830568 Homo sapiens Tumor necrosis factor alpha-induced protein 2 Proteins 0.000 claims 3
- 101000830598 Homo sapiens Tumor necrosis factor ligand superfamily member 12 Proteins 0.000 claims 3
- 102100037852 Insulin-like growth factor I Human genes 0.000 claims 3
- 102100025947 Insulin-like growth factor II Human genes 0.000 claims 3
- 102100020881 Interleukin-1 alpha Human genes 0.000 claims 3
- 108010063954 Mucins Proteins 0.000 claims 3
- 102100040919 Polycomb group RING finger protein 2 Human genes 0.000 claims 3
- 102100021669 Stromal cell-derived factor 1 Human genes 0.000 claims 3
- 102100036034 Thrombospondin-1 Human genes 0.000 claims 3
- 102100024595 Tumor necrosis factor alpha-induced protein 2 Human genes 0.000 claims 3
- 102100032100 Tumor necrosis factor ligand superfamily member 8 Human genes 0.000 claims 3
- 229960002685 biotin Drugs 0.000 claims 3
- 235000020958 biotin Nutrition 0.000 claims 3
- 239000011616 biotin Substances 0.000 claims 3
- 239000007850 fluorescent dye Substances 0.000 claims 3
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 claims 3
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 claims 3
- UCTWMZQNUQWSLP-VIFPVBQESA-N (R)-adrenaline Chemical compound CNC[C@H](O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-VIFPVBQESA-N 0.000 claims 2
- 229930182837 (R)-adrenaline Natural products 0.000 claims 2
- 102100036933 12-(S)-hydroxy-5,8,10,14-eicosatetraenoic acid receptor Human genes 0.000 claims 2
- SXXLKZCNJHJYFL-UHFFFAOYSA-N 4,5,6,7-tetrahydro-[1,2]oxazolo[4,5-c]pyridin-5-ium-3-olate Chemical compound C1CNCC2=C1ONC2=O SXXLKZCNJHJYFL-UHFFFAOYSA-N 0.000 claims 2
- WBSMIPAMAXNXFS-UHFFFAOYSA-N 5-Nitro-2-(3-phenylpropylamino)benzoic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC=C1NCCCC1=CC=CC=C1 WBSMIPAMAXNXFS-UHFFFAOYSA-N 0.000 claims 2
- 101710142585 50S ribosomal protein 6, chloroplastic Proteins 0.000 claims 2
- 102100026802 72 kDa type IV collagenase Human genes 0.000 claims 2
- 102100031901 A-kinase anchor protein 2 Human genes 0.000 claims 2
- 108010029988 AICDA (activation-induced cytidine deaminase) Proteins 0.000 claims 2
- 101150054149 ANGPTL4 gene Proteins 0.000 claims 2
- 102100028249 Acetyl-coenzyme A transporter 1 Human genes 0.000 claims 2
- 102100021886 Activin receptor type-2A Human genes 0.000 claims 2
- 102100027647 Activin receptor type-2B Human genes 0.000 claims 2
- 102100022089 Acyl-[acyl-carrier-protein] hydrolase Human genes 0.000 claims 2
- 102100035990 Adenosine receptor A2a Human genes 0.000 claims 2
- 102100036601 Aggrecan core protein Human genes 0.000 claims 2
- 108010067219 Aggrecans Proteins 0.000 claims 2
- 102100022524 Alpha-1-antichymotrypsin Human genes 0.000 claims 2
- 102100022712 Alpha-1-antitrypsin Human genes 0.000 claims 2
- 102100038910 Alpha-enolase Human genes 0.000 claims 2
- 102100022749 Aminopeptidase N Human genes 0.000 claims 2
- 102100034594 Angiopoietin-1 Human genes 0.000 claims 2
- 102100034608 Angiopoietin-2 Human genes 0.000 claims 2
- 108700042530 Angiopoietin-Like Protein 4 Proteins 0.000 claims 2
- 102100025668 Angiopoietin-related protein 3 Human genes 0.000 claims 2
- 102100025674 Angiopoietin-related protein 4 Human genes 0.000 claims 2
- 102100031936 Anterior gradient protein 2 homolog Human genes 0.000 claims 2
- 102100025511 Anti-Muellerian hormone type-2 receptor Human genes 0.000 claims 2
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 claims 2
- 101100279540 Arabidopsis thaliana EIN2 gene Proteins 0.000 claims 2
- 101100129499 Arabidopsis thaliana MAX2 gene Proteins 0.000 claims 2
- 101100351711 Arabidopsis thaliana PEX14 gene Proteins 0.000 claims 2
- 102100022717 Atypical chemokine receptor 1 Human genes 0.000 claims 2
- 102100022716 Atypical chemokine receptor 3 Human genes 0.000 claims 2
- 102100027205 B-cell antigen receptor complex-associated protein alpha chain Human genes 0.000 claims 2
- 102100027203 B-cell antigen receptor complex-associated protein beta chain Human genes 0.000 claims 2
- 102100035634 B-cell linker protein Human genes 0.000 claims 2
- 102100021631 B-cell lymphoma 6 protein Human genes 0.000 claims 2
- 108091012583 BCL2 Proteins 0.000 claims 2
- 102100024506 Bone morphogenetic protein 2 Human genes 0.000 claims 2
- 102100024505 Bone morphogenetic protein 4 Human genes 0.000 claims 2
- 102100022525 Bone morphogenetic protein 6 Human genes 0.000 claims 2
- 102100022545 Bone morphogenetic protein 8B Human genes 0.000 claims 2
- 102100025422 Bone morphogenetic protein receptor type-2 Human genes 0.000 claims 2
- 101710149814 C-C chemokine receptor type 1 Proteins 0.000 claims 2
- 102100031174 C-C chemokine receptor type 10 Human genes 0.000 claims 2
- 102100031151 C-C chemokine receptor type 2 Human genes 0.000 claims 2
- 101710149815 C-C chemokine receptor type 2 Proteins 0.000 claims 2
- 101710149862 C-C chemokine receptor type 3 Proteins 0.000 claims 2
- 101710149863 C-C chemokine receptor type 4 Proteins 0.000 claims 2
- 101710149870 C-C chemokine receptor type 5 Proteins 0.000 claims 2
- 102100036303 C-C chemokine receptor type 9 Human genes 0.000 claims 2
- 101710112613 C-C motif chemokine 13 Proteins 0.000 claims 2
- 102100023703 C-C motif chemokine 15 Human genes 0.000 claims 2
- 101710112538 C-C motif chemokine 27 Proteins 0.000 claims 2
- 102100021942 C-C motif chemokine 28 Human genes 0.000 claims 2
- 102100034871 C-C motif chemokine 8 Human genes 0.000 claims 2
- 102100028989 C-X-C chemokine receptor type 2 Human genes 0.000 claims 2
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 claims 2
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 claims 2
- 102100025277 C-X-C motif chemokine 13 Human genes 0.000 claims 2
- 102100025250 C-X-C motif chemokine 14 Human genes 0.000 claims 2
- 102100039396 C-X-C motif chemokine 16 Human genes 0.000 claims 2
- 101710085504 C-X-C motif chemokine 6 Proteins 0.000 claims 2
- 101710085500 C-X-C motif chemokine 9 Proteins 0.000 claims 2
- 102100032957 C5a anaphylatoxin chemotactic receptor 1 Human genes 0.000 claims 2
- 102100034798 CCAAT/enhancer-binding protein beta Human genes 0.000 claims 2
- 108010049990 CD13 Antigens Proteins 0.000 claims 2
- 102100027207 CD27 antigen Human genes 0.000 claims 2
- 101150116779 CD82 gene Proteins 0.000 claims 2
- 102100040531 CKLF-like MARVEL transmembrane domain-containing protein 2 Human genes 0.000 claims 2
- 102100040527 CKLF-like MARVEL transmembrane domain-containing protein 3 Human genes 0.000 claims 2
- 102100040529 CKLF-like MARVEL transmembrane domain-containing protein 4 Human genes 0.000 claims 2
- 102100040525 CKLF-like MARVEL transmembrane domain-containing protein 5 Human genes 0.000 claims 2
- 102100040528 CKLF-like MARVEL transmembrane domain-containing protein 6 Human genes 0.000 claims 2
- 102100040855 CKLF-like MARVEL transmembrane domain-containing protein 7 Human genes 0.000 claims 2
- 102100039553 CKLF-like MARVEL transmembrane domain-containing protein 8 Human genes 0.000 claims 2
- 102100028228 COUP transcription factor 1 Human genes 0.000 claims 2
- 102100028226 COUP transcription factor 2 Human genes 0.000 claims 2
- 108091011896 CSF1 Proteins 0.000 claims 2
- 108010061304 CXCR6 Receptors Proteins 0.000 claims 2
- 102000000905 Cadherin Human genes 0.000 claims 2
- 108050007957 Cadherin Proteins 0.000 claims 2
- 102100024154 Cadherin-13 Human genes 0.000 claims 2
- 102100029761 Cadherin-5 Human genes 0.000 claims 2
- 102100025331 Cadherin-8 Human genes 0.000 claims 2
- 102100025332 Cadherin-9 Human genes 0.000 claims 2
- 101100463133 Caenorhabditis elegans pdl-1 gene Proteins 0.000 claims 2
- 101100314454 Caenorhabditis elegans tra-1 gene Proteins 0.000 claims 2
- 101100293794 Canis lupus familiaris NME1 gene Proteins 0.000 claims 2
- 101001110283 Canis lupus familiaris Ras-related C3 botulinum toxin substrate 1 Proteins 0.000 claims 2
- 102100033377 Carbohydrate sulfotransferase 15 Human genes 0.000 claims 2
- 102100025597 Caspase-4 Human genes 0.000 claims 2
- 108090000712 Cathepsin B Proteins 0.000 claims 2
- 102000004225 Cathepsin B Human genes 0.000 claims 2
- 102100021633 Cathepsin B Human genes 0.000 claims 2
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 claims 2
- 108010083647 Chemokine CCL24 Proteins 0.000 claims 2
- 108010083698 Chemokine CCL26 Proteins 0.000 claims 2
- 108010055166 Chemokine CCL5 Proteins 0.000 claims 2
- 102000012286 Chitinases Human genes 0.000 claims 2
- 108010022172 Chitinases Proteins 0.000 claims 2
- 108010038447 Chromogranin A Proteins 0.000 claims 2
- 102100031186 Chromogranin-A Human genes 0.000 claims 2
- 108010044213 Class 5 Receptor-Like Protein Tyrosine Phosphatases Proteins 0.000 claims 2
- 102100038423 Claudin-3 Human genes 0.000 claims 2
- 108050007296 Claudin-7 Proteins 0.000 claims 2
- 102100033780 Collagen alpha-3(IV) chain Human genes 0.000 claims 2
- 108050006400 Cyclin Proteins 0.000 claims 2
- 108010058546 Cyclin D1 Proteins 0.000 claims 2
- 102100025191 Cyclin-A2 Human genes 0.000 claims 2
- 108010024986 Cyclin-Dependent Kinase 2 Proteins 0.000 claims 2
- 108010025464 Cyclin-Dependent Kinase 4 Proteins 0.000 claims 2
- 108010025454 Cyclin-Dependent Kinase 5 Proteins 0.000 claims 2
- 108010025468 Cyclin-Dependent Kinase 6 Proteins 0.000 claims 2
- 108010009356 Cyclin-Dependent Kinase Inhibitor p15 Proteins 0.000 claims 2
- 102000009512 Cyclin-Dependent Kinase Inhibitor p15 Human genes 0.000 claims 2
- 108010009392 Cyclin-Dependent Kinase Inhibitor p16 Proteins 0.000 claims 2
- 108010009367 Cyclin-Dependent Kinase Inhibitor p18 Proteins 0.000 claims 2
- 102000009503 Cyclin-Dependent Kinase Inhibitor p18 Human genes 0.000 claims 2
- 102100036239 Cyclin-dependent kinase 2 Human genes 0.000 claims 2
- 102100036252 Cyclin-dependent kinase 4 Human genes 0.000 claims 2
- 102100026804 Cyclin-dependent kinase 6 Human genes 0.000 claims 2
- 102100026810 Cyclin-dependent kinase 7 Human genes 0.000 claims 2
- 102100024457 Cyclin-dependent kinase 9 Human genes 0.000 claims 2
- 102100026805 Cyclin-dependent-like kinase 5 Human genes 0.000 claims 2
- 108010037462 Cyclooxygenase 2 Proteins 0.000 claims 2
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 claims 2
- 229930105110 Cyclosporin A Natural products 0.000 claims 2
- 108010036949 Cyclosporine Proteins 0.000 claims 2
- 102100031655 Cytochrome b5 Human genes 0.000 claims 2
- 102100036952 Cytoplasmic protein NCK2 Human genes 0.000 claims 2
- 102100033587 DNA topoisomerase 2-alpha Human genes 0.000 claims 2
- 102100031817 Delta-type opioid receptor Human genes 0.000 claims 2
- 102100025012 Dipeptidyl peptidase 4 Human genes 0.000 claims 2
- 102100023332 Dual specificity mitogen-activated protein kinase kinase 7 Human genes 0.000 claims 2
- 102100040565 Dynein light chain 1, cytoplasmic Human genes 0.000 claims 2
- 102100036254 E3 SUMO-protein ligase PIAS2 Human genes 0.000 claims 2
- 102100025137 Early activation antigen CD69 Human genes 0.000 claims 2
- 102100033267 Early placenta insulin-like peptide Human genes 0.000 claims 2
- 101710139422 Eotaxin Proteins 0.000 claims 2
- 108010055323 EphB4 Receptor Proteins 0.000 claims 2
- 102100031983 Ephrin type-B receptor 4 Human genes 0.000 claims 2
- 102100033940 Ephrin-A3 Human genes 0.000 claims 2
- 102100023721 Ephrin-B2 Human genes 0.000 claims 2
- 102100031939 Erythropoietin Human genes 0.000 claims 2
- 102100029951 Estrogen receptor beta Human genes 0.000 claims 2
- 102100026693 FAS-associated death domain protein Human genes 0.000 claims 2
- 102100031706 Fibroblast growth factor 1 Human genes 0.000 claims 2
- 102100028413 Fibroblast growth factor 11 Human genes 0.000 claims 2
- 102100035290 Fibroblast growth factor 13 Human genes 0.000 claims 2
- 102100035292 Fibroblast growth factor 14 Human genes 0.000 claims 2
- 102100035307 Fibroblast growth factor 16 Human genes 0.000 claims 2
- 108050002072 Fibroblast growth factor 16 Proteins 0.000 claims 2
- 102100035308 Fibroblast growth factor 17 Human genes 0.000 claims 2
- 102100035323 Fibroblast growth factor 18 Human genes 0.000 claims 2
- 102100031734 Fibroblast growth factor 19 Human genes 0.000 claims 2
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 claims 2
- 102100031361 Fibroblast growth factor 20 Human genes 0.000 claims 2
- 108090000376 Fibroblast growth factor 21 Proteins 0.000 claims 2
- 102000003973 Fibroblast growth factor 21 Human genes 0.000 claims 2
- 102100024804 Fibroblast growth factor 22 Human genes 0.000 claims 2
- 102100024802 Fibroblast growth factor 23 Human genes 0.000 claims 2
- 102100028043 Fibroblast growth factor 3 Human genes 0.000 claims 2
- 108090000381 Fibroblast growth factor 4 Proteins 0.000 claims 2
- 102100028073 Fibroblast growth factor 5 Human genes 0.000 claims 2
- 108090000382 Fibroblast growth factor 6 Proteins 0.000 claims 2
- 108090000385 Fibroblast growth factor 7 Proteins 0.000 claims 2
- 102100037680 Fibroblast growth factor 8 Human genes 0.000 claims 2
- 108090000368 Fibroblast growth factor 8 Proteins 0.000 claims 2
- 102100037665 Fibroblast growth factor 9 Human genes 0.000 claims 2
- 102100027842 Fibroblast growth factor receptor 3 Human genes 0.000 claims 2
- 101710182396 Fibroblast growth factor receptor 3 Proteins 0.000 claims 2
- 102100037362 Fibronectin Human genes 0.000 claims 2
- 108010067306 Fibronectins Proteins 0.000 claims 2
- 102100024165 G1/S-specific cyclin-D1 Human genes 0.000 claims 2
- 102100037854 G1/S-specific cyclin-E2 Human genes 0.000 claims 2
- 102000017700 GABRP Human genes 0.000 claims 2
- 101150019176 GDF10 gene Proteins 0.000 claims 2
- 101710115997 Gamma-tubulin complex component 2 Proteins 0.000 claims 2
- 102100033417 Glucocorticoid receptor Human genes 0.000 claims 2
- 102000058058 Glucose Transporter Type 2 Human genes 0.000 claims 2
- 102100030943 Glutathione S-transferase P Human genes 0.000 claims 2
- 102100033366 Glutathione hydrolase 1 proenzyme Human genes 0.000 claims 2
- 102100035379 Growth/differentiation factor 5 Human genes 0.000 claims 2
- 102100032610 Guanine nucleotide-binding protein G(s) subunit alpha isoforms XLas Human genes 0.000 claims 2
- 102100028972 HLA class I histocompatibility antigen, A alpha chain Human genes 0.000 claims 2
- 102100040505 HLA class II histocompatibility antigen, DR alpha chain Human genes 0.000 claims 2
- 108010075704 HLA-A Antigens Proteins 0.000 claims 2
- 108010067802 HLA-DR alpha-Chains Proteins 0.000 claims 2
- 102100028006 Heme oxygenase 1 Human genes 0.000 claims 2
- 108010007707 Hepatitis A Virus Cellular Receptor 2 Proteins 0.000 claims 2
- 102100034458 Hepatitis A virus cellular receptor 2 Human genes 0.000 claims 2
- 102100034676 Hepatocyte cell adhesion molecule Human genes 0.000 claims 2
- 102100038006 High affinity immunoglobulin epsilon receptor subunit alpha Human genes 0.000 claims 2
- 102000000543 Histamine Receptors Human genes 0.000 claims 2
- 108010002059 Histamine Receptors Proteins 0.000 claims 2
- 102100021454 Histone deacetylase 4 Human genes 0.000 claims 2
- 102100021453 Histone deacetylase 5 Human genes 0.000 claims 2
- 102100038719 Histone deacetylase 7 Human genes 0.000 claims 2
- 102100038720 Histone deacetylase 9 Human genes 0.000 claims 2
- 101001071349 Homo sapiens 12-(S)-hydroxy-5,8,10,14-eicosatetraenoic acid receptor Proteins 0.000 claims 2
- 101000627872 Homo sapiens 72 kDa type IV collagenase Proteins 0.000 claims 2
- 101000774738 Homo sapiens A-kinase anchor protein 2 Proteins 0.000 claims 2
- 101000970954 Homo sapiens Activin receptor type-2A Proteins 0.000 claims 2
- 101000937269 Homo sapiens Activin receptor type-2B Proteins 0.000 claims 2
- 101000824278 Homo sapiens Acyl-[acyl-carrier-protein] hydrolase Proteins 0.000 claims 2
- 101000783751 Homo sapiens Adenosine receptor A2a Proteins 0.000 claims 2
- 101000678026 Homo sapiens Alpha-1-antichymotrypsin Proteins 0.000 claims 2
- 101000823116 Homo sapiens Alpha-1-antitrypsin Proteins 0.000 claims 2
- 101000882335 Homo sapiens Alpha-enolase Proteins 0.000 claims 2
- 101000924552 Homo sapiens Angiopoietin-1 Proteins 0.000 claims 2
- 101000924533 Homo sapiens Angiopoietin-2 Proteins 0.000 claims 2
- 101000693085 Homo sapiens Angiopoietin-related protein 3 Proteins 0.000 claims 2
- 101000775021 Homo sapiens Anterior gradient protein 2 homolog Proteins 0.000 claims 2
- 101000693801 Homo sapiens Anti-Muellerian hormone type-2 receptor Proteins 0.000 claims 2
- 101000678879 Homo sapiens Atypical chemokine receptor 1 Proteins 0.000 claims 2
- 101000678890 Homo sapiens Atypical chemokine receptor 3 Proteins 0.000 claims 2
- 101000914489 Homo sapiens B-cell antigen receptor complex-associated protein alpha chain Proteins 0.000 claims 2
- 101000914491 Homo sapiens B-cell antigen receptor complex-associated protein beta chain Proteins 0.000 claims 2
- 101000803266 Homo sapiens B-cell linker protein Proteins 0.000 claims 2
- 101000971234 Homo sapiens B-cell lymphoma 6 protein Proteins 0.000 claims 2
- 101000762366 Homo sapiens Bone morphogenetic protein 2 Proteins 0.000 claims 2
- 101000762379 Homo sapiens Bone morphogenetic protein 4 Proteins 0.000 claims 2
- 101000899390 Homo sapiens Bone morphogenetic protein 6 Proteins 0.000 claims 2
- 101000899368 Homo sapiens Bone morphogenetic protein 8B Proteins 0.000 claims 2
- 101000934635 Homo sapiens Bone morphogenetic protein receptor type-2 Proteins 0.000 claims 2
- 101000980744 Homo sapiens C-C chemokine receptor type 3 Proteins 0.000 claims 2
- 101000738584 Homo sapiens C-C chemokine receptor type 4 Proteins 0.000 claims 2
- 101000716065 Homo sapiens C-C chemokine receptor type 7 Proteins 0.000 claims 2
- 101000978379 Homo sapiens C-C motif chemokine 13 Proteins 0.000 claims 2
- 101000978376 Homo sapiens C-C motif chemokine 15 Proteins 0.000 claims 2
- 101000897493 Homo sapiens C-C motif chemokine 26 Proteins 0.000 claims 2
- 101000897477 Homo sapiens C-C motif chemokine 28 Proteins 0.000 claims 2
- 101000797762 Homo sapiens C-C motif chemokine 5 Proteins 0.000 claims 2
- 101000797758 Homo sapiens C-C motif chemokine 7 Proteins 0.000 claims 2
- 101000946794 Homo sapiens C-C motif chemokine 8 Proteins 0.000 claims 2
- 101000916059 Homo sapiens C-X-C chemokine receptor type 2 Proteins 0.000 claims 2
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 claims 2
- 101000858088 Homo sapiens C-X-C motif chemokine 10 Proteins 0.000 claims 2
- 101000858064 Homo sapiens C-X-C motif chemokine 13 Proteins 0.000 claims 2
- 101000858068 Homo sapiens C-X-C motif chemokine 14 Proteins 0.000 claims 2
- 101000889133 Homo sapiens C-X-C motif chemokine 16 Proteins 0.000 claims 2
- 101000947177 Homo sapiens C-X-C motif chemokine 6 Proteins 0.000 claims 2
- 101000947172 Homo sapiens C-X-C motif chemokine 9 Proteins 0.000 claims 2
- 101000867983 Homo sapiens C5a anaphylatoxin chemotactic receptor 1 Proteins 0.000 claims 2
- 101000945963 Homo sapiens CCAAT/enhancer-binding protein beta Proteins 0.000 claims 2
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 claims 2
- 101000868215 Homo sapiens CD40 ligand Proteins 0.000 claims 2
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 claims 2
- 101000934356 Homo sapiens CD70 antigen Proteins 0.000 claims 2
- 101000749427 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 2 Proteins 0.000 claims 2
- 101000749433 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 3 Proteins 0.000 claims 2
- 101000749431 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 4 Proteins 0.000 claims 2
- 101000749437 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 5 Proteins 0.000 claims 2
- 101000749435 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 6 Proteins 0.000 claims 2
- 101000749308 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 7 Proteins 0.000 claims 2
- 101000888512 Homo sapiens CKLF-like MARVEL transmembrane domain-containing protein 8 Proteins 0.000 claims 2
- 101000860854 Homo sapiens COUP transcription factor 1 Proteins 0.000 claims 2
- 101000860860 Homo sapiens COUP transcription factor 2 Proteins 0.000 claims 2
- 101000762243 Homo sapiens Cadherin-13 Proteins 0.000 claims 2
- 101000794587 Homo sapiens Cadherin-5 Proteins 0.000 claims 2
- 101000935111 Homo sapiens Cadherin-7 Proteins 0.000 claims 2
- 101000935095 Homo sapiens Cadherin-8 Proteins 0.000 claims 2
- 101000935098 Homo sapiens Cadherin-9 Proteins 0.000 claims 2
- 101000933112 Homo sapiens Caspase-4 Proteins 0.000 claims 2
- 101000898449 Homo sapiens Cathepsin B Proteins 0.000 claims 2
- 101000882908 Homo sapiens Claudin-3 Proteins 0.000 claims 2
- 101000912652 Homo sapiens Claudin-7 Proteins 0.000 claims 2
- 101000710873 Homo sapiens Collagen alpha-3(IV) chain Proteins 0.000 claims 2
- 101000934320 Homo sapiens Cyclin-A2 Proteins 0.000 claims 2
- 101000715946 Homo sapiens Cyclin-dependent kinase 3 Proteins 0.000 claims 2
- 101000911952 Homo sapiens Cyclin-dependent kinase 7 Proteins 0.000 claims 2
- 101000980930 Homo sapiens Cyclin-dependent kinase 9 Proteins 0.000 claims 2
- 101000945639 Homo sapiens Cyclin-dependent kinase inhibitor 3 Proteins 0.000 claims 2
- 101000922386 Homo sapiens Cytochrome b5 Proteins 0.000 claims 2
- 101001024712 Homo sapiens Cytoplasmic protein NCK2 Proteins 0.000 claims 2
- 101000889276 Homo sapiens Cytotoxic T-lymphocyte protein 4 Proteins 0.000 claims 2
- 101000992305 Homo sapiens Delta-type opioid receptor Proteins 0.000 claims 2
- 101000908391 Homo sapiens Dipeptidyl peptidase 4 Proteins 0.000 claims 2
- 101000624594 Homo sapiens Dual specificity mitogen-activated protein kinase kinase 7 Proteins 0.000 claims 2
- 101000966403 Homo sapiens Dynein light chain 1, cytoplasmic Proteins 0.000 claims 2
- 101001074629 Homo sapiens E3 SUMO-protein ligase PIAS2 Proteins 0.000 claims 2
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 claims 2
- 101000998777 Homo sapiens Early placenta insulin-like peptide Proteins 0.000 claims 2
- 101000925241 Homo sapiens Ephrin-A3 Proteins 0.000 claims 2
- 101001049392 Homo sapiens Ephrin-B2 Proteins 0.000 claims 2
- 101001010910 Homo sapiens Estrogen receptor beta Proteins 0.000 claims 2
- 101000911074 Homo sapiens FAS-associated death domain protein Proteins 0.000 claims 2
- 101000917236 Homo sapiens Fibroblast growth factor 11 Proteins 0.000 claims 2
- 101000878181 Homo sapiens Fibroblast growth factor 14 Proteins 0.000 claims 2
- 101000878124 Homo sapiens Fibroblast growth factor 17 Proteins 0.000 claims 2
- 101000878128 Homo sapiens Fibroblast growth factor 18 Proteins 0.000 claims 2
- 101000846394 Homo sapiens Fibroblast growth factor 19 Proteins 0.000 claims 2
- 101000846532 Homo sapiens Fibroblast growth factor 20 Proteins 0.000 claims 2
- 101001051971 Homo sapiens Fibroblast growth factor 22 Proteins 0.000 claims 2
- 101001051973 Homo sapiens Fibroblast growth factor 23 Proteins 0.000 claims 2
- 101001060280 Homo sapiens Fibroblast growth factor 3 Proteins 0.000 claims 2
- 101001060274 Homo sapiens Fibroblast growth factor 4 Proteins 0.000 claims 2
- 101001060267 Homo sapiens Fibroblast growth factor 5 Proteins 0.000 claims 2
- 101001060265 Homo sapiens Fibroblast growth factor 6 Proteins 0.000 claims 2
- 101001060261 Homo sapiens Fibroblast growth factor 7 Proteins 0.000 claims 2
- 101001027382 Homo sapiens Fibroblast growth factor 8 Proteins 0.000 claims 2
- 101001027380 Homo sapiens Fibroblast growth factor 9 Proteins 0.000 claims 2
- 101000738575 Homo sapiens G1/S-specific cyclin-E2 Proteins 0.000 claims 2
- 101000822394 Homo sapiens Gamma-aminobutyric acid receptor subunit pi Proteins 0.000 claims 2
- 101000926939 Homo sapiens Glucocorticoid receptor Proteins 0.000 claims 2
- 101001010139 Homo sapiens Glutathione S-transferase P Proteins 0.000 claims 2
- 101000997558 Homo sapiens Glutathione hydrolase 1 proenzyme Proteins 0.000 claims 2
- 101000746373 Homo sapiens Granulocyte-macrophage colony-stimulating factor Proteins 0.000 claims 2
- 101001023988 Homo sapiens Growth/differentiation factor 5 Proteins 0.000 claims 2
- 101001014590 Homo sapiens Guanine nucleotide-binding protein G(s) subunit alpha isoforms XLas Proteins 0.000 claims 2
- 101001014594 Homo sapiens Guanine nucleotide-binding protein G(s) subunit alpha isoforms short Proteins 0.000 claims 2
- 101001079623 Homo sapiens Heme oxygenase 1 Proteins 0.000 claims 2
- 101000872875 Homo sapiens Hepatocyte cell adhesion molecule Proteins 0.000 claims 2
- 101000878611 Homo sapiens High affinity immunoglobulin epsilon receptor subunit alpha Proteins 0.000 claims 2
- 101000899259 Homo sapiens Histone deacetylase 4 Proteins 0.000 claims 2
- 101000899255 Homo sapiens Histone deacetylase 5 Proteins 0.000 claims 2
- 101001032113 Homo sapiens Histone deacetylase 7 Proteins 0.000 claims 2
- 101001032092 Homo sapiens Histone deacetylase 9 Proteins 0.000 claims 2
- 101001035752 Homo sapiens Hydroxycarboxylic acid receptor 3 Proteins 0.000 claims 2
- 101001046870 Homo sapiens Hypoxia-inducible factor 1-alpha Proteins 0.000 claims 2
- 101001019455 Homo sapiens ICOS ligand Proteins 0.000 claims 2
- 101001002508 Homo sapiens Immunoglobulin-binding protein 1 Proteins 0.000 claims 2
- 101001076604 Homo sapiens Inhibin alpha chain Proteins 0.000 claims 2
- 101000998783 Homo sapiens Insulin-like 3 Proteins 0.000 claims 2
- 101001034652 Homo sapiens Insulin-like growth factor 1 receptor Proteins 0.000 claims 2
- 101001044940 Homo sapiens Insulin-like growth factor-binding protein 2 Proteins 0.000 claims 2
- 101001044927 Homo sapiens Insulin-like growth factor-binding protein 3 Proteins 0.000 claims 2
- 101000840582 Homo sapiens Insulin-like growth factor-binding protein 6 Proteins 0.000 claims 2
- 101001078158 Homo sapiens Integrin alpha-1 Proteins 0.000 claims 2
- 101001078133 Homo sapiens Integrin alpha-2 Proteins 0.000 claims 2
- 101000994378 Homo sapiens Integrin alpha-3 Proteins 0.000 claims 2
- 101000994365 Homo sapiens Integrin alpha-6 Proteins 0.000 claims 2
- 101001046677 Homo sapiens Integrin alpha-V Proteins 0.000 claims 2
- 101001015004 Homo sapiens Integrin beta-3 Proteins 0.000 claims 2
- 101001015006 Homo sapiens Integrin beta-4 Proteins 0.000 claims 2
- 101000959820 Homo sapiens Interferon alpha-1/13 Proteins 0.000 claims 2
- 101000959794 Homo sapiens Interferon alpha-2 Proteins 0.000 claims 2
- 101000959708 Homo sapiens Interferon alpha-4 Proteins 0.000 claims 2
- 101000959704 Homo sapiens Interferon alpha-5 Proteins 0.000 claims 2
- 101000959714 Homo sapiens Interferon alpha-6 Proteins 0.000 claims 2
- 101000961126 Homo sapiens Interferon alpha-7 Proteins 0.000 claims 2
- 101001054334 Homo sapiens Interferon beta Proteins 0.000 claims 2
- 101001002470 Homo sapiens Interferon lambda-1 Proteins 0.000 claims 2
- 101001002469 Homo sapiens Interferon lambda-2 Proteins 0.000 claims 2
- 101001002466 Homo sapiens Interferon lambda-3 Proteins 0.000 claims 2
- 101000999370 Homo sapiens Interferon omega-1 Proteins 0.000 claims 2
- 101001033249 Homo sapiens Interleukin-1 beta Proteins 0.000 claims 2
- 101001076386 Homo sapiens Interleukin-1 family member 10 Proteins 0.000 claims 2
- 101000960952 Homo sapiens Interleukin-1 receptor accessory protein Proteins 0.000 claims 2
- 101000994815 Homo sapiens Interleukin-1 receptor accessory protein-like 1 Proteins 0.000 claims 2
- 101001076418 Homo sapiens Interleukin-1 receptor type 1 Proteins 0.000 claims 2
- 101001076422 Homo sapiens Interleukin-1 receptor type 2 Proteins 0.000 claims 2
- 101000852483 Homo sapiens Interleukin-1 receptor-associated kinase 1 Proteins 0.000 claims 2
- 101000852255 Homo sapiens Interleukin-1 receptor-associated kinase-like 2 Proteins 0.000 claims 2
- 101000852965 Homo sapiens Interleukin-1 receptor-like 2 Proteins 0.000 claims 2
- 101001003147 Homo sapiens Interleukin-11 receptor subunit alpha Proteins 0.000 claims 2
- 101001003142 Homo sapiens Interleukin-12 receptor subunit beta-1 Proteins 0.000 claims 2
- 101001003138 Homo sapiens Interleukin-12 receptor subunit beta-2 Proteins 0.000 claims 2
- 101001010600 Homo sapiens Interleukin-12 subunit alpha Proteins 0.000 claims 2
- 101000852992 Homo sapiens Interleukin-12 subunit beta Proteins 0.000 claims 2
- 101001003135 Homo sapiens Interleukin-13 receptor subunit alpha-1 Proteins 0.000 claims 2
- 101001003132 Homo sapiens Interleukin-13 receptor subunit alpha-2 Proteins 0.000 claims 2
- 101001003140 Homo sapiens Interleukin-15 receptor subunit alpha Proteins 0.000 claims 2
- 101001019598 Homo sapiens Interleukin-17 receptor A Proteins 0.000 claims 2
- 101000998146 Homo sapiens Interleukin-17A Proteins 0.000 claims 2
- 101000998181 Homo sapiens Interleukin-17B Proteins 0.000 claims 2
- 101000998178 Homo sapiens Interleukin-17C Proteins 0.000 claims 2
- 101000961065 Homo sapiens Interleukin-18 receptor 1 Proteins 0.000 claims 2
- 101001019615 Homo sapiens Interleukin-18 receptor accessory protein Proteins 0.000 claims 2
- 101001019591 Homo sapiens Interleukin-18-binding protein Proteins 0.000 claims 2
- 101000960946 Homo sapiens Interleukin-19 Proteins 0.000 claims 2
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 claims 2
- 101001044893 Homo sapiens Interleukin-20 receptor subunit alpha Proteins 0.000 claims 2
- 101001010626 Homo sapiens Interleukin-22 Proteins 0.000 claims 2
- 101001044887 Homo sapiens Interleukin-22 receptor subunit alpha-2 Proteins 0.000 claims 2
- 101000853009 Homo sapiens Interleukin-24 Proteins 0.000 claims 2
- 101000853000 Homo sapiens Interleukin-26 Proteins 0.000 claims 2
- 101000852998 Homo sapiens Interleukin-27 subunit alpha Proteins 0.000 claims 2
- 101000998120 Homo sapiens Interleukin-3 receptor subunit alpha Proteins 0.000 claims 2
- 101001033312 Homo sapiens Interleukin-4 receptor subunit alpha Proteins 0.000 claims 2
- 101000960936 Homo sapiens Interleukin-5 receptor subunit alpha Proteins 0.000 claims 2
- 101000599048 Homo sapiens Interleukin-6 receptor subunit alpha Proteins 0.000 claims 2
- 101000599056 Homo sapiens Interleukin-6 receptor subunit beta Proteins 0.000 claims 2
- 101001043809 Homo sapiens Interleukin-7 receptor subunit alpha Proteins 0.000 claims 2
- 101001055219 Homo sapiens Interleukin-9 receptor Proteins 0.000 claims 2
- 101000605522 Homo sapiens Kallikrein-1 Proteins 0.000 claims 2
- 101000605516 Homo sapiens Kallikrein-12 Proteins 0.000 claims 2
- 101000605514 Homo sapiens Kallikrein-13 Proteins 0.000 claims 2
- 101000605520 Homo sapiens Kallikrein-14 Proteins 0.000 claims 2
- 101000605518 Homo sapiens Kallikrein-15 Proteins 0.000 claims 2
- 101001091379 Homo sapiens Kallikrein-5 Proteins 0.000 claims 2
- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 claims 2
- 101001091356 Homo sapiens Kallikrein-9 Proteins 0.000 claims 2
- 101000998011 Homo sapiens Keratin, type I cytoskeletal 19 Proteins 0.000 claims 2
- 101001026977 Homo sapiens Keratin, type II cuticular Hb6 Proteins 0.000 claims 2
- 101001046960 Homo sapiens Keratin, type II cytoskeletal 1 Proteins 0.000 claims 2
- 101001046936 Homo sapiens Keratin, type II cytoskeletal 2 epidermal Proteins 0.000 claims 2
- 101001046952 Homo sapiens Keratin, type II cytoskeletal 2 oral Proteins 0.000 claims 2
- 101000934753 Homo sapiens Keratin, type II cytoskeletal 75 Proteins 0.000 claims 2
- 101000716729 Homo sapiens Kit ligand Proteins 0.000 claims 2
- 101001139130 Homo sapiens Krueppel-like factor 5 Proteins 0.000 claims 2
- 101001139126 Homo sapiens Krueppel-like factor 6 Proteins 0.000 claims 2
- 101001059438 Homo sapiens Leucine-rich repeat transmembrane protein FLRT1 Proteins 0.000 claims 2
- 101001017969 Homo sapiens Leukotriene B4 receptor 2 Proteins 0.000 claims 2
- 101000927946 Homo sapiens LisH domain-containing protein ARMC9 Proteins 0.000 claims 2
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 claims 2
- 101000804764 Homo sapiens Lymphotactin Proteins 0.000 claims 2
- 101000739159 Homo sapiens Mammaglobin-A Proteins 0.000 claims 2
- 101000739168 Homo sapiens Mammaglobin-B Proteins 0.000 claims 2
- 101000990902 Homo sapiens Matrix metalloproteinase-9 Proteins 0.000 claims 2
- 101000628547 Homo sapiens Metalloreductase STEAP1 Proteins 0.000 claims 2
- 101000628535 Homo sapiens Metalloreductase STEAP2 Proteins 0.000 claims 2
- 101000615613 Homo sapiens Mineralocorticoid receptor Proteins 0.000 claims 2
- 101000976899 Homo sapiens Mitogen-activated protein kinase 15 Proteins 0.000 claims 2
- 101001013159 Homo sapiens Myeloid leukemia factor 2 Proteins 0.000 claims 2
- 101000928278 Homo sapiens Natriuretic peptides B Proteins 0.000 claims 2
- 101000979293 Homo sapiens Negative elongation factor C/D Proteins 0.000 claims 2
- 101000995164 Homo sapiens Netrin-4 Proteins 0.000 claims 2
- 101001014610 Homo sapiens Neuroendocrine secretory protein 55 Proteins 0.000 claims 2
- 101000979338 Homo sapiens Nuclear factor NF-kappa-B p100 subunit Proteins 0.000 claims 2
- 101000979342 Homo sapiens Nuclear factor NF-kappa-B p105 subunit Proteins 0.000 claims 2
- 101000633503 Homo sapiens Nuclear receptor subfamily 2 group E member 1 Proteins 0.000 claims 2
- 101000633516 Homo sapiens Nuclear receptor subfamily 2 group F member 6 Proteins 0.000 claims 2
- 101001109700 Homo sapiens Nuclear receptor subfamily 4 group A member 1 Proteins 0.000 claims 2
- 101001109698 Homo sapiens Nuclear receptor subfamily 4 group A member 2 Proteins 0.000 claims 2
- 101001109689 Homo sapiens Nuclear receptor subfamily 4 group A member 3 Proteins 0.000 claims 2
- 101001109685 Homo sapiens Nuclear receptor subfamily 5 group A member 2 Proteins 0.000 claims 2
- 101001109682 Homo sapiens Nuclear receptor subfamily 6 group A member 1 Proteins 0.000 claims 2
- 101001114057 Homo sapiens P antigen family member 1 Proteins 0.000 claims 2
- 101001114052 Homo sapiens P antigen family member 4 Proteins 0.000 claims 2
- 101001098175 Homo sapiens P2X purinoceptor 7 Proteins 0.000 claims 2
- 101000613565 Homo sapiens PRKC apoptosis WT1 regulator protein Proteins 0.000 claims 2
- 101001095231 Homo sapiens Peptidyl-prolyl cis-trans isomerase D Proteins 0.000 claims 2
- 101000595751 Homo sapiens Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform Proteins 0.000 claims 2
- 101000633511 Homo sapiens Photoreceptor-specific nuclear receptor Proteins 0.000 claims 2
- 101001073422 Homo sapiens Pigment epithelium-derived factor Proteins 0.000 claims 2
- 101001091365 Homo sapiens Plasma kallikrein Proteins 0.000 claims 2
- 101001116302 Homo sapiens Platelet endothelial cell adhesion molecule Proteins 0.000 claims 2
- 101001096065 Homo sapiens Plexin domain-containing protein 1 Proteins 0.000 claims 2
- 101001056707 Homo sapiens Proepiregulin Proteins 0.000 claims 2
- 101000904173 Homo sapiens Progonadoliberin-1 Proteins 0.000 claims 2
- 101000610543 Homo sapiens Prokineticin-2 Proteins 0.000 claims 2
- 101000945496 Homo sapiens Proliferation marker protein Ki-67 Proteins 0.000 claims 2
- 101001117314 Homo sapiens Prostaglandin D2 receptor 2 Proteins 0.000 claims 2
- 101001136592 Homo sapiens Prostate stem cell antigen Proteins 0.000 claims 2
- 101000605534 Homo sapiens Prostate-specific antigen Proteins 0.000 claims 2
- 101000797903 Homo sapiens Protein ALEX Proteins 0.000 claims 2
- 101000740825 Homo sapiens Protein C10 Proteins 0.000 claims 2
- 101000994437 Homo sapiens Protein jagged-1 Proteins 0.000 claims 2
- 101000602015 Homo sapiens Protocadherin gamma-B4 Proteins 0.000 claims 2
- 101000668165 Homo sapiens RNA-binding motif, single-stranded-interacting protein 1 Proteins 0.000 claims 2
- 101001110313 Homo sapiens Ras-related C3 botulinum toxin substrate 2 Proteins 0.000 claims 2
- 101001132698 Homo sapiens Retinoic acid receptor beta Proteins 0.000 claims 2
- 101000716809 Homo sapiens Secretogranin-1 Proteins 0.000 claims 2
- 101001026870 Homo sapiens Serine/threonine-protein kinase D1 Proteins 0.000 claims 2
- 101000799194 Homo sapiens Serine/threonine-protein kinase receptor R3 Proteins 0.000 claims 2
- 101000651890 Homo sapiens Slit homolog 2 protein Proteins 0.000 claims 2
- 101000651893 Homo sapiens Slit homolog 3 protein Proteins 0.000 claims 2
- 101000689224 Homo sapiens Src-like-adapter 2 Proteins 0.000 claims 2
- 101000684994 Homo sapiens Stromal cell-derived factor 2 Proteins 0.000 claims 2
- 101000713602 Homo sapiens T-box transcription factor TBX21 Proteins 0.000 claims 2
- 101000946860 Homo sapiens T-cell surface glycoprotein CD3 epsilon chain Proteins 0.000 claims 2
- 101000738413 Homo sapiens T-cell surface glycoprotein CD3 gamma chain Proteins 0.000 claims 2
- 101000738335 Homo sapiens T-cell surface glycoprotein CD3 zeta chain Proteins 0.000 claims 2
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 claims 2
- 101000800639 Homo sapiens Teneurin-1 Proteins 0.000 claims 2
- 101000835745 Homo sapiens Teratocarcinoma-derived growth factor 1 Proteins 0.000 claims 2
- 101000799461 Homo sapiens Thrombopoietin Proteins 0.000 claims 2
- 101000633605 Homo sapiens Thrombospondin-2 Proteins 0.000 claims 2
- 101000633617 Homo sapiens Thrombospondin-4 Proteins 0.000 claims 2
- 101000845170 Homo sapiens Thymic stromal lymphopoietin Proteins 0.000 claims 2
- 101000830560 Homo sapiens Toll-interacting protein Proteins 0.000 claims 2
- 101000763537 Homo sapiens Toll-like receptor 10 Proteins 0.000 claims 2
- 101000831567 Homo sapiens Toll-like receptor 2 Proteins 0.000 claims 2
- 101000831496 Homo sapiens Toll-like receptor 3 Proteins 0.000 claims 2
- 101000669447 Homo sapiens Toll-like receptor 4 Proteins 0.000 claims 2
- 101000669460 Homo sapiens Toll-like receptor 5 Proteins 0.000 claims 2
- 101000669406 Homo sapiens Toll-like receptor 6 Proteins 0.000 claims 2
- 101000669402 Homo sapiens Toll-like receptor 7 Proteins 0.000 claims 2
- 101000800483 Homo sapiens Toll-like receptor 8 Proteins 0.000 claims 2
- 101000819111 Homo sapiens Trans-acting T-cell-specific transcription factor GATA-3 Proteins 0.000 claims 2
- 101001050288 Homo sapiens Transcription factor Jun Proteins 0.000 claims 2
- 101000795107 Homo sapiens Triggering receptor expressed on myeloid cells 1 Proteins 0.000 claims 2
- 101000795117 Homo sapiens Triggering receptor expressed on myeloid cells 2 Proteins 0.000 claims 2
- 101000801701 Homo sapiens Tropomyosin alpha-1 chain Proteins 0.000 claims 2
- 101000851892 Homo sapiens Tropomyosin beta chain Proteins 0.000 claims 2
- 101000830603 Homo sapiens Tumor necrosis factor ligand superfamily member 11 Proteins 0.000 claims 2
- 101000830600 Homo sapiens Tumor necrosis factor ligand superfamily member 13 Proteins 0.000 claims 2
- 101000597779 Homo sapiens Tumor necrosis factor ligand superfamily member 18 Proteins 0.000 claims 2
- 101000764263 Homo sapiens Tumor necrosis factor ligand superfamily member 4 Proteins 0.000 claims 2
- 101000638161 Homo sapiens Tumor necrosis factor ligand superfamily member 6 Proteins 0.000 claims 2
- 101000638255 Homo sapiens Tumor necrosis factor ligand superfamily member 8 Proteins 0.000 claims 2
- 101000638251 Homo sapiens Tumor necrosis factor ligand superfamily member 9 Proteins 0.000 claims 2
- 101000801228 Homo sapiens Tumor necrosis factor receptor superfamily member 1A Proteins 0.000 claims 2
- 101000801232 Homo sapiens Tumor necrosis factor receptor superfamily member 1B Proteins 0.000 claims 2
- 101000679857 Homo sapiens Tumor necrosis factor receptor superfamily member 3 Proteins 0.000 claims 2
- 101000611185 Homo sapiens Tumor necrosis factor receptor superfamily member 5 Proteins 0.000 claims 2
- 101000851370 Homo sapiens Tumor necrosis factor receptor superfamily member 9 Proteins 0.000 claims 2
- 101000850748 Homo sapiens Tumor necrosis factor receptor type 1-associated DEATH domain protein Proteins 0.000 claims 2
- 101000997835 Homo sapiens Tyrosine-protein kinase JAK1 Proteins 0.000 claims 2
- 101000934996 Homo sapiens Tyrosine-protein kinase JAK3 Proteins 0.000 claims 2
- 101000742579 Homo sapiens Vascular endothelial growth factor B Proteins 0.000 claims 2
- 101000742596 Homo sapiens Vascular endothelial growth factor C Proteins 0.000 claims 2
- 101000742599 Homo sapiens Vascular endothelial growth factor D Proteins 0.000 claims 2
- 101000851018 Homo sapiens Vascular endothelial growth factor receptor 1 Proteins 0.000 claims 2
- 101000994810 Homo sapiens X-linked interleukin-1 receptor accessory protein-like 2 Proteins 0.000 claims 2
- 101001059220 Homo sapiens Zinc finger protein Gfi-1 Proteins 0.000 claims 2
- 101000931371 Homo sapiens Zinc finger protein ZFPM2 Proteins 0.000 claims 2
- 101000669028 Homo sapiens Zinc phosphodiesterase ELAC protein 2 Proteins 0.000 claims 2
- 101001026578 Hordeum vulgare Ent-kaurenoic acid oxidase 1 Proteins 0.000 claims 2
- 102100039356 Hydroxycarboxylic acid receptor 3 Human genes 0.000 claims 2
- 102100022875 Hypoxia-inducible factor 1-alpha Human genes 0.000 claims 2
- 102100034980 ICOS ligand Human genes 0.000 claims 2
- 108091058536 IL1F9 Proteins 0.000 claims 2
- 102000026633 IL6 Human genes 0.000 claims 2
- 102100021042 Immunoglobulin-binding protein 1 Human genes 0.000 claims 2
- 102100025885 Inhibin alpha chain Human genes 0.000 claims 2
- 102100027004 Inhibin beta A chain Human genes 0.000 claims 2
- 102100033262 Insulin-like 3 Human genes 0.000 claims 2
- 102100039688 Insulin-like growth factor 1 receptor Human genes 0.000 claims 2
- 102100022710 Insulin-like growth factor-binding protein 2 Human genes 0.000 claims 2
- 102100022708 Insulin-like growth factor-binding protein 3 Human genes 0.000 claims 2
- 102100029180 Insulin-like growth factor-binding protein 6 Human genes 0.000 claims 2
- 102100025323 Integrin alpha-1 Human genes 0.000 claims 2
- 102100025305 Integrin alpha-2 Human genes 0.000 claims 2
- 102100032819 Integrin alpha-3 Human genes 0.000 claims 2
- 102100032816 Integrin alpha-6 Human genes 0.000 claims 2
- 102100022337 Integrin alpha-V Human genes 0.000 claims 2
- 108010008212 Integrin alpha4beta1 Proteins 0.000 claims 2
- 102100032999 Integrin beta-3 Human genes 0.000 claims 2
- 102100033000 Integrin beta-4 Human genes 0.000 claims 2
- 102100040019 Interferon alpha-1/13 Human genes 0.000 claims 2
- 102100040018 Interferon alpha-2 Human genes 0.000 claims 2
- 102100039949 Interferon alpha-4 Human genes 0.000 claims 2
- 102100039948 Interferon alpha-5 Human genes 0.000 claims 2
- 102100040007 Interferon alpha-6 Human genes 0.000 claims 2
- 102100039350 Interferon alpha-7 Human genes 0.000 claims 2
- 102100026720 Interferon beta Human genes 0.000 claims 2
- 102100020990 Interferon lambda-1 Human genes 0.000 claims 2
- 102100020989 Interferon lambda-2 Human genes 0.000 claims 2
- 102100020992 Interferon lambda-3 Human genes 0.000 claims 2
- 102100036479 Interferon omega-1 Human genes 0.000 claims 2
- 108700003107 Interleukin-1 Receptor-Like 1 Proteins 0.000 claims 2
- 102100039065 Interleukin-1 beta Human genes 0.000 claims 2
- 102100026015 Interleukin-1 family member 10 Human genes 0.000 claims 2
- 102100039880 Interleukin-1 receptor accessory protein Human genes 0.000 claims 2
- 102100034413 Interleukin-1 receptor accessory protein-like 1 Human genes 0.000 claims 2
- 102100026016 Interleukin-1 receptor type 1 Human genes 0.000 claims 2
- 102100026017 Interleukin-1 receptor type 2 Human genes 0.000 claims 2
- 102100036342 Interleukin-1 receptor-associated kinase 1 Human genes 0.000 claims 2
- 102100036433 Interleukin-1 receptor-associated kinase-like 2 Human genes 0.000 claims 2
- 102100036706 Interleukin-1 receptor-like 1 Human genes 0.000 claims 2
- 102100036697 Interleukin-1 receptor-like 2 Human genes 0.000 claims 2
- 102100030694 Interleukin-11 Human genes 0.000 claims 2
- 102100020787 Interleukin-11 receptor subunit alpha Human genes 0.000 claims 2
- 108010011429 Interleukin-12 Subunit p40 Proteins 0.000 claims 2
- 102000014158 Interleukin-12 Subunit p40 Human genes 0.000 claims 2
- 102100020790 Interleukin-12 receptor subunit beta-1 Human genes 0.000 claims 2
- 102100020792 Interleukin-12 receptor subunit beta-2 Human genes 0.000 claims 2
- 102100030698 Interleukin-12 subunit alpha Human genes 0.000 claims 2
- 102100036701 Interleukin-12 subunit beta Human genes 0.000 claims 2
- 102100020791 Interleukin-13 receptor subunit alpha-1 Human genes 0.000 claims 2
- 102100020793 Interleukin-13 receptor subunit alpha-2 Human genes 0.000 claims 2
- 102100020789 Interleukin-15 receptor subunit alpha Human genes 0.000 claims 2
- 101800003050 Interleukin-16 Proteins 0.000 claims 2
- 102000049772 Interleukin-16 Human genes 0.000 claims 2
- 108050003558 Interleukin-17 Proteins 0.000 claims 2
- 102000013691 Interleukin-17 Human genes 0.000 claims 2
- 102100035018 Interleukin-17 receptor A Human genes 0.000 claims 2
- 102100033461 Interleukin-17A Human genes 0.000 claims 2
- 102100033101 Interleukin-17B Human genes 0.000 claims 2
- 102100033105 Interleukin-17C Human genes 0.000 claims 2
- 102100039340 Interleukin-18 receptor 1 Human genes 0.000 claims 2
- 102100035010 Interleukin-18 receptor accessory protein Human genes 0.000 claims 2
- 102100035017 Interleukin-18-binding protein Human genes 0.000 claims 2
- 102100039879 Interleukin-19 Human genes 0.000 claims 2
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 claims 2
- 102100022706 Interleukin-20 receptor subunit alpha Human genes 0.000 claims 2
- 108010017411 Interleukin-21 Receptors Proteins 0.000 claims 2
- 102100030699 Interleukin-21 receptor Human genes 0.000 claims 2
- 102100030703 Interleukin-22 Human genes 0.000 claims 2
- 102100022703 Interleukin-22 receptor subunit alpha-2 Human genes 0.000 claims 2
- 102000013264 Interleukin-23 Human genes 0.000 claims 2
- 102100036671 Interleukin-24 Human genes 0.000 claims 2
- 102100036679 Interleukin-26 Human genes 0.000 claims 2
- 102100036678 Interleukin-27 subunit alpha Human genes 0.000 claims 2
- 102100033493 Interleukin-3 receptor subunit alpha Human genes 0.000 claims 2
- 102100033474 Interleukin-36 alpha Human genes 0.000 claims 2
- 102100033498 Interleukin-36 beta Human genes 0.000 claims 2
- 102100033503 Interleukin-36 gamma Human genes 0.000 claims 2
- 102100039078 Interleukin-4 receptor subunit alpha Human genes 0.000 claims 2
- 102100039881 Interleukin-5 receptor subunit alpha Human genes 0.000 claims 2
- 102100037792 Interleukin-6 receptor subunit alpha Human genes 0.000 claims 2
- 102100021593 Interleukin-7 receptor subunit alpha Human genes 0.000 claims 2
- 102100026244 Interleukin-9 receptor Human genes 0.000 claims 2
- 102100038318 Kallikrein-12 Human genes 0.000 claims 2
- 102100038315 Kallikrein-13 Human genes 0.000 claims 2
- 102100038298 Kallikrein-14 Human genes 0.000 claims 2
- 102100038301 Kallikrein-15 Human genes 0.000 claims 2
- 102100034872 Kallikrein-4 Human genes 0.000 claims 2
- 102100034868 Kallikrein-5 Human genes 0.000 claims 2
- 102100034866 Kallikrein-6 Human genes 0.000 claims 2
- 102100034876 Kallikrein-9 Human genes 0.000 claims 2
- 108700032443 Kangai-1 Proteins 0.000 claims 2
- 102000057159 Kangai-1 Human genes 0.000 claims 2
- 102100037382 Keratin, type II cuticular Hb6 Human genes 0.000 claims 2
- 102100022905 Keratin, type II cytoskeletal 1 Human genes 0.000 claims 2
- 102100022854 Keratin, type II cytoskeletal 2 epidermal Human genes 0.000 claims 2
- 102100022926 Keratin, type II cytoskeletal 2 oral Human genes 0.000 claims 2
- 102100025367 Keratin, type II cytoskeletal 75 Human genes 0.000 claims 2
- 108010066302 Keratin-19 Proteins 0.000 claims 2
- 102100020680 Krueppel-like factor 5 Human genes 0.000 claims 2
- 102100020679 Krueppel-like factor 6 Human genes 0.000 claims 2
- 102100038269 Large neutral amino acids transporter small subunit 3 Human genes 0.000 claims 2
- 102100028919 Leucine-rich repeat transmembrane protein FLRT1 Human genes 0.000 claims 2
- 102100033375 Leukotriene B4 receptor 2 Human genes 0.000 claims 2
- 102100036882 LisH domain-containing protein ARMC9 Human genes 0.000 claims 2
- 102100029193 Low affinity immunoglobulin gamma Fc region receptor III-A Human genes 0.000 claims 2
- 101150053046 MYD88 gene Proteins 0.000 claims 2
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 claims 2
- 108010031030 Mammaglobin A Proteins 0.000 claims 2
- 108010031029 Mammaglobin B Proteins 0.000 claims 2
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 claims 2
- 102100026261 Metalloproteinase inhibitor 3 Human genes 0.000 claims 2
- 102100026712 Metalloreductase STEAP1 Human genes 0.000 claims 2
- 102100026711 Metalloreductase STEAP2 Human genes 0.000 claims 2
- 108010092801 Midkine Proteins 0.000 claims 2
- 102100021316 Mineralocorticoid receptor Human genes 0.000 claims 2
- 102100023483 Mitogen-activated protein kinase 15 Human genes 0.000 claims 2
- 101000934396 Mus musculus C-C chemokine receptor-like 2 Proteins 0.000 claims 2
- 101100219997 Mus musculus Ccr1 gene Proteins 0.000 claims 2
- 101100005657 Mus musculus Ccr7 gene Proteins 0.000 claims 2
- 101100446506 Mus musculus Fgf3 gene Proteins 0.000 claims 2
- 102100026784 Myelin proteolipid protein Human genes 0.000 claims 2
- 101710094913 Myelin proteolipid protein Proteins 0.000 claims 2
- 102100024134 Myeloid differentiation primary response protein MyD88 Human genes 0.000 claims 2
- 102100029687 Myeloid leukemia factor 2 Human genes 0.000 claims 2
- 108010082695 NADPH Oxidase 5 Proteins 0.000 claims 2
- 102100021871 NADPH oxidase 5 Human genes 0.000 claims 2
- 102100036836 Natriuretic peptides B Human genes 0.000 claims 2
- 102100023069 Negative elongation factor C/D Human genes 0.000 claims 2
- 108010043296 Neurocan Proteins 0.000 claims 2
- 102100030466 Neurocan core protein Human genes 0.000 claims 2
- 108090000772 Neuropilin-1 Proteins 0.000 claims 2
- 108090000770 Neuropilin-2 Proteins 0.000 claims 2
- 108010077641 Nogo Proteins Proteins 0.000 claims 2
- 102000010410 Nogo Proteins Human genes 0.000 claims 2
- 102100023059 Nuclear factor NF-kappa-B p100 subunit Human genes 0.000 claims 2
- 102100028470 Nuclear receptor subfamily 2 group C member 1 Human genes 0.000 claims 2
- 102100028448 Nuclear receptor subfamily 2 group C member 2 Human genes 0.000 claims 2
- 102100029534 Nuclear receptor subfamily 2 group E member 1 Human genes 0.000 claims 2
- 102100029528 Nuclear receptor subfamily 2 group F member 6 Human genes 0.000 claims 2
- 102100022679 Nuclear receptor subfamily 4 group A member 1 Human genes 0.000 claims 2
- 102100022676 Nuclear receptor subfamily 4 group A member 2 Human genes 0.000 claims 2
- 102100022673 Nuclear receptor subfamily 4 group A member 3 Human genes 0.000 claims 2
- 102100022669 Nuclear receptor subfamily 5 group A member 2 Human genes 0.000 claims 2
- 102100022670 Nuclear receptor subfamily 6 group A member 1 Human genes 0.000 claims 2
- 108010042215 OX40 Ligand Proteins 0.000 claims 2
- 102100040557 Osteopontin Human genes 0.000 claims 2
- 102100023219 P antigen family member 1 Human genes 0.000 claims 2
- 102100023240 P antigen family member 4 Human genes 0.000 claims 2
- 102100037602 P2X purinoceptor 7 Human genes 0.000 claims 2
- 108091033411 PCA3 Proteins 0.000 claims 2
- 102100040853 PRKC apoptosis WT1 regulator protein Human genes 0.000 claims 2
- 101150084398 PTAFR gene Proteins 0.000 claims 2
- 108010011536 PTEN Phosphohydrolase Proteins 0.000 claims 2
- 102100037827 Peptidyl-prolyl cis-trans isomerase D Human genes 0.000 claims 2
- 102100032543 Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Human genes 0.000 claims 2
- 102100036052 Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform Human genes 0.000 claims 2
- 102100029533 Photoreceptor-specific nuclear receptor Human genes 0.000 claims 2
- 102100035846 Pigment epithelium-derived factor Human genes 0.000 claims 2
- 102100034869 Plasma kallikrein Human genes 0.000 claims 2
- 108010022233 Plasminogen Activator Inhibitor 1 Proteins 0.000 claims 2
- 102100039418 Plasminogen activator inhibitor 1 Human genes 0.000 claims 2
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 claims 2
- 108700023400 Platelet-activating factor receptors Proteins 0.000 claims 2
- 102100037596 Platelet-derived growth factor subunit A Human genes 0.000 claims 2
- 102100040990 Platelet-derived growth factor subunit B Human genes 0.000 claims 2
- 102100030477 Plectin Human genes 0.000 claims 2
- 108010054050 Plectin Proteins 0.000 claims 2
- 102100037891 Plexin domain-containing protein 1 Human genes 0.000 claims 2
- 102100025498 Proepiregulin Human genes 0.000 claims 2
- 102100024028 Progonadoliberin-1 Human genes 0.000 claims 2
- 102100040125 Prokineticin-2 Human genes 0.000 claims 2
- 102100036691 Proliferating cell nuclear antigen Human genes 0.000 claims 2
- 102100034836 Proliferation marker protein Ki-67 Human genes 0.000 claims 2
- 102100024218 Prostaglandin D2 receptor 2 Human genes 0.000 claims 2
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 claims 2
- 102100036735 Prostate stem cell antigen Human genes 0.000 claims 2
- 102100038957 Protein C10 Human genes 0.000 claims 2
- 108010015499 Protein Kinase C-theta Proteins 0.000 claims 2
- 102100032702 Protein jagged-1 Human genes 0.000 claims 2
- 102100021566 Protein kinase C theta type Human genes 0.000 claims 2
- 102100023068 Protein kinase C-binding protein NELL1 Human genes 0.000 claims 2
- 102100034433 Protein kinase C-binding protein NELL2 Human genes 0.000 claims 2
- 108010019674 Proto-Oncogene Proteins c-sis Proteins 0.000 claims 2
- 108010007100 Pulmonary Surfactant-Associated Protein A Proteins 0.000 claims 2
- 102100027773 Pulmonary surfactant-associated protein A2 Human genes 0.000 claims 2
- 102100022129 Ras-related C3 botulinum toxin substrate 2 Human genes 0.000 claims 2
- 108010038036 Receptor Activator of Nuclear Factor-kappa B Proteins 0.000 claims 2
- 102100028508 Receptor-type tyrosine-protein phosphatase zeta Human genes 0.000 claims 2
- 102100021269 Regulator of G-protein signaling 1 Human genes 0.000 claims 2
- 101710140408 Regulator of G-protein signaling 1 Proteins 0.000 claims 2
- 101710148333 Regulator of G-protein signaling 13 Proteins 0.000 claims 2
- 102100021035 Regulator of G-protein signaling 18 Human genes 0.000 claims 2
- 102100037415 Regulator of G-protein signaling 3 Human genes 0.000 claims 2
- 101710140411 Regulator of G-protein signaling 3 Proteins 0.000 claims 2
- 102100033909 Retinoic acid receptor beta Human genes 0.000 claims 2
- 108091006299 SLC2A2 Proteins 0.000 claims 2
- 108091006570 SLC33A1 Proteins 0.000 claims 2
- 108091006993 SLC43A1 Proteins 0.000 claims 2
- 101100184049 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) MID2 gene Proteins 0.000 claims 2
- 102100020867 Secretogranin-1 Human genes 0.000 claims 2
- 102100037310 Serine/threonine-protein kinase D1 Human genes 0.000 claims 2
- 102100034136 Serine/threonine-protein kinase receptor R3 Human genes 0.000 claims 2
- 108010089417 Sex Hormone-Binding Globulin Proteins 0.000 claims 2
- 102100030758 Sex hormone-binding globulin Human genes 0.000 claims 2
- 102100022433 Single-stranded DNA cytosine deaminase Human genes 0.000 claims 2
- 102100027339 Slit homolog 3 protein Human genes 0.000 claims 2
- 101710168942 Sphingosine-1-phosphate phosphatase 1 Proteins 0.000 claims 2
- 102100024510 Src-like-adapter 2 Human genes 0.000 claims 2
- 102100024471 Stabilin-1 Human genes 0.000 claims 2
- 108010048349 Steroidogenic Factor 1 Proteins 0.000 claims 2
- 102100029856 Steroidogenic factor 1 Human genes 0.000 claims 2
- 102100023184 Stromal cell-derived factor 2 Human genes 0.000 claims 2
- 102100036840 T-box transcription factor TBX21 Human genes 0.000 claims 2
- 102100035794 T-cell surface glycoprotein CD3 epsilon chain Human genes 0.000 claims 2
- 102100037911 T-cell surface glycoprotein CD3 gamma chain Human genes 0.000 claims 2
- 102100037906 T-cell surface glycoprotein CD3 zeta chain Human genes 0.000 claims 2
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 claims 2
- 102100033456 TGF-beta receptor type-1 Human genes 0.000 claims 2
- 102100033455 TGF-beta receptor type-2 Human genes 0.000 claims 2
- 102000004398 TNF receptor-associated factor 1 Human genes 0.000 claims 2
- 108090000920 TNF receptor-associated factor 1 Proteins 0.000 claims 2
- 108090000925 TNF receptor-associated factor 2 Proteins 0.000 claims 2
- 102000004399 TNF receptor-associated factor 3 Human genes 0.000 claims 2
- 108090000922 TNF receptor-associated factor 3 Proteins 0.000 claims 2
- 102000003715 TNF receptor-associated factor 4 Human genes 0.000 claims 2
- 108090000008 TNF receptor-associated factor 4 Proteins 0.000 claims 2
- 102000003718 TNF receptor-associated factor 5 Human genes 0.000 claims 2
- 108090000001 TNF receptor-associated factor 5 Proteins 0.000 claims 2
- 102000003714 TNF receptor-associated factor 6 Human genes 0.000 claims 2
- 108090000009 TNF receptor-associated factor 6 Proteins 0.000 claims 2
- 102100034779 TRAF family member-associated NF-kappa-B activator Human genes 0.000 claims 2
- 102000003623 TRPC6 Human genes 0.000 claims 2
- 102100033213 Teneurin-1 Human genes 0.000 claims 2
- 102100026404 Teratocarcinoma-derived growth factor 1 Human genes 0.000 claims 2
- 102100034195 Thrombopoietin Human genes 0.000 claims 2
- 108010046722 Thrombospondin 1 Proteins 0.000 claims 2
- 102100029529 Thrombospondin-2 Human genes 0.000 claims 2
- 102100029219 Thrombospondin-4 Human genes 0.000 claims 2
- 102100031294 Thymic stromal lymphopoietin Human genes 0.000 claims 2
- 108010031429 Tissue Inhibitor of Metalloproteinase-3 Proteins 0.000 claims 2
- 108010060818 Toll-Like Receptor 9 Proteins 0.000 claims 2
- 102100024652 Toll-interacting protein Human genes 0.000 claims 2
- 102000002689 Toll-like receptor Human genes 0.000 claims 2
- 108020000411 Toll-like receptor Proteins 0.000 claims 2
- 102100027009 Toll-like receptor 10 Human genes 0.000 claims 2
- 102100024333 Toll-like receptor 2 Human genes 0.000 claims 2
- 102100024324 Toll-like receptor 3 Human genes 0.000 claims 2
- 102100039360 Toll-like receptor 4 Human genes 0.000 claims 2
- 102100039357 Toll-like receptor 5 Human genes 0.000 claims 2
- 102100039387 Toll-like receptor 6 Human genes 0.000 claims 2
- 102100039390 Toll-like receptor 7 Human genes 0.000 claims 2
- 102100033110 Toll-like receptor 8 Human genes 0.000 claims 2
- 102100033117 Toll-like receptor 9 Human genes 0.000 claims 2
- 101710183280 Topoisomerase Proteins 0.000 claims 2
- 102100021386 Trans-acting T-cell-specific transcription factor GATA-3 Human genes 0.000 claims 2
- 102100023132 Transcription factor Jun Human genes 0.000 claims 2
- 108010011702 Transforming Growth Factor-beta Type I Receptor Proteins 0.000 claims 2
- 108010082684 Transforming Growth Factor-beta Type II Receptor Proteins 0.000 claims 2
- 102100033663 Transforming growth factor beta receptor type 3 Human genes 0.000 claims 2
- 108090000097 Transforming growth factor beta-3 Proteins 0.000 claims 2
- 102000056172 Transforming growth factor beta-3 Human genes 0.000 claims 2
- 108050001421 Transient receptor potential channel, canonical 6 Proteins 0.000 claims 2
- 102100029681 Triggering receptor expressed on myeloid cells 1 Human genes 0.000 claims 2
- 102100029678 Triggering receptor expressed on myeloid cells 2 Human genes 0.000 claims 2
- 102100033632 Tropomyosin alpha-1 chain Human genes 0.000 claims 2
- 102100036471 Tropomyosin beta chain Human genes 0.000 claims 2
- 108010078814 Tumor Suppressor Protein p53 Proteins 0.000 claims 2
- 102100024568 Tumor necrosis factor ligand superfamily member 11 Human genes 0.000 claims 2
- 102100024585 Tumor necrosis factor ligand superfamily member 13 Human genes 0.000 claims 2
- 102100035283 Tumor necrosis factor ligand superfamily member 18 Human genes 0.000 claims 2
- 102100028787 Tumor necrosis factor receptor superfamily member 11A Human genes 0.000 claims 2
- 102100033732 Tumor necrosis factor receptor superfamily member 1A Human genes 0.000 claims 2
- 102100033733 Tumor necrosis factor receptor superfamily member 1B Human genes 0.000 claims 2
- 102100022205 Tumor necrosis factor receptor superfamily member 21 Human genes 0.000 claims 2
- 102100022156 Tumor necrosis factor receptor superfamily member 3 Human genes 0.000 claims 2
- 102100036856 Tumor necrosis factor receptor superfamily member 9 Human genes 0.000 claims 2
- 102100033081 Tumor necrosis factor receptor type 1-associated DEATH domain protein Human genes 0.000 claims 2
- 108010046308 Type II DNA Topoisomerases Proteins 0.000 claims 2
- 108010089374 Type II Keratins Proteins 0.000 claims 2
- 102000007962 Type II Keratins Human genes 0.000 claims 2
- 102100033438 Tyrosine-protein kinase JAK1 Human genes 0.000 claims 2
- 102100025387 Tyrosine-protein kinase JAK3 Human genes 0.000 claims 2
- 108091008605 VEGF receptors Proteins 0.000 claims 2
- 108010073919 Vascular Endothelial Growth Factor D Proteins 0.000 claims 2
- 102100038217 Vascular endothelial growth factor B Human genes 0.000 claims 2
- 102100038232 Vascular endothelial growth factor C Human genes 0.000 claims 2
- 102100033178 Vascular endothelial growth factor receptor 1 Human genes 0.000 claims 2
- 102100034412 X-linked interleukin-1 receptor accessory protein-like 2 Human genes 0.000 claims 2
- 102100029004 Zinc finger protein Gfi-1 Human genes 0.000 claims 2
- 102100020996 Zinc finger protein ZFPM2 Human genes 0.000 claims 2
- 102100039877 Zinc phosphodiesterase ELAC protein 2 Human genes 0.000 claims 2
- 239000003263 anabolic agent Substances 0.000 claims 2
- 230000003444 anaesthetic effect Effects 0.000 claims 2
- 230000000202 analgesic effect Effects 0.000 claims 2
- 229940121369 angiogenesis inhibitor Drugs 0.000 claims 2
- 239000005557 antagonist Substances 0.000 claims 2
- 229940045799 anthracyclines and related substance Drugs 0.000 claims 2
- 230000003092 anti-cytokine Effects 0.000 claims 2
- 230000001430 anti-depressive effect Effects 0.000 claims 2
- 230000000845 anti-microbial effect Effects 0.000 claims 2
- 230000002682 anti-psoriatic effect Effects 0.000 claims 2
- 230000000561 anti-psychotic effect Effects 0.000 claims 2
- 239000000935 antidepressant agent Substances 0.000 claims 2
- 229940005513 antidepressants Drugs 0.000 claims 2
- 239000004599 antimicrobial Substances 0.000 claims 2
- 239000003080 antimitotic agent Substances 0.000 claims 2
- 229940127225 asthma medication Drugs 0.000 claims 2
- KMGARVOVYXNAOF-UHFFFAOYSA-N benzpiperylone Chemical compound C1CN(C)CCC1N1C(=O)C(CC=2C=CC=CC=2)=C(C=2C=CC=CC=2)N1 KMGARVOVYXNAOF-UHFFFAOYSA-N 0.000 claims 2
- 229940125388 beta agonist Drugs 0.000 claims 2
- 108010079292 betaglycan Proteins 0.000 claims 2
- 230000003115 biocidal effect Effects 0.000 claims 2
- 229960001265 ciclosporin Drugs 0.000 claims 2
- 229930182912 cyclosporin Natural products 0.000 claims 2
- 239000000430 cytokine receptor antagonist Substances 0.000 claims 2
- FOCAHLGSDWHSAH-UHFFFAOYSA-N difluoromethanethione Chemical compound FC(F)=S FOCAHLGSDWHSAH-UHFFFAOYSA-N 0.000 claims 2
- 229940090124 dipeptidyl peptidase 4 (dpp-4) inhibitors for blood glucose lowering Drugs 0.000 claims 2
- 230000003511 endothelial effect Effects 0.000 claims 2
- 229960005139 epinephrine Drugs 0.000 claims 2
- VLMZMRDOMOGGFA-WDBKCZKBSA-N festuclavine Chemical compound C1=CC([C@H]2C[C@H](CN(C)[C@@H]2C2)C)=C3C2=CNC3=C1 VLMZMRDOMOGGFA-WDBKCZKBSA-N 0.000 claims 2
- 108090000047 fibroblast growth factor 13 Proteins 0.000 claims 2
- 102000003684 fibroblast growth factor 13 Human genes 0.000 claims 2
- 229960001340 histamine Drugs 0.000 claims 2
- 230000001506 immunosuppresive effect Effects 0.000 claims 2
- 108010019691 inhibin beta A subunit Proteins 0.000 claims 2
- 108010024383 kallikrein 4 Proteins 0.000 claims 2
- 229940043355 kinase inhibitor Drugs 0.000 claims 2
- 102000004311 liver X receptors Human genes 0.000 claims 2
- 108090000865 liver X receptors Proteins 0.000 claims 2
- 235000015250 liver sausages Nutrition 0.000 claims 2
- 239000003589 local anesthetic agent Substances 0.000 claims 2
- 108010019677 lymphotactin Proteins 0.000 claims 2
- AEUKDPKXTPNBNY-XEYRWQBLSA-N mcp 2 Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)C1=CC=CC=C1 AEUKDPKXTPNBNY-XEYRWQBLSA-N 0.000 claims 2
- 239000003158 myorelaxant agent Substances 0.000 claims 2
- 230000003533 narcotic effect Effects 0.000 claims 2
- 239000000842 neuromuscular blocking agent Substances 0.000 claims 2
- 239000003757 phosphotransferase inhibitor Substances 0.000 claims 2
- 102000030769 platelet activating factor receptor Human genes 0.000 claims 2
- 108010017843 platelet-derived growth factor A Proteins 0.000 claims 2
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 claims 2
- XYSQXZCMOLNHOI-UHFFFAOYSA-N s-[2-[[4-(acetylsulfamoyl)phenyl]carbamoyl]phenyl] 5-pyridin-1-ium-1-ylpentanethioate;bromide Chemical compound [Br-].C1=CC(S(=O)(=O)NC(=O)C)=CC=C1NC(=O)C1=CC=CC=C1SC(=O)CCCC[N+]1=CC=CC=C1 XYSQXZCMOLNHOI-UHFFFAOYSA-N 0.000 claims 2
- 239000000932 sedative agent Substances 0.000 claims 2
- 230000001624 sedative effect Effects 0.000 claims 2
- 229960002930 sirolimus Drugs 0.000 claims 2
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 claims 2
- 150000003431 steroids Chemical class 0.000 claims 2
- 108091008744 testicular receptors 2 Proteins 0.000 claims 2
- 108091008743 testicular receptors 4 Proteins 0.000 claims 2
- HWFKCAFKXZFOQT-UHFFFAOYSA-N 1-(3,6-dibromocarbazol-9-yl)-3-piperazin-1-ylpropan-2-ol;dihydrochloride Chemical compound Cl.Cl.C12=CC=C(Br)C=C2C2=CC(Br)=CC=C2N1CC(O)CN1CCNCC1 HWFKCAFKXZFOQT-UHFFFAOYSA-N 0.000 claims 1
- WVAKRQOMAINQPU-UHFFFAOYSA-N 2-[4-[2-[5-(2,2-dimethylbutyl)-1h-imidazol-2-yl]ethyl]phenyl]pyridine Chemical compound N1C(CC(C)(C)CC)=CN=C1CCC1=CC=C(C=2N=CC=CC=2)C=C1 WVAKRQOMAINQPU-UHFFFAOYSA-N 0.000 claims 1
- 102100031912 A-kinase anchor protein 1, mitochondrial Human genes 0.000 claims 1
- 102100020979 ATP-binding cassette sub-family F member 1 Human genes 0.000 claims 1
- 102100034111 Activin receptor type-1 Human genes 0.000 claims 1
- 102100034134 Activin receptor type-1B Human genes 0.000 claims 1
- 102100032605 Adhesion G protein-coupled receptor B1 Human genes 0.000 claims 1
- 102100040121 Allograft inflammatory factor 1 Human genes 0.000 claims 1
- 102100024581 Alpha-taxilin Human genes 0.000 claims 1
- 102100022416 Aminoacyl tRNA synthase complex-interacting multifunctional protein 1 Human genes 0.000 claims 1
- 102100026468 Androgen-induced gene 1 protein Human genes 0.000 claims 1
- 102100036451 Apolipoprotein C-I Human genes 0.000 claims 1
- 101100480809 Arabidopsis thaliana TCP10 gene Proteins 0.000 claims 1
- 102100037152 BAG family molecular chaperone regulator 1 Human genes 0.000 claims 1
- 108700020463 BRCA1 Proteins 0.000 claims 1
- 101150072950 BRCA1 gene Proteins 0.000 claims 1
- 102100035388 Beta-enolase Human genes 0.000 claims 1
- 102100038495 Bile acid receptor Human genes 0.000 claims 1
- 101100396232 Bombyx mori EN03 gene Proteins 0.000 claims 1
- 108090000654 Bone morphogenetic protein 1 Proteins 0.000 claims 1
- 102000004152 Bone morphogenetic protein 1 Human genes 0.000 claims 1
- 102100025423 Bone morphogenetic protein receptor type-1A Human genes 0.000 claims 1
- 102100027052 Bone morphogenetic protein receptor type-1B Human genes 0.000 claims 1
- 102100025401 Breast cancer type 1 susceptibility protein Human genes 0.000 claims 1
- 101710149858 C-C chemokine receptor type 7 Proteins 0.000 claims 1
- 102100025074 C-C chemokine receptor-like 2 Human genes 0.000 claims 1
- 101710155857 C-C motif chemokine 2 Proteins 0.000 claims 1
- 101710155834 C-C motif chemokine 7 Proteins 0.000 claims 1
- 102100036166 C-X-C chemokine receptor type 1 Human genes 0.000 claims 1
- 108090000835 CX3C Chemokine Receptor 1 Proteins 0.000 claims 1
- 102100025805 Cadherin-1 Human genes 0.000 claims 1
- 102100024158 Cadherin-10 Human genes 0.000 claims 1
- 102100024156 Cadherin-12 Human genes 0.000 claims 1
- 102100022527 Cadherin-18 Human genes 0.000 claims 1
- 102100022529 Cadherin-19 Human genes 0.000 claims 1
- 101100355949 Caenorhabditis elegans spr-1 gene Proteins 0.000 claims 1
- 102100032145 Carbohydrate sulfotransferase 10 Human genes 0.000 claims 1
- 102100035904 Caspase-1 Human genes 0.000 claims 1
- 102100028914 Catenin beta-1 Human genes 0.000 claims 1
- 102100035888 Caveolin-1 Human genes 0.000 claims 1
- 108091007854 Cdh1/Fizzy-related Proteins 0.000 claims 1
- 102100025745 Cerberus Human genes 0.000 claims 1
- 102100031011 Chemerin-like receptor 1 Human genes 0.000 claims 1
- 102000019034 Chemokines Human genes 0.000 claims 1
- 108010012236 Chemokines Proteins 0.000 claims 1
- 102100033601 Collagen alpha-1(I) chain Human genes 0.000 claims 1
- 102100031519 Collagen alpha-1(VI) chain Human genes 0.000 claims 1
- 102100031162 Collagen alpha-1(XVIII) chain Human genes 0.000 claims 1
- 102100030291 Cornifin-B Human genes 0.000 claims 1
- 102100025176 Cyclin-A1 Human genes 0.000 claims 1
- 108010016788 Cyclin-Dependent Kinase Inhibitor p21 Proteins 0.000 claims 1
- 108010017222 Cyclin-Dependent Kinase Inhibitor p57 Proteins 0.000 claims 1
- 102000004480 Cyclin-Dependent Kinase Inhibitor p57 Human genes 0.000 claims 1
- 102100033270 Cyclin-dependent kinase inhibitor 1 Human genes 0.000 claims 1
- 101150081028 Cysltr1 gene Proteins 0.000 claims 1
- 102100038496 Cysteinyl leukotriene receptor 1 Human genes 0.000 claims 1
- 102100026234 Cytokine receptor common subunit gamma Human genes 0.000 claims 1
- 101100481404 Danio rerio tie1 gene Proteins 0.000 claims 1
- 102100028571 Disabled homolog 2-interacting protein Human genes 0.000 claims 1
- 102100024364 Disintegrin and metalloproteinase domain-containing protein 8 Human genes 0.000 claims 1
- 102100032249 Dystonin Human genes 0.000 claims 1
- 102100022183 E3 ubiquitin-protein ligase MIB1 Human genes 0.000 claims 1
- 108010043945 Ephrin-A1 Proteins 0.000 claims 1
- 102000020086 Ephrin-A1 Human genes 0.000 claims 1
- 101001039702 Escherichia coli (strain K12) Methyl-accepting chemotaxis protein I Proteins 0.000 claims 1
- 102100038595 Estrogen receptor Human genes 0.000 claims 1
- 101000837299 Euglena gracilis Trans-2-enoyl-CoA reductase Proteins 0.000 claims 1
- 102000003971 Fibroblast Growth Factor 1 Human genes 0.000 claims 1
- 102100028412 Fibroblast growth factor 10 Human genes 0.000 claims 1
- 102100037858 G1/S-specific cyclin-E1 Human genes 0.000 claims 1
- 102100028652 Gamma-enolase Human genes 0.000 claims 1
- 102100022626 Glutamate receptor ionotropic, NMDA 2D Human genes 0.000 claims 1
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 claims 1
- 101000774717 Homo sapiens A-kinase anchor protein 1, mitochondrial Proteins 0.000 claims 1
- 101000783783 Homo sapiens ATP-binding cassette sub-family F member 1 Proteins 0.000 claims 1
- 101000799140 Homo sapiens Activin receptor type-1 Proteins 0.000 claims 1
- 101000799189 Homo sapiens Activin receptor type-1B Proteins 0.000 claims 1
- 101000796780 Homo sapiens Adhesion G protein-coupled receptor B1 Proteins 0.000 claims 1
- 101000890626 Homo sapiens Allograft inflammatory factor 1 Proteins 0.000 claims 1
- 101000760787 Homo sapiens Alpha-taxilin Proteins 0.000 claims 1
- 101000755762 Homo sapiens Aminoacyl tRNA synthase complex-interacting multifunctional protein 1 Proteins 0.000 claims 1
- 101000718108 Homo sapiens Androgen-induced gene 1 protein Proteins 0.000 claims 1
- 101000928628 Homo sapiens Apolipoprotein C-I Proteins 0.000 claims 1
- 101000740062 Homo sapiens BAG family molecular chaperone regulator 1 Proteins 0.000 claims 1
- 101000877537 Homo sapiens Beta-enolase Proteins 0.000 claims 1
- 101000603876 Homo sapiens Bile acid receptor Proteins 0.000 claims 1
- 101000934638 Homo sapiens Bone morphogenetic protein receptor type-1A Proteins 0.000 claims 1
- 101000984546 Homo sapiens Bone morphogenetic protein receptor type-1B Proteins 0.000 claims 1
- 101000946926 Homo sapiens C-C chemokine receptor type 5 Proteins 0.000 claims 1
- 101000934394 Homo sapiens C-C chemokine receptor-like 2 Proteins 0.000 claims 1
- 101000947174 Homo sapiens C-X-C chemokine receptor type 1 Proteins 0.000 claims 1
- 101000762229 Homo sapiens Cadherin-10 Proteins 0.000 claims 1
- 101000762238 Homo sapiens Cadherin-12 Proteins 0.000 claims 1
- 101000899405 Homo sapiens Cadherin-18 Proteins 0.000 claims 1
- 101000710899 Homo sapiens Cannabinoid receptor 1 Proteins 0.000 claims 1
- 101000775595 Homo sapiens Carbohydrate sulfotransferase 10 Proteins 0.000 claims 1
- 101000715398 Homo sapiens Caspase-1 Proteins 0.000 claims 1
- 101000916173 Homo sapiens Catenin beta-1 Proteins 0.000 claims 1
- 101000715467 Homo sapiens Caveolin-1 Proteins 0.000 claims 1
- 101000914195 Homo sapiens Cerberus Proteins 0.000 claims 1
- 101000919756 Homo sapiens Chemerin-like receptor 1 Proteins 0.000 claims 1
- 101000941581 Homo sapiens Collagen alpha-1(VI) chain Proteins 0.000 claims 1
- 101000940068 Homo sapiens Collagen alpha-1(XVIII) chain Proteins 0.000 claims 1
- 101000702152 Homo sapiens Cornifin-B Proteins 0.000 claims 1
- 101000934314 Homo sapiens Cyclin-A1 Proteins 0.000 claims 1
- 101001055227 Homo sapiens Cytokine receptor common subunit gamma Proteins 0.000 claims 1
- 101000915396 Homo sapiens Disabled homolog 2-interacting protein Proteins 0.000 claims 1
- 101000832767 Homo sapiens Disintegrin and metalloproteinase domain-containing protein 8 Proteins 0.000 claims 1
- 101001016186 Homo sapiens Dystonin Proteins 0.000 claims 1
- 101000973503 Homo sapiens E3 ubiquitin-protein ligase MIB1 Proteins 0.000 claims 1
- 101000978392 Homo sapiens Eotaxin Proteins 0.000 claims 1
- 101000882584 Homo sapiens Estrogen receptor Proteins 0.000 claims 1
- 101000917237 Homo sapiens Fibroblast growth factor 10 Proteins 0.000 claims 1
- 101000738568 Homo sapiens G1/S-specific cyclin-E1 Proteins 0.000 claims 1
- 101001058231 Homo sapiens Gamma-enolase Proteins 0.000 claims 1
- 101000972840 Homo sapiens Glutamate receptor ionotropic, NMDA 2D Proteins 0.000 claims 1
- 101000893563 Homo sapiens Growth/differentiation factor 10 Proteins 0.000 claims 1
- 101001021527 Homo sapiens Huntingtin-interacting protein 1 Proteins 0.000 claims 1
- 101001076407 Homo sapiens Interleukin-1 receptor antagonist protein Proteins 0.000 claims 1
- 101001083151 Homo sapiens Interleukin-10 receptor subunit alpha Proteins 0.000 claims 1
- 101001003149 Homo sapiens Interleukin-10 receptor subunit beta Proteins 0.000 claims 1
- 101001055145 Homo sapiens Interleukin-2 receptor subunit beta Proteins 0.000 claims 1
- 101001010591 Homo sapiens Interleukin-20 Proteins 0.000 claims 1
- 101001044883 Homo sapiens Interleukin-22 receptor subunit alpha-1 Proteins 0.000 claims 1
- 101001008919 Homo sapiens Kallikrein-10 Proteins 0.000 claims 1
- 101001133056 Homo sapiens Mucin-1 Proteins 0.000 claims 1
- 101000978926 Homo sapiens Nuclear receptor subfamily 1 group D member 1 Proteins 0.000 claims 1
- 101000979629 Homo sapiens Nucleoside diphosphate kinase A Proteins 0.000 claims 1
- 101001021281 Homo sapiens Protein HEXIM1 Proteins 0.000 claims 1
- 101000986265 Homo sapiens Protein MTSS 1 Proteins 0.000 claims 1
- 101001116937 Homo sapiens Protocadherin alpha-4 Proteins 0.000 claims 1
- 101000655540 Homo sapiens Protransforming growth factor alpha Proteins 0.000 claims 1
- 101000650697 Homo sapiens Roundabout homolog 2 Proteins 0.000 claims 1
- 101000739195 Homo sapiens Secretoglobin family 1D member 2 Proteins 0.000 claims 1
- 101000910249 Homo sapiens Soluble calcium-activated nucleotidase 1 Proteins 0.000 claims 1
- 101000693265 Homo sapiens Sphingosine 1-phosphate receptor 1 Proteins 0.000 claims 1
- 101000716124 Homo sapiens T-cell surface glycoprotein CD1c Proteins 0.000 claims 1
- 101000659879 Homo sapiens Thrombospondin-1 Proteins 0.000 claims 1
- 101000796134 Homo sapiens Thymidine phosphorylase Proteins 0.000 claims 1
- 101000904152 Homo sapiens Transcription factor E2F1 Proteins 0.000 claims 1
- 101000635938 Homo sapiens Transforming growth factor beta-1 proprotein Proteins 0.000 claims 1
- 101000635958 Homo sapiens Transforming growth factor beta-2 proprotein Proteins 0.000 claims 1
- 101000894525 Homo sapiens Transforming growth factor-beta-induced protein ig-h3 Proteins 0.000 claims 1
- 101000830565 Homo sapiens Tumor necrosis factor ligand superfamily member 10 Proteins 0.000 claims 1
- 101000733249 Homo sapiens Tumor suppressor ARF Proteins 0.000 claims 1
- 101000666856 Homo sapiens Vasoactive intestinal polypeptide receptor 1 Proteins 0.000 claims 1
- 101000818517 Homo sapiens Zinc-alpha-2-glycoprotein Proteins 0.000 claims 1
- 102100035957 Huntingtin-interacting protein 1 Human genes 0.000 claims 1
- 102100037850 Interferon gamma Human genes 0.000 claims 1
- 102000003777 Interleukin-1 beta Human genes 0.000 claims 1
- 108090000193 Interleukin-1 beta Proteins 0.000 claims 1
- 102100026018 Interleukin-1 receptor antagonist protein Human genes 0.000 claims 1
- 102100030236 Interleukin-10 receptor subunit alpha Human genes 0.000 claims 1
- 102100020788 Interleukin-10 receptor subunit beta Human genes 0.000 claims 1
- 108010082786 Interleukin-1alpha Proteins 0.000 claims 1
- 102100030692 Interleukin-20 Human genes 0.000 claims 1
- 102100022723 Interleukin-22 receptor subunit alpha-1 Human genes 0.000 claims 1
- 108010066979 Interleukin-27 Proteins 0.000 claims 1
- 102100027613 Kallikrein-10 Human genes 0.000 claims 1
- 102100034256 Mucin-1 Human genes 0.000 claims 1
- 101100481406 Mus musculus Tie1 gene Proteins 0.000 claims 1
- 102100023170 Nuclear receptor subfamily 1 group D member 1 Human genes 0.000 claims 1
- 102100023171 Nuclear receptor subfamily 1 group D member 2 Human genes 0.000 claims 1
- 102100023252 Nucleoside diphosphate kinase A Human genes 0.000 claims 1
- 102000035195 Peptidases Human genes 0.000 claims 1
- 108091005804 Peptidases Proteins 0.000 claims 1
- 102100028951 Protein MTSS 1 Human genes 0.000 claims 1
- 102100024261 Protocadherin alpha-4 Human genes 0.000 claims 1
- 102100032350 Protransforming growth factor alpha Human genes 0.000 claims 1
- 108091008770 Rev-ErbAß Proteins 0.000 claims 1
- 102100027739 Roundabout homolog 2 Human genes 0.000 claims 1
- 108010011005 STAT6 Transcription Factor Proteins 0.000 claims 1
- 102100037279 Secretoglobin family 1D member 2 Human genes 0.000 claims 1
- 102100023980 Signal transducer and activator of transcription 6 Human genes 0.000 claims 1
- 102100024397 Soluble calcium-activated nucleotidase 1 Human genes 0.000 claims 1
- 102100025750 Sphingosine 1-phosphate receptor 1 Human genes 0.000 claims 1
- 102100030511 Stanniocalcin-1 Human genes 0.000 claims 1
- 101710142157 Stanniocalcin-1 Proteins 0.000 claims 1
- 102100036014 T-cell surface glycoprotein CD1c Human genes 0.000 claims 1
- 102100031372 Thymidine phosphorylase Human genes 0.000 claims 1
- 102100024026 Transcription factor E2F1 Human genes 0.000 claims 1
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 claims 1
- 102000011117 Transforming Growth Factor beta2 Human genes 0.000 claims 1
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 claims 1
- 102100030742 Transforming growth factor beta-1 proprotein Human genes 0.000 claims 1
- 101800000304 Transforming growth factor beta-2 Proteins 0.000 claims 1
- 102100030737 Transforming growth factor beta-2 proprotein Human genes 0.000 claims 1
- 102100021398 Transforming growth factor-beta-induced protein ig-h3 Human genes 0.000 claims 1
- 108010047933 Tumor Necrosis Factor alpha-Induced Protein 3 Proteins 0.000 claims 1
- 102100024596 Tumor necrosis factor alpha-induced protein 3 Human genes 0.000 claims 1
- 102100021144 Zinc-alpha-2-glycoprotein Human genes 0.000 claims 1
- 108010029483 alpha 1 Chain Collagen Type I Proteins 0.000 claims 1
- 230000003110 anti-inflammatory effect Effects 0.000 claims 1
- 230000003356 anti-rheumatic effect Effects 0.000 claims 1
- 239000003435 antirheumatic agent Substances 0.000 claims 1
- 230000008827 biological function Effects 0.000 claims 1
- 238000001983 electron spin resonance imaging Methods 0.000 claims 1
- 239000001963 growth medium Substances 0.000 claims 1
- 230000003472 neutralizing effect Effects 0.000 claims 1
- 108010042974 transforming growth factor beta4 Proteins 0.000 claims 1
- 102000001301 EGF receptor Human genes 0.000 description 50
- 108060006698 EGF receptor Proteins 0.000 description 50
- 210000003719 b-lymphocyte Anatomy 0.000 description 48
- 206010028980 Neoplasm Diseases 0.000 description 47
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 38
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 38
- 108090000623 proteins and genes Proteins 0.000 description 36
- 125000003275 alpha amino acid group Chemical group 0.000 description 35
- 201000011510 cancer Diseases 0.000 description 31
- 230000006907 apoptotic process Effects 0.000 description 30
- 125000000539 amino acid group Chemical group 0.000 description 27
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 24
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 21
- 238000006471 dimerization reaction Methods 0.000 description 21
- 230000006870 function Effects 0.000 description 20
- 102400000058 Neuregulin-1 Human genes 0.000 description 19
- 241000238413 Octopus Species 0.000 description 19
- 101100369992 Homo sapiens TNFSF10 gene Proteins 0.000 description 18
- 241000124008 Mammalia Species 0.000 description 17
- 239000012636 effector Substances 0.000 description 17
- 235000018102 proteins Nutrition 0.000 description 17
- 102000004169 proteins and genes Human genes 0.000 description 17
- 208000035475 disorder Diseases 0.000 description 16
- 230000000694 effects Effects 0.000 description 16
- 230000014509 gene expression Effects 0.000 description 16
- 229960004641 rituximab Drugs 0.000 description 16
- 101710100969 Receptor tyrosine-protein kinase erbB-3 Proteins 0.000 description 15
- 101710100963 Receptor tyrosine-protein kinase erbB-4 Proteins 0.000 description 15
- 238000003556 assay Methods 0.000 description 15
- 238000000338 in vitro Methods 0.000 description 15
- 229940002612 prodrug Drugs 0.000 description 15
- 239000000651 prodrug Substances 0.000 description 15
- 238000011160 research Methods 0.000 description 15
- 102100029986 Receptor tyrosine-protein kinase erbB-3 Human genes 0.000 description 14
- 102100029981 Receptor tyrosine-protein kinase erbB-4 Human genes 0.000 description 14
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 14
- 230000012010 growth Effects 0.000 description 14
- 108020004414 DNA Proteins 0.000 description 13
- 241001529936 Murinae Species 0.000 description 13
- 108090000556 Neuregulin-1 Proteins 0.000 description 13
- 210000004369 blood Anatomy 0.000 description 13
- 239000008280 blood Substances 0.000 description 13
- 230000004540 complement-dependent cytotoxicity Effects 0.000 description 13
- 210000004881 tumor cell Anatomy 0.000 description 13
- -1 wsl-1 Proteins 0.000 description 13
- 108090000542 Lymphotoxin-alpha Proteins 0.000 description 12
- 238000001994 activation Methods 0.000 description 12
- 102000004083 Lymphotoxin-alpha Human genes 0.000 description 11
- 108700020796 Oncogene Proteins 0.000 description 11
- 102100040112 Tumor necrosis factor receptor superfamily member 10B Human genes 0.000 description 11
- 102100022203 Tumor necrosis factor receptor superfamily member 25 Human genes 0.000 description 11
- 230000004913 activation Effects 0.000 description 11
- 230000002401 inhibitory effect Effects 0.000 description 11
- 206010006187 Breast cancer Diseases 0.000 description 10
- 208000026310 Breast neoplasm Diseases 0.000 description 10
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 10
- 102000010170 Death domains Human genes 0.000 description 10
- 108050001718 Death domains Proteins 0.000 description 10
- 101000610605 Homo sapiens Tumor necrosis factor receptor superfamily member 10A Proteins 0.000 description 10
- 101000610602 Homo sapiens Tumor necrosis factor receptor superfamily member 10C Proteins 0.000 description 10
- 101000610609 Homo sapiens Tumor necrosis factor receptor superfamily member 10D Proteins 0.000 description 10
- 210000001744 T-lymphocyte Anatomy 0.000 description 10
- 102100040113 Tumor necrosis factor receptor superfamily member 10A Human genes 0.000 description 10
- 102100040115 Tumor necrosis factor receptor superfamily member 10C Human genes 0.000 description 10
- 102100040110 Tumor necrosis factor receptor superfamily member 10D Human genes 0.000 description 10
- 235000001014 amino acid Nutrition 0.000 description 10
- 238000013459 approach Methods 0.000 description 10
- 230000000295 complement effect Effects 0.000 description 10
- 230000001086 cytosolic effect Effects 0.000 description 10
- 230000004044 response Effects 0.000 description 10
- NMWKYTGJWUAZPZ-WWHBDHEGSA-N (4S)-4-[[(4R,7S,10S,16S,19S,25S,28S,31R)-31-[[(2S)-2-[[(1R,6R,9S,12S,18S,21S,24S,27S,30S,33S,36S,39S,42R,47R,53S,56S,59S,62S,65S,68S,71S,76S,79S,85S)-47-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-methylbutanoyl]amino]-3-methylbutanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]-3-phenylpropanoyl]amino]-4-oxobutanoyl]amino]-3-carboxypropanoyl]amino]-18-(4-aminobutyl)-27,68-bis(3-amino-3-oxopropyl)-36,71,76-tribenzyl-39-(3-carbamimidamidopropyl)-24-(2-carboxyethyl)-21,56-bis(carboxymethyl)-65,85-bis[(1R)-1-hydroxyethyl]-59-(hydroxymethyl)-62,79-bis(1H-imidazol-4-ylmethyl)-9-methyl-33-(2-methylpropyl)-8,11,17,20,23,26,29,32,35,38,41,48,54,57,60,63,66,69,72,74,77,80,83,86-tetracosaoxo-30-propan-2-yl-3,4,44,45-tetrathia-7,10,16,19,22,25,28,31,34,37,40,49,55,58,61,64,67,70,73,75,78,81,84,87-tetracosazatetracyclo[40.31.14.012,16.049,53]heptaoctacontane-6-carbonyl]amino]-3-methylbutanoyl]amino]-7-(3-carbamimidamidopropyl)-25-(hydroxymethyl)-19-[(4-hydroxyphenyl)methyl]-28-(1H-imidazol-4-ylmethyl)-10-methyl-6,9,12,15,18,21,24,27,30-nonaoxo-16-propan-2-yl-1,2-dithia-5,8,11,14,17,20,23,26,29-nonazacyclodotriacontane-4-carbonyl]amino]-5-[[(2S)-1-[[(2S)-1-[[(2S)-3-carboxy-1-[[(2S)-1-[[(2S)-1-[[(1S)-1-carboxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-5-oxopentanoic acid Chemical compound CC(C)C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CSSC[C@H](NC(=O)[C@@H](NC(=O)[C@@H]2CSSC[C@@H]3NC(=O)[C@H](Cc4ccccc4)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](Cc4c[nH]cn4)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]4CCCN4C(=O)[C@H](CSSC[C@H](NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](Cc4c[nH]cn4)NC(=O)[C@H](Cc4ccccc4)NC3=O)[C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](Cc3ccccc3)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N3CCC[C@H]3C(=O)N[C@@H](C)C(=O)N2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](Cc2ccccc2)NC(=O)[C@H](Cc2c[nH]cn2)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)C(C)C)C(=O)N[C@@H](Cc2c[nH]cn2)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](Cc2ccc(O)cc2)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1)C(=O)N[C@@H](C)C(O)=O NMWKYTGJWUAZPZ-WWHBDHEGSA-N 0.000 description 9
- 101000610604 Homo sapiens Tumor necrosis factor receptor superfamily member 10B Proteins 0.000 description 9
- 101000679903 Homo sapiens Tumor necrosis factor receptor superfamily member 25 Proteins 0.000 description 9
- 101000597785 Homo sapiens Tumor necrosis factor receptor superfamily member 6B Proteins 0.000 description 9
- 102100035284 Tumor necrosis factor receptor superfamily member 6B Human genes 0.000 description 9
- 235000018417 cysteine Nutrition 0.000 description 9
- 208000032839 leukemia Diseases 0.000 description 9
- 210000000265 leukocyte Anatomy 0.000 description 9
- 210000004698 lymphocyte Anatomy 0.000 description 9
- 230000001404 mediated effect Effects 0.000 description 9
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 9
- 210000004180 plasmocyte Anatomy 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 8
- 101800002648 Neuregulin-1 Proteins 0.000 description 8
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 8
- 230000004069 differentiation Effects 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 238000013467 fragmentation Methods 0.000 description 8
- 238000006062 fragmentation reaction Methods 0.000 description 8
- 241000894007 species Species 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 8
- 102000009109 Fc receptors Human genes 0.000 description 7
- 108010087819 Fc receptors Proteins 0.000 description 7
- 101000871708 Homo sapiens Proheparin-binding EGF-like growth factor Proteins 0.000 description 7
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 7
- 108010035042 Osteoprotegerin Proteins 0.000 description 7
- 101710160107 Outer membrane protein A Proteins 0.000 description 7
- 102100033762 Proheparin-binding EGF-like growth factor Human genes 0.000 description 7
- 102100032236 Tumor necrosis factor receptor superfamily member 11B Human genes 0.000 description 7
- 229940024606 amino acid Drugs 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- 230000030833 cell death Effects 0.000 description 7
- 231100000433 cytotoxic Toxicity 0.000 description 7
- 230000001472 cytotoxic effect Effects 0.000 description 7
- 238000001727 in vivo Methods 0.000 description 7
- 230000001939 inductive effect Effects 0.000 description 7
- XXUPLYBCNPLTIW-UHFFFAOYSA-N octadec-7-ynoic acid Chemical compound CCCCCCCCCCC#CCCCCCC(O)=O XXUPLYBCNPLTIW-UHFFFAOYSA-N 0.000 description 7
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 7
- 230000011664 signaling Effects 0.000 description 7
- 108010008014 B-Cell Maturation Antigen Proteins 0.000 description 6
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 6
- 101800003838 Epidermal growth factor Proteins 0.000 description 6
- 102400001368 Epidermal growth factor Human genes 0.000 description 6
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 6
- 241000283984 Rodentia Species 0.000 description 6
- 102100029675 Tumor necrosis factor receptor superfamily member 13B Human genes 0.000 description 6
- 239000002246 antineoplastic agent Substances 0.000 description 6
- 238000003782 apoptosis assay Methods 0.000 description 6
- 210000003169 central nervous system Anatomy 0.000 description 6
- 229940116977 epidermal growth factor Drugs 0.000 description 6
- 229940022353 herceptin Drugs 0.000 description 6
- 229940072221 immunoglobulins Drugs 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 230000003834 intracellular effect Effects 0.000 description 6
- 210000004072 lung Anatomy 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 210000004379 membrane Anatomy 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- 210000000056 organ Anatomy 0.000 description 6
- 102000006942 B-Cell Maturation Antigen Human genes 0.000 description 5
- 102400001242 Betacellulin Human genes 0.000 description 5
- 101800001382 Betacellulin Proteins 0.000 description 5
- 201000009030 Carcinoma Diseases 0.000 description 5
- 102400001329 Epiregulin Human genes 0.000 description 5
- 101800000155 Epiregulin Proteins 0.000 description 5
- 108010073807 IgG Receptors Proteins 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 108010057466 NF-kappa B Proteins 0.000 description 5
- 108091007491 NSP3 Papain-like protease domains Proteins 0.000 description 5
- 101800000675 Neuregulin-2 Proteins 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- 102100022668 Pro-neuregulin-2, membrane-bound isoform Human genes 0.000 description 5
- 241000700159 Rattus Species 0.000 description 5
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 5
- 210000001185 bone marrow Anatomy 0.000 description 5
- 210000004899 c-terminal region Anatomy 0.000 description 5
- 230000011712 cell development Effects 0.000 description 5
- 102000051957 human ERBB2 Human genes 0.000 description 5
- 230000001900 immune effect Effects 0.000 description 5
- 210000003734 kidney Anatomy 0.000 description 5
- 230000002147 killing effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 210000001948 pro-b lymphocyte Anatomy 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 238000006467 substitution reaction Methods 0.000 description 5
- 230000008685 targeting Effects 0.000 description 5
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 4
- 101710117290 Aldo-keto reductase family 1 member C4 Proteins 0.000 description 4
- 102100038778 Amphiregulin Human genes 0.000 description 4
- 108010033760 Amphiregulin Proteins 0.000 description 4
- 208000023275 Autoimmune disease Diseases 0.000 description 4
- 208000003950 B-cell lymphoma Diseases 0.000 description 4
- 102000004634 CD30 Ligand Human genes 0.000 description 4
- 206010009944 Colon cancer Diseases 0.000 description 4
- 201000004624 Dermatitis Diseases 0.000 description 4
- 108010008177 Fd immunoglobulins Proteins 0.000 description 4
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 4
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- 101000795167 Homo sapiens Tumor necrosis factor receptor superfamily member 13B Proteins 0.000 description 4
- 101000801255 Homo sapiens Tumor necrosis factor receptor superfamily member 17 Proteins 0.000 description 4
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 4
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 4
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 4
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 101800000673 Neuregulin-3 Proteins 0.000 description 4
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 4
- 206010061535 Ovarian neoplasm Diseases 0.000 description 4
- 102100022659 Pro-neuregulin-3, membrane-bound isoform Human genes 0.000 description 4
- 102100024952 Protein CBFA2T1 Human genes 0.000 description 4
- 102000004278 Receptor Protein-Tyrosine Kinases Human genes 0.000 description 4
- 108090000873 Receptor Protein-Tyrosine Kinases Proteins 0.000 description 4
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 description 4
- 102100028785 Tumor necrosis factor receptor superfamily member 14 Human genes 0.000 description 4
- 230000001270 agonistic effect Effects 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 210000000481 breast Anatomy 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 239000000306 component Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000009089 cytolysis Effects 0.000 description 4
- 239000000539 dimer Substances 0.000 description 4
- 229960002949 fluorouracil Drugs 0.000 description 4
- 230000009036 growth inhibition Effects 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 229940127121 immunoconjugate Drugs 0.000 description 4
- 230000001976 improved effect Effects 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 230000036210 malignancy Effects 0.000 description 4
- 230000003211 malignant effect Effects 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- 210000005259 peripheral blood Anatomy 0.000 description 4
- 239000011886 peripheral blood Substances 0.000 description 4
- 238000002823 phage display Methods 0.000 description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 4
- 230000002285 radioactive effect Effects 0.000 description 4
- 210000000130 stem cell Anatomy 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 210000001685 thyroid gland Anatomy 0.000 description 4
- WYWHKKSPHMUBEB-UHFFFAOYSA-N tioguanine Chemical compound N1C(N)=NC(=S)C2=C1N=CN2 WYWHKKSPHMUBEB-UHFFFAOYSA-N 0.000 description 4
- 238000012546 transfer Methods 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 4
- BGFTWECWAICPDG-UHFFFAOYSA-N 2-[bis(4-chlorophenyl)methyl]-4-n-[3-[bis(4-chlorophenyl)methyl]-4-(dimethylamino)phenyl]-1-n,1-n-dimethylbenzene-1,4-diamine Chemical compound C1=C(C(C=2C=CC(Cl)=CC=2)C=2C=CC(Cl)=CC=2)C(N(C)C)=CC=C1NC(C=1)=CC=C(N(C)C)C=1C(C=1C=CC(Cl)=CC=1)C1=CC=C(Cl)C=C1 BGFTWECWAICPDG-UHFFFAOYSA-N 0.000 description 3
- STQGQHZAVUOBTE-UHFFFAOYSA-N 7-Cyan-hept-2t-en-4,6-diinsaeure Natural products C1=2C(O)=C3C(=O)C=4C(OC)=CC=CC=4C(=O)C3=C(O)C=2CC(O)(C(C)=O)CC1OC1CC(N)C(O)C(C)O1 STQGQHZAVUOBTE-UHFFFAOYSA-N 0.000 description 3
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 3
- IPWKGIFRRBGCJO-IMJSIDKUSA-N Ala-Ser Chemical compound C[C@H]([NH3+])C(=O)N[C@@H](CO)C([O-])=O IPWKGIFRRBGCJO-IMJSIDKUSA-N 0.000 description 3
- 102000000412 Annexin Human genes 0.000 description 3
- 108050008874 Annexin Proteins 0.000 description 3
- 108091026890 Coding region Proteins 0.000 description 3
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 3
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 description 3
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 3
- BCCRXDTUTZHDEU-VKHMYHEASA-N Gly-Ser Chemical compound NCC(=O)N[C@@H](CO)C(O)=O BCCRXDTUTZHDEU-VKHMYHEASA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 101000801234 Homo sapiens Tumor necrosis factor receptor superfamily member 18 Proteins 0.000 description 3
- 108010000521 Human Growth Hormone Proteins 0.000 description 3
- 102000002265 Human Growth Hormone Human genes 0.000 description 3
- 239000000854 Human Growth Hormone Substances 0.000 description 3
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 3
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 3
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 3
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 3
- 102000004877 Insulin Human genes 0.000 description 3
- 108090001061 Insulin Proteins 0.000 description 3
- 208000008839 Kidney Neoplasms Diseases 0.000 description 3
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 3
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 3
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 3
- 102000009151 Luteinizing Hormone Human genes 0.000 description 3
- 108010073521 Luteinizing Hormone Proteins 0.000 description 3
- 108010061593 Member 14 Tumor Necrosis Factor Receptors Proteins 0.000 description 3
- 102000003729 Neprilysin Human genes 0.000 description 3
- 108090000028 Neprilysin Proteins 0.000 description 3
- 108010025020 Nerve Growth Factor Proteins 0.000 description 3
- 206010033128 Ovarian cancer Diseases 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- 206010060862 Prostate cancer Diseases 0.000 description 3
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 3
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 3
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 3
- 206010038389 Renal cancer Diseases 0.000 description 3
- 230000018199 S phase Effects 0.000 description 3
- 208000031981 Thrombocytopenic Idiopathic Purpura Diseases 0.000 description 3
- 102000011923 Thyrotropin Human genes 0.000 description 3
- 108010061174 Thyrotropin Proteins 0.000 description 3
- 102100033728 Tumor necrosis factor receptor superfamily member 18 Human genes 0.000 description 3
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 description 3
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 230000003213 activating effect Effects 0.000 description 3
- 230000004075 alteration Effects 0.000 description 3
- 208000010668 atopic eczema Diseases 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- 210000001072 colon Anatomy 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 230000024203 complement activation Effects 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 238000010276 construction Methods 0.000 description 3
- STQGQHZAVUOBTE-VGBVRHCVSA-N daunorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(C)=O)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 STQGQHZAVUOBTE-VGBVRHCVSA-N 0.000 description 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 229960004679 doxorubicin Drugs 0.000 description 3
- 230000002357 endometrial effect Effects 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 229940028334 follicle stimulating hormone Drugs 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 230000002518 glial effect Effects 0.000 description 3
- 208000014829 head and neck neoplasm Diseases 0.000 description 3
- 210000002216 heart Anatomy 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 229940096397 interleukin-8 Drugs 0.000 description 3
- XKTZWUACRZHVAN-VADRZIEHSA-N interleukin-8 Chemical compound C([C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@@H](NC(C)=O)CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCSC)C(=O)N1[C@H](CCC1)C(=O)N1[C@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC(O)=CC=1)C(=O)N[C@H](CO)C(=O)N1[C@H](CCC1)C(N)=O)C1=CC=CC=C1 XKTZWUACRZHVAN-VADRZIEHSA-N 0.000 description 3
- 201000010982 kidney cancer Diseases 0.000 description 3
- 208000014018 liver neoplasm Diseases 0.000 description 3
- 229940040129 luteinizing hormone Drugs 0.000 description 3
- MQXVYODZCMMZEM-ZYUZMQFOSA-N mannomustine Chemical compound ClCCNC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CNCCCl MQXVYODZCMMZEM-ZYUZMQFOSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 210000003519 mature b lymphocyte Anatomy 0.000 description 3
- 102000006240 membrane receptors Human genes 0.000 description 3
- GLVAUDGFNGKCSF-UHFFFAOYSA-N mercaptopurine Chemical compound S=C1NC=NC2=C1NC=N2 GLVAUDGFNGKCSF-UHFFFAOYSA-N 0.000 description 3
- KKZJGLLVHKMTCM-UHFFFAOYSA-N mitoxantrone Chemical compound O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO KKZJGLLVHKMTCM-UHFFFAOYSA-N 0.000 description 3
- 210000001616 monocyte Anatomy 0.000 description 3
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 3
- 230000037361 pathway Effects 0.000 description 3
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 3
- 230000026731 phosphorylation Effects 0.000 description 3
- 238000006366 phosphorylation reaction Methods 0.000 description 3
- 230000005522 programmed cell death Effects 0.000 description 3
- 230000005180 public health Effects 0.000 description 3
- 108700015048 receptor decoy activity proteins Proteins 0.000 description 3
- HXCHCVDVKSCDHU-PJKCJEBCSA-N s-[(2r,3s,4s,6s)-6-[[(2r,3s,4s,5r,6r)-5-[(2s,4s,5s)-5-(ethylamino)-4-methoxyoxan-2-yl]oxy-4-hydroxy-6-[[(2s,5z,9r,13e)-9-hydroxy-12-(methoxycarbonylamino)-13-[2-(methyltrisulfanyl)ethylidene]-11-oxo-2-bicyclo[7.3.1]trideca-1(12),5-dien-3,7-diynyl]oxy]-2-m Chemical compound C1[C@H](OC)[C@@H](NCC)CO[C@H]1O[C@H]1[C@H](O[C@@H]2C\3=C(NC(=O)OC)C(=O)C[C@@](C/3=C/CSSSC)(O)C#C\C=C/C#C2)O[C@H](C)[C@@H](NO[C@@H]2O[C@H](C)[C@@H](SC(=O)C=3C(=C(OC)C(O[C@H]4[C@@H]([C@H](OC)[C@@H](O)[C@H](C)O4)O)=C(I)C=3C)OC)[C@@H](O)C2)[C@@H]1O HXCHCVDVKSCDHU-PJKCJEBCSA-N 0.000 description 3
- 210000003079 salivary gland Anatomy 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 208000011580 syndromic disease Diseases 0.000 description 3
- 238000013518 transcription Methods 0.000 description 3
- 230000035897 transcription Effects 0.000 description 3
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 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
- FDKXTQMXEQVLRF-ZHACJKMWSA-N (E)-dacarbazine Chemical compound CN(C)\N=N\c1[nH]cnc1C(N)=O FDKXTQMXEQVLRF-ZHACJKMWSA-N 0.000 description 2
- LSBDFXRDZJMBSC-UHFFFAOYSA-N 2-phenylacetamide Chemical class NC(=O)CC1=CC=CC=C1 LSBDFXRDZJMBSC-UHFFFAOYSA-N 0.000 description 2
- AOJJSUZBOXZQNB-VTZDEGQISA-N 4'-epidoxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-VTZDEGQISA-N 0.000 description 2
- 108090000672 Annexin A5 Proteins 0.000 description 2
- 102000004121 Annexin A5 Human genes 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 208000028564 B-cell non-Hodgkin lymphoma Diseases 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 206010005003 Bladder cancer Diseases 0.000 description 2
- 108010006654 Bleomycin Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 206010055113 Breast cancer metastatic Diseases 0.000 description 2
- 102000007499 CD27 Ligand Human genes 0.000 description 2
- GAGWJHPBXLXJQN-UORFTKCHSA-N Capecitabine Chemical compound C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1[C@H]1[C@H](O)[C@H](O)[C@@H](C)O1 GAGWJHPBXLXJQN-UORFTKCHSA-N 0.000 description 2
- 108010001857 Cell Surface Receptors Proteins 0.000 description 2
- 206010057248 Cell death Diseases 0.000 description 2
- 102100022641 Coagulation factor IX Human genes 0.000 description 2
- 102000007644 Colony-Stimulating Factors Human genes 0.000 description 2
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 2
- 239000004971 Cross linker Substances 0.000 description 2
- 108010092160 Dactinomycin Proteins 0.000 description 2
- 108010049207 Death Domain Receptors Proteins 0.000 description 2
- 102000009058 Death Domain Receptors Human genes 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- HTIJFSOGRVMCQR-UHFFFAOYSA-N Epirubicin Natural products COc1cccc2C(=O)c3c(O)c4CC(O)(CC(OC5CC(N)C(=O)C(C)O5)c4c(O)c3C(=O)c12)C(=O)CO HTIJFSOGRVMCQR-UHFFFAOYSA-N 0.000 description 2
- 108010076282 Factor IX Proteins 0.000 description 2
- 108010054218 Factor VIII Proteins 0.000 description 2
- 102000001690 Factor VIII Human genes 0.000 description 2
- 108010021468 Fc gamma receptor IIA Proteins 0.000 description 2
- 108010021472 Fc gamma receptor IIB Proteins 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 241000288105 Grus Species 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 108010027412 Histocompatibility Antigens Class II Proteins 0.000 description 2
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 2
- 101000840258 Homo sapiens Immunoglobulin J chain Proteins 0.000 description 2
- 101100100117 Homo sapiens TNFRSF10B gene Proteins 0.000 description 2
- 208000028622 Immune thrombocytopenia Diseases 0.000 description 2
- 102100029571 Immunoglobulin J chain Human genes 0.000 description 2
- 102000014150 Interferons Human genes 0.000 description 2
- 108010050904 Interferons Proteins 0.000 description 2
- 102000015696 Interleukins Human genes 0.000 description 2
- 108010063738 Interleukins Proteins 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 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
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 2
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 108010091221 Lymphotoxin beta Receptor Proteins 0.000 description 2
- 102000018170 Lymphotoxin beta Receptor Human genes 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 230000027311 M phase Effects 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 101100369076 Mus musculus Tdgf1 gene Proteins 0.000 description 2
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 2
- 102000015336 Nerve Growth Factor Human genes 0.000 description 2
- 102000004230 Neurotrophin 3 Human genes 0.000 description 2
- 108090000742 Neurotrophin 3 Proteins 0.000 description 2
- 102000043276 Oncogene Human genes 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- 102000003982 Parathyroid hormone Human genes 0.000 description 2
- 108090000445 Parathyroid hormone Proteins 0.000 description 2
- 108010076181 Proinsulin Proteins 0.000 description 2
- 101710100968 Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 2
- 108010039445 Stem Cell Factor Proteins 0.000 description 2
- 201000009594 Systemic Scleroderma Diseases 0.000 description 2
- 206010042953 Systemic sclerosis Diseases 0.000 description 2
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 2
- 102000036693 Thrombopoietin Human genes 0.000 description 2
- 108010041111 Thrombopoietin Proteins 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 102000009618 Transforming Growth Factors Human genes 0.000 description 2
- 108010009583 Transforming Growth Factors Proteins 0.000 description 2
- 108050002568 Tumor necrosis factor ligand superfamily member 6 Proteins 0.000 description 2
- 101710178302 Tumor necrosis factor receptor superfamily member 13B Proteins 0.000 description 2
- 102100033726 Tumor necrosis factor receptor superfamily member 17 Human genes 0.000 description 2
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 2
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- RJURFGZVJUQBHK-UHFFFAOYSA-N actinomycin D Natural products CC1OC(=O)C(C(C)C)N(C)C(=O)CN(C)C(=O)C2CCCN2C(=O)C(C(C)C)NC(=O)C1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)NC4C(=O)NC(C(N5CCCC5C(=O)N(C)CC(=O)N(C)C(C(C)C)C(=O)OC4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-UHFFFAOYSA-N 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 2
- 230000000172 allergic effect Effects 0.000 description 2
- 230000000961 alloantigen Effects 0.000 description 2
- 239000002870 angiogenesis inducing agent Substances 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 210000001106 artificial yeast chromosome Anatomy 0.000 description 2
- 230000003305 autocrine Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 210000003443 bladder cell Anatomy 0.000 description 2
- 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 2
- 230000000903 blocking effect Effects 0.000 description 2
- 239000003114 blood coagulation factor Substances 0.000 description 2
- 108010006025 bovine growth hormone Proteins 0.000 description 2
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 description 2
- 229930195731 calicheamicin Natural products 0.000 description 2
- 150000001720 carbohydrates Chemical group 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000022534 cell killing Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000002458 cell surface marker Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000036755 cellular response Effects 0.000 description 2
- 229960004630 chlorambucil Drugs 0.000 description 2
- JCKYGMPEJWAADB-UHFFFAOYSA-N chlorambucil Chemical compound OC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 JCKYGMPEJWAADB-UHFFFAOYSA-N 0.000 description 2
- 230000010428 chromatin condensation Effects 0.000 description 2
- 229960004316 cisplatin Drugs 0.000 description 2
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 2
- 229940047120 colony stimulating factors Drugs 0.000 description 2
- 238000004590 computer program Methods 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 102000003675 cytokine receptors Human genes 0.000 description 2
- 108010057085 cytokine receptors Proteins 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 229960003901 dacarbazine Drugs 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 229960000975 daunorubicin Drugs 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 230000010339 dilation Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 2
- 229960001904 epirubicin Drugs 0.000 description 2
- 210000003238 esophagus Anatomy 0.000 description 2
- VJJPUSNTGOMMGY-MRVIYFEKSA-N etoposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@H](C)OC[C@H]4O3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 VJJPUSNTGOMMGY-MRVIYFEKSA-N 0.000 description 2
- 229960004222 factor ix Drugs 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 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 description 2
- CHPZKNULDCNCBW-UHFFFAOYSA-N gallium nitrate Chemical compound [Ga+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O CHPZKNULDCNCBW-UHFFFAOYSA-N 0.000 description 2
- 208000005017 glioblastoma Diseases 0.000 description 2
- 229960002989 glutamic acid Drugs 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 230000002440 hepatic effect Effects 0.000 description 2
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 2
- 108010084091 heregulin beta1 Proteins 0.000 description 2
- 239000000833 heterodimer Substances 0.000 description 2
- 102000046935 human TNFRSF17 Human genes 0.000 description 2
- 210000005104 human peripheral blood lymphocyte Anatomy 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 229960001101 ifosfamide Drugs 0.000 description 2
- HOMGKSMUEGBAAB-UHFFFAOYSA-N ifosfamide Chemical compound ClCCNP1(=O)OCCCN1CCCl HOMGKSMUEGBAAB-UHFFFAOYSA-N 0.000 description 2
- 239000012642 immune effector Substances 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000002163 immunogen Effects 0.000 description 2
- 229940121354 immunomodulator Drugs 0.000 description 2
- 238000000099 in vitro assay Methods 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 208000027866 inflammatory disease Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 102000010681 interleukin-8 receptors Human genes 0.000 description 2
- 108010038415 interleukin-8 receptors Proteins 0.000 description 2
- 229940047122 interleukins Drugs 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 210000004153 islets of langerhan Anatomy 0.000 description 2
- 229960000310 isoleucine Drugs 0.000 description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- HAWPXGHAZFHHAD-UHFFFAOYSA-N mechlorethamine Chemical compound ClCCN(C)CCCl HAWPXGHAZFHHAD-UHFFFAOYSA-N 0.000 description 2
- 229960004961 mechlorethamine Drugs 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 210000001806 memory b lymphocyte Anatomy 0.000 description 2
- 229960001428 mercaptopurine Drugs 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 2
- 229960001156 mitoxantrone Drugs 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000000822 natural killer cell Anatomy 0.000 description 2
- QZGIWPZCWHMVQL-UIYAJPBUSA-N neocarzinostatin chromophore Chemical compound O1[C@H](C)[C@H](O)[C@H](O)[C@@H](NC)[C@H]1O[C@@H]1C/2=C/C#C[C@H]3O[C@@]3([C@@H]3OC(=O)OC3)C#CC\2=C[C@H]1OC(=O)C1=C(O)C=CC2=C(C)C=C(OC)C=C12 QZGIWPZCWHMVQL-UIYAJPBUSA-N 0.000 description 2
- 230000001613 neoplastic effect Effects 0.000 description 2
- 229940053128 nerve growth factor Drugs 0.000 description 2
- 102000020232 neurotensin type 1 receptor Human genes 0.000 description 2
- 108010016501 neurotensin type 1 receptor Proteins 0.000 description 2
- 229940032018 neurotrophin 3 Drugs 0.000 description 2
- 210000000440 neutrophil Anatomy 0.000 description 2
- 238000011580 nude mouse model Methods 0.000 description 2
- 238000011275 oncology therapy Methods 0.000 description 2
- 230000002611 ovarian Effects 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 239000000199 parathyroid hormone Substances 0.000 description 2
- 229960001319 parathyroid hormone Drugs 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 2
- 229960004622 raloxifene Drugs 0.000 description 2
- GZUITABIAKMVPG-UHFFFAOYSA-N raloxifene Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCCC3)=CC=2)C2=CC=C(O)C=C2S1 GZUITABIAKMVPG-UHFFFAOYSA-N 0.000 description 2
- 230000003252 repetitive effect Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 229910052709 silver Inorganic materials 0.000 description 2
- 239000004332 silver Substances 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- 230000004936 stimulating effect Effects 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- PVYJZLYGTZKPJE-UHFFFAOYSA-N streptonigrin Chemical compound C=1C=C2C(=O)C(OC)=C(N)C(=O)C2=NC=1C(C=1N)=NC(C(O)=O)=C(C)C=1C1=CC=C(OC)C(OC)=C1O PVYJZLYGTZKPJE-UHFFFAOYSA-N 0.000 description 2
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 2
- 229960001603 tamoxifen Drugs 0.000 description 2
- RCINICONZNJXQF-XAZOAEDWSA-N taxol® Chemical compound O([C@@H]1[C@@]2(CC(C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3(C21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-XAZOAEDWSA-N 0.000 description 2
- 229960001196 thiotepa Drugs 0.000 description 2
- 229960003087 tioguanine Drugs 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 230000005945 translocation Effects 0.000 description 2
- 238000005199 ultracentrifugation Methods 0.000 description 2
- 210000003932 urinary bladder Anatomy 0.000 description 2
- 201000005112 urinary bladder cancer Diseases 0.000 description 2
- 239000004474 valine Substances 0.000 description 2
- 229960003048 vinblastine Drugs 0.000 description 2
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 2
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 2
- 229960004528 vincristine Drugs 0.000 description 2
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 2
- NNJPGOLRFBJNIW-HNNXBMFYSA-N (-)-demecolcine Chemical compound C1=C(OC)C(=O)C=C2[C@@H](NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-HNNXBMFYSA-N 0.000 description 1
- FLWWDYNPWOSLEO-HQVZTVAUSA-N (2s)-2-[[4-[1-(2-amino-4-oxo-1h-pteridin-6-yl)ethyl-methylamino]benzoyl]amino]pentanedioic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1C(C)N(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FLWWDYNPWOSLEO-HQVZTVAUSA-N 0.000 description 1
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 1
- CGMTUJFWROPELF-YPAAEMCBSA-N (3E,5S)-5-[(2S)-butan-2-yl]-3-(1-hydroxyethylidene)pyrrolidine-2,4-dione Chemical compound CC[C@H](C)[C@@H]1NC(=O)\C(=C(/C)O)C1=O CGMTUJFWROPELF-YPAAEMCBSA-N 0.000 description 1
- TVIRNGFXQVMMGB-OFWIHYRESA-N (3s,6r,10r,13e,16s)-16-[(2r,3r,4s)-4-chloro-3-hydroxy-4-phenylbutan-2-yl]-10-[(3-chloro-4-methoxyphenyl)methyl]-6-methyl-3-(2-methylpropyl)-1,4-dioxa-8,11-diazacyclohexadec-13-ene-2,5,9,12-tetrone Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H](O)[C@@H](Cl)C=2C=CC=CC=2)C/C=C/C(=O)N1 TVIRNGFXQVMMGB-OFWIHYRESA-N 0.000 description 1
- XRBSKUSTLXISAB-XVVDYKMHSA-N (5r,6r,7r,8r)-8-hydroxy-7-(hydroxymethyl)-5-(3,4,5-trimethoxyphenyl)-5,6,7,8-tetrahydrobenzo[f][1,3]benzodioxole-6-carboxylic acid Chemical compound COC1=C(OC)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@H](O)[C@@H](CO)[C@@H]2C(O)=O)=C1 XRBSKUSTLXISAB-XVVDYKMHSA-N 0.000 description 1
- XRBSKUSTLXISAB-UHFFFAOYSA-N (7R,7'R,8R,8'R)-form-Podophyllic acid Natural products COC1=C(OC)C(OC)=CC(C2C3=CC=4OCOC=4C=C3C(O)C(CO)C2C(O)=O)=C1 XRBSKUSTLXISAB-UHFFFAOYSA-N 0.000 description 1
- AESVUZLWRXEGEX-DKCAWCKPSA-N (7S,9R)-7-[(2S,4R,5R,6R)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7H-tetracene-5,12-dione iron(3+) Chemical compound [Fe+3].COc1cccc2C(=O)c3c(O)c4C[C@@](O)(C[C@H](O[C@@H]5C[C@@H](N)[C@@H](O)[C@@H](C)O5)c4c(O)c3C(=O)c12)C(=O)CO AESVUZLWRXEGEX-DKCAWCKPSA-N 0.000 description 1
- JXVAMODRWBNUSF-KZQKBALLSA-N (7s,9r,10r)-7-[(2r,4s,5s,6s)-5-[[(2s,4as,5as,7s,9s,9ar,10ar)-2,9-dimethyl-3-oxo-4,4a,5a,6,7,9,9a,10a-octahydrodipyrano[4,2-a:4',3'-e][1,4]dioxin-7-yl]oxy]-4-(dimethylamino)-6-methyloxan-2-yl]oxy-10-[(2s,4s,5s,6s)-4-(dimethylamino)-5-hydroxy-6-methyloxan-2 Chemical compound O([C@@H]1C2=C(O)C=3C(=O)C4=CC=CC(O)=C4C(=O)C=3C(O)=C2[C@@H](O[C@@H]2O[C@@H](C)[C@@H](O[C@@H]3O[C@@H](C)[C@H]4O[C@@H]5O[C@@H](C)C(=O)C[C@@H]5O[C@H]4C3)[C@H](C2)N(C)C)C[C@]1(O)CC)[C@H]1C[C@H](N(C)C)[C@H](O)[C@H](C)O1 JXVAMODRWBNUSF-KZQKBALLSA-N 0.000 description 1
- INAUWOVKEZHHDM-PEDBPRJASA-N (7s,9s)-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-7-[(2r,4s,5s,6s)-5-hydroxy-6-methyl-4-morpholin-4-yloxan-2-yl]oxy-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound Cl.N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1 INAUWOVKEZHHDM-PEDBPRJASA-N 0.000 description 1
- RCFNNLSZHVHCEK-IMHLAKCZSA-N (7s,9s)-7-(4-amino-6-methyloxan-2-yl)oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione;hydrochloride Chemical compound [Cl-].O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)C1CC([NH3+])CC(C)O1 RCFNNLSZHVHCEK-IMHLAKCZSA-N 0.000 description 1
- NOPNWHSMQOXAEI-PUCKCBAPSA-N (7s,9s)-7-[(2r,4s,5s,6s)-4-(2,3-dihydropyrrol-1-yl)-5-hydroxy-6-methyloxan-2-yl]oxy-6,9,11-trihydroxy-9-(2-hydroxyacetyl)-4-methoxy-8,10-dihydro-7h-tetracene-5,12-dione Chemical compound N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCC=C1 NOPNWHSMQOXAEI-PUCKCBAPSA-N 0.000 description 1
- FPVKHBSQESCIEP-UHFFFAOYSA-N (8S)-3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,6,7,8-tetrahydroimidazo[4,5-d][1,3]diazepin-8-ol Natural products C1C(O)C(CO)OC1N1C(NC=NCC2O)=C2N=C1 FPVKHBSQESCIEP-UHFFFAOYSA-N 0.000 description 1
- IEXUMDBQLIVNHZ-YOUGDJEHSA-N (8s,11r,13r,14s,17s)-11-[4-(dimethylamino)phenyl]-17-hydroxy-17-(3-hydroxypropyl)-13-methyl-1,2,6,7,8,11,12,14,15,16-decahydrocyclopenta[a]phenanthren-3-one Chemical compound C1=CC(N(C)C)=CC=C1[C@@H]1C2=C3CCC(=O)C=C3CC[C@H]2[C@H](CC[C@]2(O)CCCO)[C@@]2(C)C1 IEXUMDBQLIVNHZ-YOUGDJEHSA-N 0.000 description 1
- LKJPYSCBVHEWIU-KRWDZBQOSA-N (R)-bicalutamide Chemical compound C([C@@](O)(C)C(=O)NC=1C=C(C(C#N)=CC=1)C(F)(F)F)S(=O)(=O)C1=CC=C(F)C=C1 LKJPYSCBVHEWIU-KRWDZBQOSA-N 0.000 description 1
- AGNGYMCLFWQVGX-AGFFZDDWSA-N (e)-1-[(2s)-2-amino-2-carboxyethoxy]-2-diazonioethenolate Chemical compound OC(=O)[C@@H](N)CO\C([O-])=C\[N+]#N AGNGYMCLFWQVGX-AGFFZDDWSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 1
- FONKWHRXTPJODV-DNQXCXABSA-N 1,3-bis[2-[(8s)-8-(chloromethyl)-4-hydroxy-1-methyl-7,8-dihydro-3h-pyrrolo[3,2-e]indole-6-carbonyl]-1h-indol-5-yl]urea Chemical compound C1([C@H](CCl)CN2C(=O)C=3NC4=CC=C(C=C4C=3)NC(=O)NC=3C=C4C=C(NC4=CC=3)C(=O)N3C4=CC(O)=C5NC=C(C5=C4[C@H](CCl)C3)C)=C2C=C(O)C2=C1C(C)=CN2 FONKWHRXTPJODV-DNQXCXABSA-N 0.000 description 1
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 1
- BTOTXLJHDSNXMW-POYBYMJQSA-N 2,3-dideoxyuridine Chemical compound O1[C@H](CO)CC[C@@H]1N1C(=O)NC(=O)C=C1 BTOTXLJHDSNXMW-POYBYMJQSA-N 0.000 description 1
- BOMZMNZEXMAQQW-UHFFFAOYSA-N 2,5,11-trimethyl-6h-pyrido[4,3-b]carbazol-2-ium-9-ol;acetate Chemical compound CC([O-])=O.C[N+]1=CC=C2C(C)=C(NC=3C4=CC(O)=CC=3)C4=C(C)C2=C1 BOMZMNZEXMAQQW-UHFFFAOYSA-N 0.000 description 1
- QCXJFISCRQIYID-IAEPZHFASA-N 2-amino-1-n-[(3s,6s,7r,10s,16s)-3-[(2s)-butan-2-yl]-7,11,14-trimethyl-2,5,9,12,15-pentaoxo-10-propan-2-yl-8-oxa-1,4,11,14-tetrazabicyclo[14.3.0]nonadecan-6-yl]-4,6-dimethyl-3-oxo-9-n-[(3s,6s,7r,10s,16s)-7,11,14-trimethyl-2,5,9,12,15-pentaoxo-3,10-di(propa Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N=C2C(C(=O)N[C@@H]3C(=O)N[C@H](C(N4CCC[C@H]4C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]3C)=O)[C@@H](C)CC)=C(N)C(=O)C(C)=C2O2)C2=C(C)C=C1 QCXJFISCRQIYID-IAEPZHFASA-N 0.000 description 1
- AXAVXPMQTGXXJZ-UHFFFAOYSA-N 2-aminoacetic acid;2-amino-2-(hydroxymethyl)propane-1,3-diol Chemical compound NCC(O)=O.OCC(N)(CO)CO AXAVXPMQTGXXJZ-UHFFFAOYSA-N 0.000 description 1
- FDAYLTPAFBGXAB-UHFFFAOYSA-N 2-chloro-n,n-bis(2-chloroethyl)ethanamine Chemical compound ClCCN(CCCl)CCCl FDAYLTPAFBGXAB-UHFFFAOYSA-N 0.000 description 1
- VNBAOSVONFJBKP-UHFFFAOYSA-N 2-chloro-n,n-bis(2-chloroethyl)propan-1-amine;hydrochloride Chemical compound Cl.CC(Cl)CN(CCCl)CCCl VNBAOSVONFJBKP-UHFFFAOYSA-N 0.000 description 1
- AOPRXJXHLWYPQR-UHFFFAOYSA-N 2-phenoxyacetamide Chemical class NC(=O)COC1=CC=CC=C1 AOPRXJXHLWYPQR-UHFFFAOYSA-N 0.000 description 1
- YIMDLWDNDGKDTJ-QLKYHASDSA-N 3'-deamino-3'-(3-cyanomorpholin-4-yl)doxorubicin Chemical compound N1([C@H]2C[C@@H](O[C@@H](C)[C@H]2O)O[C@H]2C[C@@](O)(CC=3C(O)=C4C(=O)C=5C=CC=C(C=5C(=O)C4=C(O)C=32)OC)C(=O)CO)CCOCC1C#N YIMDLWDNDGKDTJ-QLKYHASDSA-N 0.000 description 1
- NDMPLJNOPCLANR-UHFFFAOYSA-N 3,4-dihydroxy-15-(4-hydroxy-18-methoxycarbonyl-5,18-seco-ibogamin-18-yl)-16-methoxy-1-methyl-6,7-didehydro-aspidospermidine-3-carboxylic acid methyl ester Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 NDMPLJNOPCLANR-UHFFFAOYSA-N 0.000 description 1
- PWMYMKOUNYTVQN-UHFFFAOYSA-N 3-(8,8-diethyl-2-aza-8-germaspiro[4.5]decan-2-yl)-n,n-dimethylpropan-1-amine Chemical compound C1C[Ge](CC)(CC)CCC11CN(CCCN(C)C)CC1 PWMYMKOUNYTVQN-UHFFFAOYSA-N 0.000 description 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- 238000010600 3H thymidine incorporation assay Methods 0.000 description 1
- 102000002627 4-1BB Ligand Human genes 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- DODQJNMQWMSYGS-QPLCGJKRSA-N 4-[(z)-1-[4-[2-(dimethylamino)ethoxy]phenyl]-1-phenylbut-1-en-2-yl]phenol Chemical compound C=1C=C(O)C=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 DODQJNMQWMSYGS-QPLCGJKRSA-N 0.000 description 1
- TVZGACDUOSZQKY-LBPRGKRZSA-N 4-aminofolic acid Chemical compound C1=NC2=NC(N)=NC(N)=C2N=C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 TVZGACDUOSZQKY-LBPRGKRZSA-N 0.000 description 1
- IDPUKCWIGUEADI-UHFFFAOYSA-N 5-[bis(2-chloroethyl)amino]uracil Chemical compound ClCCN(CCCl)C1=CNC(=O)NC1=O IDPUKCWIGUEADI-UHFFFAOYSA-N 0.000 description 1
- NMUSYJAQQFHJEW-KVTDHHQDSA-N 5-azacytidine Chemical compound O=C1N=C(N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 NMUSYJAQQFHJEW-KVTDHHQDSA-N 0.000 description 1
- FHIDNBAQOFJWCA-UAKXSSHOSA-N 5-fluorouridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 FHIDNBAQOFJWCA-UAKXSSHOSA-N 0.000 description 1
- WYXSYVWAUAUWLD-SHUUEZRQSA-N 6-azauridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=N1 WYXSYVWAUAUWLD-SHUUEZRQSA-N 0.000 description 1
- 229960005538 6-diazo-5-oxo-L-norleucine Drugs 0.000 description 1
- YCWQAMGASJSUIP-YFKPBYRVSA-N 6-diazo-5-oxo-L-norleucine Chemical compound OC(=O)[C@@H](N)CCC(=O)C=[N+]=[N-] YCWQAMGASJSUIP-YFKPBYRVSA-N 0.000 description 1
- ZGXJTSGNIOSYLO-UHFFFAOYSA-N 88755TAZ87 Chemical compound NCC(=O)CCC(O)=O ZGXJTSGNIOSYLO-UHFFFAOYSA-N 0.000 description 1
- HDZZVAMISRMYHH-UHFFFAOYSA-N 9beta-Ribofuranosyl-7-deazaadenin Natural products C1=CC=2C(N)=NC=NC=2N1C1OC(CO)C(O)C1O HDZZVAMISRMYHH-UHFFFAOYSA-N 0.000 description 1
- 108010059616 Activins Proteins 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 208000026872 Addison Disease Diseases 0.000 description 1
- 206010027654 Allergic conditions Diseases 0.000 description 1
- CEIZFXOZIQNICU-UHFFFAOYSA-N Alternaria alternata Crofton-weed toxin Natural products CCC(C)C1NC(=O)C(C(C)=O)=C1O CEIZFXOZIQNICU-UHFFFAOYSA-N 0.000 description 1
- 108010005853 Anti-Mullerian Hormone Proteins 0.000 description 1
- 208000003343 Antiphospholipid Syndrome Diseases 0.000 description 1
- 101100107610 Arabidopsis thaliana ABCF4 gene Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 102100029361 Aromatase Human genes 0.000 description 1
- 108010078554 Aromatase Proteins 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 206010063836 Atrioventricular septal defect Diseases 0.000 description 1
- 208000032116 Autoimmune Experimental Encephalomyelitis Diseases 0.000 description 1
- 208000031212 Autoimmune polyendocrinopathy Diseases 0.000 description 1
- 206010050245 Autoimmune thrombocytopenia Diseases 0.000 description 1
- NOWKCMXCCJGMRR-UHFFFAOYSA-N Aziridine Chemical class C1CN1 NOWKCMXCCJGMRR-UHFFFAOYSA-N 0.000 description 1
- 108091008875 B cell receptors Proteins 0.000 description 1
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 1
- 208000023328 Basedow disease Diseases 0.000 description 1
- 208000027496 Behcet disease Diseases 0.000 description 1
- 208000009137 Behcet syndrome Diseases 0.000 description 1
- VGGGPCQERPFHOB-MCIONIFRSA-N Bestatin Chemical compound CC(C)C[C@H](C(O)=O)NC(=O)[C@@H](O)[C@H](N)CC1=CC=CC=C1 VGGGPCQERPFHOB-MCIONIFRSA-N 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- MBABCNBNDNGODA-LTGLSHGVSA-N Bullatacin Natural products O=C1C(C[C@H](O)CCCCCCCCCC[C@@H](O)[C@@H]2O[C@@H]([C@@H]3O[C@H]([C@@H](O)CCCCCCCCCC)CC3)CC2)=C[C@H](C)O1 MBABCNBNDNGODA-LTGLSHGVSA-N 0.000 description 1
- KGGVWMAPBXIMEM-ZRTAFWODSA-N Bullatacinone Chemical compound O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@@H]1[C@@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@H]2OC(=O)[C@H](CC(C)=O)C2)CC1 KGGVWMAPBXIMEM-ZRTAFWODSA-N 0.000 description 1
- KGGVWMAPBXIMEM-JQFCFGFHSA-N Bullatacinone Natural products O=C(C[C@H]1C(=O)O[C@H](CCCCCCCCCC[C@H](O)[C@@H]2O[C@@H]([C@@H]3O[C@@H]([C@@H](O)CCCCCCCCCC)CC3)CC2)C1)C KGGVWMAPBXIMEM-JQFCFGFHSA-N 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- 210000004366 CD4-positive T-lymphocyte Anatomy 0.000 description 1
- 102100027217 CD82 antigen Human genes 0.000 description 1
- 102400000113 Calcitonin Human genes 0.000 description 1
- 108060001064 Calcitonin Proteins 0.000 description 1
- 108090000312 Calcium Channels Proteins 0.000 description 1
- 102000003922 Calcium Channels Human genes 0.000 description 1
- 241000189662 Calla Species 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- GAGWJHPBXLXJQN-UHFFFAOYSA-N Capecitabine Natural products C1=C(F)C(NC(=O)OCCCCC)=NC(=O)N1C1C(O)C(O)C(C)O1 GAGWJHPBXLXJQN-UHFFFAOYSA-N 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- SHHKQEUPHAENFK-UHFFFAOYSA-N Carboquone Chemical compound O=C1C(C)=C(N2CC2)C(=O)C(C(COC(N)=O)OC)=C1N1CC1 SHHKQEUPHAENFK-UHFFFAOYSA-N 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- AOCCBINRVIKJHY-UHFFFAOYSA-N Carmofur Chemical compound CCCCCCNC(=O)N1C=C(F)C(=O)NC1=O AOCCBINRVIKJHY-UHFFFAOYSA-N 0.000 description 1
- DLGOEMSEDOSKAD-UHFFFAOYSA-N Carmustine Chemical compound ClCCNC(=O)N(N=O)CCCl DLGOEMSEDOSKAD-UHFFFAOYSA-N 0.000 description 1
- 102000011727 Caspases Human genes 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 102000016289 Cell Adhesion Molecules Human genes 0.000 description 1
- 108010067225 Cell Adhesion Molecules Proteins 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- JWBOIMRXGHLCPP-UHFFFAOYSA-N Chloditan Chemical compound C=1C=CC=C(Cl)C=1C(C(Cl)Cl)C1=CC=C(Cl)C=C1 JWBOIMRXGHLCPP-UHFFFAOYSA-N 0.000 description 1
- XCDXSSFOJZZGQC-UHFFFAOYSA-N Chlornaphazine Chemical compound C1=CC=CC2=CC(N(CCCl)CCCl)=CC=C21 XCDXSSFOJZZGQC-UHFFFAOYSA-N 0.000 description 1
- MKQWTWSXVILIKJ-LXGUWJNJSA-N Chlorozotocin Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](C=O)NC(=O)N(N=O)CCCl MKQWTWSXVILIKJ-LXGUWJNJSA-N 0.000 description 1
- 102100021809 Chorionic somatomammotropin hormone 1 Human genes 0.000 description 1
- 101710094648 Coat protein Proteins 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 108010023729 Complement 3d Receptors Proteins 0.000 description 1
- 102000011412 Complement 3d Receptors Human genes 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 229930188224 Cryptophycin Natural products 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 description 1
- 239000012624 DNA alkylating agent Substances 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- WEAHRLBPCANXCN-UHFFFAOYSA-N Daunomycin Natural products CCC1(O)CC(OC2CC(N)C(O)C(C)O2)c3cc4C(=O)c5c(OC)cccc5C(=O)c4c(O)c3C1 WEAHRLBPCANXCN-UHFFFAOYSA-N 0.000 description 1
- 208000006313 Delayed Hypersensitivity Diseases 0.000 description 1
- NNJPGOLRFBJNIW-UHFFFAOYSA-N Demecolcine Natural products C1=C(OC)C(=O)C=C2C(NC)CCC3=CC(OC)=C(OC)C(OC)=C3C2=C1 NNJPGOLRFBJNIW-UHFFFAOYSA-N 0.000 description 1
- 108010002156 Depsipeptides Proteins 0.000 description 1
- 206010012438 Dermatitis atopic Diseases 0.000 description 1
- AUGQEEXBDZWUJY-ZLJUKNTDSA-N Diacetoxyscirpenol Chemical compound C([C@]12[C@]3(C)[C@H](OC(C)=O)[C@@H](O)[C@H]1O[C@@H]1C=C(C)CC[C@@]13COC(=O)C)O2 AUGQEEXBDZWUJY-ZLJUKNTDSA-N 0.000 description 1
- AUGQEEXBDZWUJY-UHFFFAOYSA-N Diacetoxyscirpenol Natural products CC(=O)OCC12CCC(C)=CC1OC1C(O)C(OC(C)=O)C2(C)C11CO1 AUGQEEXBDZWUJY-UHFFFAOYSA-N 0.000 description 1
- 206010051392 Diapedesis Diseases 0.000 description 1
- 206010012713 Diaphragmatic hernia Diseases 0.000 description 1
- 108010053187 Diphtheria Toxin Proteins 0.000 description 1
- 102000016607 Diphtheria Toxin Human genes 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- 101100044298 Drosophila melanogaster fand gene Proteins 0.000 description 1
- 229930193152 Dynemicin Natural products 0.000 description 1
- 102000012545 EGF-like domains Human genes 0.000 description 1
- 108050002150 EGF-like domains Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010014733 Endometrial cancer Diseases 0.000 description 1
- 206010014759 Endometrial neoplasm Diseases 0.000 description 1
- AFMYMMXSQGUCBK-UHFFFAOYSA-N Endynamicin A Natural products C1#CC=CC#CC2NC(C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C3)=C3C34OC32C(C)C(C(O)=O)=C(OC)C41 AFMYMMXSQGUCBK-UHFFFAOYSA-N 0.000 description 1
- SAMRUMKYXPVKPA-VFKOLLTISA-N Enocitabine Chemical compound O=C1N=C(NC(=O)CCCCCCCCCCCCCCCCCCCCC)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 SAMRUMKYXPVKPA-VFKOLLTISA-N 0.000 description 1
- OBMLHUPNRURLOK-XGRAFVIBSA-N Epitiostanol Chemical compound C1[C@@H]2S[C@@H]2C[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 OBMLHUPNRURLOK-XGRAFVIBSA-N 0.000 description 1
- 102000056372 ErbB-3 Receptor Human genes 0.000 description 1
- 102000044591 ErbB-4 Receptor Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241001524679 Escherichia virus M13 Species 0.000 description 1
- 229930189413 Esperamicin Natural products 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 1
- 230000037057 G1 phase arrest Effects 0.000 description 1
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 1
- 208000007465 Giant cell arteritis Diseases 0.000 description 1
- 206010018364 Glomerulonephritis Diseases 0.000 description 1
- 102400000321 Glucagon Human genes 0.000 description 1
- 108060003199 Glucagon Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 description 1
- 102000006771 Gonadotropins Human genes 0.000 description 1
- 108010086677 Gonadotropins Proteins 0.000 description 1
- 208000024869 Goodpasture syndrome Diseases 0.000 description 1
- 108010069236 Goserelin Proteins 0.000 description 1
- BLCLNMBMMGCOAS-URPVMXJPSA-N Goserelin Chemical compound C([C@@H](C(=O)N[C@H](COC(C)(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1[C@@H](CCC1)C(=O)NNC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H]1NC(=O)CC1)C1=CC=C(O)C=C1 BLCLNMBMMGCOAS-URPVMXJPSA-N 0.000 description 1
- 208000015023 Graves' disease Diseases 0.000 description 1
- 239000000095 Growth Hormone-Releasing Hormone Substances 0.000 description 1
- 102100031547 HLA class II histocompatibility antigen, DO alpha chain Human genes 0.000 description 1
- 208000030836 Hashimoto thyroiditis Diseases 0.000 description 1
- 101000690301 Homo sapiens Aldo-keto reductase family 1 member C4 Proteins 0.000 description 1
- 101000914469 Homo sapiens CD82 antigen Proteins 0.000 description 1
- 101000941929 Homo sapiens Complement receptor type 2 Proteins 0.000 description 1
- 101000746783 Homo sapiens Cytochrome b-c1 complex subunit 6, mitochondrial Proteins 0.000 description 1
- 101000866278 Homo sapiens HLA class II histocompatibility antigen, DO alpha chain Proteins 0.000 description 1
- 101000984196 Homo sapiens Leukocyte immunoglobulin-like receptor subfamily A member 5 Proteins 0.000 description 1
- 101000984190 Homo sapiens Leukocyte immunoglobulin-like receptor subfamily B member 1 Proteins 0.000 description 1
- 101000980823 Homo sapiens Leukocyte surface antigen CD53 Proteins 0.000 description 1
- 101001116548 Homo sapiens Protein CBFA2T1 Proteins 0.000 description 1
- VSNHCAURESNICA-UHFFFAOYSA-N Hydroxyurea Chemical compound NC(=O)NO VSNHCAURESNICA-UHFFFAOYSA-N 0.000 description 1
- MPBVHIBUJCELCL-UHFFFAOYSA-N Ibandronate Chemical compound CCCCCN(C)CCC(O)(P(O)(O)=O)P(O)(O)=O MPBVHIBUJCELCL-UHFFFAOYSA-N 0.000 description 1
- XDXDZDZNSLXDNA-TZNDIEGXSA-N Idarubicin Chemical compound C1[C@H](N)[C@H](O)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2C[C@@](O)(C(C)=O)C1 XDXDZDZNSLXDNA-TZNDIEGXSA-N 0.000 description 1
- XDXDZDZNSLXDNA-UHFFFAOYSA-N Idarubicin Natural products C1C(N)C(O)C(C)OC1OC1C2=C(O)C(C(=O)C3=CC=CC=C3C3=O)=C3C(O)=C2CC(O)(C(C)=O)C1 XDXDZDZNSLXDNA-UHFFFAOYSA-N 0.000 description 1
- 208000010159 IgA glomerulonephritis Diseases 0.000 description 1
- 206010021263 IgA nephropathy Diseases 0.000 description 1
- 102000009490 IgG Receptors Human genes 0.000 description 1
- 102100026120 IgG receptor FcRn large subunit p51 Human genes 0.000 description 1
- 101710177940 IgG receptor FcRn large subunit p51 Proteins 0.000 description 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 description 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 102100026818 Inhibin beta E chain Human genes 0.000 description 1
- 102100025390 Integrin beta-2 Human genes 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000003996 Interferon-beta Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 102000003815 Interleukin-11 Human genes 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- SNDPXSYFESPGGJ-BYPYZUCNSA-N L-2-aminopentanoic acid Chemical compound CCC[C@H](N)C(O)=O SNDPXSYFESPGGJ-BYPYZUCNSA-N 0.000 description 1
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 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
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- SNDPXSYFESPGGJ-UHFFFAOYSA-N L-norVal-OH Natural products CCCC(N)C(O)=O SNDPXSYFESPGGJ-UHFFFAOYSA-N 0.000 description 1
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- JLERVPBPJHKRBJ-UHFFFAOYSA-N LY 117018 Chemical compound C1=CC(O)=CC=C1C1=C(C(=O)C=2C=CC(OCCN3CCCC3)=CC=2)C2=CC=C(O)C=C2S1 JLERVPBPJHKRBJ-UHFFFAOYSA-N 0.000 description 1
- 201000010743 Lambert-Eaton myasthenic syndrome Diseases 0.000 description 1
- 229920001491 Lentinan Polymers 0.000 description 1
- 201000001779 Leukocyte adhesion deficiency Diseases 0.000 description 1
- 102100025574 Leukocyte immunoglobulin-like receptor subfamily A member 5 Human genes 0.000 description 1
- 102100024221 Leukocyte surface antigen CD53 Human genes 0.000 description 1
- 108010000817 Leuprolide Proteins 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- GQYIWUVLTXOXAJ-UHFFFAOYSA-N Lomustine Chemical compound ClCCN(N=O)C(=O)NC1CCCCC1 GQYIWUVLTXOXAJ-UHFFFAOYSA-N 0.000 description 1
- 102100029204 Low affinity immunoglobulin gamma Fc region receptor II-a Human genes 0.000 description 1
- 102100029205 Low affinity immunoglobulin gamma Fc region receptor II-b Human genes 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000282567 Macaca fascicularis Species 0.000 description 1
- 101710125418 Major capsid protein Proteins 0.000 description 1
- VJRAUFKOOPNFIQ-UHFFFAOYSA-N Marcellomycin Natural products C12=C(O)C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C=C2C(C(=O)OC)C(CC)(O)CC1OC(OC1C)CC(N(C)C)C1OC(OC1C)CC(O)C1OC1CC(O)C(O)C(C)O1 VJRAUFKOOPNFIQ-UHFFFAOYSA-N 0.000 description 1
- 229930126263 Maytansine Natural products 0.000 description 1
- 108010060408 Member 25 Tumor Necrosis Factor Receptors Proteins 0.000 description 1
- 102000008166 Member 25 Tumor Necrosis Factor Receptors Human genes 0.000 description 1
- 108010061342 Member 6b Tumor Necrosis Factor Receptors Proteins 0.000 description 1
- 102000012089 Member 6b Tumor Necrosis Factor Receptors Human genes 0.000 description 1
- 108010037274 Member 9 Tumor Necrosis Factor Receptor Superfamily Proteins 0.000 description 1
- 102000011769 Member 9 Tumor Necrosis Factor Receptor Superfamily Human genes 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- IVDYZAAPOLNZKG-KWHRADDSSA-N Mepitiostane Chemical compound O([C@@H]1[C@]2(CC[C@@H]3[C@@]4(C)C[C@H]5S[C@H]5C[C@@H]4CC[C@H]3[C@@H]2CC1)C)C1(OC)CCCC1 IVDYZAAPOLNZKG-KWHRADDSSA-N 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- VFKZTMPDYBFSTM-KVTDHHQDSA-N Mitobronitol Chemical compound BrC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CBr VFKZTMPDYBFSTM-KVTDHHQDSA-N 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- 101100537545 Mus musculus Fas gene Proteins 0.000 description 1
- 206010028424 Myasthenic syndrome Diseases 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- ZDZOTLJHXYCWBA-VCVYQWHSSA-N N-debenzoyl-N-(tert-butoxycarbonyl)-10-deacetyltaxol Chemical compound O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](OC(=O)[C@H](O)[C@@H](NC(=O)OC(C)(C)C)C=4C=CC=CC=4)C[C@]1(O)C3(C)C)=O)(C)[C@@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 ZDZOTLJHXYCWBA-VCVYQWHSSA-N 0.000 description 1
- 108091008604 NGF receptors Proteins 0.000 description 1
- 102000007072 Nerve Growth Factors Human genes 0.000 description 1
- 206010029240 Neuritis Diseases 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- SYNHCENRCUAUNM-UHFFFAOYSA-N Nitrogen mustard N-oxide hydrochloride Chemical compound Cl.ClCC[N+]([O-])(C)CCCl SYNHCENRCUAUNM-UHFFFAOYSA-N 0.000 description 1
- KGTDRFCXGRULNK-UHFFFAOYSA-N Nogalamycin Natural products COC1C(OC)(C)C(OC)C(C)OC1OC1C2=C(O)C(C(=O)C3=C(O)C=C4C5(C)OC(C(C(C5O)N(C)C)O)OC4=C3C3=O)=C3C=C2C(C(=O)OC)C(C)(O)C1 KGTDRFCXGRULNK-UHFFFAOYSA-N 0.000 description 1
- 108091060545 Nonsense suppressor Proteins 0.000 description 1
- 101710141454 Nucleoprotein Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 229930187135 Olivomycin Natural products 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- VREZDOWOLGNDPW-ALTGWBOUSA-N Pancratistatin Chemical compound C1=C2[C@H]3[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O)[C@@H]3NC(=O)C2=C(O)C2=C1OCO2 VREZDOWOLGNDPW-ALTGWBOUSA-N 0.000 description 1
- VREZDOWOLGNDPW-MYVCAWNPSA-N Pancratistatin Natural products O=C1N[C@H]2[C@H](O)[C@H](O)[C@H](O)[C@H](O)[C@@H]2c2c1c(O)c1OCOc1c2 VREZDOWOLGNDPW-MYVCAWNPSA-N 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 241001504519 Papio ursinus Species 0.000 description 1
- 206010034277 Pemphigoid Diseases 0.000 description 1
- 241000721454 Pemphigus Species 0.000 description 1
- 108010057150 Peplomycin Proteins 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 208000031845 Pernicious anaemia Diseases 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 108010089430 Phosphoproteins Proteins 0.000 description 1
- 102000007982 Phosphoproteins Human genes 0.000 description 1
- KMSKQZKKOZQFFG-HSUXVGOQSA-N Pirarubicin Chemical compound O([C@H]1[C@@H](N)C[C@@H](O[C@H]1C)O[C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1CCCCO1 KMSKQZKKOZQFFG-HSUXVGOQSA-N 0.000 description 1
- 108010003044 Placental Lactogen Proteins 0.000 description 1
- 239000000381 Placental Lactogen Substances 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 101100335198 Pneumocystis carinii fol1 gene Proteins 0.000 description 1
- 206010036105 Polyneuropathy Diseases 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- HFVNWDWLWUCIHC-GUPDPFMOSA-N Prednimustine Chemical compound O=C([C@@]1(O)CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)[C@@H](O)C[C@@]21C)COC(=O)CCCC1=CC=C(N(CCCl)CCCl)C=C1 HFVNWDWLWUCIHC-GUPDPFMOSA-N 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 101710083689 Probable capsid protein Proteins 0.000 description 1
- 102000003946 Prolactin Human genes 0.000 description 1
- 108010057464 Prolactin Proteins 0.000 description 1
- 101800004937 Protein C Proteins 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- AHHFEZNOXOZZQA-ZEBDFXRSSA-N Ranimustine Chemical compound CO[C@H]1O[C@H](CNC(=O)N(CCCl)N=O)[C@@H](O)[C@H](O)[C@H]1O AHHFEZNOXOZZQA-ZEBDFXRSSA-N 0.000 description 1
- 101100501691 Rattus norvegicus Erbb2 gene Proteins 0.000 description 1
- 101100501698 Rattus norvegicus Erbb4 gene Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- 208000033464 Reiter syndrome Diseases 0.000 description 1
- 108090000103 Relaxin Proteins 0.000 description 1
- 102000003743 Relaxin Human genes 0.000 description 1
- 108090000783 Renin Proteins 0.000 description 1
- 102100028255 Renin Human genes 0.000 description 1
- 108091081062 Repeated sequence (DNA) Proteins 0.000 description 1
- OWPCHSCAPHNHAV-UHFFFAOYSA-N Rhizoxin Natural products C1C(O)C2(C)OC2C=CC(C)C(OC(=O)C2)CC2CC2OC2C(=O)OC1C(C)C(OC)C(C)=CC=CC(C)=CC1=COC(C)=N1 OWPCHSCAPHNHAV-UHFFFAOYSA-N 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- NSFWWJIQIKBZMJ-YKNYLIOZSA-N Roridin A Chemical compound C([C@]12[C@]3(C)[C@H]4C[C@H]1O[C@@H]1C=C(C)CC[C@@]13COC(=O)[C@@H](O)[C@H](C)CCO[C@H](\C=C\C=C/C(=O)O4)[C@H](O)C)O2 NSFWWJIQIKBZMJ-YKNYLIOZSA-N 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- 101100068078 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) GCN4 gene Proteins 0.000 description 1
- 206010061934 Salivary gland cancer Diseases 0.000 description 1
- 102400000827 Saposin-D Human genes 0.000 description 1
- 101800001700 Saposin-D Proteins 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 102400001107 Secretory component Human genes 0.000 description 1
- 108010029157 Sialic Acid Binding Ig-like Lectin 2 Proteins 0.000 description 1
- 206010041067 Small cell lung cancer Diseases 0.000 description 1
- 102220497176 Small vasohibin-binding protein_T47D_mutation Human genes 0.000 description 1
- 102100022831 Somatoliberin Human genes 0.000 description 1
- 101710142969 Somatoliberin Proteins 0.000 description 1
- 206010072148 Stiff-Person syndrome Diseases 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- BXFOFFBJRFZBQZ-QYWOHJEZSA-N T-2 toxin Chemical compound C([C@@]12[C@]3(C)[C@H](OC(C)=O)[C@@H](O)[C@H]1O[C@H]1[C@]3(COC(C)=O)C[C@@H](C(=C1)C)OC(=O)CC(C)C)O2 BXFOFFBJRFZBQZ-QYWOHJEZSA-N 0.000 description 1
- 208000029052 T-cell acute lymphoblastic leukemia Diseases 0.000 description 1
- 101150033527 TNF gene Proteins 0.000 description 1
- CGMTUJFWROPELF-UHFFFAOYSA-N Tenuazonic acid Natural products CCC(C)C1NC(=O)C(=C(C)/O)C1=O CGMTUJFWROPELF-UHFFFAOYSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- IWEQQRMGNVVKQW-OQKDUQJOSA-N Toremifene citrate Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O.C1=CC(OCCN(C)C)=CC=C1C(\C=1C=CC=CC=1)=C(\CCCl)C1=CC=CC=C1 IWEQQRMGNVVKQW-OQKDUQJOSA-N 0.000 description 1
- 102000050862 Transmembrane Activator and CAML Interactor Human genes 0.000 description 1
- 108700002109 Transmembrane Activator and CAML Interactor Proteins 0.000 description 1
- UMILHIMHKXVDGH-UHFFFAOYSA-N Triethylene glycol diglycidyl ether Chemical compound C1OC1COCCOCCOCCOCC1CO1 UMILHIMHKXVDGH-UHFFFAOYSA-N 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 description 1
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 101710165474 Tumor necrosis factor receptor superfamily member 5 Proteins 0.000 description 1
- 108091005906 Type I transmembrane proteins Proteins 0.000 description 1
- 108091005956 Type II transmembrane proteins Proteins 0.000 description 1
- 206010053613 Type IV hypersensitivity reaction Diseases 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 206010046851 Uveitis Diseases 0.000 description 1
- 206010047115 Vasculitis Diseases 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 241000863480 Vinca Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 206010047741 Vulval cancer Diseases 0.000 description 1
- ZYVSOIYQKUDENJ-ASUJBHBQSA-N [(2R,3R,4R,6R)-6-[[(6S,7S)-6-[(2S,4R,5R,6R)-4-[(2R,4R,5R,6R)-4-[(2S,4S,5S,6S)-5-acetyloxy-4-hydroxy-4,6-dimethyloxan-2-yl]oxy-5-hydroxy-6-methyloxan-2-yl]oxy-5-hydroxy-6-methyloxan-2-yl]oxy-7-[(3S,4R)-3,4-dihydroxy-1-methoxy-2-oxopentyl]-4,10-dihydroxy-3-methyl-5-oxo-7,8-dihydro-6H-anthracen-2-yl]oxy]-4-[(2R,4R,5R,6R)-4-hydroxy-5-methoxy-6-methyloxan-2-yl]oxy-2-methyloxan-3-yl] acetate Chemical class COC([C@@H]1Cc2cc3cc(O[C@@H]4C[C@@H](O[C@@H]5C[C@@H](O)[C@@H](OC)[C@@H](C)O5)[C@H](OC(C)=O)[C@@H](C)O4)c(C)c(O)c3c(O)c2C(=O)[C@H]1O[C@H]1C[C@@H](O[C@@H]2C[C@@H](O[C@H]3C[C@](C)(O)[C@@H](OC(C)=O)[C@H](C)O3)[C@H](O)[C@@H](C)O2)[C@H](O)[C@@H](C)O1)C(=O)[C@@H](O)[C@@H](C)O ZYVSOIYQKUDENJ-ASUJBHBQSA-N 0.000 description 1
- SPJCRMJCFSJKDE-ZWBUGVOYSA-N [(3s,8s,9s,10r,13r,14s,17r)-10,13-dimethyl-17-[(2r)-6-methylheptan-2-yl]-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1h-cyclopenta[a]phenanthren-3-yl] 2-[4-[bis(2-chloroethyl)amino]phenyl]acetate Chemical compound O([C@@H]1CC2=CC[C@H]3[C@@H]4CC[C@@H]([C@]4(CC[C@@H]3[C@@]2(C)CC1)C)[C@H](C)CCCC(C)C)C(=O)CC1=CC=C(N(CCCl)CCCl)C=C1 SPJCRMJCFSJKDE-ZWBUGVOYSA-N 0.000 description 1
- IFJUINDAXYAPTO-UUBSBJJBSA-N [(8r,9s,13s,14s,17s)-17-[2-[4-[4-[bis(2-chloroethyl)amino]phenyl]butanoyloxy]acetyl]oxy-13-methyl-6,7,8,9,11,12,14,15,16,17-decahydrocyclopenta[a]phenanthren-3-yl] benzoate Chemical compound C([C@@H]1[C@@H](C2=CC=3)CC[C@]4([C@H]1CC[C@@H]4OC(=O)COC(=O)CCCC=1C=CC(=CC=1)N(CCCl)CCCl)C)CC2=CC=3OC(=O)C1=CC=CC=C1 IFJUINDAXYAPTO-UUBSBJJBSA-N 0.000 description 1
- IHGLINDYFMDHJG-UHFFFAOYSA-N [2-(4-methoxyphenyl)-3,4-dihydronaphthalen-1-yl]-[4-(2-pyrrolidin-1-ylethoxy)phenyl]methanone Chemical compound C1=CC(OC)=CC=C1C(CCC1=CC=CC=C11)=C1C(=O)C(C=C1)=CC=C1OCCN1CCCC1 IHGLINDYFMDHJG-UHFFFAOYSA-N 0.000 description 1
- XZSRRNFBEIOBDA-CFNBKWCHSA-N [2-[(2s,4s)-4-[(2r,4s,5s,6s)-4-amino-5-hydroxy-6-methyloxan-2-yl]oxy-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-3,4-dihydro-1h-tetracen-2-yl]-2-oxoethyl] 2,2-diethoxyacetate Chemical compound O([C@H]1C[C@](CC2=C(O)C=3C(=O)C4=CC=CC(OC)=C4C(=O)C=3C(O)=C21)(O)C(=O)COC(=O)C(OCC)OCC)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 XZSRRNFBEIOBDA-CFNBKWCHSA-N 0.000 description 1
- ZOZKYEHVNDEUCO-XUTVFYLZSA-N aceglatone Chemical compound O1C(=O)[C@H](OC(C)=O)[C@@H]2OC(=O)[C@@H](OC(=O)C)[C@@H]21 ZOZKYEHVNDEUCO-XUTVFYLZSA-N 0.000 description 1
- 229950002684 aceglatone Drugs 0.000 description 1
- 229930183665 actinomycin Natural products 0.000 description 1
- RJURFGZVJUQBHK-IIXSONLDSA-N actinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=CC=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 RJURFGZVJUQBHK-IIXSONLDSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000000488 activin Substances 0.000 description 1
- 229950004955 adozelesin Drugs 0.000 description 1
- BYRVKDUQDLJUBX-JJCDCTGGSA-N adozelesin Chemical compound C1=CC=C2OC(C(=O)NC=3C=C4C=C(NC4=CC=3)C(=O)N3C[C@H]4C[C@]44C5=C(C(C=C43)=O)NC=C5C)=CC2=C1 BYRVKDUQDLJUBX-JJCDCTGGSA-N 0.000 description 1
- 208000011341 adult acute respiratory distress syndrome Diseases 0.000 description 1
- 230000009824 affinity maturation Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 229940045714 alkyl sulfonate alkylating agent Drugs 0.000 description 1
- 150000008052 alkyl sulfonates Chemical class 0.000 description 1
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 108010050122 alpha 1-Antitrypsin Proteins 0.000 description 1
- 102000015395 alpha 1-Antitrypsin Human genes 0.000 description 1
- 229940024142 alpha 1-antitrypsin Drugs 0.000 description 1
- 229960000473 altretamine Drugs 0.000 description 1
- 229960003437 aminoglutethimide Drugs 0.000 description 1
- ROBVIMPUHSLWNV-UHFFFAOYSA-N aminoglutethimide Chemical compound C=1C=C(N)C=CC=1C1(CC)CCC(=O)NC1=O ROBVIMPUHSLWNV-UHFFFAOYSA-N 0.000 description 1
- 229960002749 aminolevulinic acid Drugs 0.000 description 1
- 229960003896 aminopterin Drugs 0.000 description 1
- 210000001691 amnion Anatomy 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 229960001220 amsacrine Drugs 0.000 description 1
- XCPGHVQEEXUHNC-UHFFFAOYSA-N amsacrine Chemical compound COC1=CC(NS(C)(=O)=O)=CC=C1NC1=C(C=CC=C2)C2=NC2=CC=CC=C12 XCPGHVQEEXUHNC-UHFFFAOYSA-N 0.000 description 1
- BBDAGFIXKZCXAH-CCXZUQQUSA-N ancitabine Chemical compound N=C1C=CN2[C@@H]3O[C@H](CO)[C@@H](O)[C@@H]3OC2=N1 BBDAGFIXKZCXAH-CCXZUQQUSA-N 0.000 description 1
- 229950000242 ancitabine Drugs 0.000 description 1
- 239000003098 androgen Substances 0.000 description 1
- 229940030486 androgens Drugs 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000002280 anti-androgenic effect Effects 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 230000001455 anti-clotting effect Effects 0.000 description 1
- 229940046836 anti-estrogen Drugs 0.000 description 1
- 230000001833 anti-estrogenic effect Effects 0.000 description 1
- 239000000868 anti-mullerian hormone Substances 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000000051 antiandrogen Substances 0.000 description 1
- 229940030495 antiandrogen sex hormone and modulator of the genital system Drugs 0.000 description 1
- 238000011319 anticancer therapy Methods 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000013059 antihormonal agent Substances 0.000 description 1
- 229940045687 antimetabolites folic acid analogs Drugs 0.000 description 1
- 150000008209 arabinosides Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 201000008937 atopic dermatitis Diseases 0.000 description 1
- 229960002756 azacitidine Drugs 0.000 description 1
- VSRXQHXAPYXROS-UHFFFAOYSA-N azanide;cyclobutane-1,1-dicarboxylic acid;platinum(2+) Chemical compound [NH2-].[NH2-].[Pt+2].OC(=O)C1(C(O)=O)CCC1 VSRXQHXAPYXROS-UHFFFAOYSA-N 0.000 description 1
- 229950011321 azaserine Drugs 0.000 description 1
- 150000001541 aziridines Chemical class 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 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 1
- 229960000997 bicalutamide Drugs 0.000 description 1
- 210000000013 bile duct Anatomy 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229950008548 bisantrene Drugs 0.000 description 1
- 229950006844 bizelesin Drugs 0.000 description 1
- 201000000053 blastoma Diseases 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 229960005520 bryostatin Drugs 0.000 description 1
- MJQUEDHRCUIRLF-TVIXENOKSA-N bryostatin 1 Chemical compound C([C@@H]1CC(/[C@@H]([C@@](C(C)(C)/C=C/2)(O)O1)OC(=O)/C=C/C=C/CCC)=C\C(=O)OC)[C@H]([C@@H](C)O)OC(=O)C[C@H](O)C[C@@H](O1)C[C@H](OC(C)=O)C(C)(C)[C@]1(O)C[C@@H]1C\C(=C\C(=O)OC)C[C@H]\2O1 MJQUEDHRCUIRLF-TVIXENOKSA-N 0.000 description 1
- MUIWQCKLQMOUAT-AKUNNTHJSA-N bryostatin 20 Natural products COC(=O)C=C1C[C@@]2(C)C[C@]3(O)O[C@](C)(C[C@@H](O)CC(=O)O[C@](C)(C[C@@]4(C)O[C@](O)(CC5=CC(=O)O[C@]45C)C(C)(C)C=C[C@@](C)(C1)O2)[C@@H](C)O)C[C@H](OC(=O)C(C)(C)C)C3(C)C MUIWQCKLQMOUAT-AKUNNTHJSA-N 0.000 description 1
- MBABCNBNDNGODA-LUVUIASKSA-N bullatacin Chemical compound O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@@H]1[C@@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@@H](O)CC=2C(O[C@@H](C)C=2)=O)CC1 MBABCNBNDNGODA-LUVUIASKSA-N 0.000 description 1
- 229960002092 busulfan Drugs 0.000 description 1
- 108700002839 cactinomycin Proteins 0.000 description 1
- 229950009908 cactinomycin Drugs 0.000 description 1
- 229960004015 calcitonin Drugs 0.000 description 1
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 1
- 229950009823 calusterone Drugs 0.000 description 1
- IVFYLRMMHVYGJH-PVPPCFLZSA-N calusterone Chemical compound C1C[C@]2(C)[C@](O)(C)CC[C@H]2[C@@H]2[C@@H](C)CC3=CC(=O)CC[C@]3(C)[C@H]21 IVFYLRMMHVYGJH-PVPPCFLZSA-N 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 229960004117 capecitabine Drugs 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229960004562 carboplatin Drugs 0.000 description 1
- 229960002115 carboquone Drugs 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 229960003261 carmofur Drugs 0.000 description 1
- 229960005243 carmustine Drugs 0.000 description 1
- 229950007509 carzelesin Drugs 0.000 description 1
- BBZDXMBRAFTCAA-AREMUKBSSA-N carzelesin Chemical compound C1=2NC=C(C)C=2C([C@H](CCl)CN2C(=O)C=3NC4=CC=C(C=C4C=3)NC(=O)C3=CC4=CC=C(C=C4O3)N(CC)CC)=C2C=C1OC(=O)NC1=CC=CC=C1 BBZDXMBRAFTCAA-AREMUKBSSA-N 0.000 description 1
- 108010047060 carzinophilin Proteins 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000022131 cell cycle Effects 0.000 description 1
- 230000006369 cell cycle progression Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 229950008249 chlornaphazine Drugs 0.000 description 1
- 229960001480 chlorozotocin Drugs 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000012085 chronic inflammatory response Effects 0.000 description 1
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 1
- 208000025302 chronic primary adrenal insufficiency Diseases 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
- 201000005890 congenital diaphragmatic hernia Diseases 0.000 description 1
- 230000000139 costimulatory effect Effects 0.000 description 1
- 239000013632 covalent dimer Substances 0.000 description 1
- 230000005574 cross-species transmission Effects 0.000 description 1
- 108010089438 cryptophycin 1 Proteins 0.000 description 1
- PSNOPSMXOBPNNV-VVCTWANISA-N cryptophycin 1 Chemical compound C1=C(Cl)C(OC)=CC=C1C[C@@H]1C(=O)NC[C@@H](C)C(=O)O[C@@H](CC(C)C)C(=O)O[C@H]([C@H](C)[C@@H]2[C@H](O2)C=2C=CC=CC=2)C/C=C/C(=O)N1 PSNOPSMXOBPNNV-VVCTWANISA-N 0.000 description 1
- 108010090203 cryptophycin 8 Proteins 0.000 description 1
- PSNOPSMXOBPNNV-UHFFFAOYSA-N cryptophycin-327 Natural products C1=C(Cl)C(OC)=CC=C1CC1C(=O)NCC(C)C(=O)OC(CC(C)C)C(=O)OC(C(C)C2C(O2)C=2C=CC=CC=2)CC=CC(=O)N1 PSNOPSMXOBPNNV-UHFFFAOYSA-N 0.000 description 1
- 238000003235 crystal violet staining Methods 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960000684 cytarabine Drugs 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 description 1
- 229960000640 dactinomycin Drugs 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 229960005052 demecolcine Drugs 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229950003913 detorubicin Drugs 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- WVYXNIXAMZOZFK-UHFFFAOYSA-N diaziquone Chemical compound O=C1C(NC(=O)OCC)=C(N2CC2)C(=O)C(NC(=O)OCC)=C1N1CC1 WVYXNIXAMZOZFK-UHFFFAOYSA-N 0.000 description 1
- 229950002389 diaziquone Drugs 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- 239000003534 dna topoisomerase inhibitor Substances 0.000 description 1
- AMRJKAQTDDKMCE-UHFFFAOYSA-N dolastatin Chemical compound CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)C)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 AMRJKAQTDDKMCE-UHFFFAOYSA-N 0.000 description 1
- 229930188854 dolastatin Natural products 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- ZWAOHEXOSAUJHY-ZIYNGMLESA-N doxifluridine Chemical compound O[C@@H]1[C@H](O)[C@@H](C)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ZWAOHEXOSAUJHY-ZIYNGMLESA-N 0.000 description 1
- 229950005454 doxifluridine Drugs 0.000 description 1
- NOTIQUSPUUHHEH-UXOVVSIBSA-N dromostanolone propionate Chemical compound C([C@@H]1CC2)C(=O)[C@H](C)C[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H](OC(=O)CC)[C@@]2(C)CC1 NOTIQUSPUUHHEH-UXOVVSIBSA-N 0.000 description 1
- 229950004683 drostanolone propionate Drugs 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 229960005501 duocarmycin Drugs 0.000 description 1
- VQNATVDKACXKTF-XELLLNAOSA-N duocarmycin Chemical compound COC1=C(OC)C(OC)=C2NC(C(=O)N3C4=CC(=O)C5=C([C@@]64C[C@@H]6C3)C=C(N5)C(=O)OC)=CC2=C1 VQNATVDKACXKTF-XELLLNAOSA-N 0.000 description 1
- 229930184221 duocarmycin Natural products 0.000 description 1
- AFMYMMXSQGUCBK-AKMKHHNQSA-N dynemicin a Chemical compound C1#C\C=C/C#C[C@@H]2NC(C=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C3)=C3[C@@]34O[C@]32[C@@H](C)C(C(O)=O)=C(OC)[C@H]41 AFMYMMXSQGUCBK-AKMKHHNQSA-N 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- VLCYCQAOQCDTCN-UHFFFAOYSA-N eflornithine Chemical compound NCCCC(N)(C(F)F)C(O)=O VLCYCQAOQCDTCN-UHFFFAOYSA-N 0.000 description 1
- 238000001211 electron capture detection Methods 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- XOPYFXBZMVTEJF-PDACKIITSA-N eleutherobin Chemical compound C(/[C@H]1[C@H](C(=CC[C@@H]1C(C)C)C)C[C@@H]([C@@]1(C)O[C@@]2(C=C1)OC)OC(=O)\C=C\C=1N=CN(C)C=1)=C2\CO[C@@H]1OC[C@@H](O)[C@@H](O)[C@@H]1OC(C)=O XOPYFXBZMVTEJF-PDACKIITSA-N 0.000 description 1
- XOPYFXBZMVTEJF-UHFFFAOYSA-N eleutherobin Natural products C1=CC2(OC)OC1(C)C(OC(=O)C=CC=1N=CN(C)C=1)CC(C(=CCC1C(C)C)C)C1C=C2COC1OCC(O)C(O)C1OC(C)=O XOPYFXBZMVTEJF-UHFFFAOYSA-N 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 229950000549 elliptinium acetate Drugs 0.000 description 1
- 201000008184 embryoma Diseases 0.000 description 1
- 206010014599 encephalitis Diseases 0.000 description 1
- 201000003914 endometrial carcinoma Diseases 0.000 description 1
- 210000004696 endometrium Anatomy 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 229950011487 enocitabine Drugs 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 229950002973 epitiostanol Drugs 0.000 description 1
- 229930013356 epothilone Natural products 0.000 description 1
- 150000003883 epothilone derivatives Chemical class 0.000 description 1
- 108700021358 erbB-1 Genes Proteins 0.000 description 1
- 229950002017 esorubicin Drugs 0.000 description 1
- ITSGNOIFAJAQHJ-BMFNZSJVSA-N esorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)C[C@H](C)O1 ITSGNOIFAJAQHJ-BMFNZSJVSA-N 0.000 description 1
- LJQQFQHBKUKHIS-WJHRIEJJSA-N esperamicin Chemical compound O1CC(NC(C)C)C(OC)CC1OC1C(O)C(NOC2OC(C)C(SC)C(O)C2)C(C)OC1OC1C(\C2=C/CSSSC)=C(NC(=O)OC)C(=O)C(OC3OC(C)C(O)C(OC(=O)C=4C(=CC(OC)=C(OC)C=4)NC(=O)C(=C)OC)C3)C2(O)C#C\C=C/C#C1 LJQQFQHBKUKHIS-WJHRIEJJSA-N 0.000 description 1
- 229960001842 estramustine Drugs 0.000 description 1
- FRPJXPJMRWBBIH-RBRWEJTLSA-N estramustine Chemical compound ClCCN(CCCl)C(=O)OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 FRPJXPJMRWBBIH-RBRWEJTLSA-N 0.000 description 1
- 239000000328 estrogen antagonist Substances 0.000 description 1
- QSRLNKCNOLVZIR-KRWDZBQOSA-N ethyl (2s)-2-[[2-[4-[bis(2-chloroethyl)amino]phenyl]acetyl]amino]-4-methylsulfanylbutanoate Chemical compound CCOC(=O)[C@H](CCSC)NC(=O)CC1=CC=C(N(CCCl)CCCl)C=C1 QSRLNKCNOLVZIR-KRWDZBQOSA-N 0.000 description 1
- 229960005237 etoglucid Drugs 0.000 description 1
- 229960005420 etoposide Drugs 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 229940043168 fareston Drugs 0.000 description 1
- 108010052621 fas Receptor Proteins 0.000 description 1
- 102000018823 fas Receptor Human genes 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 229940126864 fibroblast growth factor Drugs 0.000 description 1
- 229960000961 floxuridine Drugs 0.000 description 1
- ODKNJVUHOIMIIZ-RRKCRQDMSA-N floxuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(F)=C1 ODKNJVUHOIMIIZ-RRKCRQDMSA-N 0.000 description 1
- XRECTZIEBJDKEO-UHFFFAOYSA-N flucytosine Chemical compound NC1=NC(=O)NC=C1F XRECTZIEBJDKEO-UHFFFAOYSA-N 0.000 description 1
- 229960004413 flucytosine Drugs 0.000 description 1
- 229960000390 fludarabine Drugs 0.000 description 1
- GIUYCYHIANZCFB-FJFJXFQQSA-N fludarabine phosphate Chemical compound C1=NC=2C(N)=NC(F)=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@@H]1O GIUYCYHIANZCFB-FJFJXFQQSA-N 0.000 description 1
- MKXKFYHWDHIYRV-UHFFFAOYSA-N flutamide Chemical compound CC(C)C(=O)NC1=CC=C([N+]([O-])=O)C(C(F)(F)F)=C1 MKXKFYHWDHIYRV-UHFFFAOYSA-N 0.000 description 1
- 229960002074 flutamide Drugs 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 150000002224 folic acids Chemical class 0.000 description 1
- 230000003325 follicular Effects 0.000 description 1
- 210000000285 follicular dendritic cell Anatomy 0.000 description 1
- 229960004783 fotemustine Drugs 0.000 description 1
- YAKWPXVTIGTRJH-UHFFFAOYSA-N fotemustine Chemical compound CCOP(=O)(OCC)C(C)NC(=O)N(CCCl)N=O YAKWPXVTIGTRJH-UHFFFAOYSA-N 0.000 description 1
- 230000005714 functional activity Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 229940044658 gallium nitrate Drugs 0.000 description 1
- 108010089491 gamma-heregulin Proteins 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000000762 glandular Effects 0.000 description 1
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 1
- 229960004666 glucagon Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 239000002622 gonadotropin Substances 0.000 description 1
- 229960002913 goserelin Drugs 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 201000010536 head and neck cancer Diseases 0.000 description 1
- 210000003709 heart valve Anatomy 0.000 description 1
- 208000007475 hemolytic anemia Diseases 0.000 description 1
- 230000011132 hemopoiesis Effects 0.000 description 1
- 108010034429 heregulin alpha Proteins 0.000 description 1
- 238000005734 heterodimerization reaction Methods 0.000 description 1
- UUVWYPNAQBNQJQ-UHFFFAOYSA-N hexamethylmelamine Chemical compound CN(C)C1=NC(N(C)C)=NC(N(C)C)=N1 UUVWYPNAQBNQJQ-UHFFFAOYSA-N 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 239000000710 homodimer Substances 0.000 description 1
- 102000054751 human RUNX1T1 Human genes 0.000 description 1
- 102000048638 human UQCRH Human genes 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 230000008348 humoral response Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 229960001330 hydroxycarbamide Drugs 0.000 description 1
- 230000002267 hypothalamic effect Effects 0.000 description 1
- 229940015872 ibandronate Drugs 0.000 description 1
- 229960000908 idarubicin Drugs 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- DBIGHPPNXATHOF-UHFFFAOYSA-N improsulfan Chemical compound CS(=O)(=O)OCCCNCCCOS(C)(=O)=O DBIGHPPNXATHOF-UHFFFAOYSA-N 0.000 description 1
- 229950008097 improsulfan Drugs 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006882 induction of apoptosis Effects 0.000 description 1
- 239000012678 infectious agent Substances 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 239000000893 inhibin Substances 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- UWKQSNNFCGGAFS-XIFFEERXSA-N irinotecan Chemical compound C1=C2C(CC)=C3CN(C(C4=C([C@@](C(=O)OC4)(O)CC)C=4)=O)C=4C3=NC2=CC=C1OC(=O)N(CC1)CCC1N1CCCCC1 UWKQSNNFCGGAFS-XIFFEERXSA-N 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 229940115286 lentinan Drugs 0.000 description 1
- GFIJNRVAKGFPGQ-LIJARHBVSA-N leuprolide Chemical compound CCNC(=O)[C@@H]1CCCN1C(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1N=CNC=1)NC(=O)[C@H]1NC(=O)CC1)CC1=CC=C(O)C=C1 GFIJNRVAKGFPGQ-LIJARHBVSA-N 0.000 description 1
- 229960004338 leuprorelin Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229960002247 lomustine Drugs 0.000 description 1
- 229960003538 lonidamine Drugs 0.000 description 1
- WDRYRZXSPDWGEB-UHFFFAOYSA-N lonidamine Chemical compound C12=CC=CC=C2C(C(=O)O)=NN1CC1=CC=C(Cl)C=C1Cl WDRYRZXSPDWGEB-UHFFFAOYSA-N 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 210000005210 lymphoid organ Anatomy 0.000 description 1
- 230000002934 lysing effect Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 208000026037 malignant tumor of neck Diseases 0.000 description 1
- 229950008612 mannomustine Drugs 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- WKPWGQKGSOKKOO-RSFHAFMBSA-N maytansine Chemical compound CO[C@@H]([C@@]1(O)C[C@](OC(=O)N1)([C@H]([C@@H]1O[C@@]1(C)[C@@H](OC(=O)[C@H](C)N(C)C(C)=O)CC(=O)N1C)C)[H])\C=C\C=C(C)\CC2=CC(OC)=C(Cl)C1=C2 WKPWGQKGSOKKOO-RSFHAFMBSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 229960001924 melphalan Drugs 0.000 description 1
- SGDBTWWWUNNDEQ-LBPRGKRZSA-N melphalan Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N(CCCl)CCCl)C=C1 SGDBTWWWUNNDEQ-LBPRGKRZSA-N 0.000 description 1
- 108020004084 membrane receptors Proteins 0.000 description 1
- 229950009246 mepitiostane Drugs 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 1
- VJRAUFKOOPNFIQ-TVEKBUMESA-N methyl (1r,2r,4s)-4-[(2r,4s,5s,6s)-5-[(2s,4s,5s,6s)-5-[(2s,4s,5s,6s)-4,5-dihydroxy-6-methyloxan-2-yl]oxy-4-hydroxy-6-methyloxan-2-yl]oxy-4-(dimethylamino)-6-methyloxan-2-yl]oxy-2-ethyl-2,5,7,10-tetrahydroxy-6,11-dioxo-3,4-dihydro-1h-tetracene-1-carboxylat Chemical compound O([C@H]1[C@@H](O)C[C@@H](O[C@H]1C)O[C@H]1[C@H](C[C@@H](O[C@H]1C)O[C@H]1C[C@]([C@@H](C2=CC=3C(=O)C4=C(O)C=CC(O)=C4C(=O)C=3C(O)=C21)C(=O)OC)(O)CC)N(C)C)[C@H]1C[C@H](O)[C@H](O)[C@H](C)O1 VJRAUFKOOPNFIQ-TVEKBUMESA-N 0.000 description 1
- QRMNENFZDDYDEF-GOSISDBHSA-N methyl (8s)-8-(bromomethyl)-2-methyl-4-(4-methylpiperazine-1-carbonyl)oxy-6-(5,6,7-trimethoxy-1h-indole-2-carbonyl)-7,8-dihydro-3h-pyrrolo[3,2-e]indole-1-carboxylate Chemical compound C1([C@H](CBr)CN(C1=C1)C(=O)C=2NC3=C(OC)C(OC)=C(OC)C=C3C=2)=C2C(C(=O)OC)=C(C)NC2=C1OC(=O)N1CCN(C)CC1 QRMNENFZDDYDEF-GOSISDBHSA-N 0.000 description 1
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 description 1
- 229960005485 mitobronitol Drugs 0.000 description 1
- 229960003539 mitoguazone Drugs 0.000 description 1
- MXWHMTNPTTVWDM-NXOFHUPFSA-N mitoguazone Chemical compound NC(N)=N\N=C(/C)\C=N\N=C(N)N MXWHMTNPTTVWDM-NXOFHUPFSA-N 0.000 description 1
- VFKZTMPDYBFSTM-GUCUJZIJSA-N mitolactol Chemical compound BrC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CBr VFKZTMPDYBFSTM-GUCUJZIJSA-N 0.000 description 1
- 229950010913 mitolactol Drugs 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 229960000350 mitotane Drugs 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- FOYWNSCCNCUEPU-UHFFFAOYSA-N mopidamol Chemical compound C12=NC(N(CCO)CCO)=NC=C2N=C(N(CCO)CCO)N=C1N1CCCCC1 FOYWNSCCNCUEPU-UHFFFAOYSA-N 0.000 description 1
- 229950010718 mopidamol Drugs 0.000 description 1
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 1
- 208000037890 multiple organ injury Diseases 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 206010028417 myasthenia gravis Diseases 0.000 description 1
- 229960000951 mycophenolic acid Drugs 0.000 description 1
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 208000009091 myxoma Diseases 0.000 description 1
- NJSMWLQOCQIOPE-OCHFTUDZSA-N n-[(e)-[10-[(e)-(4,5-dihydro-1h-imidazol-2-ylhydrazinylidene)methyl]anthracen-9-yl]methylideneamino]-4,5-dihydro-1h-imidazol-2-amine Chemical compound N1CCN=C1N\N=C\C(C1=CC=CC=C11)=C(C=CC=C2)C2=C1\C=N\NC1=NCCN1 NJSMWLQOCQIOPE-OCHFTUDZSA-N 0.000 description 1
- 210000004897 n-terminal region Anatomy 0.000 description 1
- 229940086322 navelbine Drugs 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 201000008383 nephritis Diseases 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- XWXYUMMDTVBTOU-UHFFFAOYSA-N nilutamide Chemical compound O=C1C(C)(C)NC(=O)N1C1=CC=C([N+]([O-])=O)C(C(F)(F)F)=C1 XWXYUMMDTVBTOU-UHFFFAOYSA-N 0.000 description 1
- 229960002653 nilutamide Drugs 0.000 description 1
- 229960001420 nimustine Drugs 0.000 description 1
- VFEDRRNHLBGPNN-UHFFFAOYSA-N nimustine Chemical compound CC1=NC=C(CNC(=O)N(CCCl)N=O)C(N)=N1 VFEDRRNHLBGPNN-UHFFFAOYSA-N 0.000 description 1
- YMVWGSQGCWCDGW-UHFFFAOYSA-N nitracrine Chemical compound C1=CC([N+]([O-])=O)=C2C(NCCCN(C)C)=C(C=CC=C3)C3=NC2=C1 YMVWGSQGCWCDGW-UHFFFAOYSA-N 0.000 description 1
- 229950008607 nitracrine Drugs 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- KGTDRFCXGRULNK-JYOBTZKQSA-N nogalamycin Chemical compound CO[C@@H]1[C@@](OC)(C)[C@@H](OC)[C@H](C)O[C@H]1O[C@@H]1C2=C(O)C(C(=O)C3=C(O)C=C4[C@@]5(C)O[C@H]([C@H]([C@@H]([C@H]5O)N(C)C)O)OC4=C3C3=O)=C3C=C2[C@@H](C(=O)OC)[C@@](C)(O)C1 KGTDRFCXGRULNK-JYOBTZKQSA-N 0.000 description 1
- 229950009266 nogalamycin Drugs 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- CZDBNBLGZNWKMC-MWQNXGTOSA-N olivomycin Chemical class O([C@@H]1C[C@@H](O[C@H](C)[C@@H]1O)OC=1C=C2C=C3C[C@H]([C@@H](C(=O)C3=C(O)C2=C(O)C=1)O[C@H]1O[C@@H](C)[C@H](O)[C@@H](OC2O[C@@H](C)[C@H](O)[C@@H](O)C2)C1)[C@H](OC)C(=O)[C@@H](O)[C@@H](C)O)[C@H]1C[C@H](O)[C@H](OC)[C@H](C)O1 CZDBNBLGZNWKMC-MWQNXGTOSA-N 0.000 description 1
- 229950011093 onapristone Drugs 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- 230000002138 osteoinductive effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 210000002741 palatine tonsil Anatomy 0.000 description 1
- VREZDOWOLGNDPW-UHFFFAOYSA-N pancratistatine Natural products C1=C2C3C(O)C(O)C(O)C(O)C3NC(=O)C2=C(O)C2=C1OCO2 VREZDOWOLGNDPW-UHFFFAOYSA-N 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 229960002340 pentostatin Drugs 0.000 description 1
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 1
- QIMGFXOHTOXMQP-GFAGFCTOSA-N peplomycin 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)NCCCN[C@@H](C)C=1C=CC=CC=1)[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=1NC=NC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C QIMGFXOHTOXMQP-GFAGFCTOSA-N 0.000 description 1
- 229950003180 peplomycin Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 201000002628 peritoneum cancer Diseases 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000000865 phosphorylative effect Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229960000952 pipobroman Drugs 0.000 description 1
- NJBFOOCLYDNZJN-UHFFFAOYSA-N pipobroman Chemical compound BrCCC(=O)N1CCN(C(=O)CCBr)CC1 NJBFOOCLYDNZJN-UHFFFAOYSA-N 0.000 description 1
- NUKCGLDCWQXYOQ-UHFFFAOYSA-N piposulfan Chemical compound CS(=O)(=O)OCCC(=O)N1CCN(C(=O)CCOS(C)(=O)=O)CC1 NUKCGLDCWQXYOQ-UHFFFAOYSA-N 0.000 description 1
- 229950001100 piposulfan Drugs 0.000 description 1
- 229960001221 pirarubicin Drugs 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 150000003057 platinum Chemical class 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 208000005987 polymyositis Diseases 0.000 description 1
- 230000007824 polyneuropathy Effects 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 229960004694 prednimustine Drugs 0.000 description 1
- XOFYZVNMUHMLCC-ZPOLXVRWSA-N prednisone Chemical compound O=C1C=C[C@]2(C)[C@H]3C(=O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 XOFYZVNMUHMLCC-ZPOLXVRWSA-N 0.000 description 1
- 229960004618 prednisone Drugs 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- CPTBDICYNRMXFX-UHFFFAOYSA-N procarbazine Chemical compound CNNCC1=CC=C(C(=O)NC(C)C)C=C1 CPTBDICYNRMXFX-UHFFFAOYSA-N 0.000 description 1
- 229960000624 procarbazine Drugs 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 229940097325 prolactin Drugs 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 108010087851 prorelaxin Proteins 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- WOLQREOUPKZMEX-UHFFFAOYSA-N pteroyltriglutamic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(=O)NC(CCC(=O)NC(CCC(O)=O)C(O)=O)C(O)=O)C(O)=O)C=C1 WOLQREOUPKZMEX-UHFFFAOYSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 229950010131 puromycin Drugs 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 229960002185 ranimustine Drugs 0.000 description 1
- BMKDZUISNHGIBY-UHFFFAOYSA-N razoxane Chemical compound C1C(=O)NC(=O)CN1C(C)CN1CC(=O)NC(=O)C1 BMKDZUISNHGIBY-UHFFFAOYSA-N 0.000 description 1
- 229960000460 razoxane Drugs 0.000 description 1
- 208000002574 reactive arthritis Diseases 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000010188 recombinant method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000026267 regulation of growth Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 229930002330 retinoic acid Natural products 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- OWPCHSCAPHNHAV-LMONGJCWSA-N rhizoxin Chemical compound C/C([C@H](OC)[C@@H](C)[C@@H]1C[C@H](O)[C@]2(C)O[C@@H]2/C=C/[C@@H](C)[C@]2([H])OC(=O)C[C@@](C2)(C[C@@H]2O[C@H]2C(=O)O1)[H])=C\C=C\C(\C)=C\C1=COC(C)=N1 OWPCHSCAPHNHAV-LMONGJCWSA-N 0.000 description 1
- 229950004892 rodorubicin Drugs 0.000 description 1
- MBABCNBNDNGODA-WPZDJQSSSA-N rolliniastatin 1 Natural products O1[C@@H]([C@@H](O)CCCCCCCCCC)CC[C@H]1[C@H]1O[C@@H]([C@H](O)CCCCCCCCCC[C@@H](O)CC=2C(O[C@@H](C)C=2)=O)CC1 MBABCNBNDNGODA-WPZDJQSSSA-N 0.000 description 1
- IMUQLZLGWJSVMV-UOBFQKKOSA-N roridin A Natural products CC(O)C1OCCC(C)C(O)C(=O)OCC2CC(=CC3OC4CC(OC(=O)C=C/C=C/1)C(C)(C23)C45CO5)C IMUQLZLGWJSVMV-UOBFQKKOSA-N 0.000 description 1
- VHXNKPBCCMUMSW-FQEVSTJZSA-N rubitecan Chemical compound C1=CC([N+]([O-])=O)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VHXNKPBCCMUMSW-FQEVSTJZSA-N 0.000 description 1
- 201000003804 salivary gland carcinoma Diseases 0.000 description 1
- 229930182947 sarcodictyin Natural products 0.000 description 1
- 201000000306 sarcoidosis Diseases 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 102000035025 signaling receptors Human genes 0.000 description 1
- 108091005475 signaling receptors Proteins 0.000 description 1
- 208000000587 small cell lung carcinoma Diseases 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 229950006315 spirogermanium Drugs 0.000 description 1
- 210000004988 splenocyte Anatomy 0.000 description 1
- ICXJVZHDZFXYQC-UHFFFAOYSA-N spongistatin 1 Natural products OC1C(O2)(O)CC(O)C(C)C2CCCC=CC(O2)CC(O)CC2(O2)CC(OC)CC2CC(=O)C(C)C(OC(C)=O)C(C)C(=C)CC(O2)CC(C)(O)CC2(O2)CC(OC(C)=O)CC2CC(=O)OC2C(O)C(CC(=C)CC(O)C=CC(Cl)=C)OC1C2C ICXJVZHDZFXYQC-UHFFFAOYSA-N 0.000 description 1
- 208000017572 squamous cell neoplasm Diseases 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 229960001052 streptozocin Drugs 0.000 description 1
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 1
- 230000004960 subcellular localization Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 229940063683 taxotere Drugs 0.000 description 1
- 206010043207 temporal arteritis Diseases 0.000 description 1
- NRUKOCRGYNPUPR-QBPJDGROSA-N teniposide Chemical compound COC1=C(O)C(OC)=CC([C@@H]2C3=CC=4OCOC=4C=C3[C@@H](O[C@H]3[C@@H]([C@@H](O)[C@@H]4O[C@@H](OC[C@H]4O3)C=3SC=CC=3)O)[C@@H]3[C@@H]2C(OC3)=O)=C1 NRUKOCRGYNPUPR-QBPJDGROSA-N 0.000 description 1
- 229960001278 teniposide Drugs 0.000 description 1
- 229960005353 testolactone Drugs 0.000 description 1
- BPEWUONYVDABNZ-DZBHQSCQSA-N testolactone Chemical compound O=C1C=C[C@]2(C)[C@H]3CC[C@](C)(OC(=O)CC4)[C@@H]4[C@@H]3CCC2=C1 BPEWUONYVDABNZ-DZBHQSCQSA-N 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 229940034208 thyroxine Drugs 0.000 description 1
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 description 1
- YFTWHEBLORWGNI-UHFFFAOYSA-N tiamiprine Chemical compound CN1C=NC([N+]([O-])=O)=C1SC1=NC(N)=NC2=C1NC=N2 YFTWHEBLORWGNI-UHFFFAOYSA-N 0.000 description 1
- 229950011457 tiamiprine Drugs 0.000 description 1
- 229940044693 topoisomerase inhibitor Drugs 0.000 description 1
- 229960000303 topotecan Drugs 0.000 description 1
- UCFGDBYHRUNTLO-QHCPKHFHSA-N topotecan Chemical compound C1=C(O)C(CN(C)C)=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 UCFGDBYHRUNTLO-QHCPKHFHSA-N 0.000 description 1
- XFCLJVABOIYOMF-QPLCGJKRSA-N toremifene Chemical compound C1=CC(OCCN(C)C)=CC=C1C(\C=1C=CC=CC=1)=C(\CCCl)C1=CC=CC=C1 XFCLJVABOIYOMF-QPLCGJKRSA-N 0.000 description 1
- 229960005026 toremifene Drugs 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 102000027257 transmembrane receptors Human genes 0.000 description 1
- 108091008578 transmembrane receptors Proteins 0.000 description 1
- 229950001353 tretamine Drugs 0.000 description 1
- IUCJMVBFZDHPDX-UHFFFAOYSA-N tretamine Chemical compound C1CN1C1=NC(N2CC2)=NC(N2CC2)=N1 IUCJMVBFZDHPDX-UHFFFAOYSA-N 0.000 description 1
- 229960001727 tretinoin Drugs 0.000 description 1
- PXSOHRWMIRDKMP-UHFFFAOYSA-N triaziquone Chemical compound O=C1C(N2CC2)=C(N2CC2)C(=O)C=C1N1CC1 PXSOHRWMIRDKMP-UHFFFAOYSA-N 0.000 description 1
- 229960004560 triaziquone Drugs 0.000 description 1
- 229930013292 trichothecene Natural products 0.000 description 1
- 150000003327 trichothecene derivatives Chemical class 0.000 description 1
- 229960001670 trilostane Drugs 0.000 description 1
- KVJXBPDAXMEYOA-CXANFOAXSA-N trilostane Chemical compound OC1=C(C#N)C[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@@]32O[C@@H]31 KVJXBPDAXMEYOA-CXANFOAXSA-N 0.000 description 1
- 239000013638 trimer Substances 0.000 description 1
- NOYPYLRCIDNJJB-UHFFFAOYSA-N trimetrexate Chemical compound COC1=C(OC)C(OC)=CC(NCC=2C(=C3C(N)=NC(N)=NC3=CC=2)C)=C1 NOYPYLRCIDNJJB-UHFFFAOYSA-N 0.000 description 1
- 229960001099 trimetrexate Drugs 0.000 description 1
- 229950000212 trioxifene Drugs 0.000 description 1
- 229960000875 trofosfamide Drugs 0.000 description 1
- UMKFEPPTGMDVMI-UHFFFAOYSA-N trofosfamide Chemical compound ClCCN(CCCl)P1(=O)OCCCN1CCCl UMKFEPPTGMDVMI-UHFFFAOYSA-N 0.000 description 1
- HDZZVAMISRMYHH-LITAXDCLSA-N tubercidin Chemical compound C1=CC=2C(N)=NC=NC=2N1[C@@H]1O[C@@H](CO)[C@H](O)[C@H]1O HDZZVAMISRMYHH-LITAXDCLSA-N 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 229950009811 ubenimex Drugs 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 229960001055 uracil mustard Drugs 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 208000012991 uterine carcinoma Diseases 0.000 description 1
- HHJUWIANJFBDHT-KOTLKJBCSA-N vindesine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(N)=O)N4C)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 HHJUWIANJFBDHT-KOTLKJBCSA-N 0.000 description 1
- 229960004355 vindesine Drugs 0.000 description 1
- GBABOYUKABKIAF-IELIFDKJSA-N vinorelbine Chemical compound C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC GBABOYUKABKIAF-IELIFDKJSA-N 0.000 description 1
- 229960002066 vinorelbine Drugs 0.000 description 1
- CILBMBUYJCWATM-PYGJLNRPSA-N vinorelbine ditartrate Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O.OC(=O)[C@H](O)[C@@H](O)C(O)=O.C1N(CC=2C3=CC=CC=C3NC=22)CC(CC)=C[C@H]1C[C@]2(C(=O)OC)C1=CC([C@]23[C@H]([C@@]([C@H](OC(C)=O)[C@]4(CC)C=CCN([C@H]34)CC2)(O)C(=O)OC)N2C)=C2C=C1OC CILBMBUYJCWATM-PYGJLNRPSA-N 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 201000005102 vulva cancer Diseases 0.000 description 1
- 229940053867 xeloda Drugs 0.000 description 1
- 229950009268 zinostatin Drugs 0.000 description 1
- 229960000641 zorubicin Drugs 0.000 description 1
- FBTUMDXHSRTGRV-ALTNURHMSA-N zorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(\C)=N\NC(=O)C=1C=CC=CC=1)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 FBTUMDXHSRTGRV-ALTNURHMSA-N 0.000 description 1
- 150000003952 β-lactams Chemical class 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- 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
-
- 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
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6875—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody being a hybrid immunoglobulin
- A61K47/6879—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody being a hybrid immunoglobulin the immunoglobulin having two or more different antigen-binding sites, e.g. bispecific or multispecific immunoglobulin
-
- 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
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- 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
- C07K16/2878—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
- C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against 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
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/64—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a combination of variable region and constant region components
-
- 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
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
-
- 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
- C07K2317/77—Internalization into the cell
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Definitions
- the present invention concerns engineered antibodies, with three or more functional antigen binding sites, and uses, such as therapeutic uses, for such engineered antibodies.
- Naturally occurring antibodies comprise two heavy chains linked together by disulfide bonds and two light chains, one light chain being linked to each of the heavy chains by disulfide bonds.
- Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains (three or four constant domains, CH1, CH2, CH3 and CH4, depending on the antibody class).
- Each light chain has a variable domain (VL) at one end and a constant domain (CL) at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. See FIG. 1 herein.
- variable domains are not involved directly in binding the antibody to an antigen, but are involved in various effector functions, such as participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC).
- the variable domains of each pair of light and heavy chains are involved directly in binding the antibody to the antigen.
- the variable domains of naturally occurring light and heavy chains have the same general structure; each comprising four framework regions (FRs), whose sequences are somewhat conserved, connected by three complementarity determining regions (CDRs) (see Kabat et al., Sequences of Proteins of Immunological Interest , National Institutes of Health, Bethesda, Md., (1991)).
- the four FRs largely adopt a beta-sheet conformation and the CDRs form loops connecting, and in some cases forming part of, the beta-sheet structure.
- the CDRs in each chain are held in close proximity by the FRs and, with the CDRs from the other chain, contribute to the formation of the antigen binding site.
- FIGS. 2A-E herein depict the structures of the five major naturally occurring immunoglobulin isotypes.
- IgG, IgD and IgE immunoglobulins possess only two antigen binding sites.
- IgA and IgM are capable of forming polymeric structures with higher valencies.
- IgM is secreted by plasma cells as a pentamer in which five monomer units are held together by disulfide bonds linking their carboxyl-terminal (CIA/CO) domains and C ⁇ 3/C ⁇ 3 domains.
- the five monomer subunits are arranged with their Fc regions in the center of the pentamer and the 10 antigen-binding sites on the periphery of the molecule.
- Each pentamer contains an additional Fc-linked polypeptide called the J (joining) chain, which is disulfide-bonded to the carboxyl-terminal cysteine residue of 2 of the 10 ⁇ chains.
- the J chain appears to be required for polymerization of the monomers to form pentameric IgM; it is added just before secretion of the pentamer.
- An IgM molecule can bind 10 small hapten molecules; however, because of steric hindrance, only 5 molecules of larger antigens can be bound simultaneously.
- the increased valency of pentameric IgM increases its capacity to bind such multi-dimensional antigens as viral particles and red blood cells (RBCs).
- IgA exists primarily as a monomer, although polymeric forms such as dimers, trimers, and even tetramers are sometimes seen.
- the IgA of external secretions consists of a dimer or tetramer, a J-chain polypeptide, and a polypeptide chain called secretory component.
- the art has attempted to overcome this problem by constructing “chimeric” antibodies in which an animal antigen binding variable domain is coupled to a human constant domain (Cabilly et al., U.S. Pat. No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984); Boulianne et al., Nature 312:643-646 (1984); and Neuberger et al., Nature 314:268-270 (1985)).
- the isotype of the human constant domain may be selected to tailor the chimeric antibody for participation in ADCC and CDC (see e.g. Bruggemann et al., J. Exp. Med.
- such chimeric antibodies contain about one third rodent (or other non-human species) sequence and thus are capable of eliciting a significant anti-globulin response in humans.
- rodent or other non-human species
- OKT3 much of the resulting anti-globulin response is directed against the variable region rather than the constant region (Jeffers et al., Transplantation 41:572-578 (1986)).
- substituting CDRs from rodent antibodies for the human CDRs in human frameworks is sufficient to transfer high antigen binding affinity (Jones et al., Nature 321:522-525 (1986); Verhoeyen et al., Science 239:1534-1536 (1988)), whereas in other cases it has been necessary to additionally replace one (Riechmann et al., Nature 332:323-327 (1988)) or several (Queen et al., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989)) framework residues. See also Co et al., Proc. Natl. Acad. Sci. USA 88:2869-2873 (1991); U.S. Pat.
- a chimeric/humanized antibody instead of a chimeric/humanized antibody, one may treat a patient with a human antibody in order to avoid human antibodies raised against a murine antibody (known as the “HAMA response”).
- HAMA response human antibodies raised against a murine antibody
- Several technologies are available for generating human antibodies.
- Human antibodies may be selected using phage display technology. Phage display has been adapted to select human antibodies from an unimmunized donor (Marks et al. J. Mol. Biol. 222:581-597 (1991)). According to this approach, PCR is used to amplify variable domain genes from mRNA prepared from human peripheral blood lymphocytes (PBLs). Primers are used such that DNA from both IgG and IgM heavy chains and both ⁇ and ⁇ chains is amplified. These genes are then randomly combined and expressed as single chain Fv (scFv) fused to the gene III coat protein of M13 phage. Human antibodies against an antigen of interest may then be identified by rounds of growth and selection by binding to that antigen (e.g. to the immobilized antigen). See Griffiths et al. EMBO J. 12:725-734 (1993).
- “Synthetic” phage-antibody repertoires have also been built from cloned human VH-gene segments.
- a repertoire (2 ⁇ 10 7 clones) was first constructed using a short H3 loop of five or eight random residues with each of 49 segments, and combined with a fixed light chain (Hoogenboom et al. J. Mol. Biol. 227:381-388 (1992)).
- H3 loops of different lengths, up to 12 residues, a single library was created from which a range of more than 20 binding specificities could be selected (Winter et al. Ann. Rev. Immuno. 12:433-55 (1994)).
- the affinity of low affinity “primary” phage-antibodies may be improved by using phage display technology.
- One approach is to use a chain-shuffling strategy in which the VH domain is held constant and then recombined with the original library of VL genes and tighter binders selected by binding to immobilized antigen. This cycle is repeated by fixing the new VL domain and recombining with the original VH library (Marks et al. Bio/Technology 10:779-783 (1992)).
- point mutations in the primary antibody may be introduced using error-prone PCR and higher affinity binders selected by using phage display. Gram et al. PNAS ( USA ) 89: 3576-3580 (1992).
- Severe combined immune deficient (SCID) mice lack the ability to produce their own immunoglobulins due to a defect in the recombinase gene.
- SCID mice Several groups have reconstituted a functional humoral immune system in these mice by transfer of human peripheral blood lymphocytes (PBLs). These hu-PBL-SCID mice can be used to raise human antibodies upon immunization with antigen. Duchosal et al. Nature 355:258-262 (1992).
- mice are turned off and then yeast artificial chromosomes (YACs) engineered with large DNA sequences containing human heavy- and light-chain genes are introduced into the mice.
- yeast artificial chromosomes YACs
- Such “XenoMice” are able to produce human antibodies upon immunization with an antigen of interest. See U.S. Pat. No. 5,434,340; U.S. Pat. No. 5,591,699; U.S. Pat. No. 5,569,825; U.S. Pat. No. 5,545,806; and U.S. Pat. No. 5,545,807.
- Human monoclonal antibodies may also be generated by immortalizing a human B lymphocyte producing an antibody of interest.
- the ethical issues surrounding immunizing humans in order to generate activated human B lymphocytes can be avoided by immunizing human lymphocytes in vitro.
- Both human PBLs (Borrebaeck et al. Proc. Natl. Acad. ScL USA 85:3995-4000 (1988)) and human splenocytes (Boerner et al. J. Immunol. 147, 86-95 (1991)) have been successfully immunized in vitro. Improvements in human hybridoma technology have been achieved by using a mouse-human heterohybrid as the fusion partner (Boerner et al.).
- Antibodies have been modified in order to increase their antigen-binding valency. For instance, Ghetie et al. homodimerized tumor-reactive monoclonal antibodies (anti-CD19, anti-CD20, anti-CD21, anti-CD22 and anti-HER2 antibodies) by chemically introducing a thioether bond between a pair of IgGs using two heterobifunctional crosslinkers . Ghetie et al. PNAS ( USA ) 94:7509-7514 (1997); and WO 99/02567. Wolff et al.
- the homodimeric IgG was said to have similar avidity to the parent IgG, but apparently showed an improved ability to internalize and retain radioisotope in target leukemia cells, and was more potent at complement-mediated leukemia cell killing and antibody-dependent cellular cytotoxicity using human effectors.
- Coloma and Morrison Nature Biotech. 15: 159-163 describe a tetravalent bispecific antibody which was engineered by fusing DNA encoding a single chain anti-dansyl antibody Fv (scFv) after the C terminus (CH3-scFv) or after the hinge (Hinge-scFv) of an IgG3 anti-dansyl antibody. See, also, WO95/09917. Smith and Morrison engineered three versions of mu-like IgG3 by engineering either (1) Cys414 of an IgM heavy chain or (2) Cys575 of an IgM heavy chain, or both (1) and (2), into the IgG3 heavy chain gene.
- U.S. Pat. No. 5,641,870 (Rinderknecht et al.) describes a bivalent, linear F(ab′) 2 fragment comprising tandem repeats of a heavy chain fragment (VH-CH1-VH-CH1) cosecreted with a light chain.
- the C-terminus of CH1 was joined directed to the N-terminus of VH without any extraneous linking protein sequences.
- the ErbB receptor tyrosine kinases are important mediators of cell growth, differentiation and survival.
- the receptor family includes at least four distinct members including Epidermal Growth Factor Receptor (EGFR or ErbB1), HER2 (ErbB2 or p185 neu ), HER3 (ErbB3) and HER4 (ErbB4 or tyro2).
- EGFR Epidermal Growth Factor Receptor
- HER2 ErbB2 or p185 neu
- HER3 ErbB3
- HER4 ErbB4 or tyro2
- EGFR encoded by the erbB1 gene
- increased expression of EGFR has been observed in breast, bladder, lung, head, neck and stomach cancer, as well as glioblastomas.
- Increased EGFR receptor expression is often associated with increased production of the EGFR ligand, Transforming Growth Factor alpha (TGF-alpha), by the same tumor cells resulting in receptor activation by an autocrine stimulatory pathway.
- TGF-alpha Transforming Growth Factor alpha
- Monoclonal antibodies directed against the EGFR or its ligands, TGF-alpha and EGF have been evaluated as therapeutic agents in the treatment of such malignancies. See, e.g., Baselga and Mendelsohn., supra; Masui et al. Cancer Research 44:1002-1007 (1984); and Wu et al. J. Clin. Invest. 95:1897-1905 (1995).
- the second member of the ErbB family, p185 neu was originally identified as the product of the transforming gene from neuroblastomas of chemically treated rats.
- the activated form of the neu proto-oncogene results from a point mutation (valine to glutamic acid) in the transmembrane region of the encoded protein.
- Amplification of the human homolog of neu is observed in breast and ovarian cancers and correlates with a poor prognosis (Slamon et al., Science, 235:177-182 (1987); Slamon et al., Science, 244:707-712 (1989); and U.S. Pat. No. 4,968,603).
- HER2 (frequently but not uniformly due to gene amplification) has also been observed in other carcinomas including carcinomas of the stomach, endometrium, salivary gland, lung, kidney, colon, thyroid, pancreas and bladder.
- Hudziak et al., Mol. Cell. Biol. 9(3):1165-1172 (1989) describe the generation of a panel of anti-HER2 antibodies which were characterized using the human breast tumor cell line SKBR3.
- Relative cell proliferation of the SKBR3 cells following exposure to the antibodies was determined by crystal violet staining of the monolayers after 72 hours.
- maximum inhibition was obtained with the antibody called 4D5 which inhibited cellular proliferation by 56%.
- Other antibodies in the panel reduced cellular proliferation to a lesser extent in this assay.
- the antibody 4D5 was further found to sensitize HER2-overexpressing breast tumor cell lines to the cytotoxic effects of TNF-alpha. See, also, U.S. Pat. No. 5,677,171, issued Oct.
- a recombinant humanized IgG1 version of the murine anti-HER2 antibody 4D5 (rhuMAb HER2 or HERCEPTIN®; commercially available from Genentech, Inc., South San Francisco) is clinically active in patients with HER2-overexpressing metastatic breast cancers that have received extensive prior anti-cancer therapy (Baselga et al., J. Clin. Oncol. 14:737-744 (1996)).
- HERCEPTIN® received marketing approval from the Food and Drug Administration Sep. 25, 1998 for the treatment of patients with metastatic breast cancer whose tumors overexpress the HER2 protein.
- ErbB receptors are generally found in various combinations in cells and heterodimerization is thought to increase the diversity of cellular responses to a variety of ErbB ligands (Earp et al. Breast Cancer Research and Treatment 35: 115-132 (1995)).
- EGFR is bound by six different ligands; Epidermal Growth Factor (EGF), Transforming Growth Factor alpha (TGF-alpha), amphiregulin, Heparin Binding Epidermal Growth Factor (HB-EGF), betacellulin and epiregulin (Groenen et al. Growth Factors 11:235-257 (1994)).
- a family of heregulin proteins resulting from alternative splicing of a single gene are ligands for HER3 and HER4.
- the heregulin family includes alpha, beta and gamma heregulins (Holmes et al., Science, 256:1205-1210 (1992); U.S. Pat. No. 5,641,869; and Schaefer et al. Oncogene 15:1385-1394 (1997)); neu differentiation factors (NDFs), glial growth factors (GGFs); acetylcholine receptor inducing activity (ARIA); and sensory and motor neuron derived factor (SMDF).
- NDFs neu differentiation factors
- GGFs glial growth factors
- ARIA acetylcholine receptor inducing activity
- SMDF sensory and motor neuron derived factor
- EGF and TGF-alpha do not bind HER2, EGF stimulates EGFR and HER2 to form a heterodimer, which activates EGFR and results in transphosphorylation of HER2 in the heterodimer. Dimerization and/or transphosphorylation appears to activate the HER2 tyrosine kinase. See Earp et al., supra.
- HER3 is co-expressed with HER2
- an active signaling complex is formed and antibodies directed against HER2 are capable of disrupting this complex (Sliwkowski et al., J. Biol. Chem., 269(20):14661-14665 (1994)).
- HER3 for heregulin (HRG) is increased to a higher affinity state when co-expressed with HER2.
- HRG heregulin
- HER4 like HER3, forms an active signaling complex with HER2 (Carraway and Cantley, Cell 78:5-8 (1994)).
- TNF-alpha Tumor Necrosis Factor-alpha
- TNF-beta Tumor Necrosis Factor-beta
- LT-alpha Lymphotoxin-alpha
- CD30 ligand CD27 ligand, CD40 ligand, OX-40 ligand, 4-1 BB ligand
- Apo-1 ligand also referred to as Fas ligand or CD95 ligand
- Apo-2 ligand also referred to as TRAIL
- Apo-3 ligand also referred to as TWEAK
- osteoprotegerin OPG
- APRIL RANK ligand
- TRANCE TALL-1
- BlyS Tumor Necrosis Factor
- TNF-alpha, TNF-beta, CD30 ligand, 4-1BB ligand, Apo-1 ligand, Apo-2 ligand (Apo2L/TRAIL) and Apo-3 ligand (TWEAK) have been reported to be involved in apoptotic cell death.
- Both TNF-alpha and TNF-beta have been reported to induce apoptotic death in susceptible tumor cells (Schmid et al., Proc. Natl. Acad. Sci., 83:1881 (1986); Dealtry et al., Eur. J. Immunol., 17:689 (1987)).
- TNF-alpha is involved in post-stimulation apoptosis of CD8-positive T cells (Zheng et al., Nature, 377:348-351 (1995)).
- CD30 ligand may be involved in deletion of self-reactive T cells in the thymus (Amakawa et al., Cold Spring Harbor Laboratory Symposium on Programmed Cell Death , Abstr. No. 10, (1995)).
- CD40 ligand activates many functions of B cells, including proliferation, immunoglobulin secretion, and survival (Renshaw et al., J. Exp. Med., 180:1889 (1994)).
- TNF family cytokine TALL-1 (BlyS)
- BlyS TNF family cytokine
- Apo-1 ligand is also reported to induce post-stimulation apoptosis in CD4-positive T lymphocytes and in B lymphocytes, and may be involved in the elimination of activated lymphocytes when their function is no longer needed (Krammer et al., supra; Nagata et al., supra).
- Agonist mouse monoclonal antibodies specifically binding to the Apo-1 receptor have been reported to exhibit cell killing activity that is comparable to or similar to that of TNF-alpha (Yonehara et al., J. Exp. Med., 169:1747-1756 (1989)).
- TNF family cytokines Induction of various cellular responses mediated by such TNF family cytokines is believed to be initiated by their binding to specific cell receptors.
- TNF receptors Two distinct TNF receptors of approximately 55-kDa (TNFR1) and 75-kDa (TNFR2) were identified (Hohman et al., J. Biol. Chem., 264:14927-14934 (1989); Brockhaus et al., Proc. Natl. Acad. Sci., 87:3127-3131 (1990); EP 417,563, published Mar.
- Those TNFRs were found to share the typical structure of cell surface receptors including extracellular, transmembrane and intracellular regions.
- TNFR1 and TNFR2 The extracellular portion of type 1 and type 2 TNFRs (TNFR1 and TNFR2) contains a repetitive amino acid sequence pattern of four cysteine-rich domains (CRDs) designated 1 through 4, starting from the NH 2 -terminus.
- CCDs cysteine-rich domains
- CRDs CRDs
- NGFR nerve growth factor receptor
- CD40 B cell antigen CD40
- OX40 T cell antigen OX40
- Fas antigen Yonehara et al., supra and Itoh et al., Cell, 66:233-243 (1991)
- CRDs are also found in the soluble TNFR (sTNFR)-like T2 proteins of the Shope and myxoma poxviruses (Upton et al., Virology, 160:20-29 (1987); Smith et al., Biochem. Biophys. Res. Commun., 176:335 (1991); Upton et al., Virology, 184:370 (1991)).
- sTNFR soluble TNFR
- Optimal alignment of these sequences indicates that the positions of the cysteine residues are well conserved.
- These receptors are sometimes collectively referred to as members of the TNF/NGF receptor superfamily.
- TNF family ligands identified to date are type II transmembrane proteins, whose C-terminus is extracellular.
- most receptors in the TNF receptor (TNFR) family identified to date are type I transmembrane proteins.
- TNFR TNF receptor
- homology identified between family members has been found mainly in the extracellular domain (“ECD”).
- ECD extracellular domain
- TNF family cytokines including TNF-alpha, Apo-1 ligand and CD40 ligand, are cleaved proteolytically at the cell surface; the resulting protein in each case typically forms a homotrimeric molecule that functions as a soluble cytokine.
- TNF receptor family proteins are also usually cleaved proteolytically to release soluble receptor ECDs that can function as inhibitors of the cognate cytokines.
- TACI Transmembrane Activator and CAML-Interactor
- BCM bovine et al.
- EMBO J., 11:3897-3904 (1992) reported identifying a new gene called “BCM” whose expression was found to coincide with B cell terminal maturation.
- the open reading frame of the BCM normal cDNA predicted a 184 amino acid long polypeptide with a single transmembrane domain.
- BCMA this gene (Laabi et al., Nucleic Acids Res., 22:1147-1154 (1994)).
- BCMA mRNA expression was reported to be absent in human malignant B cell lines which represent the pro-B lymphocyte stage, and thus, is believed to be linked to the stage of differentiation of lymphocytes (Gras et al., Int.
- Apo-3 has also been referred to by other investigators as DR3, wsl-1, TRAMP, and LARD (Chinnaiyan et al., Science, 274:990 (1996); Kitson et al., Nature, 384:372 (1996); Bodmer et al., Immunity, 6:79 (1997); Screaton et al., Proc. Natl. Acad. Sci., 94:4615-4619 (1997)).
- Pan et al. have disclosed another TNF receptor family member referred to as “DR4” (Pan et al., Science, 276:111-113 (1997); see also WO98/32856 published Jul. 30, 1998).
- the DR4 was reported to contain a cytoplasmic death domain capable of engaging the cell suicide apparatus.
- Pan et al. disclose that DR4 is believed to be a receptor for the ligand known as Apo2L/TRAIL.
- DR5 is reported to contain a cytoplasmic death domain and be capable of signaling apoptosis.
- the crystal structure of the complex formed between Apo2L/TRAIL and DR5 is described in Hymowitz et al., Molecular Cell, 4:563-571 (1999).
- DR6 death domain-containing receptor 6
- DR6 Yet another death domain-containing receptor, DR6, was recently identified (Pan et al., FEBS Letters, 431:351-356 (1998)). Aside from containing four putative extracellular cysteine rich domains and a cytoplasmic death domain, DR6 is believed to contain a putative leucine-zipper sequence that overlaps with a proline-rich motif in the cytoplasmic region. The proline-rich motif resembles sequences that bind to src-homology-3 domains, which are found in many intracellular signal-transducing molecules.
- DcR1 also referred to as TRID, LIT or TRAIL-R3
- TRID TRID
- TRAIL-R3 TRID, LIT
- McFarlane et al. J. Biol. Chem., 272:25417-25420 (1997); Schneider et al., FEBS Letters, 416:329-334 (1997); Degli-Esposti et al., J. Exp.
- TNFR family include CAR1, HVEM, GITR, ZTNFR-5, NTR-1, and TNFL1 (Brojatsch et al., Cell, 87:845-855 (1996); Montgomery et al., Cell, 87:427-436 (1996); Marsters et al., J. Biol. Chem., 272:14029-14032 (1997); Nocentini et al., Proc. Natl. Acad. Sci. USA 94:6216-6221 (1997); Emery et al., J. Biol. Chem., 273:14363-14367 (1998); WO99/04001 published Jan. 28, 1999; WO99/07738 published Feb. 18, 1999; WO99/33980 published Jul. 8, 1999).
- NF- ⁇ B is the prototype of a family of dimeric transcription factors whose subunits contain conserved Rel regions (Verma et al., Genes Develop., 9:2723-2735 (1996); Baldwin, Ann. Rev. Immunol., 14:649-681 (1996)).
- NF- ⁇ B In its latent form, NF- ⁇ B is complexed with members of the I- ⁇ B inhibitor family; upon inactivation of the I- ⁇ B in response to certain stimuli, released NF- ⁇ B translocates to the nucleus where it binds to specific DNA sequences and activates gene transcription.
- the TNFR members identified to date either include or lack an intracellular death domain region.
- Some TNFR molecules lacking a death domain, such as TNFR2, CD40, HVEM, and GITR are capable of modulating NF- ⁇ B activity. (see, e.g., Lotz et al., J. Leukocyte Biol., 60:1-7 (1996)).
- Lymphocytes are one of many types of white blood cells produced in the bone marrow during the process of hematopoiesis. There are two major populations of lymphocytes: B lymphocytes (B cells) and T lymphocytes (T cells). The lymphocytes of particular interest herein are B cells.
- B cells mature within the bone marrow and leave the marrow expressing an antigen-binding antibody on their cell surface.
- a naive B cell first encounters the antigen for which its membrane-bound antibody is specific, the cell begins to divide rapidly and its progeny differentiate into memory B cells and effector cells called “plasma cells”.
- Memory B cells have a longer life span and continue to express membrane-bound antibody with the same specificity as the original parent cell.
- Plasma cells do not produce membrane-bound antibody but instead produce the antibody in a form that can be secreted. Secreted antibodies are the major effector molecule of humoral immunity.
- the CD20 antigen also called human B-lymphocyte-restricted differentiation antigen, Bp35
- Bp35 human B-lymphocyte-restricted differentiation antigen
- CD20 regulates an early step(s) in the activation process for cell cycle initiation and differentiation (Tedder et al., supra) and possibly functions as a calcium ion channel (Tedder et al. J. Cell. Biochem. 14D:195 (1990)).
- this antigen can serve as a candidate for “targeting” of such lymphomas.
- targeting can be generalized as follows: antibodies specific to the CD20 surface antigen of B cells are administered to a patient. These anti-CD20 antibodies specifically bind to the CD20 antigen of (ostensibly) both normal and malignant B cells; the antibody bound to the CD20 surface antigen may lead to the destruction and depletion of neoplastic B cells. Additionally, chemical agents or radioactive labels having the potential to destroy the tumor can be conjugated to the anti-CD20 antibody such that the agent is specifically “delivered” to the neoplastic B cells. Irrespective of the approach, a primary goal is to destroy the tumor; the specific approach can be determined by the particular anti-CD20 antibody which is utilized and, thus, the available approaches to targeting the CD20 antigen can vary considerably.
- CD19 is another antigen that is expressed on the surface of cells of the B lineage. Like CD20, CD19 is found on cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells (Nadler, L. Lymphocyte Typing II 2: 3-37 and Appendix, Renling et al. eds. (1986) by Springer Verlag). Unlike CD20 however, antibody binding to CD19 causes internalization of the CD19 antigen. CD19 antigen is identified by the HD237-CD19 antibody (also called the “B4” antibody) (Kiesel et al. Leukemia Research II, 12: 1119 (1987)), among others.
- HD237-CD19 antibody also called the “B4” antibody
- the CD19 antigen is present on 4-8% of peripheral blood mononuclear cells and on greater than 90% of B cells isolated from peripheral blood, spleen, lymph node or tonsil. CD19 is not detected on peripheral blood T cells, monocytes or granulocytes.
- Virtually all non-T cell acute lymphoblastic leukemias (ALL), B cell chronic lymphocytic leukemias (CLL) and B cell lymphomas express CD19 detectable by the antibody B4 (Nadler et al. J. Immunol. 131:244 (1983); and Nadler et al. in Progress in Hematology Vol. XII pp. 187-206. Brown, E. ed. (1981) by Grune & Stratton, Inc).
- the rituximab (RITUXAN®) antibody is a genetically engineered chimeric murine/human monoclonal antibody directed against the CD20 antigen.
- Rituximab is the antibody called “C2B8” in U.S. Pat. No. 5,736,137 issued Apr. 7, 1998 (Anderson et al.).
- RITUXAN® is indicated for the treatment of patients with relapsed or refractory low-grade or follicular, CD20 positive, B cell non-Hodgkin's lymphoma. In vitro mechanism of action studies have demonstrated that RITUXAN® binds human complement and lyses lymphoid B cell lines through CDC (Reff et al.
- RITUXAN® has been shown to have anti-proliferative effects in tritiated thymidine incorporation assays and to induce apoptosis directly, while other anti-CD19 and CD20 antibodies do not (Maloney et al. Blood 88(10):637a (1996)). Synergy between RITUXAN® and chemotherapies and toxins has also been observed experimentally.
- RITUXAN® sensitizes drug-resistant human B cell lymphoma cell lines to the cytotoxic effects of doxorubicin, CDDP, VP-16, diphtheria toxin and ricin (Demidem et al. Cancer Chemotherapy & Radiopharmaceuticals 12(3):177-186 (1997)).
- doxorubicin doxorubicin
- CDDP CDDP
- VP-16 diphtheria toxin
- ricin diphtheria toxin
- ricin diphtheria toxin
- ricin diphtheria toxin and ricin
- the present invention provides multivalent antibodies (e.g. tetravalent antibodies) with three or more antigen binding sites, which can be readily produced by recombinant expression of nucleic acid encoding the polypeptide chains of the antibody.
- the multivalent antibody comprises a dimerization domain and three or more antigen binding sites.
- the preferred dimerization domain comprises (or consists of) an Fc region or a hinge region.
- the invention provides an isolated antibody comprising a dimerization domain and three or more antigen binding sites amino-terminal thereto.
- the invention further provides an isolated antibody comprising an Fc region and three or more antigen binding sites amino-terminal to the Fc region.
- the preferred multivalent antibody herein comprises (or consists of) three to about eight, but preferably four, antigen binding sites (which are generally all “functional”, as hereindefined). In one embodiment, the multivalent antibody comprises five or more (e.g. up to about eight) antigen binding sites.
- the multivalent antibody herein is preferably not a native sequence IgA or IgM, and may lack an Fc region or have only one Fc region.
- the multivalent antibody comprises at least one polypeptide chain (and preferably two polypeptide chains), wherein the polypeptide chain(s) comprise two or more variable domains.
- the polypeptide chain(s) may comprise VD1-(X1) n -VD2-(X2) n -Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1.
- the polypeptide chain(s) may comprise: VH-CH1-flexible linker-VH-CH1-Fc region chain; VH-CH1-VH-CH1-Fc region chain; VL-CL-flexible linker-VL-CL-Fc region chain; or VL-CL-VL-CL-Fc region chain.
- the flexible linker may comprise a peptide such as gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser, or gly-gly-gly-ser (SEQ ID NO:11).
- the multivalent antibody herein preferably further comprises at least two (and preferably four) light chain variable domain polypeptides.
- the multivalent antibody herein may, for instance, comprise from about two to about eight light chain variable domain polypeptides.
- the light chain variable domain polypeptides contemplated here comprise a light chain variable domain and, optionally, further comprise a CL domain.
- the multivalent antibodies herein have properties which are desirable, among other things, from a therapeutic standpoint.
- the multivalent antibody may (1) be internalized (and/or catabolized) faster than a bivalent antibody by a cell expressing an antigen to which the antibodies bind; (2) be an agonist antibody; and/or (3) induce cell death and/or apoptosis of a cell expressing an antigen which the multivalent antibody is capable of binding to.
- the “parent antibody” which provides at least one antigen binding specificity of the multivalent antibody may be one which is internalized (and/or catabolized) by a cell expressing an antigen to which the antibody binds; and/or may be an agonist, cell-death-inducing, and/or apoptosis-inducing antibody, and the multivalent form of the antibody as described herein may display improvement(s) in one or more of these properties.
- the parent antibody may lack any one or more of these properties, but may be endowed with them when constructed as a multivalent antibody as hereindescribed.
- the three or more antigen binding sites of the multivalent antibodies herein may all bind the same antigen; or may bind two or more (e.g. from two to about three) different antigens.
- the multivalent antibody may bind (1) a cell surface protein expressed (or overexpressed) by tumor cells, e.g. Epidermal Growth Factor Receptor (EGFR), HER2 receptor, HER3 receptor, HER4 receptor, or DcR3; (2) a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily (e.g. an Apo2L receptor, such as DR4, DR5, DcR1 or DcR2); and/or (3) a B cell surface antigen (such as CD19, CD 20 , CD22 or CD40).
- EGFR Epidermal Growth Factor Receptor
- HER2 receptor HER3 receptor
- HER4 receptor HER4 receptor
- DcR3 DcR3
- TNF Tumor Necrosis Factor
- B cell surface antigen such as CD19, CD 20 , CD22 or CD40.
- all of the functional antigen binding sites of the multivalent antibody bind the same antigen as listed above (e.g. all four antigen binding sites
- the invention also provides immunoconjugates comprising the multivalent antibody conjugated with a cytotoxic agent.
- the cytotoxic agent here may be one which is active in killing cells once internalized.
- the invention additionally pertains to a polypeptide chain comprising VD1-(X1) n -VD2 (X2) n -Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1.
- the polypeptide chain may comprise VH-CH1-flexible linker-VH-CH1-Fc region chain; VH-CH1-VH-CH1-Fc region chain; VL-CL-flexible linker-VL-CL-Fc region chain; or VL-CL-VL-CL-Fc region chain.
- the polypeptide chain comprises VH-CH1-flexible linker-VH-CH1-dimerization domain; VH-CH1-VH-CH1-dimerization domain; VL-CL-flexible linker-VL-CL-dimerization domain; or VL-CL-VL-CL-dimerization domain.
- the polypeptide chain may comprise VH-CH1-flexible linker-VH-CH 1 -hinge region; VH-CH1-VH-CH1-hinge region.
- the invention additionally provides an antibody comprising one or more (preferably two) of such polypeptide chains.
- the antibody preferably further comprises at least two (and preferably four) light chain or heavy chain variable domain polypeptides, e.g., where the light chain variable domain polypeptides comprise VL-CL and the heavy chain variable domain polypeptides comprise VH-CH1.
- the invention further provides a polypeptide chain comprising three or more heavy chain or light chain variable domains, wherein each of the variable domains is able to combine with three or more light chain or heavy chain variable domain polypeptides to form three or more antigen binding sites, each directed against the same antigen.
- the invention also provides an isolated antibody comprising the polypeptide chain.
- the polypeptide chain comprises three or more heavy chain variable domains
- the antibody preferably further comprises three or more light chain variable domain polypeptides which can combine with the heavy chain variable domains to form the three or more antigen binding sites. Examples of such antibodies are shown in FIG. 23 D (with three antigen binding sites) and FIG. 23E (with four antigen binding sites).
- the invention provides a polypeptide chain comprising the formula: (a) VL-CL-flexible linker-VL-CL-flexible linker-VL-CL; (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; (c) (VL-CL) n , wherein n is three or more; or (d) (VH-CH 1) n , wherein n is three or more.
- the invention further provides: isolated nucleic acid encoding the multivalent antibody or polypeptide chain; a vector comprising nucleic acid encoding the multimeric antibody or polypeptide chain, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising (e.g. transformed with) nucleic acid encoding the multimeric antibody or polypeptide chain; a method for producing the multivalent antibody or polypeptide chain comprising culturing the host cell so that the nucleic acid is expressed and, optionally, recovering the multivalent antibody or polypeptide chain from the host cell culture (e.g. from the host cell culture medium).
- a vector comprising nucleic acid encoding the multimeric antibody or polypeptide chain, optionally, operably linked to control sequences recognized by a host cell transformed with the vector
- a host cell comprising (e.g. transformed with) nucleic acid encoding the multimeric antibody or polypeptide chain
- a method for producing the multivalent antibody or polypeptide chain compris
- Nucleic acids encoding (1) the heavy chain variable domains and (2) the light chain variable domains of the multivalent antibody are preferrably co-expressed by a host cell transformed with both (1) and (2).
- Nucleic acids (1) and (2) may be present in the same, or different, vectors.
- the invention provides a method for determining the presence of an antigen of interest comprising exposing a sample suspected of containing the antigen to the multivalent antibody and determining binding of the multivalent antibody to the sample. Both in vitro and in vivo diagnostic methods are provided.
- the invention provides a method of treating a mammal suffering from, or predisposed to, a disease or disorder, comprising administering to the mammal a therapeutically effective amount of a multivalent antibody as disclosed herein, or of a composition comprising the multivalent antibody and a pharmaceutically acceptable carrier.
- the disorder to be treated herein may be cancer, in which case the method may further comprise administering a therapeutically effective amount of a cytotoxic agent to the mammal.
- the present invention further relates to a method of inducing apoptosis of a cancer cell comprising exposing the cell to a multivalent antibody as described herein, wherein the multivalent antibody binds a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily.
- TNF Tumor Necrosis Factor
- the method may involve killing a B cell by exposing the B cell to a multivalent antibody that binds a B cell surface antigen. Moreover, the method may relate to killing a cell which expresses (or overexpresses) an ErbB receptor comprising exposing the cell to an antibody that binds the ErbB receptor.
- FIG. 1 is a schematic representation of a native IgG and digestion thereof with (1) papain to generate two Fab fragments and an Fc region or (2) pepsin to generate a F(ab′) 2 fragment and multiple small fragments. Disulfide bonds are represented by lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain.
- FIGS. 2A-E depict the structures of the five major naturally occurring immunoglobulin isotypes; IgG ( FIG. 2A ), IgD ( FIG. 2B ), IgE ( FIG. 2C ), IgA dimer ( FIG. 2D ), and IgM pentamer ( FIG. 2E ).
- FIG. 3 depicts alignments of native sequence IgG Fc regions.
- Native sequence human IgG Fc region sequences humIgG1 (non-A and A allotypes) (SEQ ID NOs: 1 and 2, respectively), humIgG2 (SEQ ID NO:3), humIgG3 (SEQ ID NO:4) and humIgG4 (SEQ ID NO:5), are shown.
- the human IgG1 sequence is the non-A allotype, and differences between this sequence and the A allotype (at positions 356 and 358; EU numbering system) are shown below the human IgG1 sequence.
- murIgG1 SEQ ID NO:6
- murIgG2A SEQ ID NO:7
- murIgG2B SEQ ID NO:8
- murlgG3 SEQ ID NO:9
- FIGS. 4A-B depict schematically tetravalent antibodies according to the present invention.
- the four antigen binding Fabs are numbered (1 and 2 for each arm of the tetravalent antibody) and X represents a dimerization domain.
- the dimerization domain of the tetravalent antibody is an Fc region.
- FIG. 5 shows the construct used for expression of a tetravalent anti-HER2 antibody (OctHER2) in Example 1.
- FIGS. 6A-C illustrate binding of OctHER2 ( FIG. 6A ); bivalent IgG1 rhuMAb 4D5-8 expressed by 293 cells ( FIG. 6B ); and vialed HERCEPTIN® (expressed by Chinese hamster ovary (CHO) cells) ( FIG. 6C ) to HER2 extracellular domain (ECD) as determined using an enzyme-linked immunosorbent assay (ELISA).
- ELISA enzyme-linked immunosorbent assay
- FIG. 7 depicts ultracentrifugation analysis of binding of OctHER2 to HER2ECD. Average molecular weights (theoretical or experimentally determined) versus molar ratio of OctHER2 to HER2ECD are shown. Theoretical calculated average molecular weights assuming tetravalent antibody has four fully functional binding sites are shown in circles; theoretical calculated average molecular weights assuming tetravalent antibody has three fully functional binding sites are shown in squares; and triangles represent experimentally determined molecular weights.
- FIGS. 8A-D depict the growth inhibitory activity of HERCEPTIN® compared to OctHER2 using SKBR3 (3+ HER2 overexpressing) ( FIG. 8A ), MDA 361 (2+ HER2 overexpressing) ( FIG. 8B ), BT474 (3+ HER2 overexpressing) ( FIG. 8C ) and MCF7 (0+ HER2 expressing) ( FIG. 8D ) cell lines.
- FIG. 9 depicts the effect of flexible linkers on the growth inhibitory activity of tetravalent anti-HER2 antibodies with respect to MDA 231 cells (1+ HER2 overexpressing) or SKBR3 cells (3+ HER2 overexpressing).
- FIGS. 10A-B compare the rate of OctHER2 internalization/catabolism ( FIG. 10A ) to that of HERCEPTIN® ( FIG. 10B ), in relation to both MDA 453 (2+ HER2 overexpressing) and SKBR3 (3+ HER2 overexpressing) cell lines.
- FIGS. 11A-I are electron microscopy photographs showing internalization of OctHER2.
- FIGS. 11G-I show internalization at time 0 hours ( FIGS. 11G) and 5 hours ( FIGS. 11H and 11I ).
- FIGS. 12A-E depict apoptosis induced by an anti-DR5 tetravalent antibody (16E2 Octopus), an anti-DR5 bivalent IgG antibody (16E2 IgG), and Apo2L/TRAIL (Apo2L) on cancer cell lines: COLO 205 ( FIG. 12A ), SK-MES-1 ( FIG. 12B ), HCT116 ( FIG. 12C ), and HOP 92 ( FIG. 12D ), compared to a non-cancer control cell line, HUMEC ( FIG. 12E ).
- an anti-DR5 tetravalent antibody (16E2 Octopus
- an anti-DR5 bivalent IgG antibody (16E2 IgG
- Apo2L/TRAIL Apo2L/TRAIL
- FIGS. 13A-D are histology slides stained to detect apoptotic cells.
- Tumor tissues from mice treated with 16E2 Octopus or Apo2L/TRAIL were fixed in 10% formalin and then embedded into parafilm and sectioned onto slides which were then stained with hematoxylin and eosin and visualized under a 400 ⁇ magnification.
- the effect of 16E2 Octopus at 6 and 24 hours is shown in FIGS. 13A and B, respectively; control-treated cells are shown in FIG. 13C ; and Apo2L/TRAIL-treated cells are shown in FIG. 13D .
- FIG. 14 represents the in vivo activity of Apo2L/TRAIL (60 mg/kg, 5 ⁇ /week), 3H3 bivalent IgG (5 mg/kg given days 0, 3, 5 and 9), 16E2 bivalent IgG (16E2) (5 mg/kg given days 0, 3, 5 and 9), and 16E2 Octopus (5 mg/kg given days 0, 3, 5 and 9) with respect to COLO 205 tumors in athymic nude mice.
- FIG. 15 represents an alamarBlue in vitro assay confirming the apoptotic activity of the material used in the mouse studies (Apo2L/TRAIL and 16E2 Octopus) as compared to an Apo2L standard positive control.
- the anti-IgE antibody (E25) used as a negative control in the mouse studies was confirmed to have no apoptotic activity.
- FIG. 16 represents the results of a crystal violet apoptosis assay comparing anti-DR5 3H3 Octopus to various batches of the anti-DR5 16E2 Octopus.
- FIGS. 17A-B reveal the results of the alamarBlue apoptosis assay with respect to Apo2L/TRAIL (WO97/25428), anti-DR5 3H3 Octopus antibody, anti-DR5 16E2 Octopus antibody, and Apo2L/TRAIL with a FLAG epitope-tag cross linked by an anti-FLAG antibody (WO97/25428), with respect to SK-MES-1 ( FIG. 17A ) and Jurkat ( FIG. 17B ) cells in the presence of 5% fetal bovine serum (FBS).
- Apo2L/TRAIL WO97/25428
- anti-DR5 3H3 Octopus antibody anti-DR5 16E2 Octopus antibody
- Apo2L/TRAIL with a FLAG epitope-tag cross linked by an anti-FLAG antibody WO97/25428
- SK-MES-1 FIG. 17A
- Jurkat FIG. 17B
- FIGS. 18A-C depict dose response curves that show the effect of the anti-DR5 16E2 Octopus (upper graphs) compared to Apo2L/TRAIL (lower graphs) on the growth of leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer human tumor cell lines at 2 days.
- Results are from the National Cancer Institute Developmental Therapeutics Program. All samples were tested at 5 concentrations, starting at 1% of the stock solution (16E2 Octopus stock 0.2 mg/ml) and 4 ⁇ 0.5 log dilutions.
- FIGS. 19A-C depict dose response curves that show the effect of the anti-DR516E2 Octopus (upper graphs) compared to Apo2L/TRAIL (lower graphs) on the growth of leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer human tumor cell lines at 6 days.
- Results are from the National Cancer Institute Developmental Therapeutics Program. All samples were tested at 5 concentrations, starting at 1% of the stock solution (16E2 Octopus stock 0.2 mg/ml) and 4 ⁇ 0.5 log dilutions.
- FIGS. 20A-B present a quantitative summary of the 2 day in vitro results from the National Cancer Institute Developmental Therapeutics Program comparing the anti-DR5 16E2 Octopus ( FIG. 20A ) to Apo2L/TRAIL ( FIG. 20B ) analyzing growth inhibition (GI50), stasis (TGI), and toxicity (LC50).
- GI50 growth inhibition
- TGI stasis
- LC50 toxicity
- FIGS. 21A-B present a quantitative summary of the 6 day in vitro results from the National Cancer Institute Developmental Therapeutics Program comparing the anti-DR5 16E2 Octopus ( FIG. 21A ) to Apo2L/TRAIL ( FIG. 21B ) analyzing growth inhibition (G150), stasis (TGI), and toxicity (LC50).
- FIG. 22 depicts apoptosis of Wil-2 cells by the anti-CD20 antibody RITUXAN®, RITUXAN® cross-linked with anti-human IgG (RITUXAN®-IgG) and a tetravalent anti-CD20 antibody (OctCD20).
- FIGS. 23A-E are cartoons depicting the full-length Octopus/tetravalent antibody ( FIG. 23B ), the Octopus F(ab)′ 2 ( FIG. 23C ), POPoct-3 Fab ( FIG. 23D ) and POPoct-4 Fab ( FIG. 23E ) in comparison to the native IgG ( FIG. 23A ).
- a representative coomassie stained Tris-Glycine gel of anti-CD20 (C2B8) Octopus proteins compares the sizes of the intact antibodies in non-reducing conditions ( FIG. 23F ), and of the heavy chains in reducing conditions, under which disulfide bonds are disrupted resulting in separation of the heavy and light chains ( FIG. 23G ).
- FIG. 24 depicts the construction of the Octopus F(ab′) 2 backbone. Any VH/CH1 region can be substituted into the F(ab′) 2 backbone via the BamHI, NheI and BspEI restriction enzyme sites.
- FIG. 25 depicts the construction of the POPoct-3 heavy chain.
- FIG. 26 depicts the construction of the POPoct-4 heavy chain.
- FIG. 27 depicts the activity of multivalent anti-HER2 antibodies in cytostasis assays using BT474 cells.
- FIGS. 28A-B depict the activity of multivalent anti-HER2 antibodies in cytostasis assays using SKBR3 cells.
- FIGS. 29A-B show internalization capability of multivalent anti-HER2 antibodies in SKBR3 cells ( FIG. 29A ) and BT474 cells ( FIG. 29B ).
- FIGS. 30A-B reveal apoptosis of COLO205 cells by multivalent anti-DR5 antibodies
- FIGS. 31A-B demonstrate signalling of multivalent anti-DR5 antibodies through the caspase pathway.
- FIG. 32 compares apoptosis induced by IgG cross-linked RITUXAN® (RITUXAN-IgG) and IgG cross-linked OctCD20 (OctCD20-IgG).
- FIG. 33 shows apoptosis of WIL2 cells by multivalent anti-CD20 antibodies, the IF5 anti-CD20 antibody (Clark et al. PNAS ( USA ) 82: 1766-1770 (1985)) and IgG cross-linked IFS antibody (IF5+IgG-X).
- FIG. 34 depicts homotypic cell adhesion in WIL2S cells induced by IF5 anti-CD20 antibody, IgG cross-linked IF5 antibody and POPoct-3 CD20.
- FIG. 35 reflects RITUXAN® or OctCD20 internalization/catabolism on DB, WIL2 and Ramos B-cell lymphoma lines.
- the numbering of the residues in an immunoglobulin heavy chain is that of the EU index as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference.
- the “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
- ErbB receptor is a receptor protein tyrosine kinase which belongs to the ErbB receptor family and includes EGFR, HER2, ErbB3 and ErbB4 receptors as well as TEGFR (U.S. Pat. No. 5,708,156) and other members of this family to be identified in the future.
- the ErbB receptor will generally comprise an extracellular domain, which may bind an ErbB ligand; a lipophilic transmembrane domain; a conserved intracellular tyrosine kinase domain; and a carboxyl-terminal signaling domain harboring several tyrosine residues which can be phosphorylated.
- the ErbB receptor may be a native sequence ErbB receptor or an amino acid sequence variant thereof.
- the ErbB receptor is native sequence human ErbB receptor.
- ErbB ligand is meant a polypeptide which binds to and/or activates an ErbB receptor.
- the ErbB ligand of particular interest herein is a native sequence human ErbB ligand such as Epidermal Growth Factor (EGF) (Savage et al., J. Biol. Chem. 247:7612-7621 (1972)); Tansforming Growth Factor alpha (TGF-alpha) (Marquardt et al., Science 223:1079-1082 (1984)); amphiregulin also known as schwanoma or keratinocyte autocrine growth factor (Shoyab et al. Science 243:1074-1076 (1989); Kimura et al.
- EGF Epidermal Growth Factor
- TGF-alpha Tansforming Growth Factor alpha
- amphiregulin also known as schwanoma or keratinocyte autocrine growth factor (Shoyab et al. Science 243:
- ErbB ligands which bind EGFR include EGF, TGF-alpha, amphiregulin, betacellulin, HB-EGF and epiregulin.
- ErbB ligands which bind HER3 include heregulins.
- ErbB ligands capable of binding HER4 include betacellulin, epiregulin, HB-EGF, NRG-2, NRG-3 and heregulins.
- Heregulin when used herein refers to a polypeptide comprising an amino acid sequence encoded by the heregulin gene product as disclosed in U.S. Pat. No. 5,641,869 or Marchionni et al., Nature, 362:312-318 (1993), and biologically active variants of such polypeptides.
- heregulins include heregulin-alpha heregulin-beta1, heregulin-beta2 and heregulin-beta3 (Holmes et al., Science, 256:1205-1210 (1992); and U.S. Pat. No. 5,641,869); neu differentiation factor (NDF) (Peles et al.
- GGFs glial growth factors
- SMDF motor neuron derived factor
- ErbB hetero-oligomer herein is a noncovalently associated oligomer comprising at least two different ErbB receptors. Such complexes may form when a cell expressing two or more ErbB receptors is exposed to an ErbB ligand and can be isolated by immunoprecipitation and analyzed by SDS-PAGE as described in Sliwkowski et al., J. Biol. Chem., 269(20):14661-14665 (1994), for example. Examples of such ErbB hetero-oligomers include EGFR-HER2, HER2-HER3 and HER3-HER4 complexes.
- the ErbB hetero-oligomer may comprise two or more HER2 receptors combined with a different ErbB receptor, such as HER3, HER4 or EGFR.
- Other proteins such as a cytokine receptor subunit (e.g. gp130), may be included in the hetero-oligomer.
- ErbB1 refers to native sequence EGFR as disclosed, for example, in Carpenter et al. Ann. Rev. Biochem. 56:881-914 (1987), including variants thereof (e.g. a deletion mutant EGFR as in Humphrey et PNAS ( USA ) 87:4207-4211 (1990)).
- erbB1 refers to the gene encoding the EGFR protein product.
- antibodies which bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see, U.S. Pat. No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimerized 225 (C225) and reshaped human 225 (H225) (see, WO 96/40210, Imclone Systems Inc.).
- ErbB2 and HER2 are used interchangeably herein and refer to native sequence human HER2 protein described, for example, in Semba et al., PNAS ( USA ) 82:6497-6501 (1985) and Yamamoto et al. Nature 319:230-234 (1986) (Genebank accession number X03363), and variants thereof.
- the term erbB2 refers to the gene encoding human HER2 and neu refers to the gene encoding rat p185 neu .
- Preferred HER2 is native sequence human HER2.
- antibodies which bind HER2 include MAbs 4D5 (ATCC CRL 10463), 2C4 (ATCC HB-12697), 7F3 (ATCC HB-12216), and 7C2 (ATCC HB 12215) (see, U.S. Pat. No. 5,772,997; WO98/77797; and U.S. Pat. No. 5,840,525, expressly incorporated herein by reference).
- Humanized anti-HER2 antibodies include huMAb4D5-1, huMAb4D5-2, huMAb4D5-3, huMAb4D5-4, huMAb4D5-5, huMAb4D5-6, huMAb4D5-7 and huMAb4D5-8 (HERCEPTIN®) as described in Table 3 of U.S. Pat. No. 5,821,337 expressly incorporated herein by reference; humanized 520C9 (WO93/21319). Human anti-HER2 antibodies are described in U.S. Pat. No. 5,772,997 issued Jun. 30, 1998 and WO 97/00271 published Jan. 3, 1997.
- ErbB3 and “HER3” refer to the receptor polypeptide as disclosed, for example, in U.S. Pat. Nos. 5,183,884 and 5,480,968 as well as Kraus et al. PNAS ( USA ) 86:9193-9197 (1989), including variants thereof.
- Examples of antibodies which bind HER3 are described in U.S. Pat. No. 5,968,511 (Akita and Sliwkowski), e.g. the 8B8 antibody (ATCC HB 12070) or a humanized variant thereof.
- ErbB4 and HER4 herein refer to the receptor polypeptide as disclosed, for example, in EP Pat Appin No 599,274; Plowman et al., Proc. Natl. Acad. ScL USA, 90:1746-1750 (1993); and Plowman et al., Nature, 366:473-475 (1993), including variants thereof such as the HER4 isoforms disclosed in WO 99/19488.
- a “B cell surface marker” herein is an antigen expressed on the surface of a B cell which can be targeted with an antibody which binds thereto.
- Exemplary B cell surface markers include the CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD40, CD53, CD72, CD73, CD74, CDw75, CDw76, CD77, CDw78, CD79a, CD79b, CD80, CD81, CD82, CD83, CDw84, CD85 and CD86 leukocyte surface markers.
- the B cell surface marker of particular interest is preferentially expressed on B cells compared to other non-B cell tissues of a mammal and may be expressed on both precursor B cells and mature B cells.
- the marker is one, like CD20 or CD19, which is found on B cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells.
- the preferred B cell surface markers herein are CD19, CD20, CD22 and CD40.
- the “CD20” antigen is an about 35 kDa, non-glycosylated phosphoprotein found on the surface of greater than 90% of B cells from peripheral blood or lymphoid organs. CD20 is expressed during early pre-B cell development and remains until plasma cell differentiation. CD20 is present on both normal B cells as well as malignant B cells. Other names for CD20 in the literature include “B-lymphocyte-restricted antigen” and “Bp35”. The CD20 antigen is described in Clark et al. PNAS ( USA ) 82:1766 (1985), for example. Examples of antibodies which bind the CD20 antigen include: “C2B8” which is now called “rituximab” (“RITUXAN®”) (U.S. Pat. No.
- the “CD19” antigen refers to the about 90 kDa antigen identified, for example, by the HD237-CD19 or B4 antibody (Kiesel et al. Leukemia Research II, 12: 1119 (1987)). Like CD 20 , CD19 is found on cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells. Binding of an antibody to CD19 may cause internalization of the CD19 antigen. Examples of antibodies which bind the CD19 antigen include the anti-CD19 antibodies in Hekman et al. Cancer Immunol. Immunother. 32:364-372 (1991) and Vlasveld et al. Cancer Immunol. Immunother. 40:37-47 (1995); and the B4 antibody in Kiesel et al. Leukemia Research II, 12: 1119 (1987).
- CD22 has a molecular weight of about 140,000 kD. CD22 is expressed in the cytoplasm of early pre-B and progenitor cells, appears on the surface of only mature B cells and on the majority of non-Hodgkin's lymphoma (NHL) cells, and is then lost during terminal differentiation prior to the plasma cell stage from both the surface and cytoplasm.
- An example of an anti-CD22 antibody is the LL2 antibody described in Juweid et al. Cancer Research 55:5899-5907 (1995), including chimeric/humanized variants thereof.
- CD40 antigen is a cell surface phosphorylated glycoprotein that is expressed on a variety of cell types, including B cells, B cell malignancies, follicular dendritic cells, basal epithelial cells and carcinomas.
- CD40 binds CD40 ligand (CD40L). Aside from being a B cell surface antigen, CD40 is also a member of the TNF receptor superfamily. Examples of antibodies that bind CD40 include those which (1) block CD40/CD40L interaction and have anti-neoplastic properties (Armitage et al., U.S. Pat. No. 5,674,492); (2) antagonize signaling through CD40 (deBoer et al., U.S. Pat. No.
- TNF receptor superfamily refers to receptor polypeptides bound by cytokines in the TNF family. Generally, these receptors are Type I transmembrane receptors with one or more cysteine rich repeat sequences in their extracellular domain.
- the TNF receptor superfamily may be further subdivided into (1) death receptors; (2) decoy receptors; and (3) signaling receptors that lack death domains.
- the “death receptors” contain in their cytoplasmic or intracellular region a “death domain”, i.e., a region or sequence which acts to transduce signals in the cell which can result in apoptosis or in induction of certain genes.
- the “decoy receptors” lack a functional death domain and are incapable of transducing signals which result in apoptosis.
- cytokines in the TNF gene family include Tumor Necrosis Factor-alpha (TNF-alpha), Tumor Necrosis Factor-beta (TNF-beta or lymphotoxin), CD30 ligand, CD27 ligand, CD40 ligand, OX-40 ligand, 4-1 BB ligand, Apo-1 ligand (also referred to as Fas ligand or CD95 ligand), Apo-2 ligand (also referred to as TRAIL), Apo-3 ligand (also referred to as TWEAK), osteoprotegerin (OPG), APRIL, RANK ligand (also referred to as TRANCE), and TALL-1 (also referred to as BlyS, BAFF or THANK).
- TNF-alpha Tumor Necrosis Factor-alpha
- TNF receptor superfamily examples include: type 1 Tumor Necrosis Factor Receptor (TNFR1), type 2 Tumor Necrosis Factor Receptor (TNFR2), p75 Nerve Growth Factor receptor (NGFR), the B cell surface antigen CD40, the T cell antigen OX-40, Apo-1 receptor (also called Fas or CD95), Apo-3 receptor (also called DR3, swl-1, TRAMP and LARD), the receptor called “Transmembrane Activator and CAML-Interactor” or “TACI”, BCMA protein, DR4, DR5 (alternatively referred to as Apo-2; TRAIL-R2, TR6, Tango-63, hAPO8, TRICK2 or KILLER), DR6, DcR1 (also referred to as TRID, LIT or TRAIL-R3), DcR2 (also called TRAIL-R4 or TRUNDD), OPG, DcR3 (also called TR6 or M68), CAR1, HVE
- Apo-2 ligand or “Apo2L” refer to the Apo2L polypeptides disclosed in WO97/25428, published 17 Jul. 1997 and expressly incorporated herein by reference. For purposes of the present application, these terms also refer to the polypeptides referred to as TRAIL disclosed in WO97/01633, published 16 Jan., 1997 and U.S. Pat. No. 5,763,223, issued Jun. 9, 1998 and expressly incorporated herein by reference.
- An “Apo2L receptor” is a polypeptide to which Apo2L can specifically bind.
- the term “Apo2L receptor” when used herein encompasses native sequence Apo2L receptors and variants thereof. These terms encompass Apo2L receptor from a variety of mammals, including humans.
- the Apo2L receptor may be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. Examples of “native sequence” Apo2L receptors include Apo-2 polypeptide or DR5 (WO98/51793, expressly incorporated herein by reference), native sequence DR4 as described in Pan et al.
- anti-DR5 antibodies examples include 3F11.39.7 (ATCC HB-12456), 3H3.14.5 (ATCC HB-12534), 3D5.1.10 (HB-12536) and 3H1.18.10 (HB-12535), 16E2 and 20E6 (see WO 98/51793, expressly incorporated herein by reference).
- anti-DR4 antibodies examples include 4E7.24.3 (ATCC HB-12454) and 4H6.17.8 (ATCC HB-12455) (see, WO 99/37684, expressly incorporated herein by reference).
- DcR3 Native sequence “DcR3” is described in WO99/14330, expressly incorporated herein by reference. That patent publication describes the following mAbs directed against DcR3: 4C4.1.4 (ATCC HB-12573); 5C4.14.7 (ATCC HB-12574); 11C5.2.8 (ATCC HB-12572); 8D3.1.5 (ATCC HB-12571); and 4B7.1.1 (ATCC HB-12575).
- a “native sequence” polypeptide comprises a polypeptide having the same amino acid sequence as a polypeptide derived from nature.
- a native sequence polypeptide can have the amino acid sequence of naturally-occurring polypeptide from any mammal.
- Such native sequence polypeptide can be isolated from nature or can be produced by recombinant or synthetic means.
- the term “native sequence” polypeptide specifically encompasses naturally-occurring truncated or secreted forms of the polypeptide (e.g., an extracellular domain sequence), naturally-occurring variant forms (e.g., alternatively spliced forms) and naturally-occurring allelic variants of the polypeptide.
- a polypeptide “variant” means a biologically active polypeptide having at least about 80% amino acid sequence identity with the native sequence polypeptide.
- variants include, for instance, polypeptides wherein one or more amino acid residues are added, or deleted, at the N- or C-terminus of the polypeptide.
- a variant will have at least about 80% amino acid sequence identity, more preferably at least about 90% amino acid sequence identity, and even more preferably at least about 95% amino acid sequence identity with the native sequence polypeptide.
- Apoptosis refers to programmed cell death. Physiological events often indicative of the occurrence of apoptosis include: fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies). Various methods are available for evaluating the cellular events associated with apoptosis. For example, phosphatidyl serine (PS) translocation can be measured by annexin V binding; DNA fragmentation can be evaluated through DNA laddering or propidium-iodine staining; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells.
- PS phosphatidyl serine
- antibody is used in the broadest sense and includes monoclonal antibodies (including full length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments (see below) so long as they exhibit the desired biological activity.
- multivalent antibody is used throughout this specification to denote an antibody comprising three or more antigen binding sites.
- the multivalent antibody is preferably engineered to have the three or more antigen binding sites and is generally not a native sequence IgM or IgA antibody.
- Antibody fragments comprise only a portion of an intact antibody, generally including an antigen binding site of the intact antibody and thus retaining the ability to bind antigen.
- Examples of antibody fragments encompassed by the present definition include: (i) the Fab fragment, having VL, CL, VH and CH1 domains; (ii) the Fab′ fragment, which is a Fab fragment having one or more cysteine residues at the C-terminus of the CH1 domain; (iii) the Fd fragment having VH and CH1 domains; (iv) the Fd′ fragment having VH and CH1 domains and one or more cysteine residues at the C-terminus of the CH1 domain; (v) the Fv fragment having the VL and VH domains of a single arm of an antibody; (vi) the dAb fragment (Ward et al., Nature 341, 544-546 (1989)) which consists of a VH domain; (vii) isolated CDR regions; (viii) F(ab′) 2 fragments,
- the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigen. Furthermore, in contrast to polyclonal antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
- the modifier “monoclonal” is not to be construed as requiring production of the antibody by any particular method.
- the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567).
- the “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) or Marks et al., J. Mol. Biol. 222:581-597 (1991), for example.
- the monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. ScL USA 81:6851-6855 (1984)).
- “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin.
- humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
- donor antibody such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
- framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
- humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance.
- the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence.
- the humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
- Fc immunoglobulin constant region
- a “human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues.
- Human antibodies can be produced using various techniques known in the art. In one embodiment, the human antibody is selected from a phage library, where that phage library expresses human antibodies (Vaughan et al. Nature Biotechnology 14:309-314 (1996): Sheets et al. PNAS ( USA ) 95:6157-6162 (1998)); Hoogenboom and Winter, J. Mol.
- Human antibodies can also be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos.
- the human antibody may be prepared via immortalization of human B lymphocytes producing an antibody directed against a target antigen (such B lymphocytes may be recovered from an individual or may have been immunized in vitro). See, e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy , Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147 (1):86-95 (1991); and U.S. Pat. No. 5,750,373.
- variable refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called hypervariable regions both in the light chain and the heavy chain variable domains. The more highly conserved portions of variable domains are called the framework regions (FRs).
- the variable domains of native heavy and light chains each comprise four FRs, largely adopting a beta-sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the beta-sheet structure.
- the hypervariable regions in each chain are held together in close proximity by the FRs and, with the hypervariable regions from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).
- the constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC).
- hypervariable region when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding.
- the hypervariable region generally comprises amino acid residues from a “complementarity determining region” or “CDR” (e.g. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and/or those residues from a “hypervariable loop” (e.g.
- “Framework Region” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
- intact antibodies can be assigned to different “classes”. There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into “subclasses” (isotypes), e.g., IgG1 (including non-A and A allotypes), IgG2, IgG3, IgG4, IgA, and IgA2.
- the heavy-chain constant domains that correspond to the different classes of antibodies are called ⁇ , ⁇ , ⁇ , ⁇ and ⁇ , respectively.
- the subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.
- the light chains of antibodies from any vertebrate species can be assigned to one of two clearly distinct types, called kappa ( ⁇ ) and lambda ( ⁇ ), based on the amino acid sequences of their constant domains.
- Fc region is used to define the C-terminal region of an immunoglobulin heavy chain which may be generated by papain digestion of an intact antibody.
- the Fc region may be a native sequence Fc region or a variant Fc region.
- the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at about position Cys226, or from about position Pro230, to the carboxyl-terminus of the Fc region.
- the Fc region of an immunoglobulin generally comprises two constant domains, a CH2 domain and a CH3 domain, and optionally comprises a CH4 domain.
- Fc region chain herein is meant one of the two polypeptide chains of an Fc region.
- the “CH2 domain” of a human IgG Fc region usually extends from an amino acid residue at about position 231 to an amino acid residue at about position 340.
- the CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain.
- the CH2 domain herein may be a native sequence CH2 domain or variant CH2 domain.
- the “CH3 domain” comprises the stretch of residues C-terminal to a CH2 domain in an Fc region (i.e. from an amino acid residue at about position 341 to an amino acid residue at about position 447 of an IgG).
- the CH3 region herein may be a native sequence CH3 domain or a variant CH3 domain (e.g. a CH3 domain with an introduced “protroberance” in one chain thereof and a corresponding introduced “cavity” in the other chain thereof; see U.S. Pat. No. 5,821,333, expressly incorporated herein by reference).
- Such variant CH3 domains may be used to make multispecific (e.g. bispecific) antibodies as herein described.
- “Hinge region” is generally defined as stretching from about Glu216, or about Cys226, to about Pro230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S—S bonds in the same positions.
- the hinge region herein may be a native sequence hinge region or a variant hinge region.
- the two polypeptide chains of a variant hinge region generally retain at least one cysteine residue per polypeptide chain, so that the two polypeptide chains of the variant hinge region can form a disulfide bond between the two chains.
- the preferred hinge region herein is a native sequence human hinge region, e.g. a native sequence human IgG1 hinge region.
- a “functional Fc region” possesses at least one “effector function” of a native sequence Fc region.
- effector functions include C1 q binding; complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g. B cell receptor; BCR), etc.
- Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody variable domain) and can be assessed using various assays known in the art for evaluating such antibody effector functions.
- a “native sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature.
- FIG. 3 provides amino acid sequences of native sequence human and murine IgG Fc regions.
- a “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification.
- the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, e.g. from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide.
- the variant Fc region herein will preferably possess at least about 80% sequence identity with a native sequence Fc region and/or with an Fc region of a parent polypeptide, and most preferably at least about 90% sequence identity therewith, more preferably at least about 95% sequence identity therewith.
- Antibody-dependent cell-mediated cytotoxicity and “ADCC” refer to a cell-mediated reaction in which nonspecific cytotoxic cells that express Fc receptors (FcRs) (e.g. Natural Killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and subsequently cause lysis of the target cell.
- FcRs Fc receptors
- FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991).
- ADCC activity of a molecule of interest may be assessed in vitro, such as that described in U.S. Pat. No. 5,500,362 or 5,821,337.
- Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.
- PBMC peripheral blood mononuclear cells
- NK Natural Killer
- ADCC activity of the molecule of interest may be assessed in vivo, e.g., in a animal model such as that disclosed in Clynes et al. PNAS ( USA ) 95:652-656 (1998).
- Human effector cells are leukocytes which express one or more FcRs and perform effector functions. Preferably, the cells express at least Fc ⁇ RIII and perform ADCC effector function. Examples of human leukocytes which mediate ADCC include peripheral blood mononuclear cells (PBMC), natural killer (NK) cells, monocytes, cytotoxic T cells and neutrophils; with PBMCs and NK cells being preferred.
- PBMC peripheral blood mononuclear cells
- NK natural killer cells
- monocytes cytotoxic T cells and neutrophils
- the effector cells may be isolated from a native source thereof, e.g. from blood or PBMCs as described herein.
- Fc receptor and “FcR” are used to describe a receptor that binds to the Fc region of an antibody.
- the preferred FcR is a native sequence human FcR.
- a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII subclasses, including allelic variants and alternatively spliced forms of these receptors.
- Fc ⁇ RII receptors include Fc ⁇ RIIA (an “activating receptor”) and Fc ⁇ RIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof.
- Activating receptor Fc ⁇ RIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain.
- Inhibiting receptor Fc ⁇ RIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)).
- FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995).
- FcR FcR
- FcRn neonatal receptor
- “Complement dependent cytotoxicity” and “CDC” refer to the lysing of a target in the presence of complement.
- the complement activation pathway is initiated by the binding of the first component of the complement system (C1q) to a molecule (e.g. an antibody) complexed with a cognate antigen.
- a CDC assay e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996), may be performed.
- affinity matured antibody is one with one or more alterations in one or more CDRs thereof which result an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess those alteration(s).
- Preferred affinity matured antibodies will have nanomolar or even picomolar affinities for the target antigen.
- Affinity matured antibodies are produced by procedures known in the art. Marks et al. Bio/Technology 10:779-783 (1992) describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of CDR and/or framework residues is described by: Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al.
- Percent (%) amino acid sequence identity herein is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a selected sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared.
- % amino acid sequence identity values are obtained as described below by using the sequence comparison computer program ALIGN-2.
- the ALIGN-2 sequence comparison computer program was authored by Genentech, Inc. has been filed with user documentation in the U.S. Copyright Office, Washington D.C., 20559, where it is registered under U.S. Copyright Registration No. TXU510087, and is publicly available through Genentech, Inc., South San Francisco, Calif.
- the ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.
- % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B is calculated as follows:
- a “polypeptide chain” is a polypeptide wherein each of the domains thereof is joined to other domain(s) by peptide bond(s), as opposed to non-covalent interactions or disulfide bonds.
- a “flexible linker” herein refers to a peptide comprising two or more amino acid residues joined by peptide bond(s), and provides more rotational freedom for two polypeptides (such as two Fd regions) linked thereby. Such rotational freedom allows two or more antigen binding sites joined by the flexible linker to each access target antigen(s) more efficiently.
- suitable flexible linker peptide sequences include gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser, and gly-gly-gly-ser (SEQ ID NO:11).
- the flexible linker comprises 2 to about 10 amino acid residues, and most preferably four or less residues.
- a “dimerization domain” is formed by the association of at least two amino acid residues (generally cysteine residues) or of at least two peptides or polypeptides (which may have the same, or different, amino acid sequences).
- the peptides or polypeptides may interact with each other through covalent and/or non-covalent association(s).
- dimerization domains herein include an Fc region; a hinge region; a CH3 domain; a CH4 domain; a CH1-CL pair; an “interface” with an engineered “knob” and/or “protruberance” as described in U.S. Pat. No. 5,821,333, expressly incorporated herein by reference; a leucine zipper (e.g.
- a jun/fos leucine zipper see Kostelney et al., J. Immunol., 148: 1547-1553 (1992); or a yeast GCN4 leucine zipper); an isoleucine zipper; a receptor dimer pair (e.g., interleukin-8 receptor (IL-8R); and integrin heterodimers such as LFA-1 and GPIIIb/IIIa), or the dimerization region(s) thereof; dimeric ligand polypeptides (e.g. nerve growth factor (NGF), neurotrophin-3 (NT-3), interleukin-8 (IL-8), vascular endothelial growth factor (VEGF), and brain-derived neurotrophic factor (BDNF); see Arakawa et al. J.
- NGF nerve growth factor
- NT-3 neurotrophin-3
- IL-8 interleukin-8
- VEGF vascular endothelial growth factor
- BDNF brain-derived neurotrophic factor
- dimerization region(s) thereof a pair of cysteine residues able to form a disulfide bond; a pair of peptides or polypeptides, each comprising at least one cysteine residue (e.g. from about one, two or three to about ten cysteine residues) such that disulfide bond(s) can form between the peptides or polypeptides (hereinafter “a synthetic hinge”); and antibody variable domains.
- the most preferred dimerization domain herein is an Fc region or a hinge region.
- “Naturally occurring amino acid residues” may be selected from the group consisting of: alanine (Ala); arginine (Arg); asparagine (Asn); aspartic acid (Asp); cysteine (Cys); glutamine (Gin); glutamic acid (Glu); glycine (Gly); histidine (H is); isoleucine (Ile): leucine (Leu); lysine (Lys); methionine (Met); phenylalanine (Phe); proline (Pro); serine (Ser); threonine (Thr); tryptophan (Trp); tyrosine (Tyr); and valine (Val).
- non-naturally occurring amino acid residue refers to a residue, other than those naturally occurring amino acid residues listed above, which is able to covalently bind adjacent amino acid residues(s) in a polypeptide chain.
- non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine and other amino acid residue analogues such as those described in Ellman et al. Meth. Enzym. 202:301-336 (1991).
- the procedures of Noren et al. Science 244:182 (1989) and Ellman et al., supra can be used. Briefly, these procedures involve chemically activating a suppressor tRNA with a non-naturally occurring amino acid residue followed by in vitro transcription and translation of the RNA.
- an “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.
- the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain.
- Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.
- a “functional antigen binding site” of an antibody is one which is capable of binding a target antigen.
- the antigen binding affinity of the antigen binding site is not necessarily as strong as the parent antibody from which the antigen binding site is derived, but the ability to bind antigen must be measurable using any one of a variety of methods known for evaluating antibody binding to an antigen.
- the antigen binding affinity of each of the antigen binding sites of a multivalent antibody herein need not be quantitatively the same.
- the number of functional antigen binding sites can be evaluated using ultracentrifugation analysis as described in Example 2 below.
- ligand activation of a receptor is meant signal transduction (e.g. for a tyrosine kinase receptor, that caused by an intracellular kinase domain of a tyrosine kinase receptor phosphorylating tyrosine residues in the receptor or a substrate polypeptide) mediated by ligand binding to the receptor (or a receptor complex comprising the receptor of interest).
- an ErbB receptor In the case of an ErbB receptor, generally, this will involve binding of an ErbB ligand to an ErbB hetero-oligomer which activates a kinase domain of one or more of the ErbB receptors in the hetero-oligomer and thereby results in phosphorylation of tyrosine residues in one or more of the ErbB receptors and/or phosphorylation of tyrosine residues in additional substrate polypeptides(s).
- An antibody which “blocks” ligand activation of an receptor is one which reduces or prevents such activation as hereinabove defined. Such blocking can occur by any means, e.g. by interfering with: ligand binding to the receptor, receptor complex formation, tyrosine kinase activity of a tyrosine kinase receptor in a receptor complex and/or phosphorylation of tyrosine kinase residue(s) in or by the receptor.
- antibodies which block ligand activation of an ErbB receptor include monoclonal antibodies 2C4 and 7F3 (which block HRG activation of HER2/HER3 and HER2/HER4 hetero-oligomers; and EGF, TGF-beta or amphiregulin activation of an EGFR/HER2 hetero-oligomer); and L26, L96 and L288 antibodies (Klapper et al. Oncogene 14:2099-2109 (1997)), which block EGF and NDF binding to T47D cells which express EGFR, HER2, HER3 and HER4.
- An antibody having a “biological characteristic” of a designated antibody is one which possesses one or more of the biological characteristics of that antibody which distinguish it from other antibodies that bind to the same antigen.
- a “growth inhibitory agent” when used herein refers to a compound or composition which inhibits growth of a cell in vitro and/or in vivo.
- the growth inhibitory agent may be one which significantly reduces the percentage of cells in S phase.
- growth inhibitory agents include agents that block cell cycle progression (at a place other than S phase), such as agents that induce G1 arrest and M-phase arrest.
- Classical M-phase blockers include the vincas (vincristine and vinblastine), TAXOL®, and topo II inhibitors such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin.
- DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C. Further information can be found in The Molecular Basis of Cancer , Mendelsohn and Israel, eds., Chapter 1, entitled “Cell cycle regulation, oncogenes, and antineoplastic drugs” by Murakami et al. (WB Saunders: Philadelphia, 1995), especially p. 13.
- growth inhibitory anti-HER2 antibodies are those which bind to HER2 and inhibit the growth of cancer cells overexpressing HER2.
- Preferred growth inhibitory anti-HER2 antibodies inhibit growth of SKBR3 breast tumor cells in cell culture by greater than 20%, and preferably greater than 50% (e.g. from about 50% to about 100%) at an antibody concentration of about 0.5 to 30 ⁇ g/ml, where the growth inhibition is determined six days after exposure of the SKBR3 cells to the antibody (see U.S. Pat. No. 5,677,171 issued Oct. 14, 1997).
- an antibody which “induces cell death” is one which causes a viable cell to become nonviable.
- the cell is generally one which expresses the antigen to which the antibody binds, especially where the cell overexpresses the antigen.
- the cell is a cancer cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell.
- the cell may be a SKBR3, BT474, Calu 3, MDA-MB-453, MDA-MB-361 or SKOV3 cell.
- Cell death in vitro may be determined in the absence of complement and immune effector cells to distinguish cell death induced by antibody dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC).
- ADCC antibody dependent cell-mediated cytotoxicity
- CDC complement dependent cytotoxicity
- the assay for cell death may be performed using heat inactivated serum (i.e. in the absence of complement) and in the absence of immune effector cells.
- loss of membrane integrity as evaluated by uptake of propidium iodide (P1), trypan blue (see Moore et al. Cytotechnology 17:1-11 (1995)) or 7AAD can be assessed relative to untreated cells.
- P1 propidium iodide
- trypan blue see Moore et al. Cytotechnology 17:1-11 (1995)
- 7AAD can be assessed relative to untreated cells.
- An antibody which “induces apoptosis” is one which induces programmed cell death as determined by binding of annexin V, fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies).
- the cell is one which expresses the antigen to which the antibody binds and may be one which overexpresses the antigen.
- the cell may be a tumor cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell.
- the cell may be a SKBR3, BT474, Calu 3 cell, MDA-MB-453, MDA-MB-361 or SKOV3 cell.
- Various methods are available for evaluating the cellular events associated with apoptosis. For example, phosphatidyl serine (PS) translocation can be measured by annexin binding; DNA fragmentation can be evaluated through DNA laddering as disclosed in the example herein; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells.
- PS phosphatidyl serine
- the antibody which induces apoptosis is one which results in about 2 to 50 fold, preferably about 5 to 50 fold, and most preferably about 10 to 50 fold, induction of annexin binding relative to untreated cell in an annexin binding assay using cells expressing the antigen to which the antibody binds.
- antibodies which induce apoptosis include the anti-HER2 monoclonal antibodies 7F3 (ATCC HB-12216), and 7C2 (ATCC HB 12215), including humanized and/or affinity matured variants thereof; the anti-DR5 antibodies 3F11.39.7 (ATCC HB-12456); 3H3.14.5 (ATCC HB-12534); 3D5.1.10 (ATCC HB-12536); and 3H3.14.5 (ATCC HB-12534), including humanized and/or affinity matured variants thereof; the human anti-DR5 receptor antibodies 16E2 and 20E6, including affinity matured variants thereof (WO98/51793, expressly incorporated herein by reference); the anti-DR4 antibodies 4E7.24.3 (ATCC HB-12454); 4H6.17.8 (ATCC HB-12455); 1H5.25.9 (ATCC HB-12695); 4G7.18.8 (ATCC PTA-99); and 5G11.17.1 (ATCC HB-12694), including humanized and/or affinity
- an “agonist antibody” is an antibody which binds to and activates a receptor.
- the receptor activation capability of the agonist antibody will be at least qualitatively similar (and may be essentially quantitatively similar) to a native agonist ligand of the receptor.
- An example of an agonist antibody is one which binds to a receptor in the TNF receptor superfamily and induces apoptosis of cells expressing the TNF receptor. Assays for determining induction of apoptosis are described in WO98/51793 and WO99/37684, both of which are expressly incorporated herein by reference.
- a “disorder” is any condition that would benefit from treatment with the antibody. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question.
- disorders to be treated herein include benign and malignant tumors; leukemias and lymphoid malignancies; neuronal, glial, astrocytal, hypothalamic and other glandular, macrophagal, epithelial, stromal and blastocoelic disorders; and inflammatory, angiogenic and immunologic disorders.
- the term “therapeutically effective amount” refers to an amount of a drug effective to treat a disease or disorder in a mammal.
- the therapeutically effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the disorder.
- the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic.
- efficacy in vivo can, for example, be measured by assessing the time to disease progression (TTP) and/or determining the response rates (RR).
- Treatment refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
- cancer and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth.
- Examples of cancer include but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia.
- cancers include squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma and various types of head and neck cancer.
- autoimmune disease herein is a non-malignant disease or disorder arising from and directed against an individual's own tissues.
- autoimmune diseases or disorders include, but are not limited to, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g.
- atopic dermatitis atopic dermatitis
- systemic scleroderma and sclerosis responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g.
- Type I diabetes mellitus or insulin dependent diabetes mellitis multiple sclerosis; Reynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjorgen's syndrome; juvenile onset diabetes; and immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis; pernicious anemia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder; multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemia or Coombs positive anemia); myasthenia gravis; antigen-antibody complex mediated diseases; anti-glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyen
- foreign antigen is meant a molecule or molecules which is/are not endogenous or native to a mammal which is exposed to it.
- the foreign antigen may elicit an immune response, e.g. a humoral and/or T cell mediated response in the mammal. Generally, the foreign antigen will provoke the production of antibodies thereagainst.
- immunogenic therapeutic agents e.g. proteins such as antibodies, particularly antibodies comprising non-human amino acid residues (e.g.
- rodent, chimeric/humanized, and primatized antibodies include toxins (optionally conjugated to a targeting molecule such as an antibody, wherein the targeting molecule may also be immunogenic); gene therapy viral vectors, such as retroviruses and adenoviruses; grafts; infectious agents (e.g. bacteria and virus); alloantigens (i.e. an antigen that occurs in some, but not in other members of the same species) such as differences in blood types, human lymphocyte antigens (HLA), platelet antigens, antigens expressed on transplanted organs, blood components, pregnancy (Rh), and hemophilic factors (e.g. Factor VIII and Factor IX).
- HLA human lymphocyte antigens
- platelet antigens antigens expressed on transplanted organs
- blood components blood components
- pregnancy (Rh) pregnancy
- hemophilic factors e.g. Factor VIII and Factor IX
- blocking an immune response to a foreign antigen is meant reducing or preventing at least one immune-mediated response resulting from exposure to a foreign antigen.
- a foreign antigen For example, one may dampen a humoral response to the foreign antigen, i.e., by preventing or reducing the production of antibodies directed against the antigen in the mammal.
- graft refers to biological material derived from a donor for transplantation into a recipient. Grafts include such diverse material as, for example, isolated cells such as islet cells; tissue such as the amniotic membrane of a newborn, bone marrow, hematopoietic precursor cells, and ocular tissue, such as corneal tissue; and organs such as skin, heart, liver, spleen, pancreas, thyroid lobe, lung, kidney, tubular organs (e.g., intestine, blood vessels, or esophagus), etc. The tubular organs can be used to replace damaged portions of esophagus, blood vessels, or bile duct.
- isolated cells such as islet cells
- tissue such as the amniotic membrane of a newborn, bone marrow, hematopoietic precursor cells, and ocular tissue, such as corneal tissue
- organs such as skin, heart, liver, spleen, pancreas, thyroid lobe, lung, kidney, tubular
- the skin grafts can be used not only for burns, but also as a dressing to damaged intestine or to close certain defects such as diaphragmatic hernia.
- the graft is derived from any mammalian source, including human, whether from cadavers or living donors.
- the graft is bone marrow or an organ such as heart and the donor of the graft and the host are matched for HLA class II antigens.
- mammalian host refers to any compatible transplant recipient.
- compatible is meant a mammalian host that will accept the donated graft.
- the host is human. If both the donor of the graft and the host are human, they are preferably matched for HLA class II antigens so as to improve histocompatibility.
- donor refers to the mammalian species, dead or alive, from which the graft is derived.
- the donor is human.
- Human donors are preferably volunteer blood-related donors that are normal on physical examination and of the same major ABO blood group, because crossing major blood group barriers possibly prejudices survival of the allograft. It is, however, possible to transplant, for example, a kidney of a type O donor into an A, B or AB recipient.
- transplant refers to the insertion of a graft into a host, whether the transplantation is syngeneic (where the donor and recipient are genetically identical), allogeneic (where the donor and recipient are of different genetic origins but of the same species), or xenogeneic (where the donor and recipient are from different species).
- the host is human and the graft is an isograft, derived from a human of the same or different genetic origins.
- the graft is derived from a species different from that into which it is transplanted, such as a baboon heart transplanted into a human recipient host, and including animals from phylogenically widely separated species, for example, a pig heart valve, or animal beta islet cells or neuronal cells transplanted into a human host.
- a species different from that into which it is transplanted such as a baboon heart transplanted into a human recipient host, and including animals from phylogenically widely separated species, for example, a pig heart valve, or animal beta islet cells or neuronal cells transplanted into a human host.
- the expression “desensitizing a mammal awaiting transplantation” refers to reducing or abolishing allergic sensitivity or reactivity to a transplant, prior to administration of the transplant to the mammal. This may be achieved by any mechanism, such as a reduction in anti-donor antibodies in the desensitized mammal, e.g. where such anti-donor antibodies are directed against human lymphocyte antigen (HLA).
- HLA human lymphocyte antigen
- cytotoxic agent refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells.
- the term is intended to include radioactive isotopes (e.g. At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 and radioactive isotopes of Lu), chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.
- radioactive isotopes e.g. At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 and radioactive isotopes of Lu
- chemotherapeutic agents e.g. At 211 , I 131 , I 125 , Y 90 , Re 186
- chemotherapeutic agent is a chemical compound useful in the treatment of cancer.
- examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXANTM); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycin
- calicheamicin especially calicheamicin ⁇ 1 I and calicheamicin ⁇ I 1 , see, e.g., Agnew Chem. Intl. Ed. Engl. 33:183-186 (1994); dynemicin, including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromomophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin,
- paclitaxel TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.
- doxetaxel TAXOTERE®, Rhone-Poulenc Rorer, Antony, France
- chlorambucil gemcitabine
- 6-thioguanine mercaptopurine
- methotrexate platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above.
- DMFO diflu
- anti-hormonal agents that act to regulate or inhibit hormone action on tumors
- anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above.
- cytokine is a generic term for proteins released by one cell population which act on another cell as intercellular mediators.
- cytokines are lymphokines, monokines, and traditional polypeptide hormones. Included among the cytokines are growth hormone such as human growth hormone, N-methionyl human growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), and luteinizing hormone (LH); hepatic growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor-alpha and -beta; mullerian-inhibiting substance; mouse gonadotropin-associated peptide; inhibin; activin; vascular endothelial growth factor; integrin; thrombopoietin (TPO); nerve growth factors such as NGF-alpha; platelet
- prodrug refers to a precursor or derivative form of a pharmaceutically active substance that is less cytotoxic to tumor cells compared to the parent drug and is capable of being enzymatically activated or converted into the more active parent form. See, e.g., Wilman, “Prodrugs in Cancer Chemotherapy” Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Harbor (1986) and Stella et al., “Prodrugs: A Chemical Approach to Targeted Drug Delivery,” Directed Drug Delivery , Borchardt et al., (ed.), pp. 247-267, Humana Press (1985).
- the prodrugs of this invention include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, beta-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs or optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs which can be converted into the more active cytotoxic free drug.
- cytotoxic drugs that can be derivatized into a prodrug form for use in this invention include, but are not limited to, those chemotherapeutic agents described above.
- angiogenic factor is a growth factor which stimulates the development of blood vessels.
- the preferred angiogenic factor herein is Vascular Endothelial Growth Factor (VEGF).
- label when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide.
- the label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.
- an “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid.
- An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells.
- an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells.
- control sequences refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism.
- the control sequences that are suitable for prokaryotes include a promoter, optionally an operator sequence, and a ribosome binding site.
- Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.
- Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence.
- DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide;
- a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or
- a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation.
- “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
- the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny.
- the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context.
- the invention herein relates to a method for making a multivalent antibody.
- Various techniques for generating the “parent” or “starting” antibody from which the variable domain(s) of the multivalent antibody may be derived will be described later in this application.
- the multivalent antibody of particular interest herein is one which comprises at least three (and preferably four, or more, e.g. four or five to about eight) antigen binding sites. Generally, all of the antigen binding sites are “functional” as defined hereinabove.
- the multivalent antibody does not exist in nature and is not a native sequence IgM or IgA antibody.
- the multivalent antibody herein is preferably not produced in vitro by chemically cross-linking a pair antibodies (e.g. as in Ghetie et al. (1997), supra or Wolff et al. (1993), supra).
- the present application also provides multivalent antibodies which do not require introduced cysteine residue(s) in a parent antibody in order to make the multivalent antibody via disulfide bond(s) between a pair of Fc regions (e.g. as in Shopes et al. (1992), supra or Caron et al. (1992), supra).
- the multivalent antibody comprises a first polypeptide chain comprising at least two heavy chain (or light chain) variable domains and a second polypeptide chain comprising at least two heavy chain (or light chain) variable domains.
- the first polypeptide chain comprises two heavy chain variable domains and the second polypeptide chain also comprises two heavy chain variable domains, which can be combined with corresponding light chain variable domains (at least two for each polypeptide chain) to generate four (or more) antigen binding sites.
- the multivalent antibody comprises a dimerization domain which combines (1) two (or more) antigen binding sites with (2) one, two (or more) antigen binding sites.
- dimerization domains are contemplated herein, but the preferred dimerization domain is an Fc region or a hinge region.
- the multivalent antibody comprises an Fc region (e.g. a native sequence or variant Fc region)
- the Fc region is preferably “functional” as defined hereinabove and thus is capable of performing one or more antibody effector functions, such as ADCC or CDC.
- the multivalent antibody has only one Fc region or lacks an Fc region.
- the multivalent antibody comprises an Fc region
- the three or more antigen binding sites are provided amino terminal to the Fc region (rather than at the carboxy terminus of the Fc region as in Coloma and Morrison, (1997) supra).
- This may be achieved by providing a first polypeptide chain represented by the formula VD1-X1-VD2-X2-Fc, wherein (1) VD1 is a first heavy or light chain variable domain (preferably a heavy chain variable domain), (2) VD2 is a second heavy or light chain variable domain (preferably a heavy chain variable domain), (3) Fc comprises one chain of an Fc region, and (4) X1 and X2 represent an optional intervening amino acid or polypeptide.
- X1 and X2 comprise, or consist of, a CH1 domain (where VD1 or VD2 is a heavy chain variable domain) or a CL domain (where VD1 or VD2 is a light chain variable domain).
- X1 further comprises a flexible linker which is generally C-terminal to VD1 (or C-terminal to CH1 or CL, if present).
- the flexible linker may comprise a peptide such as gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser or gly-gly-gly-ser (SEQ ID NO:11).
- the multivalent antibody of particular interest herein comprises three or more (e.g. four or five to about eight) Fab polypeptides, each capable of binding antigen.
- the Fab fragments are preferably provided amino terminal to the Fc region (where the multivalent antibody has an Fc region).
- two or more Fd fragments may be fused to the amino terminus of one chain of an Fc region.
- the polypeptide chain thus engineered may be combined with (1) another polypeptide chain formed by two or more Fd fragments fused to the amino terminus of the other chain of the Fc region, as well as (2) complementary VL domains (e.g. four or more VL domains which each, optionally, are fused to a CL domain).
- the antibody comprises a flexible linker between the two or more Fd fragments.
- the multivalent antibody may, for example, comprise a pair of polypeptide chains with the formula (1) VH-CH1-flexible linker-VH-CH1-Fc chain, or (2) VH-CH1-VH-CH1-Fc chain (i.e. where there is no flexible linker between the two Fd fragments).
- the three or more functional antigen binding sites of the multivalent antibody herein are each preferably formed by a heavy and light chain variable domain.
- two or more heavy chain variable domains are fused together (optionally with intervening amino acid residue(s) as noted above)
- two or more complementary light chain variable domain-containing polypeptides are combined with the heavy chain variable domains (for instance by co-expressing the fusion protein and the light chain variable domain polypeptide(s) in the same host cell).
- the antibody comprises four, or five, or more (e.g. up to about eight) light chain variable domain polypeptides, which each, optionally, comprise a CL domain.
- the antibody with three or more more (e.g. three to about ten, but preferably three or four) antigen binding sites may comprise a polypeptide chain comprising three or more (e.g. three to about ten, but preferably three or four) heavy chain or light chain variable domains, wherein each of the variable domains is combined with, or associated with, three or more (e.g. three to about ten, but preferably three or four) light chain or heavy chain variable domain polypeptides in such a way as to form the antigen binding sites.
- the polypeptide chain comprises three or more heavy chain variable domains, it is combined or associated with three or more corresponding light chain variable domain polypeptides (e.g. with VL-CL polypeptides).
- the polypeptide chain comprises three or more light chain variable domains
- it is combined or associated with three or more corresponding heavy chain variable domain polypeptides (e.g. with VH-CH1 polypeptides).
- each of the three or more antigen binding sites is directed against the same antigen.
- antigens bound by such antibodies include (1) a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily (such receptors may be ‘trimeric receptors’, hence the antibody need only include only three antigen binding sites as desired) such as DR4 and DRS; (2) a B cell surface antigen such as CD20; (3) an ErbB receptor exemplified by the HER2 receptor; or (4) a cell surface protein expressed by tumor cells.
- TNF Tumor Necrosis Factor
- the polypeptide chain may comprise three (or four) heavy chain variable domains which are able to combine with three (or four) light chain variable domain polypeptides (preferably VL-CL polypeptides) to generate three (or four) antigen binding sites directed against the same antigen.
- VL-CL polypeptides preferably VL-CL polypeptides
- the multivalent antibody may also comprise a polypeptide chain comprising the formula: (a) VL-CL-flexible linker-VL-CL-flexible linker-VL-CL; In this embodiment, the polypeptide may comprise three to about eight VL-CL polypeptides joined by flexible linkers.
- the polypeptide may comprise three to about eight VH-CH1 polypeptides joined by flexible linkers.
- (c) (VL-CL) n wherein n is three or more more (e.g. three to about eight, but preferably three or four); or (d) (VH-CH1) n , wherein n is three or more more (e.g. three to about eight, but preferably three or four).
- the polypeptide chain comprises the formula: (a) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; or (c) (VH-CH1) n , wherein n is three or four.
- the multivalent antibodies herein have desirable properties particularly for in vivo therapy and diagnosis.
- the multivalent antibody may be internalized and catabolized by a cell expressing an antigen, to which the antibody binds, faster than a bivalent antibody.
- the invention provides an immunoconjugate comprising the multivalent antibody conjugated with a cytotoxic agent (e.g. one which is active in killing cells once internalized).
- a cytotoxic agent e.g. one which is active in killing cells once internalized.
- cytotoxic agents for generating an immunoconjugate are described herein, but the preferred cytotoxic agent is a radioactive isotope, a maytansinoid or a calecheamicin.
- the multivalent antibody, and/or a parent antibody from which at least one of the multivalent antibody's antigen binding specificities is derived may have certain properties.
- the multivalent antibody and/or parent antibody may (1) be an agonist antibody (e.g. where an antigen bound by the antibody is a receptor in the TNF receptor family or a B cell surface antigen); (2) induce apoptosis (for instance, where an antigen bound by the antibody is an ErbB receptor or a receptor in the TNF receptor superfamily); (3) bind a cell surface protein (such as a B cell surface antigen or an ErbB receptor) expressed on tumor cells; (4) bind a cell surface protein (e.g. Epidermal Growth Factor Receptor (EGFR), HER2 receptor, ErbB3 receptor, ErbB4 receptor, or DcR3 receptor) overexpressed by tumor cells; and/or (5) be a growth inhibitory antibody .
- EGFR Epidermal Growth Factor Receptor
- HER2 receptor ErbB3 receptor
- the multivalent antibody herein may have specificity for only one antigen, or more than one antigens (e.g. from two to about three antigens).
- the three or more functional antigen binding sites of the multivalent antibody may all bind the same antigen (preferably the same epitope on that antigen, in which case the multivalent antibody would be considered to be “monospecific”).
- This application also provides “multispecific” antibodies.
- the three or more functional antigen binding sites may bind two or more (e.g. from two to about three) different antigens or epitopes.
- a multivalent antibody directed against a receptor antigen can be engineered which, surprisingly, has agonistic and/or apoptosis-inducing capability which is quantitatively similar to that of the native ligand.
- quantitatively similar here is meant that the multivalent antibody has an activity in an assay which determines agonistic and/or apoptosis-inducing activity, within about ten fold, and preferably within about five fold of the agonistic and/or apoptosis-inducing activity of the native ligand.
- the antibody with agonistic and/or apoptosis-inducing activity may be one with specificity for a receptor in the TNF receptor superfamily, e.g.
- an Apo2L receptor such as DR4, DR5, DcR1 and DcR2 (preferably DR4 or DR5), in which case the activity of the antibody in an apoptosis assay such as those described in Example 3 below is within about ten fold, e.g. within about five fold, of the activity of Apo2L in that assay.
- the multivalent antibody herein may, in one embodiment of the invention, bind a B cell surface antigen.
- Preferred B cell surface antigens include CD19, CD20, CD22 and CD40, and most preferably CD20.
- multivalent antibodies herein possesses one or more functional Fc regions, it is anticipated to have the ability to mediate effector functions (such as ADCC and CDC) and have a longer half-life than multivalent antibodies lacking an Fc region.
- Such multivalent antibodies may be used where killing of cells, such as tumor or cancer cells, is desired.
- Other forms of the multivalent antibodies herein which lack a Fc region may be desirable where a shorter half-life is desired (e.g. for treating cardiovascular or inflammatory diseases or disorders, or where the antibody is conjugated with a cytotoxic agent); where internalization of the antibody is desired (e.g.
- an immunoconjugate comprising the antibody and a cytotoxic agent
- a cytotoxic agent for improved penetration of a solid tumor
- expression of the multivalent antibody in a non-mammalian host cell e.g. a prokaryotic host cell such as an E. coli host cell
- a non-mammalian host cell e.g. a prokaryotic host cell such as an E. coli host cell
- Such forms of the antibody may comprise a multivalent antibody including a dimerization domain, wherein the dimerization domain comprises an antibody hinge region fused to a leucine zipper domain (the leucine zipper domain facilitates association of the polypeptides which form the dimerization domain, but may be subsequently proteolytically removed prior to administration to a patient) (see FIG. 23C ); a multivalent antibody with three antigen binding sites such as that shown in FIG. 23D ; or a multivalent antibody with four antigen binding sites such as that depicted in FIG. 23E .
- the multivalent antibody herein is directed against, or binds specifically to, one or more target antigen(s).
- at least one of the antigens bound by the multivalent antibody is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal.
- antibodies directed against nonpolypeptide antigens are also contemplated.
- the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor.
- exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clotting factors such as factor VIIIC, factor IX, tissue factor (TF), and von Willebrands factor; anti-clotting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and
- Preferred molecular targets for antibodies encompassed by the present invention include leukocyte surface markers or CD proteins such as CD1a-c, CD2, CD2R, CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CDw12, CD13, CD14, CD15, CD15s, CD16, CD16b, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, C41, CD42a-d, CD43, CD44, CD44R, CD45, CD45A, CD45B, CD450, CD46-CD48, CD49a-f, CD50, CD51, CD52, CD53-CD59, CDw60, CD61, CD62E, CD62L, CD62P, CD63, CD64, CDw65
- anti-CD11a, anti-CD18 or anti-CD11b antibodies growth factors such as VEGF; tissue factor (TF); alpha interferon ( ⁇ -IFN); an interleukin, such as IL-8; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; c-mpl receptor; CTLA-4; protein C etc.
- growth factors such as VEGF; tissue factor (TF); alpha interferon ( ⁇ -IFN); an interleukin, such as IL-8; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; c-mpl receptor; CTLA-4; protein C etc.
- Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies.
- immunogens for transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen.
- transmembrane molecules such as receptors
- fragments of these e.g. the extracellular domain of a receptor
- cells expressing the transmembrane molecule can be used as the immunogen.
- Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule.
- Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art.
- Preferred target antigens for the multivalent antibodies herein include (1) ErbB receptors, including EGFR, HER2, HER3 and HER4; (2) receptors in the TNF receptor superfamily, e.g. Apo2L receptors, such as DR4, DR5, DcR1 and DcR2; (3) B cell surface antigens, especially CD19, CD20, CD22 and CD40; (4) antigens expressed by tumor cells; (5) antigens overexpressed by tumor cells (e.g. ErbB receptors; DcR3 receptors); (6) receptors activated by multimeric (e.g. dimeric or trimeric) ligands (e.g. receptors in the TNF receptor superfamily; VEGF receptors, etc.).
- three or more (e.g. four to about eight) of the antigen binding sites of the multivalent antibody may all be directed against the same antigenic determinant or epitope on one of the above antigens.
- the present application also provides multispecific antibodies, i.e., antibodies that have binding specificities for at least two different epitopes or antigenic determinants.
- Multispecific antibodies e.g. bispecific antibodies; BsAbs
- BsAbs bispecific antibodies
- Bispecific antibodies have been very useful in probing the functional properties of cell surface molecules and in defining the ability of the different Fc receptors to mediate cytotoxicity (Fanger et al., Crit. Rev. Immunol. 12:101-124 (1992)). Nolan et al., Biochem. Biophys. Acta. 1040:1-11 (1990) describe other diagnostic applications for BsAbs.
- BsAbs can be constructed to immobilize enzymes for use in enzyme immunoassays.
- one arm of the BsAb can be designed to bind to a specific epitope on the enzyme so that binding does not cause enzyme inhibition, the other arm of the BsAb binds to the immobilizing matrix ensuring a high enzyme density at the desired site.
- BsAbs examples include the rabbit anti-IgG/anti-ferritin BsAb described by Hammerling et al., J. Exp. Med. 128:1461-1473 (1968) which was used to locate surface antigens. BsAbs having binding specificities for Horse Radish Peroxidase (HRP) as well as a hormone have also been developed.
- HRP Horse Radish Peroxidase
- Another potential immunochemical application for BsAbs involves their use in two-site immunoassays. For example, two BsAbs are produced binding to two separate epitopes on the analyte protein—one BsAb binds the complex to an insoluble matrix, the other binds an indicator enzyme (see Nolan et al., supra).
- Multispecific antibodies can also be used for in vitro or in vivo immunodiagnosis of various diseases such as cancer (Songsivilai et al., Clin. Exp. Immunol. 79:315 (1990)).
- one arm of the BsAb can bind a tumor associated antigen and the other arm can bind a detectable marker such as a chelator which tightly binds a radionuclide.
- a detectable marker such as a chelator which tightly binds a radionuclide.
- Multispecific antibodies can also be used for human therapy in redirected cytotoxicity by providing one arm which binds a target (e.g. pathogen or tumor cell) and another arm which binds a cytotoxic trigger molecule, such as the T-cell receptor or an Fc gamma receptor. Accordingly, multispecific antibodies can be used to direct a patient's cellular immune defense mechanisms specifically to the tumor cell or infectious agent. Using this strategy, it has been demonstrated that bispecific antibodies which bind to the Fc gamma RIII (i.e. CD16) can mediate tumor cell killing by natural killer (NK) cell/large granular lymphocyte (LGL) cells in vitro and are effective in preventing tumor growth in vivo.
- NK natural killer
- LGL large granular lymphocyte
- the bispecific antibodies link the CD3 complex on T cells to a tumor-associated antigen.
- a fully humanized F(ab′) 2 BsAb consisting of anti-CD3 linked to anti-p185 HER2 has been used to target T cells to kill tumor cells overexpressing the HER2 receptor.
- Bispecific antibodies have been tested in several early phase clinical trials with encouraging results. In one trial, 12 patients with lung, ovarian or breast cancer were treated with infusions of activated T-lymphocytes targeted with an anti-CD3/anti-tumor (MOC31) bispecific antibody. deLeij et al.
- Multispecific antibodies may also be used as fibrinolytic agents or vaccine adjuvants. Furthermore, these antibodies may be used in the treatment of infectious diseases (e.g. for targeting of effector cells to virally infected cells such as HIV or influenza virus or protozoa such as Toxoplasma gondii ), used to deliver immunotoxins to tumor cells, or target immune complexes to cell surface receptors (see Fanger et al., supra).
- infectious diseases e.g. for targeting of effector cells to virally infected cells such as HIV or influenza virus or protozoa such as Toxoplasma gondii
- Toxoplasma gondii protozoa
- the multispecific antibody may bind two or more different epitopes on an antigen of interest.
- the multispecfic antibody may have specificity for (1) an antigen expressed by a target cell (e.g. where the target cell is a tumor cell) and (2) a triggering molecule on a leukocyte, such as a T-cell receptor molecule (e.g. CD2 or CD3), or Fc receptors for IgG (Fc gamma R), such as Fc gamma RI (CD64), Fc gamma RII (CD32) and Fc gamma RIII (CD16) so as to focus cellular defense mechanisms to the antigen-expressing cell.
- a target cell e.g. where the target cell is a tumor cell
- a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2 or CD3), or Fc receptors for IgG (Fc gamma R), such as Fc gamma RI (CD
- Multispecific antibodies may also be used to localize cytotoxic agents to cells which express the target antigen. These antibodies possess an target antigen-binding arm and an arm which binds the cytotoxic agent (e.g. saporin, interferon-alpha, vinca alkaloid, ricin A chain, methotrexate or radioactive isotope hapten).
- cytotoxic agent e.g. saporin, interferon-alpha, vinca alkaloid, ricin A chain, methotrexate or radioactive isotope hapten.
- WO 96/16673 describes a bispecific anti-HER2/anti-Fc gamma RIII antibody and U.S. Pat. No. 5,837,234 discloses a bispecific anti-HER2/anti-Fc gamma RI antibody. A bispecific anti-HER2/Fc alpha antibody is shown in WO98/02463. U.S. Pat. No. 5,821,337 teaches a bispecific anti-HER2/anti-CD3 antibody.
- a “parent” or “starting” antibody with variable domains directed against an antigen may be prepared using various methodologies for making antibodies, such as those described hereinbelow.
- the sequences of the variable domains of the starting or parent antibody may be used in the design of the multivalent antibody herein.
- Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl 2 , or R 1 N ⁇ C ⁇ NR, where R and R 1 are different alkyl groups.
- a protein that is immunogenic in the species to be immunized e.g., keyhole limpet hemocyanin, serum albumin, bovine thy
- Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 ⁇ g or 5 ⁇ g of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites.
- the animals are boosted with 1 ⁇ 5 to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites.
- Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus.
- the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent.
- Conjugates also can be made in recombinant cell culture as protein fusions.
- aggregating agents such as alum are suitably used to enhance the immune response.
- Monoclonal antibodies are obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts.
- the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies.
- the monoclonal antibodies may be made using the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567).
- a mouse or other appropriate host animal such as a hamster
- lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization.
- lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , pp. 59-103 (Academic Press, 1986)).
- the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
- a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
- the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
- Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium.
- preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA.
- Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); and Brodeur et al., Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
- Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen.
- the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
- RIA radioimmunoassay
- ELISA enzyme-linked immunoabsorbent assay
- the binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980).
- the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice , pp. 59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
- the hybridoma cells may be grown in vivo as ascites tumors in an animal.
- the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional antibody purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
- DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies).
- the hybridoma cells serve as a preferred source of such DNA.
- the DNA may be placed into expression vectors, which are then transfected into host cells such as E. coli cells, simian COS cells, Chinese Hamster Ovary (CHO) cells, or myeloma cells that do not otherwise produce antibody protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells.
- host cells such as E. coli cells, simian COS cells, Chinese Hamster Ovary (CHO) cells, or myeloma cells that do not otherwise produce antibody protein.
- Recombinant expression of antibodies is described in more detail below.
- monoclonal antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al., Nature, 348:552-554 (1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biot., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries.
- the DNA also may be modified, for example, by substituting the coding sequence for human heavy chain and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; and Morrison, et al., Proc. Natl. Acad. Sci. USA, 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
- non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen.
- Human monoclonal antibodies may be made via an adaptation of the hybridoma method first described by Kohler and Milstein by using human B lymphocytes as the fusion partner.
- Human B lymphocytes producing an antibody of interest may, for example, be isolated from a human individual, after obtaining informed consent.
- the individual may be producing antibodies against an autoantigen as occurs with certain disorders such as systemic lupus erythematosus (Shoenfeld et al. J. Clin. Invest. 70:205 (1982)), immune-mediated thrombocytopenic purpura (ITP) (Nugent et al. Blood 70(1):16-22 (1987)), or cancer.
- lymphocytes may be immunized in vitro.
- a lysomotrophic agent e.g. L-leucine-O-methyl ester, L-glutamic acid dimethly ester or L-leucyl-L-leucine-O-methyl ester
- adjuvants such as 8-mercaptoguanosine and cytokines
- the B lymphocytes recovered from the subject or immunized in vitro are then generally immortalized in order to generate a human monoclonal antibody.
- Techniques for immortalizing the B lymphocyte include, but are not limited to: (a) fusion of the human B lymphocyte with human, murine myelomas or mouse-human heteromyeloma cells; (b) viral transformation (e.g. with an Epstein-Barr virus; see Nugent et al., supra, for example); (c) fusion with a lymphoblastoid cell line; or (d) fusion with lymphoma cells.
- Lymphocytes may be fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice , pp. 59-103 (Academic Press, 1986)).
- a suitable fusing agent such as polyethylene glycol
- the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
- the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
- HGPRT medium hypoxanthine, aminopterin, and thymidine
- Suitable human myeloma and mouse-human heteromyeloma cell lines have been described (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications , pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
- Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen.
- the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
- RIA radioimmunoassay
- ELISA enzyme-linked immunoabsorbent assay
- the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice , pp. 59-103 (Academic Press, 1986)).
- Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
- the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A chromatography, gel electrophoresis, dialysis, or affinity chromatography.
- Human antibodies may also be generated using a non-human host, such as a mouse, which is capable of producing human antibodies.
- a non-human host such as a mouse
- transgenic mice are now available that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production.
- J H antibody heavy-chain joining region
- the human antibody may be selected from a human antibody phage display library.
- the preparation of libraries of antibodies or fragments thereof is well known in the art and any of the known methods may be used to construct a family of transformation vectors which may be introduced into host cells.
- Libraries of antibody light and heavy chains in phage (Huse et al., Science, 246:1275 (1989)) or of fusion proteins in phage or phagemid can be prepared according to known procedures. See, for example, Vaughan et al., Nature Biotechnology 14:309-314 (1996); Barbas et al., Proc. Natl. Acad. Sci., USA, 88:7978-7982 (1991); Marks et al., J. Mol.
- a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain.
- Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting hypervariable region sequences for the corresponding sequences of a human antibody.
- humanized antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567) wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species.
- humanized antibodies are typically human antibodies in which some hypervariable region residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
- variable domains both light and heavy
- sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences.
- the human sequence which is closest to that of the rodent is then accepted as the human framework for the humanized antibody (Sims et al., J. Immunol., 151:2296 (1993); Chothia et al., J. Mol. Biol., 196:901 (1987)).
- Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains.
- the same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623 (1993)).
- humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences.
- Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art.
- Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, L e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen.
- FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved.
- the hypervariable region residues are directly and most substantially involved in influencing antigen binding.
- antibody fragments Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see, e.g., Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992); and Brennan et al., Science, 229:81 (1985)). However, these fragments can now be produced directly by recombinant host cells. For example, the antibody fragments can be isolated from the antibody phage libraries discussed above. Alternatively, Fab′-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab′) 2 fragments (Carter et al., Bio/Technology 10:163-167 (1992)).
- F(ab′) 2 fragments can be isolated directly from recombinant host cell culture.
- the antibody of choice is a single chain Fv fragment (scFv). See WO 93/16185; U.S. Pat. No. 5,571,894; and U.S. Pat. No. 5,587,458.
- the antibody fragment may also be a “linear antibody”, e.g., as described in U.S. Pat. No. 5,641,870 for example. Such linear antibody fragments may be monospecific or bispecific.
- Amino acid sequence modification(s) of the antibodies described herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody.
- Amino acid sequence variants of the antibody are prepared by introducing appropriate nucleotide changes into the antibody nucleic acid, or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of, residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics.
- the amino acid changes also may alter post-translational processes of the antibody, such as changing the number or position of glycosylation sites. Such alterations may be made to the parent antibody and/or multivalent antibody and/or may be introduced in the multivalent antibody amino acid sequence at the time that sequence is made.
- a useful method for identification of certain residues or regions of the antibody that are preferred locations for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells Science, 244:1081-1085 (1989).
- a residue or group of target residues are identified (e.g., charged residues such as arg, asp, his, lys, and glu) and replaced by a neutral or negatively charged amino acid (most preferably alanine or polyalanine) to affect the interaction of the amino acids with antigen.
- Those amino acid locations demonstrating functional sensitivity to the substitutions then are refined by introducing further or other variants at, or for, the sites of substitution.
- the site for introducing an amino acid sequence variation is predetermined, the nature of the mutation per se need not be predetermined. For example, to analyze the performance of a mutation at a given site, ala scanning or random mutagenesis is conducted at the target codon or region and the expressed multivalent antibodies are screened for the desired activity.
- Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues.
- terminal insertions include an antibody with an N-terminal methionyl residue or the antibody fused to a cytotoxic polypeptide.
- Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody to an enzyme (e.g. for ADEPT) or a polypeptide which increases the serum half-life of the antibody.
- variants are an amino acid substitution variant. These variants have at least one amino acid residue in the antibody molecule replaced by a different residue.
- the sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but FR alterations are also contemplated.
- Conservative substitutions are shown in Table 1 under the heading of “preferred substitutions”. If such substitutions result in a change in biological activity, then more substantial changes, denominated “exemplary substitutions” in Table 1, or as further described below in reference to amino acid classes, may be introduced and the products screened.
- Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
- Naturally occurring residues are divided into groups based on common side-chain properties:
- hydrophobic norleucine, met, ala, val, leu, ile
- Non-conservative substitutions will entail exchanging a member of one of these classes for another class.
- cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability.
- a particularly preferred type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human antibody).
- a parent antibody e.g. a humanized or human antibody
- the resulting variant(s) selected for further development will have improved biological properties relative to the parent antibody from which they are generated.
- a convenient way for generating such substitutional variants involves affinity maturation using phage display. Briefly, several hypervariable region sites (e.g. 6-7 sites) are mutated to generate all possible amino substitutions at each site.
- the multivalent antibodies thus generated are displayed in a monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage-displayed variants are then screened for their biological activity (e.g. binding affinity) as herein disclosed.
- alanine scanning mutagenesis can be performed to identify hypervariable region residues contributing significantly to antigen binding.
- the panel of variants is subjected to screening as described herein and antibodies with superior properties in one or more relevant assays may be selected for further development.
- Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. By altering is meant deleting one or more carbohydrate moieties found in the antibody, and/or adding one or more glycosylation sites that are not present in the antibody.
- N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue.
- the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain.
- X is any amino acid except proline
- O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.
- glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites).
- the alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).
- Nucleic acid molecules encoding amino acid sequence variants of the antibody are prepared by a variety of methods known in the art. These methods include, but are not limited to, isolation from a natural source (in the case of naturally occurring amino acid sequence variants) or preparation by oligonucleotide-mediated (or site-directed) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared variant or a non-variant version of the antibody.
- the antibody of the invention may be desirable to modify the antibody of the invention with respect to effector function, e.g. so as to enhance antigen-dependent cell-mediated cyotoxicity (ADCC) and/or complement dependent cytotoxicity (CDC) of the antibody.
- ADCC antigen-dependent cell-mediated cyotoxicity
- CDC complement dependent cytotoxicity
- This may be achieved by introducing one or more amino acid modifications in an Fc region of the antibody, thereby generating a variant Fc region.
- the Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.
- the variant Fc region may mediate antibody-dependent cell-mediated cytotoxicity (ADCC) in the presence of human effector cells more effectively, or bind an Fc gamma receptor (Fc ⁇ R) with better affinity, than a native sequence Fc region.
- ADCC antibody-dependent cell-mediated cytotoxicity
- Such Fc region variants may comprise an amino acid modification at any one or more of positions 256, 290, 298, 312, 326, 330, 333, 334, 360, 378 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant with reduced binding to an Fc ⁇ R may comprise an amino acid modification at any one or more of amino acid positions 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 292, 293, 294, 295, 296, 298, 301, 303, 322, 324, 327, 329, 333, 335, 338, 340, 373, 376, 382, 388, 389, 414, 416, 419, 434, 435, 437, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant may display reduced binding to an Fc ⁇ RI and comprise an amino acid modification at any one or more of amino acid positions 238, 265, 269, 270, 327 or 329 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant may display reduced binding to an Fc ⁇ RII and comprise an amino acid modification at any one or more of amino acid positions 238, 265, 269, 270, 292, 294, 295, 298, 303, 324, 327, 329, 333, 335, 338, 373, 376, 414, 416, 419, 435, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant of interest may display reduced binding to an Fc ⁇ RIII and comprise an amino acid modification at one or more of amino acid positions 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 293, 294, 295, 296, 301, 303, 322, 327, 329, 338, 340, 373, 376, 382, 388, 389, 416, 434, 435 or 437 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant displays improved binding to the Fc ⁇ R and comprises an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 298, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 333, 334, 337, 340, 360, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant may display increased binding to an Fc ⁇ RIII and, optionally, may further display decreased binding to an Fc ⁇ RII.
- An exemplary such variant comprises amino acid modification(s) at position(s) 298 and/or 333 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the Fc region variant may display increased binding to an Fc ⁇ RII and comprise an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 337, 340, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- Such Fc region variants with increased binding to an Fc ⁇ RII may optionally further display decreased binding to an Fc ⁇ RIII and may, for example, comprise an amino acid modification at any one or more of amino acid positions 268, 272, 298, 301, 322 or 340 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the variant Fc region may alternatively or additionally have altered neonatal Fc receptor (FcRn) binding affinity.
- Such variant Fc regions may comprise an amino acid modification at any one or more of amino acid positions 238, 252, 253, 254, 255, 256, 265, 272, 286, 288, 303, 305, 307, 309, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 386, 388, 400, 413, 415, 424, 433, 434, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- Fc region variants with reduced binding to an FcRn may comprise an amino acid modification at any one or more of amino acid positions 252, 253, 254, 255, 288, 309, 386, 388, 400, 415, 433, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- the above-mentioned Fc region variants may, alternatively, display increased binding to FcRn and comprise an amino acid modification at any one or more of amino acid positions 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- Fc region variants with altered (i.e. improved or diminished) C1q binding and/or Complement Dependent Cytotoxicity (CDC) are described in WO99/51642.
- Such variants may comprise an amino acid substitution at one or more of amino acid positions 270, 322, 326, 327, 329, 331, 333 or 334 of the Fc region. See, also, Duncan & Winter Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO94/29351 concerning Fc region variants.
- a salvage receptor binding epitope refers to an epitope of the Fc region of an IgG molecule (e.g., IgG 1 , IgG 2 , IgG 3 , or IgG 4 ) that is responsible for increasing the in vivo serum half-life of the IgG molecule.
- the invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g. a small molecule toxin or an enzymatically active toxin of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof), or a radioactive isotope (i.e., a radioconjugate).
- a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g. a small molecule toxin or an enzymatically active toxin of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof), or a radioactive isotope (i.e., a radioconjugate).
- Conjugates of an antibody and one or more small molecule toxins such as a calicheamicin, a maytansine (U.S. Pat. No. 5,208,020), a trichothene, and CC1065 are also contemplated herein.
- the antibody is conjugated to one or more maytansine molecules (e.g. about 1 to about 10 maytansine molecules per antibody molecule).
- Maytansine may, for example, be converted to May-SS-Me which may be reduced to May-SH3 and reacted with modified antibody (Chari et al. Cancer Research 52: 127-131 (1992)) to generate a maytansinoid-antibody immunoconjugate.
- Another immunoconjugate of interest comprises an antibody conjugated to one or more calicheamicin molecules.
- the calicheamicin family of antibiotics are capable of producing double-stranded DNA breaks at sub-picomolar concentrations.
- Structural analogues of calicheamicin which may be used include, but are not limited to, ⁇ 1 I , ⁇ 2 I , ⁇ 3 I , N-acetyl- ⁇ 1 I , PSAG and ⁇ I 1 (Hinman et al. Cancer Research 53: 3336-3342 (1993) and Lode et al. Cancer Research 58: 2925-2928 (1998)). See, also, U.S. Pat. Nos. 5,714,586; 5,712,374; 5,264,586; and 5,773,001 expressly incorporated herein by reference.
- Enzymatically active toxins and fragments thereof which can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa ), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and the tricothecenes. See, for example, WO 93/21232 published Oct. 28, 1993.
- the present invention further contemplates an immunoconjugate formed between an antibody and a compound with nucleolytic activity (e.g. a ribonuclease or a DNA endonuclease such as a deoxyribonuclease; DNase).
- a compound with nucleolytic activity e.g. a ribonuclease or a DNA endonuclease such as a deoxyribonuclease; DNase.
- radioactive isotopes are available for the production of radioconjugated antibodies. Examples include At 211 , I 131 , I 125 , Y 90 , Re 186 , Re 188 , Sm 153 , Bi 212 , P 32 and radioactive isotopes of Lu.
- Conjugates of the antibody and cytotoxic agent may be made using a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate, iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoylyethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro
- a ricin immunotoxin can be prepared as described in Vitetta et al. Science 238: 1098 (1987).
- Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026.
- the linker may be a “cleavable linker” facilitating release of the cytotoxic drug in the cell.
- an acid-labile linker, peptidase-sensitive linker, dimethyl linker or disulfide-containing linker (Chari et al. Cancer Research 52: 127-131 (1992)) may be used.
- a fusion protein comprising the antibody and cytotoxic agent may be made, e.g. by recombinant techniques or peptide synthesis.
- the antibody may be conjugated to a “receptor” (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a “ligand” (e.g. avidin) which is conjugated to a cytotoxic agent (e.g. a radionucleotide).
- a “receptor” such streptavidin
- a ligand e.g. avidin
- cytotoxic agent e.g. a radionucleotide
- the antibodies of the present invention may also be used in ADEPT by conjugating the antibody to a prodrug-activating enzyme which converts a prodrug (e.g. a peptidyl chemotherapeutic agent, see WO81/01145) to an active anti-cancer drug.
- a prodrug e.g. a peptidyl chemotherapeutic agent, see WO81/01145
- WO 88/07378 and U.S. Pat. No. 4,975,278 See, for example, WO 88/07378 and U.S. Pat. No. 4,975,278.
- the enzyme component of the immunoconjugate useful for ADEPT includes any enzyme capable of acting on a prodrug in such a way so as to covert it into its more active, cytotoxic form.
- Enzymes that are useful in the method of this invention include, but are not limited to, alkaline phosphatase useful for converting phosphate-containing prodrugs into free drugs; arylsulfatase useful for converting sulfate-containing prodrugs into free drugs; cytosine deaminase useful for converting non-toxic 5-fluorocytosine into the anti-cancer drug, 5-fluorouracil; proteases, such as serratia protease, thermolysin, subtilisin, carboxypeptidases and cathepsins (such as cathepsins B and L), that are useful for converting peptide-containing prodrugs into free drugs; D-alanylcarboxypeptidases, useful for converting prodrugs that contain D-amino acid substituents; carbohydrate-cleaving enzymes such as beta-galactosidase and neuraminidase useful for converting glycosylated prodrugs into free drugs; beta-lac
- antibodies with enzymatic activity can be used to convert the prodrugs of the invention into free active drugs (see, e.g., Massey, Nature 328: 457-458 (1987)).
- Antibody-abzyme conjugates can be prepared as described herein for delivery of the abzyme to a tumor cell population.
- the enzymes of this invention can be covalently bound to the antibodies by techniques well known in the art such as the use of the heterobifunctional crosslinking reagents discussed above.
- fusion proteins comprising at least the antigen binding region of an antibody of the invention linked to at least a functionally active portion of an enzyme of the invention can be constructed using recombinant DNA techniques well known in the art (see, e.g., Neuberger et al., Nature, 312: 604-608 (1984).
- the antibody may be linked to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol.
- the antibody also may be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules), or in macroemulsions.
- colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules
- the antibodies disclosed herein may also be formulated as immunoliposomes.
- Liposomes containing the antibody are prepared by methods known in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA, 82:3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA, 77:4030 (1980); U.S. Pat. Nos. 4,485,045 and 4,544,545; and WO97/38731 published Oct. 23, 1997. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556.
- Particularly useful liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter.
- Fab′ fragments of the antibody of the present invention can be conjugated to the liposomes as described in Martin et al. J. Biol. Chem. 257: 286-288 (1982) via a disulfide interchange reaction. A chemotherapeutic agent is optionally contained within the liposome. See Gabizon et al. J. National Cancer Inst. 81(19)1484 (1989).
- the invention also provides isolated nucleic acid encoding an antibody as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the antibody.
- the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression.
- DNA encoding the antibody is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the antibody).
- Many vectors are available.
- the vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
- the multivalent antibody of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
- a heterologous polypeptide which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
- the heterologous signal sequence selected preferably is one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell.
- the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
- a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders.
- yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, ⁇ factor leader (including Saccharomyces and Kluyveromyces ⁇ -factor leaders), or acid phosphatase leader, the C. albicans glucoamylase leader, or the signal described in WO 90/13646.
- mammalian signal sequences as well as viral secretory leaders for example, the herpes simplex gD signal, are available.
- the DNA for such precursor region is ligated in reading frame to DNA encoding the multivalent antibody.
- Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells.
- this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences.
- origins of replication or autonomously replicating sequences are well known for a variety of bacteria, yeast, and viruses.
- the origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2 ⁇ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells.
- the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter).
- Selection genes may contain a selection gene, also termed a selectable marker.
- Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli.
- One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin.
- Suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the multivalent antibody nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc.
- cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR.
- Mtx methotrexate
- An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity.
- host cells particularly wild-type hosts that contain endogenous DHFR transformed or co-transformed with DNA sequences encoding multivalent antibody, wild-type DHFR protein, and another selectable marker such as aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199.
- APH aminoglycoside 3′-phosphotransferase
- a suitable selection gene for use in yeast is the trp1 gene present in the yeast plasmid YRp7 (Stinchcomb et al., Nature, 282:39 (1979)).
- the trp1 gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977).
- the presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan.
- Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
- vectors derived from the 1.6 ⁇ m circular plasmid pKD1 can be used for transformation of Kluyveromyces yeasts.
- an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis . Van den Berg, Bio/Technology, 8:135 (1990).
- Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology, 9:968-975 (1991).
- Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the multivalent antibody nucleic acid.
- Promoters suitable for use with prokaryotic hosts include the phoA promoter, ⁇ -lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter.
- phoA promoter phoA promoter
- ⁇ -lactamase and lactose promoter systems alkaline phosphatase
- trp tryptophan
- hybrid promoters such as the tac promoter.
- Other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the multivalent antibody.
- S.D. Shine-Dalgarno
- Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors.
- suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase.
- 3-phosphoglycerate kinase or other glycolytic enzymes such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate
- yeast promoters which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization.
- Suitable vectors and promoters for use in yeast expression are further described in EP 73,657.
- Yeast enhancers also are advantageously used with yeast promoters.
- Multivalent antibody transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems.
- viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV
- the early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication.
- the immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment.
- a system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978. See also Reyes et al., Nature 297:598-601 (1982) on expression of human ⁇ -interferon cDNA in mouse cells under the control of a thymidine kinase promoter from herpes simplex virus. Alternatively, the rous sarcoma virus long terminal repeat can be used as the promoter.
- Enhancer sequences are now known from mammalian genes (globin, elastase, albumin, ⁇ -fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
- the enhancer may be spliced into the vector at a position 5′ or 3′ to the multivalent antibody-encoding sequence, but is preferably located at a site 5′ from the promoter.
- Expression vectors used in eukaryotic host cells will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′, untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the multivalent antibody.
- One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein.
- Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above.
- Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia , e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella , e.g., Salmonella typhimurium, Serratia , e.g., Serratia marcescans , and Shigella , as well as Bacilli such as B. subtilis and B.
- Enterobacteriaceae such as Escherichia , e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus
- Salmonella e.g., Salmonella typhimurium
- Serratia
- E. coli cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coli X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
- eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for multivalent antibody-encoding vectors.
- Saccharomyces cerevisiae or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms.
- a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K.
- waltii ATCC 56,500
- K. drosophilarum ATCC 36,906
- K. thermotolerans K. marxianus
- yarrowia EP 402,226
- Pichia pastoris EP 183,070
- Candida Trichoderma reesia
- Neurospora crassa Schwanniomyces such as Schwanniomyces occidentalis
- filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium , and Aspergillus hosts such as A. nidulans and A. niger.
- Suitable host cells for the expression of glycosylated multivalent antibody are derived from multicellular organisms.
- invertebrate cells include plant and insect cells.
- Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruitfly), and Bombyx mori have been identified.
- a variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
- Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
- vertebrate cells have been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure.
- useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR(CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod.
- monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals N.Y. Acad. Sci.
- MRC 5 cells MRC 5 cells; FS4 cells; a human hepatoma line (Hep G2); and myeloma or lymphoma cells (e.g. Y0, J558L, P3 and NS0 cells) (see U.S. Pat. No. 5,807,715).
- Hep G2 human hepatoma line
- myeloma or lymphoma cells e.g. Y0, J558L, P3 and NS0 cells
- Host cells are transformed with the above-described expression or cloning vectors for multivalent antibody production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
- the host cells used to produce the multivalent antibody of this invention may be cultured in a variety of media.
- Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells.
- any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCINTM drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art.
- the culture conditions such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
- the multivalent antibody can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the multivalent antibody is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/Technology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E. coli . Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min.
- sodium acetate pH 3.5
- EDTA EDTA
- PMSF phenylmethylsulfonylfluoride
- Cell debris can be removed by centrifugation.
- supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit.
- a protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
- the multivalent antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique.
- affinity chromatography is the preferred purification technique.
- the suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the multivalent antibody.
- Protein A can be used to purify antibodies that are based on human ⁇ 1, ⁇ 2, or ⁇ 4 heavy chains (Lindmark et al., J. Immunol. Meth. 62:1-13 (1983)). Protein G is recommended for all mouse isotypes and for human ⁇ 3 (Guss et al., EMBO J.
- the matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose.
- the Bakerbond ABXTM resin J. T. Baker, Phillipsburg, N.J. is useful for purification.
- Therapeutic formulations of the multivalent antibody are prepared for storage by mixing the multivalent antibody having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers ( Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of aqueous solutions, lyophilized or other dried formulations.
- Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine,
- the formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other.
- active compounds are provided in Section G below entitled “In Vivo Uses for the Multivalent Antibody”. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
- the active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions.
- colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules
- the formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
- Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the multivalent antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No.
- copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate copolymers of L-glutamic acid and ⁇ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOTTM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-( ⁇ )-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
- encapsulated antibodies When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37° C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S—S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
- the multivalent antibody of the invention may be used as an affinity purification agent.
- the multivalent antibody is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art.
- the immobilized multivalent antibody is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized multivalent antibody. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the multivalent antibody.
- the multivalent antibody may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum.
- the multivalent antibody typically will be labeled with a detectable moiety.
- a detectable moiety Numerous labels are available which can be generally grouped into the following categories:
- Radioisotopes such as 35 S, 14 C, 125 I, 3 H, and 131 I.
- the multivalent antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology , Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, N.Y., Pubs. (1991) for example and radioactivity can be measured using scintillation counting.
- Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available.
- the fluorescent labels can be conjugated to the multivalent antibody using the techniques disclosed in Current Protocols in Immunology , supra, for example. Fluorescence can be quantified using a fluorimeter.
- the enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above.
- the chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor.
- enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, ⁇ -galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.
- luciferases e.g., firefly luciferase and bacterial lucifera
- enzyme-substrate combinations include, for example
- HRPO Horseradish peroxidase
- HPO horseradish peroxidase
- OPD orthophenylene diamine
- TMB 3,3′,5,5′-tetramethyl benzidine hydrochloride
- ⁇ -D-galactosidase ( ⁇ -D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl- ⁇ -D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase.
- a chromogenic substrate e.g., p-nitrophenyl- ⁇ -D-galactosidase
- fluorogenic substrate 4-methylumbelliferyl- ⁇ -D-galactosidase
- the label is indirectly conjugated with the multivalent antibody.
- the multivalent antibody can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the multivalent antibody in this indirect manner.
- the multivalent antibody is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten multivalent antibody (e.g., anti-digoxin antibody).
- a small hapten e.g., digoxin
- an anti-hapten multivalent antibody e.g., anti-digoxin antibody
- the multivalent antibody need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the multivalent antibody.
- the multivalent antibody of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies: A Manual of Techniques , pp. 147-158 (CRC Press, Inc. 1987).
- the multivalent antibody may also be used for in vivo diagnostic assays.
- the multivalent antibody is labeled with a radionuclide (such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S) so that the antigen or cells expressing it can be localized using immunoscintiography.
- a radionuclide such as 111 In, 99 Tc, 14 C, 131 I, 125 I, 3 H, 32 P or 35 S
- the multivalent antibody of the present invention may be used to treat a mammal e.g. a patient suffering from, or predisposed to, a disease or disorder who could benefit from administration of the multivalent antibody.
- conditions to be treated therewith include benign or malignant tumors; leukemias and lymphoid malignancies; other disorders such as neuronal, glial, astrocytal, hypothalamic, glandular, macrophagal, epithelial, stromal, blastocoelic, inflammatory, angiogenic and immunologic disorders.
- the disease or disorder to be treated with the antibody that binds an ErbB receptor is cancer.
- cancers to be treated herein include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include squamous cell cancer, lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma as well as head and neck cancer.
- lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of
- the cancer will generally comprise cells that express an antigen bound by the antibody, such that the antibody is able to bind to the cancer.
- the cancer may be characterized by overexpression of the antigen (e.g. overexpression of an ErbB receptor).
- overexpression may be analyzed by INC, e.g. using the HERCEPTEST® (Dako) where the antigen is HER2.
- HERCEPTEST® Dako
- parrafin embedded tissue sections from a tumor biopsy may be subjected to the IHC assay and accorded a HER2 protein staining intensity criteria as follows:
- Those tumors with 0 or 1+ scores for HER2 overexpression assessment may be characterized as not overexpressing HER2, whereas those tumors with 2+ or 3+ scores may be characterized as overexpressing HER2.
- FISH assays such as the INFORMTM (sold by Ventana, Ariz.) or PATHVISIONTM (Vysis, Ill.) may be carried out on formalin-fixed, paraffin-embedded tumor tissue to determine the extent (if any) of antigen overexpression by the tumor.
- the cancer will be one which expresses (and may overexpress) an ErbB receptor selected from the group consisting of EGFR, ErbB3 and ErbB4.
- Examples of cancers which may express/overexpress EGFR, ErbB3 or ErbB4 include squamous cell cancer, lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma as well as head and neck cancer as well as glioblastomas.
- the cancer to be treated herein may be one characterized by excessive activation of an ErbB receptor, e.g. EGFR. Such excessive activation may be attributable to overexpression or increased production of the ErbB receptor or an ErbB ligand.
- a diagnostic or prognostic assay will be performed to determine whether the patient's cancer is characterized by excessive activation of an ErbB receptor. For example, ErbB gene amplification and/or overexpression of an ErbB receptor in the cancer may be determined.
- Assays for determining such amplification/overexpression are available in the art and include the IHC, FISH and shed antigen assays described above.
- levels of an ErbB ligand, such as TGF-alpha, in or associated with the tumor may be determined according to known procedures. Such assays may detect protein and/or nucleic acid encoding it in the sample to be tested. In one embodiment, ErbB ligand levels in the tumor may be determined using immunohistochemistry (1HC); see, for example, Scher et al. Clin. Cancer Research 1:545-550 (1995). Alternatively, or additionally, one may evaluate levels of ErbB ligand-encoding nucleic acid in the sample to be tested; e.g. via fluorescent in situ hybridization or FISH, southern blotting, or polymerase chain reaction (PCR) techniques.
- PCR polymerase chain reaction
- ErbB receptor or ErbB ligand overexpression or amplification may be evaluated using an in vivo diagnostic assay, e.g. by administering a molecule (such as an antibody) which binds the molecule to be detected and is tagged with a detectable label (e.g. a radioactive isotope) and externally scanning the patient for localization of the label.
- a detectable label e.g. a radioactive isotope
- the antibody may be used to treat a B cell lymphoma (including low grade/follicular non-Hodkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; and chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD).
- B cell lymphoma including low grade/follicular non-Hodkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade
- the antibody e.g. the anti-B cell surface antigen antibody
- autoimmune diseases or disorders include, but are not limited to, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g.
- atopic dermatitis atopic dermatitis
- systemic scleroderma and sclerosis responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g.
- Type I diabetes mellitus or insulin dependent diabetes mellitis multiple sclerosis; Reynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjorgen's syndrome; juvenile onset diabetes; and immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis; pernicious anemia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder; multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemia or Coombs positive anemia); myasthenia gravis; antigen-antibody complex mediated diseases; anti-glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyen
- Antibodies directed against B cell surface antigens may also be used to block an immune response to a foreign antigen.
- foreign antigen here is meant a molecule or molecules which is/are not endogenous or native to a mammal which is exposed to it.
- the foreign antigen may elicit an immune response, e.g. a humoral and/or T cell mediated response in the mammal. Generally, the foreign antigen will provoke the production of antibodies thereagainst.
- foreign antigens contemplated herein include immunogenic therapeutic agents, e.g. proteins such as antibodies, particularly antibodies comprising non-human amino acid residues (e.g.
- rodent, chimeric/humanized, and primatized antibodies include toxins (optionally conjugated to a targeting molecule such as an antibody, wherein the targeting molecule may also be immunogenic); gene therapy viral vectors, such as retroviruses and adenoviruses; grafts; infectious agents (e.g. bacteria and virus); alloantigens (i.e. an antigen that occurs in some, but not in other members of the same species) such as differences in blood types, human lymphocyte antigens (HLA), platelet antigens, antigens expressed on transplanted organs, blood components, pregnancy (Rh), and hemophilic factors (e.g. Factor VIII and Factor IX).
- HLA human lymphocyte antigens
- platelet antigens antigens expressed on transplanted organs
- blood components blood components
- pregnancy (Rh) pregnancy
- hemophilic factors e.g. Factor VIII and Factor IX
- the anti-B cell surface antigen antibody may also be used to desenzitize a mammal awaiting transplantation.
- Antibodies directed against a receptor in the TNF receptor superfamily may be employed to activate or stimulate apoptosis in cancer cells.
- an immunoconjugate comprising the antibody conjugated with a cytotoxic agent is administered to the patient.
- the immunoconjugate and/or antigen to which it is bound is/are internalized by the cell, resulting in increased therapeutic efficacy of the immunoconjugate in killing the cancer cell to which it binds.
- the cytotoxic agent targets or interferes with nucleic acid in the cancer cell. Examples of such cytotoxic agents include any of the chemotherapeutic agents noted herein (such as a maytansinoid or a calicheamicin), a radioactive isotope, or a ribonuclease or a DNA endonuclease.
- the multivalent antibody may also be used for ADEPT.
- the present application contemplates combining the multivalent antibody (or immunoconjugate thereof) with one or more other therapeutic agent(s), especially for treating cancer.
- the multivalent antibody may be co-administered with another multivalent antibody (or multivalent antibodies), a monovalent or bivalent antibody (or antibodies), chemotherapeutic agent(s) (including cocktails of chemotherapeutic agents), other cytotoxic agent(s), anti-angiogenic agent(s), cytokines, and/or growth inhibitory agent(s).
- the multivalent antibody induces apoptosis
- it may be particularly desirable to combine the multivalent antibody with one or more other therapeutic agent(s) which also induce apoptosis.
- pro-apoptotic antibodies e.g.
- bivalent or multivalent antibodies directed against B cell surface antigens may be combined with (1) pro-apoptotic antibodies (e.g. bivalent or multivalent antibodies directed against a receptor in the TNF receptor superfamily, such as anti-DR4 or anti-DR5 antibodies) or (2) with cytokines in the TNF family of cytokines (e.g. Apo2L).
- pro-apoptotic antibodies e.g. bivalent or multivalent antibodies directed against a receptor in the TNF receptor superfamily, such as anti-DR4 or anti-DR5 antibodies
- cytokines in the TNF family of cytokines e.g. Apo2L
- anti-ErbB antibodies e.g. HERCEPTIN® anti-HER2 antibody
- the patient may receive combined radiation therapy (e.g.
- Such combined therapies noted above include combined administration (where the two or more agents are included in the same or separate formulations), and separate administration, in which case, administration of the multivalent antibody can occur prior to, and/or following, administration of the adjunct therapy or therapies.
- the multivalent antibody (and adjunct therapeutic agent) is/are administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration.
- Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
- the multivalent antibody is suitably administered by pulse infusion, particularly with declining doses of the multivalent antibody.
- the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic.
- the present application contemplates administration of the antibody by gene therapy.
- administration of nucleic acid encoding the antibody is encompassed by the expression “administering a therapeutically effective amount of an antibody”. See, for example, WO96/07321 published Mar. 14, 1996 concerning the use of gene therapy to generate intracellular antibodies.
- nucleic acid (optionally contained in a vector) into the patient's cells
- in vivo and ex vivo the nucleic acid is injected directly into the patient, usually at the site where the antibody is required.
- ex vivo treatment the patient's cells are removed, the nucleic acid is introduced into these isolated cells and the modified cells are administered to the patient either directly or, for example, encapsulated within porous membranes which are implanted into the patient (see, e.g. U.S. Pat. Nos. 4,892,538 and 5,283,187).
- techniques available for introducing nucleic acids into viable cells There are a variety of techniques available for introducing nucleic acids into viable cells.
- the techniques vary depending upon whether the nucleic acid is transferred into cultured cells in vitro, or in vivo in the cells of the intended host.
- Techniques suitable for the transfer of nucleic acid into mammalian cells in vitro include the use of liposomes, electroporation, microinjection, cell fusion, DEAE-dextran, the calcium phosphate precipitation method, etc.
- a commonly used vector for ex vivo delivery of the gene is a retrovirus.
- the currently preferred in vivo nucleic acid transfer techniques include transfection with viral vectors (such as adenovirus, Herpes simplex I virus, or adeno-associated virus) and lipid-based systems (useful lipids for lipid-mediated transfer of the gene are DOTMA, DOPE and DC-Chol, for example).
- viral vectors such as adenovirus, Herpes simplex I virus, or adeno-associated virus
- lipid-based systems useful lipids for lipid-mediated transfer of the gene are DOTMA, DOPE and DC-Chol, for example.
- an agent that targets the target cells such as an antibody specific for a cell surface membrane protein or the target cell, a ligand for a receptor on the target cell, etc.
- proteins which bind to a cell surface membrane protein associated with endocytosis may be used for targeting and/or to facilitate uptake, e.g.
- capsid proteins or fragments thereof tropic for a particular cell type antibodies for proteins which undergo internalization in cycling, and proteins that target intracellular localization and enhance intracellular half-life.
- the technique of receptor-mediated endocytosis is described, for example, by Wu et al., J. Biol. Chem. 262:4429-4432 (1987); and Wagner et al., Proc. Natl. Acad. Sci. USA 87:3410-3414 (1990).
- Wu et al. J. Biol. Chem. 262:4429-4432 (1987); and Wagner et al., Proc. Natl. Acad. Sci. USA 87:3410-3414 (1990).
- the appropriate dosage of multivalent antibody will depend on the type of disease to be treated, the severity and course of the disease, whether the multivalent antibody is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the multivalent antibody, and the discretion of the attending physician.
- the multivalent antibody is suitably administered to the patient at one time or over a series of treatments.
- ⁇ g/kg to 15 mg/kg (e.g., 0.1-20 mg/kg) of multivalent antibody is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion.
- a typical daily dosage might range from about 1 ⁇ g/kg to 100 mg/kg or more, depending on the factors mentioned above.
- the treatment is sustained until a desired suppression of disease symptoms occurs.
- other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.
- the multivalent antibody composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
- the “therapeutically effective amount” of the multivalent antibody to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder.
- the multivalent antibody need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question.
- the effective amount of such other agents depends on the amount of multivalent antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages.
- an article of manufacture containing materials useful for the treatment of the disorders described above comprises a container and a label or package insert on or associated with the container.
- Suitable containers include, for example, bottles, vials, syringes, etc.
- the containers may be formed from a variety of materials such as glass or plastic.
- the container holds a composition which is effective for treating the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).
- At least one active agent in the composition is a multivalent antibody.
- the label or package insert indicates that the composition is used for treating the condition of choice, such as cancer.
- the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises a multivalent antibody; and (b) a second container with a composition contained therein, wherein the composition comprises a further cytotoxic agent.
- the article of manufacture in this embodiment of the invention may further comprises a package insert indicating that the first and second antibody compositions can be used to treat cancer.
- the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.
- BWFI bacteriostatic water for injection
- phosphate-buffered saline such as bacteriostatic water for injection (BWFI), phosphate-buffered saline,
- hybridoma cell lines have been deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, USA (ATCC):
- Antibody Designation ATCC No. Deposit Date 7C2 (anti-HER2) ATCC HB-12215 Oct. 17, 1996 7F3 (anti-HER2) ATCC HB-12216 Oct. 17, 1996 4D5 (anti-HER2) ATCC CRL 10463 May 24, 1990 2C4 (anti-HER2) ATCC HB-12697 Apr. 8, 1999 3F11.39.7 (anti-DR5) HB-12456 Jan. 13, 1998 3H3.14.5 (anti-DR5) HB-12534 Jun. 2, 1998 3D5.1.10 (anti-DR5) HB-12536 Jun. 2, 1998 3H1.18.10 (anti-DR5) HB-12535 Jun. 2, 1998 4E7.24.3 (anti-DR4) HB-12454 Jan. 13, 1998 4H6.17.8 (anti-DR4) HB-12455 Jan. 13, 1998
- FIG. 5 The construct used to generate a tetravalent anti-HER2 antibody, called an “Octopus antibody” (OctHER2), is illustrated in FIG. 5 herein.
- the backbone of this Octopus antibody is the recombinant, humanized monoclonal antibody 4D5 variant 8 (rhuMAb 4D5-8) (U.S. Pat. No. 5,821,337, Carter et al., expressly incorporated herein by reference).
- the heavy chain of rhuMAb 4D5-8 was subcloned into the pRK5 vector (EP 307,247, published Mar. 15, 1989).
- the VH-CH1 region of the heavy chain was removed by mutagenesis, and three unique restriction sites (BamHI; NheI; BspEI) were inserted. These sites were incorporated into PCR primers designed to amplify the VH-CH1 region from different antibodies. The resulting fragments were subcloned into the vector to create the Octopus heavy chain. Co-expression of the Octopus heavy chain with the appropriate light chain in a pRK5 vector in mammalian cell transfections results in the completed Octopus antibody ( FIG. 4 ).
- Octopus constructs containing flexible linkers inserted between the tandem Fd regions were also engineered.
- DNA encoding either “gly-ser” (flex 1 linker) or “gly-ser-gly-ser” (SEQ ID NO:10) (flex 2 linker) was inserted between the DNA encoding the VH-CH1 regions of the heavy chain.
- OctHER2 was expressed in transiently transfected 293 cells (Graham et al. J. Gen. Virol. 36:59-72 (1977)) and purified over a Protein A sepharose column.
- the complete antibody is approximately 245 kDa, as compared to the 150 kDa molecular weight of the parent antibody.
- the Octopus heavy chain is 75 kDa (without carbohydrate), and the light chain is 30 kDa.
- HER2 extracellular domain HER2 extracellular domain
- OctHER2 binds the HER2ECD similar to HERCEPTIN® when analyzed in an ELISA assay.
- the rhuMAb 4D5-8 expressed by 293 cells binds identically to the vialed HERCEPTIN® (produced by Chinese Hamster Ovary (CHO) cells), indicating that 293 cells do not substantially alter the antigen binding capability of the antibodies.
- OctHER2 was compared to HERCEPTIN® in functional assays measuring growth inhibition of HER2 overexpressing tumor cell lines.
- OctHER2 was similar to slightly better at inhibiting growth of SKBR3 cells ( FIG. 8A ), however was not as effective on BT474 cells ( FIG. 8B ). Interestingly, OctHER2 inhibited more effectively than HERCEPTIN® a 2+ overexpressing cell line, MDA 361 ( FIG. 8B ).
- the flexible linker Octopus constructs (OctHER2flex1, OctHER2flex2) inhibited cell growth more effectively than HERCEPTIN®.
- the antibody was radioiodinated, and incubated for varying times with the cells. This was followed by measurements of the amount of intact, unbound antibody in the supernatant, the amount bound to the cell surface, the amount internalized, and finally, the amount catabolized and degraded.
- Electron Microscopy Autoradiography To confirm that the Octopus antibody was being internalized and degraded in the appropriate vesicles, and not just nonspecifically, Electron Microscopy (EM) autoradiography was used. The Octopus antibody was iodinated and incubated with the cells in the same fashion as in the internalization assays. The results depicted in FIGS. 11A-C confirm that the Octopus antibody was being internalized into the correct vesicles (early endosome, FIG. 11B ; and lysosome, FIG. 11C ). Additionally, the percentage of internalization observed with OctHER2 and HERCEPTIN® in these assays matched with the measurements in the internalization assays.
- DR5 a member of the TNF receptor superfamily that binds the trimeric Apo2L/TRAIL (Apo2L). After Apo2L receptors bind Apo2L and are clustered, death domains in the cytosolic region of the receptors induce caspases to trigger cellular apoptosis.
- anti-DR5 Octopus constructs Two versions of anti-DR5 Octopus constructs were made: one from 16E2, an anti-DR5 cloned from a single-chain human Fv phage library (see WO98/51793, expressly incorporated herein by reference); the second anti-DR5 Octopus antibody was made from Mab 3H3.14.5 (the “3H3” antibody; ATCC HB-12534, WO99/64461), a murine anti-DR5MAb that induces apoptosis when it is crosslinked. Since anti-Death receptor monoclonal antibodies may require crosslinking to trigger apoptosis, they are candidates for the Octopus antibody construct.
- the anti-DR5 Octopus antibodies were prepared by replacing the variable domains of the OctHER2 construct described above with the VL and VH domains from 16E2 or 3H3.
- the anti-DR5 Octopus antibodies were analyzed in apoptosis assays using either crystal violet or alamarBlue staining. Briefly, serial dilutions of the Octopus antibody or Apo2L were added to the media of plated cells which were then allowed to continue growing for 24 hours. After this time, the media was either removed and the cells were stained with crystal violet, or alamarBlue was added to the media and incubated briefly with the cells. Crystal violet stains the cells, whereas alamarBlue detects metabolic activity in the culture media, thus these dyes allow for measurement of cells that survive treatment. Staining by both colorimetric dyes, crystal violet and alamarBlue, was quantitated by spectrophotometry.
- the 16E2 Octopus surprisingly, induces apoptosis with comparable potency to Apo2L in lung (SK-MES-1; HOP 92) and colon (HCT116; COLO 205) tumor cell lines, however does not cause apoptosis on normal control cell line (HUMEC).
- HUMEC normal control cell line
- the anti-DR516E2 Octopus was also effective in vivo in inducing apoptosis and shrinking a colon tumor, human COLO205, in athymic nude mice.
- FIG. 13A-D histology slides of tumor tissues stained with hematoxylin and eosin from mice treated with the 16E2 Octopus or Apo2L induced similar levels of apoptotic cells.
- mice also demonstrated significant decrease in tumor volume, similar to that measured for the Apo2L and two bivalent anti-DR5 mAbs, 16E2 and 3H3, as shown in FIG. 14 .
- Mice that did not receive any anti-DR5 antibodies or Apo2L (Vehicle) showed dramatic increase in their tumor volume due to uncontrolled growth.
- the apoptotic activity of the material used in the mouse studies was confirmed in an in vitro apoptotic assay in FIG. 15 .
- the anti-DR516E2 Octopus and the Apo2L used in the study were compared to an Apo2L standard positive control and an anti-IgE MAb (E25) negative control in an alamarBlue apoptosis assay.
- FIG. 16 demonstrates that another anti-DR5 Octopus, 3H3 Octopus, is capable of inducing apoptosis similar to the 16E2 Octopus. Additionally, FIG. 16 shows that the apoptotic activity of the Octopus antibody is not lot dependent, as several 16E2 Octopus antibodies prepared on different dates retain similar function.
- the apoptotic activity of both the 16E2 and 3H3 Octopus antibodies is better than Apo2L on a lung tumor cell line, SK-MES-1 ( FIG. 17A ), and a T cell tumor line, Jurkat ( FIG. 17B ).
- the anti-DR5 Octopus antibodies may be more effective at clustering DR5 on the tumor cell surface than Apo2L.
- FIGS. 18A-C depict the 2-day dose response curves showing the effects of the 16E2 Octopus and Apo2L on the growth of several human leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer tumor cell lines, while FIGS. 19A-C show dose response curves from the 6 day screens. Comparable results were observed for 16E2 Octopus and Apo2L against most of the tumor cell lines, again indicating that the anti-DR5 Octopus functions similar to Apo2L.
- FIGS. 20 A and B 2-day results
- FIGS. 21A and B 6-day results
- 16E2 Octopus may be effective against more types of cancer than previously observed.
- RITUXAN® alone does not trigger much apoptosis of a non-Hodgkins lymphoma B cell line, Wil-2, unless it is crosslinked with anti-human IgG (RITUXAN®-IgG).
- OctCD20 is capable of inducing apoptosis in Wil-2 cells independent of crosslinking.
- the level of apoptosis observed with OctCD20 is lower than that of crosslinked RITUXAN®, however, suggesting that the apoptotic activity of OctCD20 could be improved, perhaps through the use of the flexible linkers.
- Octopus antibodies of Example 2 (anti-HER2), Example 3 (anti-DR5) and Example 4 (anti-CD20) with an antibody hing region dimerization domain (designated “Octopus F(ab′) 2 ” herein) were engineered.
- the anti-HER2 Octopus F(ab′) 2 construct was engineered by replacing the Fc region of the heavy chain cDNA with sequence encoding a leucine-zipper motif which, when expressed as protein, dimerizes to effectively join the Octopus Fab arms ( FIG. 23C ).
- the octopus F(ab′) 2 can maintain the leucine zipper motif, or that motif can, e.g., be proteolytically removed as desired. As depicted in FIG.
- PCR was used to amplify the duplicate VH/CH1 domains and to insert a restriction site onto the end of the Octopus heavy chain cDNA (NotI) to permit in-frame subcloning into a vector (VG15) containing a leucine-zipper motif. PCR was again utilized to add another restriction site downstream of the heavy chain termination codon (XhoI) to allow subcloning into the pRK vector for expression in mammalian cells.
- the VH/CH1 domains of anti-DR5Mab16E2 and anti-CD20 Mab C2B8 were substituted into the Oct F(ab)′ 2 heavy chain backbone using the unique restriction sites BamHI, NheI, and BspEI.
- POPoctopus antibodies were created by linking together Fab domains in tandem repeats to form linear Fab multimers.
- POPoct-3 contains three linked Fab domains ( FIG. 23D ), while “POPoct-4” has four Fab repeats ( FIG. 23E ).
- Anti-HER2 (rhuMab 4D5), anti-DR5 (16E2), and anti-CD20 (C2B8) POPoct-3 constructs were generated, as were anti-HER2 (rhuMab 4D5) and anti-DR5 (16E2) POPoct-4 constructs.
- POPoct-3 antibodies were engineered both with and without flex 1 linkers.
- FIG. 25 depicts the construction of the POPoct-3 heavy chain cDNA.
- PCR was used to amplify the VH/CH1 domain adding a 5′-BspEI site and a 3′-NotI site. This sequence was digested and along with BamHI/BspEI digested Octopus heavy chain, ligated into a pRK vector to yield an Octopus heavy chain containing sequence for three VH/CH1 domains.
- the BspEI site encodes for a serine and a glycine residue.
- the different Octopus heavy chains were transiently cotransfected with the appropriate light chain cDNAs into 293 mammalian cells to express antibodies containing either three Fab domains (POPoct-3 Fab) or four Fab domains (full-length Octopus; Octopus F(ab)′ 2 ; POPoct-4 Fab). While native IgG Mabs and full-length Octopus antibodies were purified over Protein A sepharose, Octopus F(ab)′ 2 and POPoct-3 and -4 were purified over Protein G sepharose columns.
- the Octopus F(ab)′ 2 is approximately 200 kDa ( FIG. 23F , lane 4), smaller than the 240 kDa of the full-length Octopus antibody ( FIG. 23F , lane 3), but larger than the 150 kDa native IgG Mab ( FIG. 23F , lanes 1 and 2).
- POPoct-3 is slightly smaller than native IgG Mab
- POPoct-4 is slightly larger at 190 kDa.
- the heavy chain of the Octopus F(ab)′ 2 ( FIG. 23G , lane 4) is approximately the same size as the native IgG Mab heavy chain ( FIG.
- the POPoct-3 heavy chain ( FIG. 23G , lane 5) is similar in size to the full-length Octopus heavy chain ( FIG. 23G , lane 3), while at approximately 97 kDa the POPoct-4 has the largest heavy chain
- POPoct-3HER2 was compared to OctHER and HERCEPTIN® in internalization assays on two 3+HER2 over-expressing tumor cell lines, SKBR3 and BT474, to assess its candidacy for applications in immunotoxin therapies. Although structurally different than the full-length OctHER2 antibody, POPoct-3HER2 was internalized and catabolized identically to OctHER2 by both cell lines ( FIGS. 29A and B) and at twice the rate of HERCEPTIN®.
- OctDR5flex-1 showed increased potency compared to OctDR5, especially at lower concentrations ( FIG. 30A ), indicating that flexibility between the Fab arms improves efficacy.
- POPoct-3flex-1DR5 induced equivalent levels of apoptosis as OctHER ( FIG. 30A ) and showed similar efficacy to POPoct16-3 and POPoct16-4 ( FIG. 30B ).
- Apo2L binds to the death receptors and triggers cellular apoptosis through the caspase signaling pathway.
- the anti-DR5 Octopus antibodies were shown to induce apoptosis through the same signaling pathway as Apo2L.
- Oct16E2 triggered similar levels of apoptosis as APO2L on the lung tumor cell line SK-MES-1 ( FIG. 31A , dashed lines), but after the addition of ZVAD, an inhibitor of caspase 3 and 9, cellular apoptosis triggered by both Apo2L and Oct16E2 was inhibited ( FIG. 31B solid lines).
- FIG. 31B Further evidence that the anti-DR5 Octopus antibodies signaled through the same pathway as Apo2L was obtained by DISC (Death Induced Signaling Complex) analyses ( FIG. 31B ).
- BJAB cells a B-cell lymphoma line that expresses DR5
- Purification of the antibody-DR5 complexes was followed Western blot analysis to identify the signaling molecules that copurified with the complexes.
- Apo2L the signaling molecules caspase 8 and FADD associated with DR5 after the receptor was bound by both Oct16E2 and Oct3H3 ( FIG. 31B ).
- RITUXAN® did not efficiently trigger apoptosis in vitro on the B-cell lymphoma cell line WIL-2 unless first crosslinked by anti-IgG antibody.
- OctCD20 was capable of inducing apoptosis of WIL-2 cells independent of crosslinking, at levels higher than RITUXAN® alone, yet slightly lower than anti-IgG-crosslinked RITUXAN®.
- When crosslinked with anti-IgG antibody OctCD20 induced more apoptosis of the WIL-2 cells than crosslinked RITUXAN® ( FIG. 32 ). Since one potential explanation for the efficacy of RITUXAN® in vivo is that the antibody is being crosslinked by either complement or Fc ⁇ R bearing cells, this observation suggests that OctCD20 will be even more efficacious in vivo.
- OctCD20 F(ab)′ 2 , POPoct-3CD20 and POPoct-3CD20flex-1 were tested at various concentrations in apoptosis assays with WIL-2 cells, and the optimal doses are shown in the maximum response curves in FIG. 33 .
- the Octopus antibodies were compared to the anti-CD20 antibody 1F5 (Clark et al. supra), which functions similar to RITUXAN® in that it does not induce apoptosis unless crosslinked with anti-IgG antibody.
- Octopus antibodies tested induced either similar (OctCD20 F(ab)′ 2 ) or higher (POPoct-3CD20, POPoct-3CD20flex-1) levels of apoptosis than crosslinked IF5 anti-CD20. Additionally, the Octopus antibodies were efficacious at considerably lower concentrations than the crosslinked anti-CD20.
- Apoptosis induction by the various anti-CD20 antibodies was further assessed using blood from a patient with chronic lymphocytic leukemia (CLL). PBL's were separated out using dextran sedimentation, washed and plated in serum-free lymphocyte medium treated overnight with no sample, 1F5 (20 ⁇ g/ml), 1F5+cross-linking mouse anti-IgG (100 ⁇ g/ml), OctCD20 F(ab′) 2 at approx 0.5 or 1.0 ⁇ g/mland POPoct-3 CD20 at 0.5 ⁇ g/ml.
- CLL chronic lymphocytic leukemia
- apoptosis assay was performed using annexin and PI staining. The percentage of apoptotic cells were:
- the data indicate that multivalent anti-CD20 antibodies (especially POPoct-3 CD20) enhance apoptosis in a dose-dependent manner.
- OctCD20 was also evaluated as a candidate for immunotoxin therapy in internalization assays on three B-cell lymphoma lines, DB, WIL-2, and Ramos, and compared to RITUXAN®. As shown in FIG. 35 , twice as much OctCD20 was internalized by the cells as compared to RITUXAN®, which was not internalized by the cells at appreciable levels. The higher avidity that would be expected for the multivalent antibodies due to the increased number of binding sites is evident in the fact that more OctCD20 remains bound to the cell surface of the cells over time as compared to RITUXAN®.
Abstract
The present application describes engineered antibodies, with three or more functional antigen binding sites, and uses, such as therapeutic applications, for such engineered antibodies.
Description
- This application is a continuation application claiming priority to application Ser. No. 11/218,821, filed Sep. 2, 2005, which is a continuation application claiming priority to application Ser. No. 09/813,341, filed Mar. 20, 2001, which claims priority under 35 U.S.C. §119(e) to provisional application No. 60/195,819, filed Apr. 11, 2000, the entire contents of which are hereby incorporated by reference.
- 1. Field of the Invention
- The present invention concerns engineered antibodies, with three or more functional antigen binding sites, and uses, such as therapeutic uses, for such engineered antibodies.
- 2. Description of Related Art
- Naturally occurring antibodies (immunoglobulins) comprise two heavy chains linked together by disulfide bonds and two light chains, one light chain being linked to each of the heavy chains by disulfide bonds. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains (three or four constant domains, CH1, CH2, CH3 and CH4, depending on the antibody class). Each light chain has a variable domain (VL) at one end and a constant domain (CL) at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. See
FIG. 1 herein. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains, see e.g. Chothia et al., J. Mol. Biol. 186:651-663 (1985); and Novotny and Haber, Proc. Natl. Acad. Sci. USA 82:4592-4596 (1985). - The constant domains are not involved directly in binding the antibody to an antigen, but are involved in various effector functions, such as participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC). The variable domains of each pair of light and heavy chains are involved directly in binding the antibody to the antigen. The variable domains of naturally occurring light and heavy chains have the same general structure; each comprising four framework regions (FRs), whose sequences are somewhat conserved, connected by three complementarity determining regions (CDRs) (see Kabat et al., Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., (1991)). The four FRs largely adopt a beta-sheet conformation and the CDRs form loops connecting, and in some cases forming part of, the beta-sheet structure. The CDRs in each chain are held in close proximity by the FRs and, with the CDRs from the other chain, contribute to the formation of the antigen binding site.
-
FIGS. 2A-E herein depict the structures of the five major naturally occurring immunoglobulin isotypes. IgG, IgD and IgE immunoglobulins possess only two antigen binding sites. IgA and IgM, on the other hand, are capable of forming polymeric structures with higher valencies. - IgM is secreted by plasma cells as a pentamer in which five monomer units are held together by disulfide bonds linking their carboxyl-terminal (CIA/CO) domains and Cμ3/Cμ3 domains. The five monomer subunits are arranged with their Fc regions in the center of the pentamer and the 10 antigen-binding sites on the periphery of the molecule. Each pentamer contains an additional Fc-linked polypeptide called the J (joining) chain, which is disulfide-bonded to the carboxyl-terminal cysteine residue of 2 of the 10μ chains. The J chain appears to be required for polymerization of the monomers to form pentameric IgM; it is added just before secretion of the pentamer. An IgM molecule can bind 10 small hapten molecules; however, because of steric hindrance, only 5 molecules of larger antigens can be bound simultaneously. The increased valency of pentameric IgM increases its capacity to bind such multi-dimensional antigens as viral particles and red blood cells (RBCs).
- IgA exists primarily as a monomer, although polymeric forms such as dimers, trimers, and even tetramers are sometimes seen. The IgA of external secretions consists of a dimer or tetramer, a J-chain polypeptide, and a polypeptide chain called secretory component.
- Widespread use has been made of monoclonal antibodies, particularly those derived from rodents including mice, however they are frequently antigenic in human clinical use. For example, a major limitation in the clinical use of rodent monoclonal antibodies is an anti-globulin response during therapy (Miller at al., Blood 62:988-995 (1983); and Schroff, R. W. et al., Cancer Res. 45:879-885 (1985)).
- The art has attempted to overcome this problem by constructing “chimeric” antibodies in which an animal antigen binding variable domain is coupled to a human constant domain (Cabilly et al., U.S. Pat. No. 4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984); Boulianne et al., Nature 312:643-646 (1984); and Neuberger et al., Nature 314:268-270 (1985)). The isotype of the human constant domain may be selected to tailor the chimeric antibody for participation in ADCC and CDC (see e.g. Bruggemann et al., J. Exp. Med. 166:1351-1361 (1987); Riechmann et al., Nature 332:323-327 (1988); Love et al., Methods in Enzymology 178:515-527 (1989); and Bindon et al., J. Exp. Med. 168:127-142 (1988)). In the typical embodiment, such chimeric antibodies contain about one third rodent (or other non-human species) sequence and thus are capable of eliciting a significant anti-globulin response in humans. For example, in the case of the murine anti-CD3 antibody, OKT3, much of the resulting anti-globulin response is directed against the variable region rather than the constant region (Jeffers et al., Transplantation 41:572-578 (1986)).
- In a further effort to resolve the antigen binding functions of antibodies and to minimize the use of heterologous sequences in human antibodies, Winter and colleagues (Jones et al., Nature 321:522-525 (1986); Riechmann et al., i Nature 332:323-327 (1988); and Verhoeyen et al., Science 239:1534-1536 (1988)) have substituted rodent CDRs or CDR sequences for the corresponding segments of a human antibody.
- The therapeutic promise of this approach is supported by the clinical efficacy of a humanized antibody specific for the CAMPATH-1 antigen with two non-Hodgkin lymphoma patients, one of whom had previously developed an anti-globulin response to the parental rat antibody (Riechmann et al., Nature 332:323-327 (1988); and Hale et al., Lancet i:1394-1399 (1988)).
- In some cases, substituting CDRs from rodent antibodies for the human CDRs in human frameworks is sufficient to transfer high antigen binding affinity (Jones et al., Nature 321:522-525 (1986); Verhoeyen et al., Science 239:1534-1536 (1988)), whereas in other cases it has been necessary to additionally replace one (Riechmann et al., Nature 332:323-327 (1988)) or several (Queen et al., Proc. Natl. Acad. Sci. USA 86:10029-10033 (1989)) framework residues. See also Co et al., Proc. Natl. Acad. Sci. USA 88:2869-2873 (1991); U.S. Pat. No. 5,821,337 (Carter et al.); and U.S. Pat. No. 5,530,101 (Queen et al.). Additional references relating to humanization of antibodies include Gorman et al., Proc. Natl. Acad. Sci. USA 88:4181-4185 (1991); Daugherty et al., Nucleic Acids Research 19(9):2471-2476 (1991); Brown et al., Proc. Natl. Acad. Sci. USA 88:2663-2667 (1991); and Junghans et al., Cancer Research 50:1495-1502 (1990).
- Instead of a chimeric/humanized antibody, one may treat a patient with a human antibody in order to avoid human antibodies raised against a murine antibody (known as the “HAMA response”). Several technologies are available for generating human antibodies.
- Human antibodies may be selected using phage display technology. Phage display has been adapted to select human antibodies from an unimmunized donor (Marks et al. J. Mol. Biol. 222:581-597 (1991)). According to this approach, PCR is used to amplify variable domain genes from mRNA prepared from human peripheral blood lymphocytes (PBLs). Primers are used such that DNA from both IgG and IgM heavy chains and both κ and λ chains is amplified. These genes are then randomly combined and expressed as single chain Fv (scFv) fused to the gene III coat protein of M13 phage. Human antibodies against an antigen of interest may then be identified by rounds of growth and selection by binding to that antigen (e.g. to the immobilized antigen). See Griffiths et al. EMBO J. 12:725-734 (1993).
- “Synthetic” phage-antibody repertoires have also been built from cloned human VH-gene segments. A repertoire (2×107 clones) was first constructed using a short H3 loop of five or eight random residues with each of 49 segments, and combined with a fixed light chain (Hoogenboom et al. J. Mol. Biol. 227:381-388 (1992)). By adding a range of H3 loops of different lengths, up to 12 residues, a single library was created from which a range of more than 20 binding specificities could be selected (Winter et al. Ann. Rev. Immuno. 12:433-55 (1994)). Other synthetic libraries have been built from the framework of a single antibody by randomizing CDRs of the human antibody (Garrard and Henner Gene 128:103-109 (1993)). Antibodies derived from such synthetic phage-antibody repertoires are also considered to be “human” antibodies herein.
- The affinity of low affinity “primary” phage-antibodies may be improved by using phage display technology. One approach is to use a chain-shuffling strategy in which the VH domain is held constant and then recombined with the original library of VL genes and tighter binders selected by binding to immobilized antigen. This cycle is repeated by fixing the new VL domain and recombining with the original VH library (Marks et al. Bio/Technology 10:779-783 (1992)). Alternatively, point mutations in the primary antibody may be introduced using error-prone PCR and higher affinity binders selected by using phage display. Gram et al. PNAS (USA) 89: 3576-3580 (1992).
- One may also produce human antibodies by immunizing mice which have been genetically engineered to express human antibodies. Severe combined immune deficient (SCID) mice lack the ability to produce their own immunoglobulins due to a defect in the recombinase gene. Several groups have reconstituted a functional humoral immune system in these mice by transfer of human peripheral blood lymphocytes (PBLs). These hu-PBL-SCID mice can be used to raise human antibodies upon immunization with antigen. Duchosal et al. Nature 355:258-262 (1992). Using another approach, the heavy- and light-chain genes within mice are turned off and then yeast artificial chromosomes (YACs) engineered with large DNA sequences containing human heavy- and light-chain genes are introduced into the mice. Such “XenoMice” are able to produce human antibodies upon immunization with an antigen of interest. See U.S. Pat. No. 5,434,340; U.S. Pat. No. 5,591,699; U.S. Pat. No. 5,569,825; U.S. Pat. No. 5,545,806; and U.S. Pat. No. 5,545,807.
- Human monoclonal antibodies may also be generated by immortalizing a human B lymphocyte producing an antibody of interest. The ethical issues surrounding immunizing humans in order to generate activated human B lymphocytes can be avoided by immunizing human lymphocytes in vitro. Both human PBLs (Borrebaeck et al. Proc. Natl. Acad. ScL USA 85:3995-4000 (1988)) and human splenocytes (Boerner et al. J. Immunol. 147, 86-95 (1991)) have been successfully immunized in vitro. Improvements in human hybridoma technology have been achieved by using a mouse-human heterohybrid as the fusion partner (Boerner et al.).
- Antibodies have been modified in order to increase their antigen-binding valency. For instance, Ghetie et al. homodimerized tumor-reactive monoclonal antibodies (anti-CD19, anti-CD20, anti-CD21, anti-CD22 and anti-HER2 antibodies) by chemically introducing a thioether bond between a pair of IgGs using two heterobifunctional crosslinkers . Ghetie et al. PNAS (USA) 94:7509-7514 (1997); and WO 99/02567. Wolff et al. Cancer Research 53: 2560-2565 (1993) also chemically linked an IgG monoclonal antibody (CHiBR96) using heterobifunctional cross-linkers to generate a monoclonal antibody homodimer with enhanced anti-tumor activity in nude mice.
- Shopes et al., replaced a serine residue near the carboxyl terminus of a human IgG1 heavy chain (Ser444) with a cysteine. The introduced intermolecular disulfide bonds between Cys444 residues linked pairs of immunoglobulins “tail-to-tail” to form covalent dimers (H2L2)2. The anti-dansyl dimers were said to be more efficient than monomeric human IgG1 at antibody-dependent complement-mediated cytolysis of hapten-bearing erythrocytes. Shopes, B. J. Immunol. 148(9): 2918-2922 (1992); and WO 91/19515. This approach, involving introduction of cysteine residues, has also been used to generate a homodimeric form of the CAMPATH-1H antibody. The homodimeric CAMPATH-1H antibody exhibited improved lysis using target cells expressing antigen at low density, but no improvement in lysis was observed using cells expressing antigen at high density. Greenwood et al. Ther. Immunol. 1:247-255 (1994). See, also, Caron et al. J. Exp. Med. 176:1191-1195 (1992), concerning an engineered anti-CD33 antibody with a serine to cysteine substitution at position 444 of the heavy chain allowing interchain disulfide bond formation at the COOH terminus of the IgG. The homodimeric IgG was said to have similar avidity to the parent IgG, but apparently showed an improved ability to internalize and retain radioisotope in target leukemia cells, and was more potent at complement-mediated leukemia cell killing and antibody-dependent cellular cytotoxicity using human effectors.
- Coloma and Morrison Nature Biotech. 15: 159-163 (1997) describe a tetravalent bispecific antibody which was engineered by fusing DNA encoding a single chain anti-dansyl antibody Fv (scFv) after the C terminus (CH3-scFv) or after the hinge (Hinge-scFv) of an IgG3 anti-dansyl antibody. See, also, WO95/09917. Smith and Morrison engineered three versions of mu-like IgG3 by engineering either (1) Cys414 of an IgM heavy chain or (2) Cys575 of an IgM heavy chain, or both (1) and (2), into the IgG3 heavy chain gene. All three mutant constructs were expressed by Sp2/0 cells and assembled into polymers containing up to six H2L2 subunits. The thus-produced ‘IgM-like’ polymers of IgG were considered to possess both the Fc gamma receptor binding properties of IgG and the more potent complement activity of IgM. See, Smith and Morrison Bio/Technology 12:683-688 (1994).
- Shuford and collegues isolated a human IgG1 anti-group B streptococci antibody oligomer from a transfected myeloma cell line. Shuford et al. Science 252:724-727 (1991). Immunochemical analysis and DNA sequencing indicated that the cell line produced both a normal kappa light chain and a 37 kD V-V-C variant light chain (L37). Contransfection of vectors encoding the heavy chain and L37 resulted in the production of oligomeric IgG.
- U.S. Pat. No. 5,641,870 (Rinderknecht et al.) describes a bivalent, linear F(ab′)2 fragment comprising tandem repeats of a heavy chain fragment (VH-CH1-VH-CH1) cosecreted with a light chain. The C-terminus of CH1 was joined directed to the N-terminus of VH without any extraneous linking protein sequences.
- Other publications on antibody variants include WO 00/06605; U.S. Pat. No. 5,591,828; U.S. Pat. No. 5,959,083; U.S. Pat. No. 6,027,725; WO98/58965; WO94/13804; Tutt et al. J. Immunol. 147:60-69 (1991); WO99/37791; U.S. Pat. No. 5,989,830; WO94/15642; EP 628,078B1; WO97/14719; Stevenson et al. Anti-Cancer Drug Design 3:219-230 (1989).
- The ErbB receptor tyrosine kinases are important mediators of cell growth, differentiation and survival. The receptor family includes at least four distinct members including Epidermal Growth Factor Receptor (EGFR or ErbB1), HER2 (ErbB2 or p185neu), HER3 (ErbB3) and HER4 (ErbB4 or tyro2).
- EGFR, encoded by the erbB1 gene, has been causally implicated in human malignancy. In particular, increased expression of EGFR has been observed in breast, bladder, lung, head, neck and stomach cancer, as well as glioblastomas. Increased EGFR receptor expression is often associated with increased production of the EGFR ligand, Transforming Growth Factor alpha (TGF-alpha), by the same tumor cells resulting in receptor activation by an autocrine stimulatory pathway. Baselga and Mendelsohn Pharmac. Ther. 64:127-154 (1994). Monoclonal antibodies directed against the EGFR or its ligands, TGF-alpha and EGF, have been evaluated as therapeutic agents in the treatment of such malignancies. See, e.g., Baselga and Mendelsohn., supra; Masui et al. Cancer Research 44:1002-1007 (1984); and Wu et al. J. Clin. Invest. 95:1897-1905 (1995).
- The second member of the ErbB family, p185neu, was originally identified as the product of the transforming gene from neuroblastomas of chemically treated rats. The activated form of the neu proto-oncogene results from a point mutation (valine to glutamic acid) in the transmembrane region of the encoded protein. Amplification of the human homolog of neu is observed in breast and ovarian cancers and correlates with a poor prognosis (Slamon et al., Science, 235:177-182 (1987); Slamon et al., Science, 244:707-712 (1989); and U.S. Pat. No. 4,968,603). To date, no point mutation analogous to that in the neu proto-oncogene has been reported for human tumors. Overexpression of HER2 (frequently but not uniformly due to gene amplification) has also been observed in other carcinomas including carcinomas of the stomach, endometrium, salivary gland, lung, kidney, colon, thyroid, pancreas and bladder.
- Antibodies directed against the rat p185neu and human HER2 protein products have been described. Drebin and colleagues have raised antibodies against the rat neu gene product, p185neu. See, for example, Drebin et al., Cell 41:695-706 (1985); Myers et al., Meth. Enzym. 198:277-290 (1991); and WO94/22478. Drebin et al. Oncogene 2:273-277 (1988) report that mixtures of antibodies reactive with two distinct regions of p185neu result in synergistic anti-tumor effects on neu-transformed NIH-3T3 cells implanted into nude mice. See also U.S. Pat. No. 5,824,311 issued Oct. 20, 1998.
- Hudziak et al., Mol. Cell. Biol. 9(3):1165-1172 (1989) describe the generation of a panel of anti-HER2 antibodies which were characterized using the human breast tumor cell line SKBR3. Relative cell proliferation of the SKBR3 cells following exposure to the antibodies was determined by crystal violet staining of the monolayers after 72 hours. Using this assay, maximum inhibition was obtained with the antibody called 4D5 which inhibited cellular proliferation by 56%. Other antibodies in the panel reduced cellular proliferation to a lesser extent in this assay. The antibody 4D5 was further found to sensitize HER2-overexpressing breast tumor cell lines to the cytotoxic effects of TNF-alpha. See, also, U.S. Pat. No. 5,677,171, issued Oct. 14, 1997. The anti-HER2 antibodies discussed in Hudziak et al. were further characterized in Fendly et al. Cancer Research 50:1550-1558 (1990); Kotts et al. In Vitro 26(3):59A (1990); Sarup et al. Growth Regulation 1:72-82 (1991); Shepard et al. J. Clin. Immunol. 11(3):117-127 (1991); Kumar et al. Mol. Cell. Biol. 11(2):979-986 (1991); Lewis et al. Cancer Immunol. Immunother. 37:255-263 (1993); Pietras et al. Oncogene 9:1829-1838 (1994); Vitetta et al. Cancer Research 54:5301-5309 (1994); Sliwkowski et al. J. Biol. Chem. 269(20):14661-14665 (1994); Scott et al. J. Biol. Chem. 266:14300-5 (1991); D'souza et al. Proc. Natl. Acad. Sci. 91:7202-7206 (1994); Lewis et al. Cancer Research 56:1457-1465 (1996); and Schaefer et al. Oncogene 15:1385-1394 (1997).
- A recombinant humanized IgG1 version of the murine anti-HER2 antibody 4D5 (rhuMAb HER2 or HERCEPTIN®; commercially available from Genentech, Inc., South San Francisco) is clinically active in patients with HER2-overexpressing metastatic breast cancers that have received extensive prior anti-cancer therapy (Baselga et al., J. Clin. Oncol. 14:737-744 (1996)). HERCEPTIN® received marketing approval from the Food and Drug Administration Sep. 25, 1998 for the treatment of patients with metastatic breast cancer whose tumors overexpress the HER2 protein.
- Other anti-HER2 antibodies with various properties have been described in Tagliabue et al. Int. J. Cancer 47:933-937 (1991); McKenzie et al. Oncogene 4:543-548 (1989); Maier et al. Cancer Res. 51:5361-5369 (1991); Bacus et al. Molecular Carcinogenesis 3:350-362 (1990); Stancovski et al. PNAS (USA) 88:8691-8695 (1991); Bacus et al. Cancer Research 52:2580-2589 (1992); Xu et al. Int. J. Cancer 53:401-408 (1993); WO94/00136; Kasprzyk et al. Cancer Research 52:2771-2776 (1992); Hancock et al. Cancer Res. 51:4575-4580 (1991); Shawver et al. Cancer Res. 54:1367-1373 (1994); Arteaga et al. Cancer Res. 54:3758-3765 (1994); Harwerth et al. J. Biol. Chem. 267:15160-15167 (1992); U.S. Pat. No. 5,783,186; Klapper et al. Oncogene 14:2099-2109 (1997); WO 98/77797; and U.S. Pat. No. 5,783,186. Homology screening has resulted in the identification of two other ErbB receptor family members; HER3 (U.S. Pat. Nos. 5,183,884 and 5,480,968 as well as Kraus et al. PNAS (USA) 86:9193-9197 (1989)) and HER4 (EP Pat Appln No 599,274; Plowman et al., Proc. Natl. Acad. Sci. USA, 90:1746-1750 (1993); and Plowman et al., Nature, 366:473-475 (1993)). Both of these receptors display increased expression on at least some breast cancer cell lines.
- The ErbB receptors are generally found in various combinations in cells and heterodimerization is thought to increase the diversity of cellular responses to a variety of ErbB ligands (Earp et al. Breast Cancer Research and Treatment 35: 115-132 (1995)). EGFR is bound by six different ligands; Epidermal Growth Factor (EGF), Transforming Growth Factor alpha (TGF-alpha), amphiregulin, Heparin Binding Epidermal Growth Factor (HB-EGF), betacellulin and epiregulin (Groenen et al. Growth Factors 11:235-257 (1994)). A family of heregulin proteins resulting from alternative splicing of a single gene are ligands for HER3 and HER4. The heregulin family includes alpha, beta and gamma heregulins (Holmes et al., Science, 256:1205-1210 (1992); U.S. Pat. No. 5,641,869; and Schaefer et al. Oncogene 15:1385-1394 (1997)); neu differentiation factors (NDFs), glial growth factors (GGFs); acetylcholine receptor inducing activity (ARIA); and sensory and motor neuron derived factor (SMDF). For a review, see Groenen et al. Growth Factors 11:235-257 (1994); Lemke, G. Molec. & Cell. Neurosci. 7:247-262 (1996) and Lee et al. Pharm. Rev. 47:51-85 (1995). Recently, two additional ErbB ligands were identified; neuregulin-2 (NRG-2) which is reported to bind either HER3 or HER4 (Chang et al. Nature 387 509-512 (1997); and Carraway et al Nature 387:512-516 (1997)) and neuregulin-3 which binds HER4 (Zhang et al. PNAS (USA) 94(18):9562-7 (1997)). HB-EGF, betacellulin and epiregulin also bind to HER4.
- While EGF and TGF-alpha do not bind HER2, EGF stimulates EGFR and HER2 to form a heterodimer, which activates EGFR and results in transphosphorylation of HER2 in the heterodimer. Dimerization and/or transphosphorylation appears to activate the HER2 tyrosine kinase. See Earp et al., supra. Likewise, when HER3 is co-expressed with HER2, an active signaling complex is formed and antibodies directed against HER2 are capable of disrupting this complex (Sliwkowski et al., J. Biol. Chem., 269(20):14661-14665 (1994)). Additionally, the affinity of HER3 for heregulin (HRG) is increased to a higher affinity state when co-expressed with HER2. See also, Levi et al., Journal of Neuroscience 15: 1329-1340 (1995); Morrissey et al., Proc. Natl. Acad. Sci. USA 92: 1431-1435 (1995); and Lewis et al., Cancer Res., 56:1457-1465 (1996) with respect to the HER2-HER3 protein complex. HER4, like HER3, forms an active signaling complex with HER2 (Carraway and Cantley, Cell 78:5-8 (1994)).
- Various molecules, such as Tumor Necrosis Factor-alpha (“TNF-alpha”), Tumor Necrosis Factor-beta (“TNF-beta”), Lymphotoxin-alpha (“LT-alpha”), CD30 ligand, CD27 ligand, CD40 ligand, OX-40 ligand, 4-1 BB ligand, Apo-1 ligand (also referred to as Fas ligand or CD95 ligand), Apo-2 ligand (also referred to as TRAIL), Apo-3 ligand (also referred to as TWEAK), osteoprotegerin (OPG), APRIL, RANK ligand (also referred to as TRANCE), and TALL-1 (also referred to as BlyS, BAFF or THANK) have been identified as members of the Tumor Necrosis Factor (“TNF”) family of cytokines (See, e.g., Gruss and Dower, Blood, 85:3378-3404 (1995); Pitti et al., J. Biol. Chem., 271:12687-12690 (1996); Wiley et al., Immunity, 3:673-682 (1995); Browning et al., Cell, 72:847-856 (1993); Armitage et al. Nature, 357:80-82 (1992); WO 97/01633 published Jan. 16, 1997; WO 97/25428 published Jul. 17, 1997; Marsters et al., Curr. Biol., 8:525-528 (1998); Simonet et al., Cell, 89:309-319 (1997); Chicheportiche et al., Biol. Chem., 272:32401-32410 (1997); Hahne et al., J. Exp. Med., 188:1185-1190 (1998); WO98/28426 published Jul. 2, 1998; WO98/46751 published Oct. 22, 1998; WO/98/18921 published May 7, 1998; Moore et al., Science, 285:260-263 (1999); Shu et al., J. Leukocyte Biol., 65:680 (1999); Schneider et al., J. Exp. Med., 189:1747-1756 (1999); and Mukhopadhyay et al., J. Biol. Chem., 274:15978-15981 (1999)). Among these molecules, TNF-alpha, TNF-beta, CD30 ligand, 4-1BB ligand, Apo-1 ligand, Apo-2 ligand (Apo2L/TRAIL) and Apo-3 ligand (TWEAK) have been reported to be involved in apoptotic cell death. Both TNF-alpha and TNF-beta have been reported to induce apoptotic death in susceptible tumor cells (Schmid et al., Proc. Natl. Acad. Sci., 83:1881 (1986); Dealtry et al., Eur. J. Immunol., 17:689 (1987)).
- Various molecules in the TNF family also have purported role(s) in the function or development of the immune system (Gruss et al., Blood, 85:3378 (1995)). Zheng et al. have reported that TNF-alpha is involved in post-stimulation apoptosis of CD8-positive T cells (Zheng et al., Nature, 377:348-351 (1995)). Other investigators have reported that CD30 ligand may be involved in deletion of self-reactive T cells in the thymus (Amakawa et al., Cold Spring Harbor Laboratory Symposium on Programmed Cell Death, Abstr. No. 10, (1995)). CD40 ligand activates many functions of B cells, including proliferation, immunoglobulin secretion, and survival (Renshaw et al., J. Exp. Med., 180:1889 (1994)). Another recently identified TNF family cytokine, TALL-1 (BlyS), has been reported, under certain conditions, to induce B cell proliferation and immunoglobulin secretion. (Moore et al., supra; Schneider et al., supra; Mackay et al., J. Exp. Med., 190:1697 (1999)).
- Mutations in the mouse Fas/Apo-1 receptor or ligand genes (called lpr and gld, respectively) have been associated with some autoimmune disorders, indicating that Apo-1 ligand may play a role in regulating the clonal deletion of self-reactive lymphocytes in the periphery (Krammer et al., Curr. Op. Immunol., 6:279-289 (1994); Nagata et al., Science, 267:1449-1456 (1995)). Apo-1 ligand is also reported to induce post-stimulation apoptosis in CD4-positive T lymphocytes and in B lymphocytes, and may be involved in the elimination of activated lymphocytes when their function is no longer needed (Krammer et al., supra; Nagata et al., supra). Agonist mouse monoclonal antibodies specifically binding to the Apo-1 receptor have been reported to exhibit cell killing activity that is comparable to or similar to that of TNF-alpha (Yonehara et al., J. Exp. Med., 169:1747-1756 (1989)).
- Induction of various cellular responses mediated by such TNF family cytokines is believed to be initiated by their binding to specific cell receptors. Previously, two distinct TNF receptors of approximately 55-kDa (TNFR1) and 75-kDa (TNFR2) were identified (Hohman et al., J. Biol. Chem., 264:14927-14934 (1989); Brockhaus et al., Proc. Natl. Acad. Sci., 87:3127-3131 (1990); EP 417,563, published Mar. 20, 1991; Loetscher et al., Cell, 61:351 (1990); Schall et al., Cell, 61:361 (1990); Smith et al., Science, 248:1019-1023 (1990); Lewis et al., Proc. Natl. Acad. Sci., 88:2830-2834 (1991); Goodwin et al., Mol. Cell. Biol., 11:3020-3026 (1991)). Those TNFRs were found to share the typical structure of cell surface receptors including extracellular, transmembrane and intracellular regions. The extracellular portions of both receptors were found naturally also as soluble TNF-binding proteins (Nophar et al., EMBO J., 9:3269 (1990); and Kohno et al., Proc. Natl. Acad. Sci. U.S.A., 87:8331 (1990); Hale et al., J. Cell. Biochem. Supplement 15F, 1991, p. 113 (P424)).
- The extracellular portion of
type 1 andtype 2 TNFRs (TNFR1 and TNFR2) contains a repetitive amino acid sequence pattern of four cysteine-rich domains (CRDs) designated 1 through 4, starting from the NH2-terminus. (Schall et al., supra; Loetscher et al., supra; Smith et al., supra; Nophar et al., supra; Kohno et al., supra; Banner et al., Cell, 73:431-435 (1993)). A similar repetitive pattern of CRDs exists in several other cell-surface proteins, including the p75 nerve growth factor receptor (NGFR) (Johnson et al., Cell, 47:545 (1986); Radeke et al., Nature, 325:593 (1987)), the B cell antigen CD40 (Stamenkovic et al., EMBO J., 8:1403 (1989)), the T cell antigen OX40 (Mallet et al., EMBO J., 9:1063 (1990)) and the Fas antigen (Yonehara et al., supra and Itoh et al., Cell, 66:233-243 (1991)). CRDs are also found in the soluble TNFR (sTNFR)-like T2 proteins of the Shope and myxoma poxviruses (Upton et al., Virology, 160:20-29 (1987); Smith et al., Biochem. Biophys. Res. Commun., 176:335 (1991); Upton et al., Virology, 184:370 (1991)). Optimal alignment of these sequences indicates that the positions of the cysteine residues are well conserved. These receptors are sometimes collectively referred to as members of the TNF/NGF receptor superfamily. - The TNF family ligands identified to date, with the exception of Lymphotoxin-alpha, are type II transmembrane proteins, whose C-terminus is extracellular. In contrast, most receptors in the TNF receptor (TNFR) family identified to date are type I transmembrane proteins. In both the TNF ligand and receptor families, however, homology identified between family members has been found mainly in the extracellular domain (“ECD”). Several of the TNF family cytokines, including TNF-alpha, Apo-1 ligand and CD40 ligand, are cleaved proteolytically at the cell surface; the resulting protein in each case typically forms a homotrimeric molecule that functions as a soluble cytokine. TNF receptor family proteins are also usually cleaved proteolytically to release soluble receptor ECDs that can function as inhibitors of the cognate cytokines.
- More recently, other members of the TNFR family have been identified. In von Bulow et al., Science, 278:138-141 (1997), investigators describe a plasma membrane receptor referred to as Transmembrane Activator and CAML-Interactor or “TACI”. The TACI receptor is reported to contain a cysteine-rich motif characteristic of the TNFR family. In an in vitro assay, cross linking of TACI on the surface of transfected Jurkat cells with TACI-specific antibodies led to activation of NF-KB (see also, WO 98/39361 published Sep. 18, 1998).
- Laabi et al., EMBO J., 11:3897-3904 (1992) reported identifying a new gene called “BCM” whose expression was found to coincide with B cell terminal maturation. The open reading frame of the BCM normal cDNA predicted a 184 amino acid long polypeptide with a single transmembrane domain. These investigators later termed this gene “BCMA.” (Laabi et al., Nucleic Acids Res., 22:1147-1154 (1994)). BCMA mRNA expression was reported to be absent in human malignant B cell lines which represent the pro-B lymphocyte stage, and thus, is believed to be linked to the stage of differentiation of lymphocytes (Gras et al., Int. Immunology, 7:1093-1106 (1995)). In Madry et al., Int. Immunology, 10:1693-1702 (1998), the cloning of murine BCMA cDNA was described. The murine BCMA cDNA is reported to encode a 185 amino acid long polypeptide having 62% identity to the human BCMA polypeptide. Alignment of the murine and human BCMA protein sequences revealed a conserved motif of six cysteines in the N-terminal region, suggesting that the BCMA protein belongs to the TNFR superfamily (Madry et al., supra).
- In Marsters et al., Curr. Biol., 6:750 (1996), investigators describe a full length native sequence human polypeptide, called Apo-3, which exhibits similarity to the TNFR family in its extracellular cysteine-rich repeats and resembles TNFR1 and CD95 in that it contains a cytoplasmic death domain sequence (see also Marsters et al., Curr. Biol., 6:1669 (1996)). Apo-3 has also been referred to by other investigators as DR3, wsl-1, TRAMP, and LARD (Chinnaiyan et al., Science, 274:990 (1996); Kitson et al., Nature, 384:372 (1996); Bodmer et al., Immunity, 6:79 (1997); Screaton et al., Proc. Natl. Acad. Sci., 94:4615-4619 (1997)).
- Pan et al. have disclosed another TNF receptor family member referred to as “DR4” (Pan et al., Science, 276:111-113 (1997); see also WO98/32856 published Jul. 30, 1998). The DR4 was reported to contain a cytoplasmic death domain capable of engaging the cell suicide apparatus. Pan et al. disclose that DR4 is believed to be a receptor for the ligand known as Apo2L/TRAIL.
- In Sheridan et al., Science, 277:818-821 (1997) and Pan et al., Science, 277:815-818 (1997), another molecule believed to be a receptor for Apo2L/TRAIL is described (see also, WO98/51793 published Nov. 19, 1998; and WO98/41629 published Sep. 24, 1998). That molecule is referred to as DR5 (it has also been alternatively referred to as Apo-2; TRAIL-R, TR6, Tango-63, hAPO8, TRICK2 or KILLER (Screaton et al., Curr. Biol., 7:693-696 (1997); Walczak et al., EMBO J., 16:5386-5387 (1997); Wu et al., Nature Genetics, 17:141-143 (1997); WO98/35986 published Aug. 20, 1998; EP870,827 published Oct. 14, 1998; WO98/46643 published Oct. 22, 1998; WO99/02653 published Jan. 21, 1999; WO99/09165 published Feb. 25, 1999; and WO99/11791 published Mar. 11, 1999). Like DR4, DR5 is reported to contain a cytoplasmic death domain and be capable of signaling apoptosis. The crystal structure of the complex formed between Apo2L/TRAIL and DR5 is described in Hymowitz et al., Molecular Cell, 4:563-571 (1999).
- Yet another death domain-containing receptor, DR6, was recently identified (Pan et al., FEBS Letters, 431:351-356 (1998)). Aside from containing four putative extracellular cysteine rich domains and a cytoplasmic death domain, DR6 is believed to contain a putative leucine-zipper sequence that overlaps with a proline-rich motif in the cytoplasmic region. The proline-rich motif resembles sequences that bind to src-homology-3 domains, which are found in many intracellular signal-transducing molecules.
- A further group of recently identified receptors are referred to as “decoy receptors,” which are believed to function as inhibitors, rather than transducers of signaling. This group includes DcR1 (also referred to as TRID, LIT or TRAIL-R3) (Pan et al., Science, 276:111-113 (1997); Sheridan et al., Science, 277:818-821 (1997); McFarlane et al., J. Biol. Chem., 272:25417-25420 (1997); Schneider et al., FEBS Letters, 416:329-334 (1997); Degli-Esposti et al., J. Exp. Med., 186:1165-1170 (1997); and Mongkolsapaya et al., J. Immunol., 160:3-6 (1998)) and DcR2 (also called TRUNDD or TRAIL-R4) (Marsters et al., Curr. Biol., 7:1003-1006 (1997); Pan et al., FEBS Letters, 424:41-45 (1998); Degli-Esposti et al., Immunity, 7:813-820 (1997)), both cell surface molecules, as well as OPG (Simonet et al., supra; Emery et al., infra) and DcR3 (Pitti et al., Nature, 396:699-703 (1998)), both of which are secreted, soluble proteins.
- Additional newly identified members of the TNFR family include CAR1, HVEM, GITR, ZTNFR-5, NTR-1, and TNFL1 (Brojatsch et al., Cell, 87:845-855 (1996); Montgomery et al., Cell, 87:427-436 (1996); Marsters et al., J. Biol. Chem., 272:14029-14032 (1997); Nocentini et al., Proc. Natl. Acad. Sci. USA 94:6216-6221 (1997); Emery et al., J. Biol. Chem., 273:14363-14367 (1998); WO99/04001 published Jan. 28, 1999; WO99/07738 published Feb. 18, 1999; WO99/33980 published Jul. 8, 1999).
- As reviewed recently by Tewari et al., TNFR1, TNFR2 and CD40 modulate the expression of proinflammatory and costimulatory cytokines, cytokine receptors, and cell adhesion molecules through activation of the transcription factor, NF-κB (Tewari et al., Curr. Op. Genet. Develop., 6:39-44 (1996)). NF-κB is the prototype of a family of dimeric transcription factors whose subunits contain conserved Rel regions (Verma et al., Genes Develop., 9:2723-2735 (1996); Baldwin, Ann. Rev. Immunol., 14:649-681 (1996)). In its latent form, NF-κB is complexed with members of the I-κB inhibitor family; upon inactivation of the I-κB in response to certain stimuli, released NF-κB translocates to the nucleus where it binds to specific DNA sequences and activates gene transcription. As described above, the TNFR members identified to date either include or lack an intracellular death domain region. Some TNFR molecules lacking a death domain, such as TNFR2, CD40, HVEM, and GITR, are capable of modulating NF-κB activity. (see, e.g., Lotz et al., J. Leukocyte Biol., 60:1-7 (1996)).
- For a review of the TNF family of cytokines and their receptors, see Ashkenazi and Dixit, Science, 281:1305-1308 (1998); Golstein, Curr. Biol., 7:750-753 (1997); Gruss and Dower, supra, and Nagata, Cell, 88:355-365 (1997).
- Lymphocytes are one of many types of white blood cells produced in the bone marrow during the process of hematopoiesis. There are two major populations of lymphocytes: B lymphocytes (B cells) and T lymphocytes (T cells). The lymphocytes of particular interest herein are B cells.
- B cells mature within the bone marrow and leave the marrow expressing an antigen-binding antibody on their cell surface. When a naive B cell first encounters the antigen for which its membrane-bound antibody is specific, the cell begins to divide rapidly and its progeny differentiate into memory B cells and effector cells called “plasma cells”. Memory B cells have a longer life span and continue to express membrane-bound antibody with the same specificity as the original parent cell. Plasma cells do not produce membrane-bound antibody but instead produce the antibody in a form that can be secreted. Secreted antibodies are the major effector molecule of humoral immunity.
- The CD20 antigen (also called human B-lymphocyte-restricted differentiation antigen, Bp35) is a hydrophobic transmembrane protein with a molecular weight of approximately 35 kD located on pre-B and mature B lymphocytes (Valentine et al. J. Biol. Chem. 264(19):11282-11287 (1989); and Einfeld et al. EMBO J. 7(3):711-717 (1988)). The antigen is also expressed on greater than 90% of B cell non-Hodgkin's lymphomas (NHL) (Anderson et al. Blood 63(6):1424-1433 (1984)), but is not found on hematopoietic stem cells, pro-B cells, normal plasma cells or other normal tissues (Tedder et al. J. Immunol. 135(2):973-979 (1985)). CD20 regulates an early step(s) in the activation process for cell cycle initiation and differentiation (Tedder et al., supra) and possibly functions as a calcium ion channel (Tedder et al. J. Cell. Biochem. 14D:195 (1990)).
- Given the expression of CD20 in B cell lymphomas, this antigen can serve as a candidate for “targeting” of such lymphomas. In essence, such targeting can be generalized as follows: antibodies specific to the CD20 surface antigen of B cells are administered to a patient. These anti-CD20 antibodies specifically bind to the CD20 antigen of (ostensibly) both normal and malignant B cells; the antibody bound to the CD20 surface antigen may lead to the destruction and depletion of neoplastic B cells. Additionally, chemical agents or radioactive labels having the potential to destroy the tumor can be conjugated to the anti-CD20 antibody such that the agent is specifically “delivered” to the neoplastic B cells. Irrespective of the approach, a primary goal is to destroy the tumor; the specific approach can be determined by the particular anti-CD20 antibody which is utilized and, thus, the available approaches to targeting the CD20 antigen can vary considerably.
- CD19 is another antigen that is expressed on the surface of cells of the B lineage. Like CD20, CD19 is found on cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells (Nadler, L. Lymphocyte Typing II 2: 3-37 and Appendix, Renling et al. eds. (1986) by Springer Verlag). Unlike CD20 however, antibody binding to CD19 causes internalization of the CD19 antigen. CD19 antigen is identified by the HD237-CD19 antibody (also called the “B4” antibody) (Kiesel et al. Leukemia Research II, 12: 1119 (1987)), among others. The CD19 antigen is present on 4-8% of peripheral blood mononuclear cells and on greater than 90% of B cells isolated from peripheral blood, spleen, lymph node or tonsil. CD19 is not detected on peripheral blood T cells, monocytes or granulocytes. Virtually all non-T cell acute lymphoblastic leukemias (ALL), B cell chronic lymphocytic leukemias (CLL) and B cell lymphomas express CD19 detectable by the antibody B4 (Nadler et al. J. Immunol. 131:244 (1983); and Nadler et al. in Progress in Hematology Vol. XII pp. 187-206. Brown, E. ed. (1981) by Grune & Stratton, Inc).
- Additional antibodies which recognize differentiation stage-specific antigens expressed by cells of the B cell lineage have been identified. Among these are the B2 antibody directed against the CD21 antigen; B3 antibody directed against the CD22 antigen; and the J5 antibody directed against the CD10 antigen (also called CALLA). See U.S. Pat. No. 5,595,721 issued Jan. 21, 1997 (Kaminski et al.).
- The rituximab (RITUXAN®) antibody is a genetically engineered chimeric murine/human monoclonal antibody directed against the CD20 antigen. Rituximab is the antibody called “C2B8” in U.S. Pat. No. 5,736,137 issued Apr. 7, 1998 (Anderson et al.). RITUXAN® is indicated for the treatment of patients with relapsed or refractory low-grade or follicular, CD20 positive, B cell non-Hodgkin's lymphoma. In vitro mechanism of action studies have demonstrated that RITUXAN® binds human complement and lyses lymphoid B cell lines through CDC (Reff et al. Blood 83(2):435-445 (1994)). Additionally, it has significant activity in assays for ADCC. More recently, RITUXAN® has been shown to have anti-proliferative effects in tritiated thymidine incorporation assays and to induce apoptosis directly, while other anti-CD19 and CD20 antibodies do not (Maloney et al. Blood 88(10):637a (1996)). Synergy between RITUXAN® and chemotherapies and toxins has also been observed experimentally. In particular, RITUXAN® sensitizes drug-resistant human B cell lymphoma cell lines to the cytotoxic effects of doxorubicin, CDDP, VP-16, diphtheria toxin and ricin (Demidem et al. Cancer Chemotherapy & Radiopharmaceuticals 12(3):177-186 (1997)). In vivo preclinical studies have shown that RITUXAN® depletes B cells from the peripheral blood, lymph nodes, and bone marrow of cynomolgus monkeys, presumably through complement and cell-mediated processes (Reff et al. Blood 83(2):435-445 (1994)).
- The present invention provides multivalent antibodies (e.g. tetravalent antibodies) with three or more antigen binding sites, which can be readily produced by recombinant expression of nucleic acid encoding the polypeptide chains of the antibody. In one embodiment, the multivalent antibody comprises a dimerization domain and three or more antigen binding sites. The preferred dimerization domain comprises (or consists of) an Fc region or a hinge region. In one embodiment, the invention provides an isolated antibody comprising a dimerization domain and three or more antigen binding sites amino-terminal thereto. The invention further provides an isolated antibody comprising an Fc region and three or more antigen binding sites amino-terminal to the Fc region. The preferred multivalent antibody herein comprises (or consists of) three to about eight, but preferably four, antigen binding sites (which are generally all “functional”, as hereindefined). In one embodiment, the multivalent antibody comprises five or more (e.g. up to about eight) antigen binding sites. The multivalent antibody herein is preferably not a native sequence IgA or IgM, and may lack an Fc region or have only one Fc region.
- In the preferred embodiment, the multivalent antibody comprises at least one polypeptide chain (and preferably two polypeptide chains), wherein the polypeptide chain(s) comprise two or more variable domains. For instance, the polypeptide chain(s) may comprise VD1-(X1)n-VD2-(X2)n-Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1. For instance, the polypeptide chain(s) may comprise: VH-CH1-flexible linker-VH-CH1-Fc region chain; VH-CH1-VH-CH1-Fc region chain; VL-CL-flexible linker-VL-CL-Fc region chain; or VL-CL-VL-CL-Fc region chain. Where the polypeptide chain (or polypeptide chains) comprise Fd-flexible linker-Fd, the flexible linker may comprise a peptide such as gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser, or gly-gly-gly-ser (SEQ ID NO:11).
- The multivalent antibody herein preferably further comprises at least two (and preferably four) light chain variable domain polypeptides. The multivalent antibody herein may, for instance, comprise from about two to about eight light chain variable domain polypeptides. The light chain variable domain polypeptides contemplated here comprise a light chain variable domain and, optionally, further comprise a CL domain.
- The multivalent antibodies herein have properties which are desirable, among other things, from a therapeutic standpoint. For instance, the multivalent antibody may (1) be internalized (and/or catabolized) faster than a bivalent antibody by a cell expressing an antigen to which the antibodies bind; (2) be an agonist antibody; and/or (3) induce cell death and/or apoptosis of a cell expressing an antigen which the multivalent antibody is capable of binding to. The “parent antibody” which provides at least one antigen binding specificity of the multivalent antibody may be one which is internalized (and/or catabolized) by a cell expressing an antigen to which the antibody binds; and/or may be an agonist, cell-death-inducing, and/or apoptosis-inducing antibody, and the multivalent form of the antibody as described herein may display improvement(s) in one or more of these properties. Moreover, the parent antibody may lack any one or more of these properties, but may be endowed with them when constructed as a multivalent antibody as hereindescribed.
- The three or more antigen binding sites of the multivalent antibodies herein may all bind the same antigen; or may bind two or more (e.g. from two to about three) different antigens.
- The multivalent antibody may bind (1) a cell surface protein expressed (or overexpressed) by tumor cells, e.g. Epidermal Growth Factor Receptor (EGFR), HER2 receptor, HER3 receptor, HER4 receptor, or DcR3; (2) a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily (e.g. an Apo2L receptor, such as DR4, DR5, DcR1 or DcR2); and/or (3) a B cell surface antigen (such as CD19, CD20, CD22 or CD40). In the preferred embodiment of the invention, all of the functional antigen binding sites of the multivalent antibody bind the same antigen as listed above (e.g. all four antigen binding sites of a tetravalent antibody bind either (1), (2) or (3)).
- The invention also provides immunoconjugates comprising the multivalent antibody conjugated with a cytotoxic agent. The cytotoxic agent here may be one which is active in killing cells once internalized.
- The invention additionally pertains to a polypeptide chain comprising VD1-(X1)n-VD2 (X2)n-Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1. For instance, the polypeptide chain may comprise VH-CH1-flexible linker-VH-CH1-Fc region chain; VH-CH1-VH-CH1-Fc region chain; VL-CL-flexible linker-VL-CL-Fc region chain; or VL-CL-VL-CL-Fc region chain. In another embodiment, the polypeptide chain comprises VH-CH1-flexible linker-VH-CH1-dimerization domain; VH-CH1-VH-CH1-dimerization domain; VL-CL-flexible linker-VL-CL-dimerization domain; or VL-CL-VL-CL-dimerization domain. For instance, the polypeptide chain may comprise VH-CH1-flexible linker-VH-CH1-hinge region; VH-CH1-VH-CH1-hinge region. The invention additionally provides an antibody comprising one or more (preferably two) of such polypeptide chains. The antibody preferably further comprises at least two (and preferably four) light chain or heavy chain variable domain polypeptides, e.g., where the light chain variable domain polypeptides comprise VL-CL and the heavy chain variable domain polypeptides comprise VH-CH1.
- The invention further provides a polypeptide chain comprising three or more heavy chain or light chain variable domains, wherein each of the variable domains is able to combine with three or more light chain or heavy chain variable domain polypeptides to form three or more antigen binding sites, each directed against the same antigen. The invention also provides an isolated antibody comprising the polypeptide chain. In the preferred embodiment, where the polypeptide chain comprises three or more heavy chain variable domains, the antibody preferably further comprises three or more light chain variable domain polypeptides which can combine with the heavy chain variable domains to form the three or more antigen binding sites. Examples of such antibodies are shown in
FIG. 23 D (with three antigen binding sites) andFIG. 23E (with four antigen binding sites). In addition, the invention provides a polypeptide chain comprising the formula: (a) VL-CL-flexible linker-VL-CL-flexible linker-VL-CL; (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; (c) (VL-CL)n, wherein n is three or more; or (d) (VH-CH 1)n, wherein n is three or more. - The invention further provides: isolated nucleic acid encoding the multivalent antibody or polypeptide chain; a vector comprising nucleic acid encoding the multimeric antibody or polypeptide chain, optionally, operably linked to control sequences recognized by a host cell transformed with the vector; a host cell comprising (e.g. transformed with) nucleic acid encoding the multimeric antibody or polypeptide chain; a method for producing the multivalent antibody or polypeptide chain comprising culturing the host cell so that the nucleic acid is expressed and, optionally, recovering the multivalent antibody or polypeptide chain from the host cell culture (e.g. from the host cell culture medium). Nucleic acids encoding (1) the heavy chain variable domains and (2) the light chain variable domains of the multivalent antibody are preferrably co-expressed by a host cell transformed with both (1) and (2). Nucleic acids (1) and (2) may be present in the same, or different, vectors.
- Diagnostic and therapeutic uses for the multivalent antibodies disclosed herein are contemplated. In one diagnostic application, the invention provides a method for determining the presence of an antigen of interest comprising exposing a sample suspected of containing the antigen to the multivalent antibody and determining binding of the multivalent antibody to the sample. Both in vitro and in vivo diagnostic methods are provided.
- In one therapeutic application, the invention provides a method of treating a mammal suffering from, or predisposed to, a disease or disorder, comprising administering to the mammal a therapeutically effective amount of a multivalent antibody as disclosed herein, or of a composition comprising the multivalent antibody and a pharmaceutically acceptable carrier. The disorder to be treated herein may be cancer, in which case the method may further comprise administering a therapeutically effective amount of a cytotoxic agent to the mammal. The present invention further relates to a method of inducing apoptosis of a cancer cell comprising exposing the cell to a multivalent antibody as described herein, wherein the multivalent antibody binds a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily. The method may involve killing a B cell by exposing the B cell to a multivalent antibody that binds a B cell surface antigen. Moreover, the method may relate to killing a cell which expresses (or overexpresses) an ErbB receptor comprising exposing the cell to an antibody that binds the ErbB receptor.
-
FIG. 1 is a schematic representation of a native IgG and digestion thereof with (1) papain to generate two Fab fragments and an Fc region or (2) pepsin to generate a F(ab′)2 fragment and multiple small fragments. Disulfide bonds are represented by lines between CH1 and CL domains and the two CH2 domains. V is variable domain; C is constant domain; L stands for light chain and H stands for heavy chain. -
FIGS. 2A-E depict the structures of the five major naturally occurring immunoglobulin isotypes; IgG (FIG. 2A ), IgD (FIG. 2B ), IgE (FIG. 2C ), IgA dimer (FIG. 2D ), and IgM pentamer (FIG. 2E ). -
FIG. 3 depicts alignments of native sequence IgG Fc regions. Native sequence human IgG Fc region sequences, humIgG1 (non-A and A allotypes) (SEQ ID NOs: 1 and 2, respectively), humIgG2 (SEQ ID NO:3), humIgG3 (SEQ ID NO:4) and humIgG4 (SEQ ID NO:5), are shown. The human IgG1 sequence is the non-A allotype, and differences between this sequence and the A allotype (at positions 356 and 358; EU numbering system) are shown below the human IgG1 sequence. Native sequence murine IgG Fc region sequences, murIgG1 (SEQ ID NO:6), murIgG2A (SEQ ID NO:7), murIgG2B (SEQ ID NO:8) and murlgG3 (SEQ ID NO:9), are also shown. -
FIGS. 4A-B depict schematically tetravalent antibodies according to the present invention. InFIG. 4A , the four antigen binding Fabs are numbered (1 and 2 for each arm of the tetravalent antibody) and X represents a dimerization domain. InFIG. 4B , the dimerization domain of the tetravalent antibody is an Fc region. -
FIG. 5 shows the construct used for expression of a tetravalent anti-HER2 antibody (OctHER2) in Example 1. -
FIGS. 6A-C illustrate binding of OctHER2 (FIG. 6A ); bivalent IgG1 rhuMAb 4D5-8 expressed by 293 cells (FIG. 6B ); and vialed HERCEPTIN® (expressed by Chinese hamster ovary (CHO) cells) (FIG. 6C ) to HER2 extracellular domain (ECD) as determined using an enzyme-linked immunosorbent assay (ELISA). -
FIG. 7 depicts ultracentrifugation analysis of binding of OctHER2 to HER2ECD. Average molecular weights (theoretical or experimentally determined) versus molar ratio of OctHER2 to HER2ECD are shown. Theoretical calculated average molecular weights assuming tetravalent antibody has four fully functional binding sites are shown in circles; theoretical calculated average molecular weights assuming tetravalent antibody has three fully functional binding sites are shown in squares; and triangles represent experimentally determined molecular weights. -
FIGS. 8A-D depict the growth inhibitory activity of HERCEPTIN® compared to OctHER2 using SKBR3 (3+ HER2 overexpressing) (FIG. 8A ), MDA 361 (2+ HER2 overexpressing) (FIG. 8B ), BT474 (3+ HER2 overexpressing) (FIG. 8C ) and MCF7 (0+ HER2 expressing) (FIG. 8D ) cell lines. -
FIG. 9 depicts the effect of flexible linkers on the growth inhibitory activity of tetravalent anti-HER2 antibodies with respect toMDA 231 cells (1+ HER2 overexpressing) or SKBR3 cells (3+ HER2 overexpressing). -
FIGS. 10A-B compare the rate of OctHER2 internalization/catabolism (FIG. 10A ) to that of HERCEPTIN® (FIG. 10B ), in relation to both MDA 453 (2+ HER2 overexpressing) and SKBR3 (3+ HER2 overexpressing) cell lines. -
FIGS. 11A-I are electron microscopy photographs showing internalization of OctHER2.FIGS. 11A-F show subcellular localization of 125I-OctHER2 in SKBR3 cells. Autoradiographic silver grains were observed associated with the villi of the apical cell membrane (FIG. 11A ), in close proximity with a forming coated pit (FIG. 11B , arrow), with smooth cytosolic vesicles (FIGS. 11C and D) and endosomes (FIGS. 11E and F). Bars=0.25 μM.FIGS. 11G-I show internalization attime 0 hours (FIGS. 11G) and 5 hours (FIGS. 11H and 11I ). -
FIGS. 12A-E depict apoptosis induced by an anti-DR5 tetravalent antibody (16E2 Octopus), an anti-DR5 bivalent IgG antibody (16E2 IgG), and Apo2L/TRAIL (Apo2L) on cancer cell lines: COLO 205 (FIG. 12A ), SK-MES-1 (FIG. 12B ), HCT116 (FIG. 12C ), and HOP 92 (FIG. 12D ), compared to a non-cancer control cell line, HUMEC (FIG. 12E ). -
FIGS. 13A-D are histology slides stained to detect apoptotic cells. Tumor tissues from mice treated with 16E2 Octopus or Apo2L/TRAIL were fixed in 10% formalin and then embedded into parafilm and sectioned onto slides which were then stained with hematoxylin and eosin and visualized under a 400× magnification. The effect of 16E2 Octopus at 6 and 24 hours is shown inFIGS. 13A and B, respectively; control-treated cells are shown inFIG. 13C ; and Apo2L/TRAIL-treated cells are shown inFIG. 13D . -
FIG. 14 represents the in vivo activity of Apo2L/TRAIL (60 mg/kg, 5×/week), 3H3 bivalent IgG (5 mg/kg givendays days days COLO 205 tumors in athymic nude mice. -
FIG. 15 represents an alamarBlue in vitro assay confirming the apoptotic activity of the material used in the mouse studies (Apo2L/TRAIL and 16E2 Octopus) as compared to an Apo2L standard positive control. The anti-IgE antibody (E25) used as a negative control in the mouse studies was confirmed to have no apoptotic activity. -
FIG. 16 represents the results of a crystal violet apoptosis assay comparing anti-DR5 3H3 Octopus to various batches of the anti-DR5 16E2 Octopus. -
FIGS. 17A-B reveal the results of the alamarBlue apoptosis assay with respect to Apo2L/TRAIL (WO97/25428), anti-DR5 3H3 Octopus antibody, anti-DR5 16E2 Octopus antibody, and Apo2L/TRAIL with a FLAG epitope-tag cross linked by an anti-FLAG antibody (WO97/25428), with respect to SK-MES-1 (FIG. 17A ) and Jurkat (FIG. 17B ) cells in the presence of 5% fetal bovine serum (FBS). -
FIGS. 18A-C depict dose response curves that show the effect of the anti-DR5 16E2 Octopus (upper graphs) compared to Apo2L/TRAIL (lower graphs) on the growth of leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer human tumor cell lines at 2 days. Results are from the National Cancer Institute Developmental Therapeutics Program. All samples were tested at 5 concentrations, starting at 1% of the stock solution (16E2 Octopus stock 0.2 mg/ml) and 4×0.5 log dilutions. -
FIGS. 19A-C depict dose response curves that show the effect of the anti-DR516E2 Octopus (upper graphs) compared to Apo2L/TRAIL (lower graphs) on the growth of leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer human tumor cell lines at 6 days. Results are from the National Cancer Institute Developmental Therapeutics Program. All samples were tested at 5 concentrations, starting at 1% of the stock solution (16E2 Octopus stock 0.2 mg/ml) and 4×0.5 log dilutions. -
FIGS. 20A-B present a quantitative summary of the 2 day in vitro results from the National Cancer Institute Developmental Therapeutics Program comparing the anti-DR5 16E2 Octopus (FIG. 20A ) to Apo2L/TRAIL (FIG. 20B ) analyzing growth inhibition (GI50), stasis (TGI), and toxicity (LC50). -
FIGS. 21A-B present a quantitative summary of the 6 day in vitro results from the National Cancer Institute Developmental Therapeutics Program comparing the anti-DR5 16E2 Octopus (FIG. 21A ) to Apo2L/TRAIL (FIG. 21B ) analyzing growth inhibition (G150), stasis (TGI), and toxicity (LC50). -
FIG. 22 depicts apoptosis of Wil-2 cells by the anti-CD20 antibody RITUXAN®, RITUXAN® cross-linked with anti-human IgG (RITUXAN®-IgG) and a tetravalent anti-CD20 antibody (OctCD20). -
FIGS. 23A-E are cartoons depicting the full-length Octopus/tetravalent antibody (FIG. 23B ), the Octopus F(ab)′2 (FIG. 23C ), POPoct-3 Fab (FIG. 23D ) and POPoct-4 Fab (FIG. 23E ) in comparison to the native IgG (FIG. 23A ). A representative coomassie stained Tris-Glycine gel of anti-CD20 (C2B8) Octopus proteins compares the sizes of the intact antibodies in non-reducing conditions (FIG. 23F ), and of the heavy chains in reducing conditions, under which disulfide bonds are disrupted resulting in separation of the heavy and light chains (FIG. 23G ). -
FIG. 24 depicts the construction of the Octopus F(ab′)2 backbone. Any VH/CH1 region can be substituted into the F(ab′)2 backbone via the BamHI, NheI and BspEI restriction enzyme sites. -
FIG. 25 depicts the construction of the POPoct-3 heavy chain. -
FIG. 26 depicts the construction of the POPoct-4 heavy chain. -
FIG. 27 depicts the activity of multivalent anti-HER2 antibodies in cytostasis assays using BT474 cells. -
FIGS. 28A-B depict the activity of multivalent anti-HER2 antibodies in cytostasis assays using SKBR3 cells. The figures are representative plots of n=4 cytostasis assays. -
FIGS. 29A-B show internalization capability of multivalent anti-HER2 antibodies in SKBR3 cells (FIG. 29A ) and BT474 cells (FIG. 29B ). -
FIGS. 30A-B reveal apoptosis of COLO205 cells by multivalent anti-DR5 antibodies -
FIGS. 31A-B demonstrate signalling of multivalent anti-DR5 antibodies through the caspase pathway. -
FIG. 32 compares apoptosis induced by IgG cross-linked RITUXAN® (RITUXAN-IgG) and IgG cross-linked OctCD20 (OctCD20-IgG). -
FIG. 33 shows apoptosis of WIL2 cells by multivalent anti-CD20 antibodies, the IF5 anti-CD20 antibody (Clark et al. PNAS (USA) 82: 1766-1770 (1985)) and IgG cross-linked IFS antibody (IF5+IgG-X). -
FIG. 34 depicts homotypic cell adhesion in WIL2S cells induced by IF5 anti-CD20 antibody, IgG cross-linked IF5 antibody and POPoct-3 CD20. -
FIG. 35 reflects RITUXAN® or OctCD20 internalization/catabolism on DB, WIL2 and Ramos B-cell lymphoma lines. - Throughout the present specification and claims, the numbering of the residues in an immunoglobulin heavy chain is that of the EU index as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991), expressly incorporated herein by reference. The “EU index as in Kabat” refers to the residue numbering of the human IgG1 EU antibody.
- An “ErbB receptor” is a receptor protein tyrosine kinase which belongs to the ErbB receptor family and includes EGFR, HER2, ErbB3 and ErbB4 receptors as well as TEGFR (U.S. Pat. No. 5,708,156) and other members of this family to be identified in the future. The ErbB receptor will generally comprise an extracellular domain, which may bind an ErbB ligand; a lipophilic transmembrane domain; a conserved intracellular tyrosine kinase domain; and a carboxyl-terminal signaling domain harboring several tyrosine residues which can be phosphorylated. The ErbB receptor may be a native sequence ErbB receptor or an amino acid sequence variant thereof. Preferably the ErbB receptor is native sequence human ErbB receptor.
- By “ErbB ligand” is meant a polypeptide which binds to and/or activates an ErbB receptor. The ErbB ligand of particular interest herein is a native sequence human ErbB ligand such as Epidermal Growth Factor (EGF) (Savage et al., J. Biol. Chem. 247:7612-7621 (1972)); Tansforming Growth Factor alpha (TGF-alpha) (Marquardt et al., Science 223:1079-1082 (1984)); amphiregulin also known as schwanoma or keratinocyte autocrine growth factor (Shoyab et al. Science 243:1074-1076 (1989); Kimura et al. Nature 348:257-260 (1990); and Cook et al. Mol. Cell. Biol. 11:2547-2557 (1991)); betacellulin (Shing et al., Science 259:1604-1607 (1993); and Sasada et al. Biochem. Biophys. Res. Commun. 190:1173 (1993)); heparin-binding epidermal growth factor (HB-EGF) (Higashiyama et al., Science 251:936-939 (1991)); epiregulin (Toyoda et al., J. Biol. Chem. 270:7495-7500 (1995); and Komurasaki et al. Oncogene 15:2841-2848 (1997)), a heregulin (see below); neuregulin-2 (NRG-2) (Carraway et al., Nature 387:512-516 (1997)); neuregulin-3 (NRG-3) (Zhang et al., Proc. Natl. Acad. Sci. 94:9562-9567 (1997)); or cripto (CR-1) (Kannan et al. J. Biol. Chem. 272(6):3330-3335 (1997)). ErbB ligands which bind EGFR include EGF, TGF-alpha, amphiregulin, betacellulin, HB-EGF and epiregulin. ErbB ligands which bind HER3 include heregulins. ErbB ligands capable of binding HER4 include betacellulin, epiregulin, HB-EGF, NRG-2, NRG-3 and heregulins.
- “Heregulin” (HRG) when used herein refers to a polypeptide comprising an amino acid sequence encoded by the heregulin gene product as disclosed in U.S. Pat. No. 5,641,869 or Marchionni et al., Nature, 362:312-318 (1993), and biologically active variants of such polypeptides. Examples of heregulins include heregulin-alpha heregulin-beta1, heregulin-beta2 and heregulin-beta3 (Holmes et al., Science, 256:1205-1210 (1992); and U.S. Pat. No. 5,641,869); neu differentiation factor (NDF) (Peles et al. Cell 69: 205-216 (1992)); acetylcholine receptor-inducing activity (ARIA) (Falls et al. Cell 72:801-815 (1993)); glial growth factors (GGFs) (Marchionni et al., Nature, 362:312-318 (1993)); sensory and motor neuron derived factor (SMDF) (Ho et al. J. Biol. Chem. 270:14523-14532 (1995)); gamma-heregulin (Schaefer et al. Oncogene 15:1385-1394 (1997)). An example of a biologically active fragment/amino acid sequence variant of a native sequence HRG polypeptide, is an EGF-like domain fragment (e.g. HRGbeta1177-244).
- An “ErbB hetero-oligomer” herein is a noncovalently associated oligomer comprising at least two different ErbB receptors. Such complexes may form when a cell expressing two or more ErbB receptors is exposed to an ErbB ligand and can be isolated by immunoprecipitation and analyzed by SDS-PAGE as described in Sliwkowski et al., J. Biol. Chem., 269(20):14661-14665 (1994), for example. Examples of such ErbB hetero-oligomers include EGFR-HER2, HER2-HER3 and HER3-HER4 complexes. Moreover, the ErbB hetero-oligomer may comprise two or more HER2 receptors combined with a different ErbB receptor, such as HER3, HER4 or EGFR. Other proteins, such as a cytokine receptor subunit (e.g. gp130), may be included in the hetero-oligomer.
- The terms “ErbB1”, “epidermal growth factor receptor” and “EGFR” are used interchangeably herein and refer to native sequence EGFR as disclosed, for example, in Carpenter et al. Ann. Rev. Biochem. 56:881-914 (1987), including variants thereof (e.g. a deletion mutant EGFR as in Humphrey et PNAS (USA) 87:4207-4211 (1990)). erbB1 refers to the gene encoding the EGFR protein product. Examples of antibodies which bind to EGFR include MAb 579 (ATCC CRL HB 8506), MAb 455 (ATCC CRL HB8507), MAb 225 (ATCC CRL 8508), MAb 528 (ATCC CRL 8509) (see, U.S. Pat. No. 4,943,533, Mendelsohn et al.) and variants thereof, such as chimerized 225 (C225) and reshaped human 225 (H225) (see, WO 96/40210, Imclone Systems Inc.).
- The expressions “ErbB2” and “HER2” are used interchangeably herein and refer to native sequence human HER2 protein described, for example, in Semba et al., PNAS (USA) 82:6497-6501 (1985) and Yamamoto et al. Nature 319:230-234 (1986) (Genebank accession number X03363), and variants thereof. The term erbB2 refers to the gene encoding human HER2 and neu refers to the gene encoding rat p185neu. Preferred HER2 is native sequence human HER2. Examples of antibodies which bind HER2 include MAbs 4D5 (ATCC CRL 10463), 2C4 (ATCC HB-12697), 7F3 (ATCC HB-12216), and 7C2 (ATCC HB 12215) (see, U.S. Pat. No. 5,772,997; WO98/77797; and U.S. Pat. No. 5,840,525, expressly incorporated herein by reference). Humanized anti-HER2 antibodies include huMAb4D5-1, huMAb4D5-2, huMAb4D5-3, huMAb4D5-4, huMAb4D5-5, huMAb4D5-6, huMAb4D5-7 and huMAb4D5-8 (HERCEPTIN®) as described in Table 3 of U.S. Pat. No. 5,821,337 expressly incorporated herein by reference; humanized 520C9 (WO93/21319). Human anti-HER2 antibodies are described in U.S. Pat. No. 5,772,997 issued Jun. 30, 1998 and WO 97/00271 published Jan. 3, 1997.
- “ErbB3” and “HER3” refer to the receptor polypeptide as disclosed, for example, in U.S. Pat. Nos. 5,183,884 and 5,480,968 as well as Kraus et al. PNAS (USA) 86:9193-9197 (1989), including variants thereof. Examples of antibodies which bind HER3 are described in U.S. Pat. No. 5,968,511 (Akita and Sliwkowski), e.g. the 8B8 antibody (ATCC HB 12070) or a humanized variant thereof.
- The terms “ErbB4” and “HER4” herein refer to the receptor polypeptide as disclosed, for example, in EP Pat Appin No 599,274; Plowman et al., Proc. Natl. Acad. ScL USA, 90:1746-1750 (1993); and Plowman et al., Nature, 366:473-475 (1993), including variants thereof such as the HER4 isoforms disclosed in WO 99/19488.
- A “B cell surface marker” herein is an antigen expressed on the surface of a B cell which can be targeted with an antibody which binds thereto. Exemplary B cell surface markers include the CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD40, CD53, CD72, CD73, CD74, CDw75, CDw76, CD77, CDw78, CD79a, CD79b, CD80, CD81, CD82, CD83, CDw84, CD85 and CD86 leukocyte surface markers. The B cell surface marker of particular interest is preferentially expressed on B cells compared to other non-B cell tissues of a mammal and may be expressed on both precursor B cells and mature B cells. In one embodiment, the marker is one, like CD20 or CD19, which is found on B cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells. The preferred B cell surface markers herein are CD19, CD20, CD22 and CD40.
- The “CD20” antigen is an about 35 kDa, non-glycosylated phosphoprotein found on the surface of greater than 90% of B cells from peripheral blood or lymphoid organs. CD20 is expressed during early pre-B cell development and remains until plasma cell differentiation. CD20 is present on both normal B cells as well as malignant B cells. Other names for CD20 in the literature include “B-lymphocyte-restricted antigen” and “Bp35”. The CD20 antigen is described in Clark et al. PNAS (USA) 82:1766 (1985), for example. Examples of antibodies which bind the CD20 antigen include: “C2B8” which is now called “rituximab” (“RITUXAN®”) (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); the yttrium-[90]-labeled 2B8 murine antibody designated “Y2B8” (U.S. Pat. No. 5,736,137, expressly incorporated herein by reference); murine IgG2a “B1” optionally labeled with 131I to generate the “131I-B1” antibody (BEXXAR™) (U.S. Pat. No. 5,595,721, expressly incorporated herein by reference); murine monoclonal antibody “1F5” (Press et al. Blood 69(2):584-591 (1987)); “chimeric 2H7” antibody (U.S. Pat. No. 5,677,180, expressly incorporated herein by reference); and monoclonal antibodies L27, G28-2, 93-1B3, B-C1 or NU-B2 available from the International Leukocyte Typing Workshop (Valentine et al., In: Leukocyte Typing III (McMichael, Ed., p. 440, Oxford University Press (1987)).
- The “CD19” antigen refers to the about 90 kDa antigen identified, for example, by the HD237-CD19 or B4 antibody (Kiesel et al. Leukemia Research II, 12: 1119 (1987)). Like CD20, CD19 is found on cells throughout differentiation of the lineage from the stem cell stage up to a point just prior to terminal differentiation into plasma cells. Binding of an antibody to CD19 may cause internalization of the CD19 antigen. Examples of antibodies which bind the CD19 antigen include the anti-CD19 antibodies in Hekman et al. Cancer Immunol. Immunother. 32:364-372 (1991) and Vlasveld et al. Cancer Immunol. Immunother. 40:37-47 (1995); and the B4 antibody in Kiesel et al. Leukemia Research II, 12: 1119 (1987).
- The “CD22” antigen has a molecular weight of about 140,000 kD. CD22 is expressed in the cytoplasm of early pre-B and progenitor cells, appears on the surface of only mature B cells and on the majority of non-Hodgkin's lymphoma (NHL) cells, and is then lost during terminal differentiation prior to the plasma cell stage from both the surface and cytoplasm. An example of an anti-CD22 antibody is the LL2 antibody described in Juweid et al. Cancer Research 55:5899-5907 (1995), including chimeric/humanized variants thereof.
- The “CD40” antigen is a cell surface phosphorylated glycoprotein that is expressed on a variety of cell types, including B cells, B cell malignancies, follicular dendritic cells, basal epithelial cells and carcinomas. CD40 binds CD40 ligand (CD40L). Aside from being a B cell surface antigen, CD40 is also a member of the TNF receptor superfamily. Examples of antibodies that bind CD40 include those which (1) block CD40/CD40L interaction and have anti-neoplastic properties (Armitage et al., U.S. Pat. No. 5,674,492); (2) antagonize signaling through CD40 (deBoer et al., U.S. Pat. No. 5,677,165); (3) deliver a stimulatory signal through CD40 but do not increase the interaction between CD40 and CD40L, e.g., G28-5 (Ledbetter et al., U.S. Pat. No. 5,182,368); (4) increase the interaction between CD40 and CD40L, e.g., CD40.4 (5C3) (PharMingen, San Diego, Calif.) and S2C6 (deposited with the American Type Culture Collection (ATCC), Manassass, Va. on May 25, 1999 under accession number PTA-110).
- The “tumor necrosis factor receptor superfamily” or “TNF receptor superfamily” herein refers to receptor polypeptides bound by cytokines in the TNF family. Generally, these receptors are Type I transmembrane receptors with one or more cysteine rich repeat sequences in their extracellular domain. The TNF receptor superfamily may be further subdivided into (1) death receptors; (2) decoy receptors; and (3) signaling receptors that lack death domains. The “death receptors” contain in their cytoplasmic or intracellular region a “death domain”, i.e., a region or sequence which acts to transduce signals in the cell which can result in apoptosis or in induction of certain genes. The “decoy receptors” lack a functional death domain and are incapable of transducing signals which result in apoptosis. Examples of cytokines in the TNF gene family include Tumor Necrosis Factor-alpha (TNF-alpha), Tumor Necrosis Factor-beta (TNF-beta or lymphotoxin), CD30 ligand, CD27 ligand, CD40 ligand, OX-40 ligand, 4-1 BB ligand, Apo-1 ligand (also referred to as Fas ligand or CD95 ligand), Apo-2 ligand (also referred to as TRAIL), Apo-3 ligand (also referred to as TWEAK), osteoprotegerin (OPG), APRIL, RANK ligand (also referred to as TRANCE), and TALL-1 (also referred to as BlyS, BAFF or THANK). Examples of receptors in the TNF receptor superfamily include:
type 1 Tumor Necrosis Factor Receptor (TNFR1),type 2 Tumor Necrosis Factor Receptor (TNFR2), p75 Nerve Growth Factor receptor (NGFR), the B cell surface antigen CD40, the T cell antigen OX-40, Apo-1 receptor (also called Fas or CD95), Apo-3 receptor (also called DR3, swl-1, TRAMP and LARD), the receptor called “Transmembrane Activator and CAML-Interactor” or “TACI”, BCMA protein, DR4, DR5 (alternatively referred to as Apo-2; TRAIL-R2, TR6, Tango-63, hAPO8, TRICK2 or KILLER), DR6, DcR1 (also referred to as TRID, LIT or TRAIL-R3), DcR2 (also called TRAIL-R4 or TRUNDD), OPG, DcR3 (also called TR6 or M68), CAR1, HVEM (also called ATAR or TR2), GITR, ZTNFR-5, NTR-1, TNFL1, CD30, Lymphotoxin beta receptor (LTBr), 4-1BB receptor and TR9 (EP988, 371 A1). - The terms “Apo-2 ligand” or “Apo2L” refer to the Apo2L polypeptides disclosed in WO97/25428, published 17 Jul. 1997 and expressly incorporated herein by reference. For purposes of the present application, these terms also refer to the polypeptides referred to as TRAIL disclosed in WO97/01633, published 16 Jan., 1997 and U.S. Pat. No. 5,763,223, issued Jun. 9, 1998 and expressly incorporated herein by reference.
- An “Apo2L receptor” is a polypeptide to which Apo2L can specifically bind. The term “Apo2L receptor” when used herein encompasses native sequence Apo2L receptors and variants thereof. These terms encompass Apo2L receptor from a variety of mammals, including humans. The Apo2L receptor may be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. Examples of “native sequence” Apo2L receptors include Apo-2 polypeptide or DR5 (WO98/51793, expressly incorporated herein by reference), native sequence DR4 as described in Pan et al. Science 276:111-113 (1997); native
sequence decoy receptor 1 or DcR1 as in Sheridan et al., Science 277:818-821 (1997); and nativesequence decoy receptor 2 or DcR2 as in Marsters et al. Curr. Biol. 7:1003-1006 (1997); native sequence osteoprotegerin (see Simonet et al. Cell 89:309-319 (1997); and Emery et al. J. Interferon and Cytokine Research 18(5): A47 Abstract 2.17 (1998)). Examples of anti-DR5 antibodies include 3F11.39.7 (ATCC HB-12456), 3H3.14.5 (ATCC HB-12534), 3D5.1.10 (HB-12536) and 3H1.18.10 (HB-12535), 16E2 and 20E6 (see WO 98/51793, expressly incorporated herein by reference). Examples of anti-DR4 antibodies include 4E7.24.3 (ATCC HB-12454) and 4H6.17.8 (ATCC HB-12455) (see, WO 99/37684, expressly incorporated herein by reference). - Native sequence “DcR3” is described in WO99/14330, expressly incorporated herein by reference. That patent publication describes the following mAbs directed against DcR3: 4C4.1.4 (ATCC HB-12573); 5C4.14.7 (ATCC HB-12574); 11C5.2.8 (ATCC HB-12572); 8D3.1.5 (ATCC HB-12571); and 4B7.1.1 (ATCC HB-12575).
- A “native sequence” polypeptide comprises a polypeptide having the same amino acid sequence as a polypeptide derived from nature. Thus, a native sequence polypeptide can have the amino acid sequence of naturally-occurring polypeptide from any mammal. Such native sequence polypeptide can be isolated from nature or can be produced by recombinant or synthetic means. The term “native sequence” polypeptide specifically encompasses naturally-occurring truncated or secreted forms of the polypeptide (e.g., an extracellular domain sequence), naturally-occurring variant forms (e.g., alternatively spliced forms) and naturally-occurring allelic variants of the polypeptide.
- A polypeptide “variant” means a biologically active polypeptide having at least about 80% amino acid sequence identity with the native sequence polypeptide. Such variants include, for instance, polypeptides wherein one or more amino acid residues are added, or deleted, at the N- or C-terminus of the polypeptide. Ordinarily, a variant will have at least about 80% amino acid sequence identity, more preferably at least about 90% amino acid sequence identity, and even more preferably at least about 95% amino acid sequence identity with the native sequence polypeptide.
- “Apoptosis” refers to programmed cell death. Physiological events often indicative of the occurrence of apoptosis include: fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies). Various methods are available for evaluating the cellular events associated with apoptosis. For example, phosphatidyl serine (PS) translocation can be measured by annexin V binding; DNA fragmentation can be evaluated through DNA laddering or propidium-iodine staining; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells.
- The term “antibody” is used in the broadest sense and includes monoclonal antibodies (including full length or intact monoclonal antibodies), polyclonal antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments (see below) so long as they exhibit the desired biological activity.
- Unless indicated otherwise, the expression “multivalent antibody” is used throughout this specification to denote an antibody comprising three or more antigen binding sites. The multivalent antibody is preferably engineered to have the three or more antigen binding sites and is generally not a native sequence IgM or IgA antibody.
- “Antibody fragments” comprise only a portion of an intact antibody, generally including an antigen binding site of the intact antibody and thus retaining the ability to bind antigen. Examples of antibody fragments encompassed by the present definition include: (i) the Fab fragment, having VL, CL, VH and CH1 domains; (ii) the Fab′ fragment, which is a Fab fragment having one or more cysteine residues at the C-terminus of the CH1 domain; (iii) the Fd fragment having VH and CH1 domains; (iv) the Fd′ fragment having VH and CH1 domains and one or more cysteine residues at the C-terminus of the CH1 domain; (v) the Fv fragment having the VL and VH domains of a single arm of an antibody; (vi) the dAb fragment (Ward et al., Nature 341, 544-546 (1989)) which consists of a VH domain; (vii) isolated CDR regions; (viii) F(ab′)2 fragments, a bivalent fragment including two Fab′ fragments linked by a disulphide bridge at the hinge region; (ix) single chain antibody molecules (e.g. single chain Fv; scFv) (Bird et al., Science 242:423-426 (1988); and Huston et al., PNAS (USA) 85:5879-5883 (1988)); (x) “diabodies” with two antigen binding sites, comprising a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (see, e.g., EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); (xi) “linear antibodies” comprising a pair of tandem Fd segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen binding regions (Zapata et al. Protein Eng. 8(10):1057-1062 (1995); and U.S. Pat. No. 5,641,870).
- The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigen. Furthermore, in contrast to polyclonal antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) or Marks et al., J. Mol. Biol. 222:581-597 (1991), for example.
- The monoclonal antibodies herein specifically include “chimeric” antibodies in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567; and Morrison et al., Proc. Natl. Acad. ScL USA 81:6851-6855 (1984)).
- “Humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region of the recipient are replaced by residues from a hypervariable region of a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity. In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).
- A “human antibody” is one which possesses an amino acid sequence which corresponds to that of an antibody produced by a human and/or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art. In one embodiment, the human antibody is selected from a phage library, where that phage library expresses human antibodies (Vaughan et al. Nature Biotechnology 14:309-314 (1996): Sheets et al. PNAS (USA) 95:6157-6162 (1998)); Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991)). Human antibodies can also be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the following scientific publications: Marks et aL, Bio/Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368:812-13 (1994); Fishwild et al., Nature Biotechnology 14: 845-51 (1996); Neuberger, Nature Biotechnology 14: 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol. 13:65-93 (1995). Alternatively, the human antibody may be prepared via immortalization of human B lymphocytes producing an antibody directed against a target antigen (such B lymphocytes may be recovered from an individual or may have been immunized in vitro). See, e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147 (1):86-95 (1991); and U.S. Pat. No. 5,750,373.
- The term “variable” refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called hypervariable regions both in the light chain and the heavy chain variable domains. The more highly conserved portions of variable domains are called the framework regions (FRs). The variable domains of native heavy and light chains each comprise four FRs, largely adopting a beta-sheet configuration, connected by three hypervariable regions, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The hypervariable regions in each chain are held together in close proximity by the FRs and, with the hypervariable regions from the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC).
- The term “hypervariable region” when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable region generally comprises amino acid residues from a “complementarity determining region” or “CDR” (e.g. residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)) and/or those residues from a “hypervariable loop” (e.g. residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). “Framework Region” or “FR” residues are those variable domain residues other than the hypervariable region residues as herein defined.
- Depending on the amino acid sequence of the constant domain of their heavy chains, intact antibodies can be assigned to different “classes”. There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into “subclasses” (isotypes), e.g., IgG1 (including non-A and A allotypes), IgG2, IgG3, IgG4, IgA, and IgA2. The heavy-chain constant domains that correspond to the different classes of antibodies are called α, δ, ε, γ and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.
- The light chains of antibodies from any vertebrate species can be assigned to one of two clearly distinct types, called kappa (κ) and lambda (λ), based on the amino acid sequences of their constant domains.
- The term “Fc region” is used to define the C-terminal region of an immunoglobulin heavy chain which may be generated by papain digestion of an intact antibody. The Fc region may be a native sequence Fc region or a variant Fc region. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy chain Fc region is usually defined to stretch from an amino acid residue at about position Cys226, or from about position Pro230, to the carboxyl-terminus of the Fc region. The Fc region of an immunoglobulin generally comprises two constant domains, a CH2 domain and a CH3 domain, and optionally comprises a CH4 domain.
- By “Fc region chain” herein is meant one of the two polypeptide chains of an Fc region.
- The “CH2 domain” of a human IgG Fc region (also referred to as “Cγ2” domain) usually extends from an amino acid residue at about
position 231 to an amino acid residue at aboutposition 340. The CH2 domain is unique in that it is not closely paired with another domain. Rather, two N-linked branched carbohydrate chains are interposed between the two CH2 domains of an intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for the domain-domain pairing and help stabilize the CH2 domain. Burton, Molec. Immunol. 22:161-206 (1985). The CH2 domain herein may be a native sequence CH2 domain or variant CH2 domain. - The “CH3 domain” comprises the stretch of residues C-terminal to a CH2 domain in an Fc region (i.e. from an amino acid residue at about position 341 to an amino acid residue at about position 447 of an IgG). The CH3 region herein may be a native sequence CH3 domain or a variant CH3 domain (e.g. a CH3 domain with an introduced “protroberance” in one chain thereof and a corresponding introduced “cavity” in the other chain thereof; see U.S. Pat. No. 5,821,333, expressly incorporated herein by reference). Such variant CH3 domains may be used to make multispecific (e.g. bispecific) antibodies as herein described.
- “Hinge region” is generally defined as stretching from about Glu216, or about Cys226, to about Pro230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). Hinge regions of other IgG isotypes may be aligned with the IgG1 sequence by placing the first and last cysteine residues forming inter-heavy chain S—S bonds in the same positions. The hinge region herein may be a native sequence hinge region or a variant hinge region. The two polypeptide chains of a variant hinge region generally retain at least one cysteine residue per polypeptide chain, so that the two polypeptide chains of the variant hinge region can form a disulfide bond between the two chains. The preferred hinge region herein is a native sequence human hinge region, e.g. a native sequence human IgG1 hinge region.
- A “functional Fc region” possesses at least one “effector function” of a native sequence Fc region. Exemplary “effector functions” include C1 q binding; complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g. B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding domain (e.g. an antibody variable domain) and can be assessed using various assays known in the art for evaluating such antibody effector functions.
- A “native sequence Fc region” comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature.
FIG. 3 provides amino acid sequences of native sequence human and murine IgG Fc regions. - A “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification. Preferably, the variant Fc region has at least one amino acid substitution compared to a native sequence Fc region or to the Fc region of a parent polypeptide, e.g. from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region herein will preferably possess at least about 80% sequence identity with a native sequence Fc region and/or with an Fc region of a parent polypeptide, and most preferably at least about 90% sequence identity therewith, more preferably at least about 95% sequence identity therewith.
- “Antibody-dependent cell-mediated cytotoxicity” and “ADCC” refer to a cell-mediated reaction in which nonspecific cytotoxic cells that express Fc receptors (FcRs) (e.g. Natural Killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and subsequently cause lysis of the target cell. The primary cells for mediating ADCC, NK cells, express FcγRIII only, whereas monocytes express FcγRI, FcγRII and FcγRIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess ADCC activity of a molecule of interest, an in vitro ADCC assay, such as that described in U.S. Pat. No. 5,500,362 or 5,821,337 may be performed. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in a animal model such as that disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).
- “Human effector cells” are leukocytes which express one or more FcRs and perform effector functions. Preferably, the cells express at least FcγRIII and perform ADCC effector function. Examples of human leukocytes which mediate ADCC include peripheral blood mononuclear cells (PBMC), natural killer (NK) cells, monocytes, cytotoxic T cells and neutrophils; with PBMCs and NK cells being preferred. The effector cells may be isolated from a native source thereof, e.g. from blood or PBMCs as described herein.
- The terms “Fc receptor” and “FcR” are used to describe a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcγRII receptors include FcγRIIA (an “activating receptor”) and FcγRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)).
- “Complement dependent cytotoxicity” and “CDC” refer to the lysing of a target in the presence of complement. The complement activation pathway is initiated by the binding of the first component of the complement system (C1q) to a molecule (e.g. an antibody) complexed with a cognate antigen. To assess complement activation, a CDC assay, e.g. as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996), may be performed.
- An “affinity matured” antibody is one with one or more alterations in one or more CDRs thereof which result an improvement in the affinity of the antibody for antigen, compared to a parent antibody which does not possess those alteration(s). Preferred affinity matured antibodies will have nanomolar or even picomolar affinities for the target antigen. Affinity matured antibodies are produced by procedures known in the art. Marks et al. Bio/Technology 10:779-783 (1992) describes affinity maturation by VH and VL domain shuffling. Random mutagenesis of CDR and/or framework residues is described by: Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al, J. Mol. Biol. 226:889-896 (1992).
- “Percent (%) amino acid sequence identity” herein is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a selected sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared. For purposes herein, however, % amino acid sequence identity values are obtained as described below by using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was authored by Genentech, Inc. has been filed with user documentation in the U.S. Copyright Office, Washington D.C., 20559, where it is registered under U.S. Copyright Registration No. TXU510087, and is publicly available through Genentech, Inc., South San Francisco, Calif. The ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.
- For purposes herein, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be phrased as a given amino acid sequence A that has or comprises a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows:
-
100 times the fraction X/Y - where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A.
- A “polypeptide chain” is a polypeptide wherein each of the domains thereof is joined to other domain(s) by peptide bond(s), as opposed to non-covalent interactions or disulfide bonds.
- A “flexible linker” herein refers to a peptide comprising two or more amino acid residues joined by peptide bond(s), and provides more rotational freedom for two polypeptides (such as two Fd regions) linked thereby. Such rotational freedom allows two or more antigen binding sites joined by the flexible linker to each access target antigen(s) more efficiently. Examples of suitable flexible linker peptide sequences include gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser, and gly-gly-gly-ser (SEQ ID NO:11). Preferably the flexible linker comprises 2 to about 10 amino acid residues, and most preferably four or less residues.
- A “dimerization domain” is formed by the association of at least two amino acid residues (generally cysteine residues) or of at least two peptides or polypeptides (which may have the same, or different, amino acid sequences). The peptides or polypeptides may interact with each other through covalent and/or non-covalent association(s). Examples of dimerization domains herein include an Fc region; a hinge region; a CH3 domain; a CH4 domain; a CH1-CL pair; an “interface” with an engineered “knob” and/or “protruberance” as described in U.S. Pat. No. 5,821,333, expressly incorporated herein by reference; a leucine zipper (e.g. a jun/fos leucine zipper, see Kostelney et al., J. Immunol., 148: 1547-1553 (1992); or a yeast GCN4 leucine zipper); an isoleucine zipper; a receptor dimer pair (e.g., interleukin-8 receptor (IL-8R); and integrin heterodimers such as LFA-1 and GPIIIb/IIIa), or the dimerization region(s) thereof; dimeric ligand polypeptides (e.g. nerve growth factor (NGF), neurotrophin-3 (NT-3), interleukin-8 (IL-8), vascular endothelial growth factor (VEGF), and brain-derived neurotrophic factor (BDNF); see Arakawa et al. J. Biol. Chem. 269(45): 27833-27839 (1994) and Radziejewski et al. Biochem. 32(48): 1350 (1993)), or the dimerization region(s) thereof; a pair of cysteine residues able to form a disulfide bond; a pair of peptides or polypeptides, each comprising at least one cysteine residue (e.g. from about one, two or three to about ten cysteine residues) such that disulfide bond(s) can form between the peptides or polypeptides (hereinafter “a synthetic hinge”); and antibody variable domains. The most preferred dimerization domain herein is an Fc region or a hinge region.
- “Naturally occurring amino acid residues” (i.e. amino acid residues encoded by the genetic code) may be selected from the group consisting of: alanine (Ala); arginine (Arg); asparagine (Asn); aspartic acid (Asp); cysteine (Cys); glutamine (Gin); glutamic acid (Glu); glycine (Gly); histidine (H is); isoleucine (Ile): leucine (Leu); lysine (Lys); methionine (Met); phenylalanine (Phe); proline (Pro); serine (Ser); threonine (Thr); tryptophan (Trp); tyrosine (Tyr); and valine (Val). A “non-naturally occurring amino acid residue” refers to a residue, other than those naturally occurring amino acid residues listed above, which is able to covalently bind adjacent amino acid residues(s) in a polypeptide chain. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine and other amino acid residue analogues such as those described in Ellman et al. Meth. Enzym. 202:301-336 (1991). To generate such non-naturally occurring amino acid residues, the procedures of Noren et al. Science 244:182 (1989) and Ellman et al., supra, can be used. Briefly, these procedures involve chemically activating a suppressor tRNA with a non-naturally occurring amino acid residue followed by in vitro transcription and translation of the RNA.
- An “isolated” polypeptide is one that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. In preferred embodiments, the polypeptide will be purified (1) to greater than 95% by weight of polypeptide as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain. Isolated polypeptide includes the polypeptide in situ within recombinant cells since at least one component of the polypeptide's natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.
- A “functional antigen binding site” of an antibody is one which is capable of binding a target antigen. The antigen binding affinity of the antigen binding site is not necessarily as strong as the parent antibody from which the antigen binding site is derived, but the ability to bind antigen must be measurable using any one of a variety of methods known for evaluating antibody binding to an antigen. Moreover, the antigen binding affinity of each of the antigen binding sites of a multivalent antibody herein need not be quantitatively the same. For the multimeric antibodies herein, the number of functional antigen binding sites can be evaluated using ultracentrifugation analysis as described in Example 2 below. According to this method of analysis, different ratios of target antigen to multimeric antibody are combined and the average molecular weight of the complexes is calculated assuming differing numbers of functional binding sites. These theoretical values are compared to the actual experimental values obtained in order to evaluate the number of functional binding sites.
- By “ligand activation of a receptor” is meant signal transduction (e.g. for a tyrosine kinase receptor, that caused by an intracellular kinase domain of a tyrosine kinase receptor phosphorylating tyrosine residues in the receptor or a substrate polypeptide) mediated by ligand binding to the receptor (or a receptor complex comprising the receptor of interest). In the case of an ErbB receptor, generally, this will involve binding of an ErbB ligand to an ErbB hetero-oligomer which activates a kinase domain of one or more of the ErbB receptors in the hetero-oligomer and thereby results in phosphorylation of tyrosine residues in one or more of the ErbB receptors and/or phosphorylation of tyrosine residues in additional substrate polypeptides(s).
- An antibody which “blocks” ligand activation of an receptor is one which reduces or prevents such activation as hereinabove defined. Such blocking can occur by any means, e.g. by interfering with: ligand binding to the receptor, receptor complex formation, tyrosine kinase activity of a tyrosine kinase receptor in a receptor complex and/or phosphorylation of tyrosine kinase residue(s) in or by the receptor. Examples of antibodies which block ligand activation of an ErbB receptor include monoclonal antibodies 2C4 and 7F3 (which block HRG activation of HER2/HER3 and HER2/HER4 hetero-oligomers; and EGF, TGF-beta or amphiregulin activation of an EGFR/HER2 hetero-oligomer); and L26, L96 and L288 antibodies (Klapper et al. Oncogene 14:2099-2109 (1997)), which block EGF and NDF binding to T47D cells which express EGFR, HER2, HER3 and HER4.
- An antibody having a “biological characteristic” of a designated antibody is one which possesses one or more of the biological characteristics of that antibody which distinguish it from other antibodies that bind to the same antigen.
- A “growth inhibitory agent” when used herein refers to a compound or composition which inhibits growth of a cell in vitro and/or in vivo. Thus, the growth inhibitory agent may be one which significantly reduces the percentage of cells in S phase. Examples of growth inhibitory agents include agents that block cell cycle progression (at a place other than S phase), such as agents that induce G1 arrest and M-phase arrest. Classical M-phase blockers include the vincas (vincristine and vinblastine), TAXOL®, and topo II inhibitors such as doxorubicin, epirubicin, daunorubicin, etoposide, and bleomycin. Those agents that arrest G1 also spill over into S-phase arrest, for example, DNA alkylating agents such as tamoxifen, prednisone, dacarbazine, mechlorethamine, cisplatin, methotrexate, 5-fluorouracil, and ara-C. Further information can be found in The Molecular Basis of Cancer, Mendelsohn and Israel, eds.,
Chapter 1, entitled “Cell cycle regulation, oncogenes, and antineoplastic drugs” by Murakami et al. (WB Saunders: Philadelphia, 1995), especially p. 13. - Examples of “growth inhibitory” anti-HER2 antibodies are those which bind to HER2 and inhibit the growth of cancer cells overexpressing HER2. Preferred growth inhibitory anti-HER2 antibodies inhibit growth of SKBR3 breast tumor cells in cell culture by greater than 20%, and preferably greater than 50% (e.g. from about 50% to about 100%) at an antibody concentration of about 0.5 to 30 μg/ml, where the growth inhibition is determined six days after exposure of the SKBR3 cells to the antibody (see U.S. Pat. No. 5,677,171 issued Oct. 14, 1997).
- An antibody which “induces cell death” is one which causes a viable cell to become nonviable. The cell is generally one which expresses the antigen to which the antibody binds, especially where the cell overexpresses the antigen. Preferably, the cell is a cancer cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell. In vitro, the cell may be a SKBR3, BT474,
Calu 3, MDA-MB-453, MDA-MB-361 or SKOV3 cell. Cell death in vitro may be determined in the absence of complement and immune effector cells to distinguish cell death induced by antibody dependent cell-mediated cytotoxicity (ADCC) or complement dependent cytotoxicity (CDC). Thus, the assay for cell death may be performed using heat inactivated serum (i.e. in the absence of complement) and in the absence of immune effector cells. To determine whether the antibody is able to induce cell death, loss of membrane integrity as evaluated by uptake of propidium iodide (P1), trypan blue (see Moore et al. Cytotechnology 17:1-11 (1995)) or 7AAD can be assessed relative to untreated cells. - An antibody which “induces apoptosis” is one which induces programmed cell death as determined by binding of annexin V, fragmentation of DNA, cell shrinkage, dilation of endoplasmic reticulum, cell fragmentation, and/or formation of membrane vesicles (called apoptotic bodies). The cell is one which expresses the antigen to which the antibody binds and may be one which overexpresses the antigen. The cell may be a tumor cell, e.g. a breast, ovarian, stomach, endometrial, salivary gland, lung, kidney, colon, thyroid, pancreatic or bladder cell. In vitro, the cell may be a SKBR3, BT474,
Calu 3 cell, MDA-MB-453, MDA-MB-361 or SKOV3 cell. Various methods are available for evaluating the cellular events associated with apoptosis. For example, phosphatidyl serine (PS) translocation can be measured by annexin binding; DNA fragmentation can be evaluated through DNA laddering as disclosed in the example herein; and nuclear/chromatin condensation along with DNA fragmentation can be evaluated by any increase in hypodiploid cells. Preferably, the antibody which induces apoptosis is one which results in about 2 to 50 fold, preferably about 5 to 50 fold, and most preferably about 10 to 50 fold, induction of annexin binding relative to untreated cell in an annexin binding assay using cells expressing the antigen to which the antibody binds. - Examples of antibodies which induce apoptosis include the anti-HER2 monoclonal antibodies 7F3 (ATCC HB-12216), and 7C2 (ATCC HB 12215), including humanized and/or affinity matured variants thereof; the anti-DR5 antibodies 3F11.39.7 (ATCC HB-12456); 3H3.14.5 (ATCC HB-12534); 3D5.1.10 (ATCC HB-12536); and 3H3.14.5 (ATCC HB-12534), including humanized and/or affinity matured variants thereof; the human anti-DR5 receptor antibodies 16E2 and 20E6, including affinity matured variants thereof (WO98/51793, expressly incorporated herein by reference); the anti-DR4 antibodies 4E7.24.3 (ATCC HB-12454); 4H6.17.8 (ATCC HB-12455); 1H5.25.9 (ATCC HB-12695); 4G7.18.8 (ATCC PTA-99); and 5G11.17.1 (ATCC HB-12694), including humanized and/or affinity matured variants thereof.
- In order to screen for antibodies which bind to an epitope on an antigen bound by an antibody of interest, a routine cross-blocking assay such as that described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed.
- An “agonist antibody” is an antibody which binds to and activates a receptor. Generally, the receptor activation capability of the agonist antibody will be at least qualitatively similar (and may be essentially quantitatively similar) to a native agonist ligand of the receptor. An example of an agonist antibody is one which binds to a receptor in the TNF receptor superfamily and induces apoptosis of cells expressing the TNF receptor. Assays for determining induction of apoptosis are described in WO98/51793 and WO99/37684, both of which are expressly incorporated herein by reference.
- A “disorder” is any condition that would benefit from treatment with the antibody. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question. Non-limiting examples of disorders to be treated herein include benign and malignant tumors; leukemias and lymphoid malignancies; neuronal, glial, astrocytal, hypothalamic and other glandular, macrophagal, epithelial, stromal and blastocoelic disorders; and inflammatory, angiogenic and immunologic disorders.
- The term “therapeutically effective amount” refers to an amount of a drug effective to treat a disease or disorder in a mammal. In the case of cancer, the therapeutically effective amount of the drug may reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the disorder. To the extent the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy in vivo can, for example, be measured by assessing the time to disease progression (TTP) and/or determining the response rates (RR).
- “Treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
- The terms “cancer” and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More particular examples of such cancers include squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma and various types of head and neck cancer.
- An “autoimmune disease” herein is a non-malignant disease or disorder arising from and directed against an individual's own tissues. Examples of autoimmune diseases or disorders include, but are not limited to, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g. atopic dermatitis); systemic scleroderma and sclerosis; responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g. Type I diabetes mellitus or insulin dependent diabetes mellitis); multiple sclerosis; Reynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjorgen's syndrome; juvenile onset diabetes; and immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis; pernicious anemia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder; multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemia or Coombs positive anemia); myasthenia gravis; antigen-antibody complex mediated diseases; anti-glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyendocrinopathies; Reiter's disease; stiff-man syndrome; Behcet disease; giant cell arteritis; immune complex nephritis; IgA nephropathy; IgM polyneuropathies; immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia etc.
- By “foreign antigen” is meant a molecule or molecules which is/are not endogenous or native to a mammal which is exposed to it. The foreign antigen may elicit an immune response, e.g. a humoral and/or T cell mediated response in the mammal. Generally, the foreign antigen will provoke the production of antibodies thereagainst. Examples of foreign antigens contemplated herein include immunogenic therapeutic agents, e.g. proteins such as antibodies, particularly antibodies comprising non-human amino acid residues (e.g. rodent, chimeric/humanized, and primatized antibodies); toxins (optionally conjugated to a targeting molecule such as an antibody, wherein the targeting molecule may also be immunogenic); gene therapy viral vectors, such as retroviruses and adenoviruses; grafts; infectious agents (e.g. bacteria and virus); alloantigens (i.e. an antigen that occurs in some, but not in other members of the same species) such as differences in blood types, human lymphocyte antigens (HLA), platelet antigens, antigens expressed on transplanted organs, blood components, pregnancy (Rh), and hemophilic factors (e.g. Factor VIII and Factor IX).
- By “blocking an immune response” to a foreign antigen is meant reducing or preventing at least one immune-mediated response resulting from exposure to a foreign antigen. For example, one may dampen a humoral response to the foreign antigen, i.e., by preventing or reducing the production of antibodies directed against the antigen in the mammal. Alternatively, or additionally, one may suppress idiotype; “pacify” the removal of cells coated with alloantibody; and/or affect alloantigen presentation through depletion of antigen-presenting cells.
- The term “graft” as used herein refers to biological material derived from a donor for transplantation into a recipient. Grafts include such diverse material as, for example, isolated cells such as islet cells; tissue such as the amniotic membrane of a newborn, bone marrow, hematopoietic precursor cells, and ocular tissue, such as corneal tissue; and organs such as skin, heart, liver, spleen, pancreas, thyroid lobe, lung, kidney, tubular organs (e.g., intestine, blood vessels, or esophagus), etc. The tubular organs can be used to replace damaged portions of esophagus, blood vessels, or bile duct. The skin grafts can be used not only for burns, but also as a dressing to damaged intestine or to close certain defects such as diaphragmatic hernia. The graft is derived from any mammalian source, including human, whether from cadavers or living donors. Preferably the graft is bone marrow or an organ such as heart and the donor of the graft and the host are matched for HLA class II antigens.
- The term “mammalian host” as used herein refers to any compatible transplant recipient. By “compatible” is meant a mammalian host that will accept the donated graft. Preferably, the host is human. If both the donor of the graft and the host are human, they are preferably matched for HLA class II antigens so as to improve histocompatibility.
- The term “donor” as used herein refers to the mammalian species, dead or alive, from which the graft is derived. Preferably, the donor is human. Human donors are preferably volunteer blood-related donors that are normal on physical examination and of the same major ABO blood group, because crossing major blood group barriers possibly prejudices survival of the allograft. It is, however, possible to transplant, for example, a kidney of a type O donor into an A, B or AB recipient.
- The term “transplant” and variations thereof refers to the insertion of a graft into a host, whether the transplantation is syngeneic (where the donor and recipient are genetically identical), allogeneic (where the donor and recipient are of different genetic origins but of the same species), or xenogeneic (where the donor and recipient are from different species). Thus, in a typical scenario, the host is human and the graft is an isograft, derived from a human of the same or different genetic origins. In another scenario, the graft is derived from a species different from that into which it is transplanted, such as a baboon heart transplanted into a human recipient host, and including animals from phylogenically widely separated species, for example, a pig heart valve, or animal beta islet cells or neuronal cells transplanted into a human host.
- The expression “desensitizing a mammal awaiting transplantation” refers to reducing or abolishing allergic sensitivity or reactivity to a transplant, prior to administration of the transplant to the mammal. This may be achieved by any mechanism, such as a reduction in anti-donor antibodies in the desensitized mammal, e.g. where such anti-donor antibodies are directed against human lymphocyte antigen (HLA).
- The term “cytotoxic agent” as used herein refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes (e.g. At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32 and radioactive isotopes of Lu), chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof.
- A “chemotherapeutic agent” is a chemical compound useful in the treatment of cancer. Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN™); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CBI-TMI); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as the enediyne antibiotics (e.g. calicheamicin, especially calicheamicin γ1 I and calicheamicin θI 1, see, e.g., Agnew Chem. Intl. Ed. Engl. 33:183-186 (1994); dynemicin, including dynemicin A; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromomophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone; 2,2′,2″-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g. paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) and doxetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above.
- The term “cytokine” is a generic term for proteins released by one cell population which act on another cell as intercellular mediators. Examples of such cytokines are lymphokines, monokines, and traditional polypeptide hormones. Included among the cytokines are growth hormone such as human growth hormone, N-methionyl human growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin; proinsulin; relaxin; prorelaxin; glycoprotein hormones such as follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH), and luteinizing hormone (LH); hepatic growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor-alpha and -beta; mullerian-inhibiting substance; mouse gonadotropin-associated peptide; inhibin; activin; vascular endothelial growth factor; integrin; thrombopoietin (TPO); nerve growth factors such as NGF-alpha; platelet-growth factor; transforming growth factors (TGFs) such as TGF-alpha and TGF-beta; insulin-like growth factor-I and -II; erythropoietin (EPO); osteoinductive factors; interferons such as interferon-alpha, -beta and -gamma colony stimulating factors (CSFs) such as macrophage-CSF (M-CSF); granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (G-CSF); interleukins (ILs) such as IL-1, IL-1alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12; a tumor necrosis factor such as TNF-alpha or TNF-beta; and other polypeptide factors including LIF and kit ligand (KL). As used herein, the term cytokine includes proteins from natural sources or from recombinant cell culture and biologically active equivalents of the native sequence cytokines.
- The term “prodrug” as used in this application refers to a precursor or derivative form of a pharmaceutically active substance that is less cytotoxic to tumor cells compared to the parent drug and is capable of being enzymatically activated or converted into the more active parent form. See, e.g., Wilman, “Prodrugs in Cancer Chemotherapy” Biochemical Society Transactions, 14, pp. 375-382, 615th Meeting Belfast (1986) and Stella et al., “Prodrugs: A Chemical Approach to Targeted Drug Delivery,” Directed Drug Delivery, Borchardt et al., (ed.), pp. 247-267, Humana Press (1985). The prodrugs of this invention include, but are not limited to, phosphate-containing prodrugs, thiophosphate-containing prodrugs, sulfate-containing prodrugs, peptide-containing prodrugs, D-amino acid-modified prodrugs, glycosylated prodrugs, beta-lactam-containing prodrugs, optionally substituted phenoxyacetamide-containing prodrugs or optionally substituted phenylacetamide-containing prodrugs, 5-fluorocytosine and other 5-fluorouridine prodrugs which can be converted into the more active cytotoxic free drug. Examples of cytotoxic drugs that can be derivatized into a prodrug form for use in this invention include, but are not limited to, those chemotherapeutic agents described above.
- An “angiogenic factor” is a growth factor which stimulates the development of blood vessels. The preferred angiogenic factor herein is Vascular Endothelial Growth Factor (VEGF).
- The word “label” when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the polypeptide. The label may be itself be detectable (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.
- An “isolated” nucleic acid molecule is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide nucleic acid. An isolated nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated nucleic acid molecules therefore are distinguished from the nucleic acid molecule as it exists in natural cells. However, an isolated nucleic acid molecule includes a nucleic acid molecule contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells.
- The expression “control sequences” refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. The control sequences that are suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, and a ribosome binding site. Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.
- Nucleic acid is “operably linked” when it is placed into a functional relationship with another nucleic acid sequence. For example, DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, “operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
- As used herein, the expressions “cell,” “cell line,” and “cell culture” are used interchangeably and all such designations include progeny. Thus, the words “transformants” and “transformed cells” include the primary subject cell and cultures derived therefrom without regard for the number of transfers. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Mutant progeny that have the same function or biological activity as screened for in the originally transformed cell are included. Where distinct designations are intended, it will be clear from the context.
- A. Multivalent Antibodies
- The invention herein relates to a method for making a multivalent antibody. Various techniques for generating the “parent” or “starting” antibody from which the variable domain(s) of the multivalent antibody may be derived will be described later in this application.
- The multivalent antibody of particular interest herein is one which comprises at least three (and preferably four, or more, e.g. four or five to about eight) antigen binding sites. Generally, all of the antigen binding sites are “functional” as defined hereinabove. Preferably, the multivalent antibody does not exist in nature and is not a native sequence IgM or IgA antibody. The multivalent antibody herein is preferably not produced in vitro by chemically cross-linking a pair antibodies (e.g. as in Ghetie et al. (1997), supra or Wolff et al. (1993), supra). The present application also provides multivalent antibodies which do not require introduced cysteine residue(s) in a parent antibody in order to make the multivalent antibody via disulfide bond(s) between a pair of Fc regions (e.g. as in Shopes et al. (1992), supra or Caron et al. (1992), supra).
- In one embodiment, the multivalent antibody comprises a first polypeptide chain comprising at least two heavy chain (or light chain) variable domains and a second polypeptide chain comprising at least two heavy chain (or light chain) variable domains. Preferably, the first polypeptide chain comprises two heavy chain variable domains and the second polypeptide chain also comprises two heavy chain variable domains, which can be combined with corresponding light chain variable domains (at least two for each polypeptide chain) to generate four (or more) antigen binding sites.
- In one preferred embodiment of the invention, the multivalent antibody comprises a dimerization domain which combines (1) two (or more) antigen binding sites with (2) one, two (or more) antigen binding sites. Various dimerization domains are contemplated herein, but the preferred dimerization domain is an Fc region or a hinge region. Where the multivalent antibody comprises an Fc region (e.g. a native sequence or variant Fc region), the Fc region is preferably “functional” as defined hereinabove and thus is capable of performing one or more antibody effector functions, such as ADCC or CDC. Preferably, the multivalent antibody has only one Fc region or lacks an Fc region.
- Where the multivalent antibody comprises an Fc region, preferably, the three or more antigen binding sites are provided amino terminal to the Fc region (rather than at the carboxy terminus of the Fc region as in Coloma and Morrison, (1997) supra). This may be achieved by providing a first polypeptide chain represented by the formula VD1-X1-VD2-X2-Fc, wherein (1) VD1 is a first heavy or light chain variable domain (preferably a heavy chain variable domain), (2) VD2 is a second heavy or light chain variable domain (preferably a heavy chain variable domain), (3) Fc comprises one chain of an Fc region, and (4) X1 and X2 represent an optional intervening amino acid or polypeptide. Preferably X1 and X2 comprise, or consist of, a CH1 domain (where VD1 or VD2 is a heavy chain variable domain) or a CL domain (where VD1 or VD2 is a light chain variable domain). Optionally, X1 further comprises a flexible linker which is generally C-terminal to VD1 (or C-terminal to CH1 or CL, if present). The flexible linker may comprise a peptide such as gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser or gly-gly-gly-ser (SEQ ID NO:11).
- The multivalent antibody of particular interest herein comprises three or more (e.g. four or five to about eight) Fab polypeptides, each capable of binding antigen. The Fab fragments are preferably provided amino terminal to the Fc region (where the multivalent antibody has an Fc region). For instance, two or more Fd fragments may be fused to the amino terminus of one chain of an Fc region. The polypeptide chain thus engineered may be combined with (1) another polypeptide chain formed by two or more Fd fragments fused to the amino terminus of the other chain of the Fc region, as well as (2) complementary VL domains (e.g. four or more VL domains which each, optionally, are fused to a CL domain). Optionally, the antibody comprises a flexible linker between the two or more Fd fragments. The multivalent antibody may, for example, comprise a pair of polypeptide chains with the formula (1) VH-CH1-flexible linker-VH-CH1-Fc chain, or (2) VH-CH1-VH-CH1-Fc chain (i.e. where there is no flexible linker between the two Fd fragments).
- The three or more functional antigen binding sites of the multivalent antibody herein are each preferably formed by a heavy and light chain variable domain. Thus, where two or more heavy chain variable domains are fused together (optionally with intervening amino acid residue(s) as noted above), two or more complementary light chain variable domain-containing polypeptides are combined with the heavy chain variable domains (for instance by co-expressing the fusion protein and the light chain variable domain polypeptide(s) in the same host cell). Preferably, the antibody comprises four, or five, or more (e.g. up to about eight) light chain variable domain polypeptides, which each, optionally, comprise a CL domain.
- In one embodiment herein, the antibody with three or more more (e.g. three to about ten, but preferably three or four) antigen binding sites may comprise a polypeptide chain comprising three or more (e.g. three to about ten, but preferably three or four) heavy chain or light chain variable domains, wherein each of the variable domains is combined with, or associated with, three or more (e.g. three to about ten, but preferably three or four) light chain or heavy chain variable domain polypeptides in such a way as to form the antigen binding sites. Thus, where the polypeptide chain comprises three or more heavy chain variable domains, it is combined or associated with three or more corresponding light chain variable domain polypeptides (e.g. with VL-CL polypeptides). Alternatively, where the polypeptide chain comprises three or more light chain variable domains, it is combined or associated with three or more corresponding heavy chain variable domain polypeptides (e.g. with VH-CH1 polypeptides). Preferably each of the three or more antigen binding sites is directed against the same antigen. Examples of antigens bound by such antibodies include (1) a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily (such receptors may be ‘trimeric receptors’, hence the antibody need only include only three antigen binding sites as desired) such as DR4 and DRS; (2) a B cell surface antigen such as CD20; (3) an ErbB receptor exemplified by the HER2 receptor; or (4) a cell surface protein expressed by tumor cells. For instance, the polypeptide chain may comprise three (or four) heavy chain variable domains which are able to combine with three (or four) light chain variable domain polypeptides (preferably VL-CL polypeptides) to generate three (or four) antigen binding sites directed against the same antigen. Such antibodies are exemplified by those depicted in
FIG. 23D (with three antigen binding sites) andFIG. 23E (with four antigen binding sites). The multivalent antibody may also comprise a polypeptide chain comprising the formula: (a) VL-CL-flexible linker-VL-CL-flexible linker-VL-CL; In this embodiment, the polypeptide may comprise three to about eight VL-CL polypeptides joined by flexible linkers. (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; In this embodiment, the polypeptide may comprise three to about eight VH-CH1 polypeptides joined by flexible linkers. (c) (VL-CL)n, wherein n is three or more more (e.g. three to about eight, but preferably three or four); or (d) (VH-CH1)n, wherein n is three or more more (e.g. three to about eight, but preferably three or four). Preferably, the polypeptide chain comprises the formula: (a) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; or (c) (VH-CH1)n, wherein n is three or four. - The multivalent antibodies herein have desirable properties particularly for in vivo therapy and diagnosis. For instance, the multivalent antibody may be internalized and catabolized by a cell expressing an antigen, to which the antibody binds, faster than a bivalent antibody. Thus, the invention provides an immunoconjugate comprising the multivalent antibody conjugated with a cytotoxic agent (e.g. one which is active in killing cells once internalized). Various cytotoxic agents for generating an immunoconjugate are described herein, but the preferred cytotoxic agent is a radioactive isotope, a maytansinoid or a calecheamicin.
- The multivalent antibody, and/or a parent antibody from which at least one of the multivalent antibody's antigen binding specificities is derived, may have certain properties. For instance, the multivalent antibody and/or parent antibody may (1) be an agonist antibody (e.g. where an antigen bound by the antibody is a receptor in the TNF receptor family or a B cell surface antigen); (2) induce apoptosis (for instance, where an antigen bound by the antibody is an ErbB receptor or a receptor in the TNF receptor superfamily); (3) bind a cell surface protein (such as a B cell surface antigen or an ErbB receptor) expressed on tumor cells; (4) bind a cell surface protein (e.g. Epidermal Growth Factor Receptor (EGFR), HER2 receptor, ErbB3 receptor, ErbB4 receptor, or DcR3 receptor) overexpressed by tumor cells; and/or (5) be a growth inhibitory antibody .
- The multivalent antibody herein may have specificity for only one antigen, or more than one antigens (e.g. from two to about three antigens). In one embodiment, the three or more functional antigen binding sites of the multivalent antibody may all bind the same antigen (preferably the same epitope on that antigen, in which case the multivalent antibody would be considered to be “monospecific”). This application also provides “multispecific” antibodies. Thus, the three or more functional antigen binding sites may bind two or more (e.g. from two to about three) different antigens or epitopes.
- The present application shows that a multivalent antibody directed against a receptor antigen can be engineered which, surprisingly, has agonistic and/or apoptosis-inducing capability which is quantitatively similar to that of the native ligand. By “quantitatively similar” here is meant that the multivalent antibody has an activity in an assay which determines agonistic and/or apoptosis-inducing activity, within about ten fold, and preferably within about five fold of the agonistic and/or apoptosis-inducing activity of the native ligand. In this embodiment, the antibody with agonistic and/or apoptosis-inducing activity may be one with specificity for a receptor in the TNF receptor superfamily, e.g. an Apo2L receptor such as DR4, DR5, DcR1 and DcR2 (preferably DR4 or DR5), in which case the activity of the antibody in an apoptosis assay such as those described in Example 3 below is within about ten fold, e.g. within about five fold, of the activity of Apo2L in that assay.
- The multivalent antibody herein may, in one embodiment of the invention, bind a B cell surface antigen. Preferred B cell surface antigens include CD19, CD20, CD22 and CD40, and most preferably CD20.
- Various applications for the multivalent antibodies herein are contemplated and described in more detail below. Where the multivalent antibody possesses one or more functional Fc regions, it is anticipated to have the ability to mediate effector functions (such as ADCC and CDC) and have a longer half-life than multivalent antibodies lacking an Fc region. Such multivalent antibodies may be used where killing of cells, such as tumor or cancer cells, is desired. Other forms of the multivalent antibodies herein which lack a Fc region may be desirable where a shorter half-life is desired (e.g. for treating cardiovascular or inflammatory diseases or disorders, or where the antibody is conjugated with a cytotoxic agent); where internalization of the antibody is desired (e.g. for therapy with an immunoconjugate comprising the antibody and a cytotoxic agent); for improved penetration of a solid tumor; where expression of the multivalent antibody in a non-mammalian host cell (e.g. a prokaryotic host cell such as an E. coli host cell) is desired; for therapy of nononcological diseases or disorders; and/or to avoid the ‘first dose affect’ observed upon administration of certain antibodies possessing effector function(s) to patients. Such forms of the antibody may comprise a multivalent antibody including a dimerization domain, wherein the dimerization domain comprises an antibody hinge region fused to a leucine zipper domain (the leucine zipper domain facilitates association of the polypeptides which form the dimerization domain, but may be subsequently proteolytically removed prior to administration to a patient) (see
FIG. 23C ); a multivalent antibody with three antigen binding sites such as that shown inFIG. 23D ; or a multivalent antibody with four antigen binding sites such as that depicted inFIG. 23E . - B. Antigen Binding Specificity
- The multivalent antibody herein is directed against, or binds specifically to, one or more target antigen(s). Preferably, at least one of the antigens bound by the multivalent antibody is a biologically important polypeptide and administration of the antibody to a mammal suffering from a disease or disorder can result in a therapeutic benefit in that mammal. However, antibodies directed against nonpolypeptide antigens (such as tumor-associated glycolipid antigens; see U.S. Pat. No. 5,091,178) are also contemplated.
- Where the antigen is a polypeptide, it may be a transmembrane molecule (e.g. receptor) or ligand such as a growth factor. Exemplary antigens include molecules such as renin; a growth hormone, including human growth hormone and bovine growth hormone; growth hormone releasing factor; parathyroid hormone; thyroid stimulating hormone; lipoproteins; alpha-1-antitrypsin; insulin A-chain; insulin B-chain; proinsulin; follicle stimulating hormone; calcitonin; luteinizing hormone; glucagon; clotting factors such as factor VIIIC, factor IX, tissue factor (TF), and von Willebrands factor; anti-clotting factors such as Protein C; atrial natriuretic factor; lung surfactant; a plasminogen activator, such as urokinase or human urine or tissue-type plasminogen activator (t-PA); bombesin; thrombin; hemopoietic growth factor; tumor necrosis factor-alpha and -beta; enkephalinase; RANTES (regulated on activation normally T-cell expressed and secreted); human macrophage inflammatory protein (MIP-1-alpha); a serum albumin such as human serum albumin; Muellerian-inhibiting substance; relaxin A-chain; relaxin B-chain; prorelaxin; mouse gonadotropin-associated peptide; a microbial protein, such as beta-lactamase; DNase; IgE; a cytotoxic T-lymphocyte associated antigen (CTLA), such as CTLA-4; inhibin; activin; vascular endothelial growth factor (VEGF); receptors for hormones or growth factors; protein A or D; rheumatoid factors; a neurotrophic factor such as bone-derived neurotrophic factor (BDNF), neurotrophin-3, -4, -5, or -6 (NT-3, NT-4, NT-5, or NT-6), or a nerve growth factor such as NGF-β; platelet-derived growth factor (PDGF); fibroblast growth factor such as aFGF and bFGF; epidermal growth factor (EGF); transforming growth factor (TGF) such as TGF-alpha and TGF-beta, including TGF-β1, TGF-β2, TGF-β3, TGF-β4, or TGF-β5; insulin-like growth factor-1 and -II (IGF-1 and IGF-II); des(1-3)-IGF-I (brain IGF-I), insulin-like growth factor binding proteins; CD proteins such as CD3, CD4, CD8, CD19, CD20 and CD25 (Tac subunit of the IL-2 receptor); erythropoietin; osteoinductive factors; immunotoxins; a bone morphogenetic protein (BMP); an interferon such as interferon-alpha, -beta, and -gamma; colony stimulating factors (CSFs), e.g., M-CSF, GM-CSF, and G-CSF; interleukins (ILs), e.g., IL-1 to IL-10; superoxide dismutase; T-cell receptors; surface membrane proteins; decay accelerating factor; viral antigen such as, for example, a portion of the AIDS envelope; transport proteins; homing receptors; addressins; regulatory proteins; integrins such as CD11a, CD11b, CD11c, CD18, an ICAM, VLA-4 or VCAM; a tumor associated antigen such as HER2, HER3 or HER4 receptor; and fragments of any of the above-listed polypeptides.
- Preferred molecular targets for antibodies encompassed by the present invention include leukocyte surface markers or CD proteins such as CD1a-c, CD2, CD2R, CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CDw12, CD13, CD14, CD15, CD15s, CD16, CD16b, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, C41, CD42a-d, CD43, CD44, CD44R, CD45, CD45A, CD45B, CD450, CD46-CD48, CD49a-f, CD50, CD51, CD52, CD53-CD59, CDw60, CD61, CD62E, CD62L, CD62P, CD63, CD64, CDw65, CD66a-e, CD68-CD74, CDw75, CDw76, CD77, CDw78, CD79a-b, CD80-CD83, CDw84, CD85-CD89, CDw90, CD91, CDw92, CD93-CD98, CD99, CD99R, CD100, CDw101, CD102-CD106, CD107a-b, CDw108, CDw109, CD115, CDw116, CD117, CD119, CD120a-b, CD121a-b, CD122, CDw124, CD126-CD129, and CD130; members of the ErbB receptor family such as the EGF receptor, HER2 receptor, HER3 receptor or HER4 receptor; prostate specific antigen(s); cell adhesion molecules such as IIb/IIIa, LFA-1, Mac1, p150.95, VLA-4, ICAM-1, VCAM, α4/β7 integrin, and αv/β3 integrin including either α or β subunits thereof (e.g. anti-CD11a, anti-CD18 or anti-CD11b antibodies); growth factors such as VEGF; tissue factor (TF); alpha interferon (α-IFN); an interleukin, such as IL-8; IgE; blood group antigens; flk2/flt3 receptor; obesity (OB) receptor; c-mpl receptor; CTLA-4; protein C etc.
- Soluble antigens or fragments thereof, optionally conjugated to other molecules, can be used as immunogens for generating antibodies. For transmembrane molecules, such as receptors, fragments of these (e.g. the extracellular domain of a receptor) can be used as the immunogen. Alternatively, cells expressing the transmembrane molecule can be used as the immunogen. Such cells can be derived from a natural source (e.g. cancer cell lines) or may be cells which have been transformed by recombinant techniques to express the transmembrane molecule. Other antigens and forms thereof useful for preparing antibodies will be apparent to those in the art.
- Preferred target antigens for the multivalent antibodies herein include (1) ErbB receptors, including EGFR, HER2, HER3 and HER4; (2) receptors in the TNF receptor superfamily, e.g. Apo2L receptors, such as DR4, DR5, DcR1 and DcR2; (3) B cell surface antigens, especially CD19, CD20, CD22 and CD40; (4) antigens expressed by tumor cells; (5) antigens overexpressed by tumor cells (e.g. ErbB receptors; DcR3 receptors); (6) receptors activated by multimeric (e.g. dimeric or trimeric) ligands (e.g. receptors in the TNF receptor superfamily; VEGF receptors, etc.). In one embodiment, three or more (e.g. four to about eight) of the antigen binding sites of the multivalent antibody may all be directed against the same antigenic determinant or epitope on one of the above antigens.
- The present application also provides multispecific antibodies, i.e., antibodies that have binding specificities for at least two different epitopes or antigenic determinants. Multispecific antibodies (e.g. bispecific antibodies; BsAbs) have significant potential in a wide range of clinical applications as targeting agents for in vitro and in vivo immunodiagnosis and therapy, and for diagnostic immunoassays.
- Bispecific antibodies have been very useful in probing the functional properties of cell surface molecules and in defining the ability of the different Fc receptors to mediate cytotoxicity (Fanger et al., Crit. Rev. Immunol. 12:101-124 (1992)). Nolan et al., Biochem. Biophys. Acta. 1040:1-11 (1990) describe other diagnostic applications for BsAbs. In particular, BsAbs can be constructed to immobilize enzymes for use in enzyme immunoassays. To achieve this, one arm of the BsAb can be designed to bind to a specific epitope on the enzyme so that binding does not cause enzyme inhibition, the other arm of the BsAb binds to the immobilizing matrix ensuring a high enzyme density at the desired site. Examples of such diagnostic BsAbs include the rabbit anti-IgG/anti-ferritin BsAb described by Hammerling et al., J. Exp. Med. 128:1461-1473 (1968) which was used to locate surface antigens. BsAbs having binding specificities for Horse Radish Peroxidase (HRP) as well as a hormone have also been developed. Another potential immunochemical application for BsAbs involves their use in two-site immunoassays. For example, two BsAbs are produced binding to two separate epitopes on the analyte protein—one BsAb binds the complex to an insoluble matrix, the other binds an indicator enzyme (see Nolan et al., supra).
- Multispecific antibodies can also be used for in vitro or in vivo immunodiagnosis of various diseases such as cancer (Songsivilai et al., Clin. Exp. Immunol. 79:315 (1990)). To facilitate this diagnostic use of the BsAb, one arm of the BsAb can bind a tumor associated antigen and the other arm can bind a detectable marker such as a chelator which tightly binds a radionuclide. Using this approach, Le Doussal et al. made a BsAb useful for radioimmunodetection of colorectal and thryoid carcinomas which had one arm which bound a carcinoembryonic antigen (CEA) and another arm which bound diethylenetriaminepentacetic acid (DPTA). See Le Doussal et al., Int. J. Cancer Suppl. 7:58-62 (1992) and Le Doussal et al., J. Nucl. Med. 34:1662-1671 (1993). Stickney et al. similarly describe a strategy for detecting colorectal cancers expressing CEA using radioimmunodetection. These investigators describe a BsAb which binds CEA as well as hydroxyethylthiourea-benzyl-EDTA (EOTUBE). See Stickney et al., Cancer Res. 51:6650-6655 (1991).
- Multispecific antibodies can also be used for human therapy in redirected cytotoxicity by providing one arm which binds a target (e.g. pathogen or tumor cell) and another arm which binds a cytotoxic trigger molecule, such as the T-cell receptor or an Fc gamma receptor. Accordingly, multispecific antibodies can be used to direct a patient's cellular immune defense mechanisms specifically to the tumor cell or infectious agent. Using this strategy, it has been demonstrated that bispecific antibodies which bind to the Fc gamma RIII (i.e. CD16) can mediate tumor cell killing by natural killer (NK) cell/large granular lymphocyte (LGL) cells in vitro and are effective in preventing tumor growth in vivo. Segal et al., Chem. Immunol. 47:179 (1989) and Segal et al., Biologic Therapy of Cancer 2(4) DeVita et al. eds. J. B. Lippincott, Philadelphia (1992) p. 1. Similarly, a bispecific antibody having one arm which binds Fc gamma RIII and another which binds to the HER2 receptor has been developed for therapy of ovarian and breast tumors that overexpress the HER2 antigen. (Hseih-Ma et al. Cancer Research 52:6832-6839 (1992) and Weiner et al. Cancer Research 53:94-100 (1993)). Bispecific antibodies can also mediate killing by T cells. Normally, the bispecific antibodies link the CD3 complex on T cells to a tumor-associated antigen. A fully humanized F(ab′)2 BsAb consisting of anti-CD3 linked to anti-p185HER2 has been used to target T cells to kill tumor cells overexpressing the HER2 receptor. Shalaby et al., J. Exp. Med. 175(1):217 (1992). Bispecific antibodies have been tested in several early phase clinical trials with encouraging results. In one trial, 12 patients with lung, ovarian or breast cancer were treated with infusions of activated T-lymphocytes targeted with an anti-CD3/anti-tumor (MOC31) bispecific antibody. deLeij et al. Bispecific Antibodies and Targeted Cellular Cytotoxicity, Romet-Lemonne, Fanger and Segal Eds., Lienhart (1991) p. 249. The targeted cells induced considerable local lysis of tumor cells, a mild inflammatory reaction, but no toxic side effects or anti-mouse antibody responses. In a very preliminary trial of an anti-CD3/anti-CD19 bispecific antibody in a patient with B cell malignancy, significant reduction in peripheral tumor cell counts was also achieved. Clark et al. Bispecific Antibodies and Targeted Cellular Cytotoxicity, Romet-Lemonne, Fanger and Segal Eds., Lienhart (1991) p. 243. See also Kroesen et al., Cancer Immunol. Immunother. 37:400-407 (1993), Kroesen et al., Br. J. Cancer 70:652-661 (1994) and Weiner et al., J. Immunol. 152:2385 (1994) concerning therapeutic applications for multispecific antibodies.
- Multispecific antibodies may also be used as fibrinolytic agents or vaccine adjuvants. Furthermore, these antibodies may be used in the treatment of infectious diseases (e.g. for targeting of effector cells to virally infected cells such as HIV or influenza virus or protozoa such as Toxoplasma gondii), used to deliver immunotoxins to tumor cells, or target immune complexes to cell surface receptors (see Fanger et al., supra).
- Various multispecific antibodies are contemplated herein. For instance, the multispecific antibody may bind two or more different epitopes on an antigen of interest. Alternatively, the multispecfic antibody may have specificity for (1) an antigen expressed by a target cell (e.g. where the target cell is a tumor cell) and (2) a triggering molecule on a leukocyte, such as a T-cell receptor molecule (e.g. CD2 or CD3), or Fc receptors for IgG (Fc gamma R), such as Fc gamma RI (CD64), Fc gamma RII (CD32) and Fc gamma RIII (CD16) so as to focus cellular defense mechanisms to the antigen-expressing cell. Multispecific antibodies may also be used to localize cytotoxic agents to cells which express the target antigen. These antibodies possess an target antigen-binding arm and an arm which binds the cytotoxic agent (e.g. saporin, interferon-alpha, vinca alkaloid, ricin A chain, methotrexate or radioactive isotope hapten).
- WO 96/16673 describes a bispecific anti-HER2/anti-Fc gamma RIII antibody and U.S. Pat. No. 5,837,234 discloses a bispecific anti-HER2/anti-Fc gamma RI antibody. A bispecific anti-HER2/Fc alpha antibody is shown in WO98/02463. U.S. Pat. No. 5,821,337 teaches a bispecific anti-HER2/anti-CD3 antibody.
- C. Preparation of the Parent Antibody
- In order to generate the multivalent antibody, a “parent” or “starting” antibody with variable domains directed against an antigen may be prepared using various methodologies for making antibodies, such as those described hereinbelow. The sequences of the variable domains of the starting or parent antibody may be used in the design of the multivalent antibody herein.
- (i) Polyclonal Antibodies
- Polyclonal antibodies are preferably raised in animals by multiple subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N-hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOCl2, or R1N═C═NR, where R and R1 are different alkyl groups.
- Animals are immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 μg or 5 μg of the protein or conjugate (for rabbits or mice, respectively) with 3 volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites. One month later the animals are boosted with ⅕ to 1/10 the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites. Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus. Preferably, the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent. Conjugates also can be made in recombinant cell culture as protein fusions. Also, aggregating agents such as alum are suitably used to enhance the immune response.
- (ii) Monoclonal Antibodies
- Monoclonal antibodies are obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Thus, the modifier “monoclonal” indicates the character of the antibody as not being a mixture of discrete antibodies.
- For example, the monoclonal antibodies may be made using the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by recombinant DNA methods (U.S. Pat. No. 4,816,567).
- In the hybridoma method, a mouse or other appropriate host animal, such as a hamster, is immunized as hereinabove described to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)).
- The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
- Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these, preferred myeloma cell lines are murine myeloma lines, such as those derived from MOPC-21 and MPC-11 mouse tumors available from the Salk Institute Cell Distribution Center, San Diego, Calif. USA, and SP-2 or X63-Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); and Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
- Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
- The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980).
- After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and/or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal.
- The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional antibody purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
- DNA encoding the monoclonal antibodies is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells serve as a preferred source of such DNA. Once isolated, the DNA may be placed into expression vectors, which are then transfected into host cells such as E. coli cells, simian COS cells, Chinese Hamster Ovary (CHO) cells, or myeloma cells that do not otherwise produce antibody protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. Review articles on recombinant expression in bacteria of DNA encoding the antibody include Skerra et al., Curr. Opinion in Immunol., 5:256-262 (1993) and Plückthun, Immunol. Revs., 130:151-188 (1992). Recombinant expression of antibodies is described in more detail below.
- In a further embodiment, monoclonal antibodies or antibody fragments can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al., Nature, 348:552-554 (1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biot., 222:581-597 (1991) describe the isolation of murine and human antibodies, respectively, using phage libraries. Subsequent publications describe the production of high affinity (nM range) human antibodies by chain shuffling (Marks et al., Bio/Technology, 10:779-783 (1992)), as well as combinatorial infection and in vivo recombination as a strategy for constructing very large phage libraries (Waterhouse et al., Nuc. Acids. Res., 21:2265-2266 (1993)). Thus, these techniques are viable alternatives to traditional monoclonal antibody hybridoma techniques for isolation of monoclonal antibodies.
- The DNA also may be modified, for example, by substituting the coding sequence for human heavy chain and light chain constant domains in place of the homologous murine sequences (U.S. Pat. No. 4,816,567; and Morrison, et al., Proc. Natl. Acad. Sci. USA, 81:6851 (1984)), or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
- Typically such non-immunoglobulin polypeptides are substituted for the constant domains of an antibody, or they are substituted for the variable domains of one antigen-combining site of an antibody to create a chimeric bivalent antibody comprising one antigen-combining site having specificity for an antigen and another antigen-combining site having specificity for a different antigen.
- (iii) Human Antibodies
- Human monoclonal antibodies may be made via an adaptation of the hybridoma method first described by Kohler and Milstein by using human B lymphocytes as the fusion partner. Human B lymphocytes producing an antibody of interest may, for example, be isolated from a human individual, after obtaining informed consent. For instance, the individual may be producing antibodies against an autoantigen as occurs with certain disorders such as systemic lupus erythematosus (Shoenfeld et al. J. Clin. Invest. 70:205 (1982)), immune-mediated thrombocytopenic purpura (ITP) (Nugent et al. Blood 70(1):16-22 (1987)), or cancer. Alternatively, or additionally, lymphocytes may be immunized in vitro. For instance, one may expose isolated human periperal blood lymphocytes in vitro to a lysomotrophic agent (e.g. L-leucine-O-methyl ester, L-glutamic acid dimethly ester or L-leucyl-L-leucine-O-methyl ester) (U.S. Pat. No. 5,567,610, Borrebaeck et al.); and/or T-cell depleted human peripheral blood lymphocytes may be treated in vitro with adjuvants such as 8-mercaptoguanosine and cytokines (U.S. Pat. No. 5,229,275, Goroff et al.).
- The B lymphocytes recovered from the subject or immunized in vitro, are then generally immortalized in order to generate a human monoclonal antibody. Techniques for immortalizing the B lymphocyte include, but are not limited to: (a) fusion of the human B lymphocyte with human, murine myelomas or mouse-human heteromyeloma cells; (b) viral transformation (e.g. with an Epstein-Barr virus; see Nugent et al., supra, for example); (c) fusion with a lymphoblastoid cell line; or (d) fusion with lymphoma cells.
- Lymphocytes may be fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells. Suitable human myeloma and mouse-human heteromyeloma cell lines have been described (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)). Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
- After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and/or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A chromatography, gel electrophoresis, dialysis, or affinity chromatography.
- Human antibodies may also be generated using a non-human host, such as a mouse, which is capable of producing human antibodies. As noted above, transgenic mice are now available that are capable, upon immunization, of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production. For example, it has been described that the homozygous deletion of the antibody heavy-chain joining region (JH) gene in chimeric and germ-line mutant mice results in complete inhibition of endogenous antibody production. Transfer of the human germ-line immunoglobulin gene array in such germ-line mutant mice will result in the production of human antibodies upon antigen challenge. See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al., Year in Immuno., 7:33 (1993); U.S. Pat. No. 5,591,669; U.S. Pat. No. 5,589,369; and U.S. Pat. No. 5,545,807. Human antibodies may also be prepared using SCID-hu mice (Duchosal et al. Nature 355:258-262 (1992)).
- In another embodiment, the human antibody may be selected from a human antibody phage display library. The preparation of libraries of antibodies or fragments thereof is well known in the art and any of the known methods may be used to construct a family of transformation vectors which may be introduced into host cells. Libraries of antibody light and heavy chains in phage (Huse et al., Science, 246:1275 (1989)) or of fusion proteins in phage or phagemid can be prepared according to known procedures. See, for example, Vaughan et al., Nature Biotechnology 14:309-314 (1996); Barbas et al., Proc. Natl. Acad. Sci., USA, 88:7978-7982 (1991); Marks et al., J. Mol. Biol., 222:581-597 (1991); Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992); Barbas et al., Proc. Natl. Acad. Sci., USA, 89:4457-4461 (1992); Griffiths et al., EMBO Journal, 13:3245-3260 (1994); de Kruif et al., J. Mol. Biol., 248:97-105 (1995); WO 98/05344; WO 98/15833; WO 97/47314; WO 97/44491; WO 97/35196; WO 95/34648; U.S. Pat. No. 5,712,089; U.S. Pat. No. 5,702,892; U.S. Pat. No. 5,427,908; U.S. Pat. No. 5,403,484; U.S. Pat. No. 5,432,018; U.S. Pat. No. 5,270,170; WO 92/06176; WO 99/06587; U.S. Pat. No. 5,514,548; WO97/08320; and U.S. Pat. No. 5,702,892. The antigen of interest is panned against the phage library using procedures known in the field for selecting phage-antibodies which bind to the target antigen
- (iv) Humanized Antibodies
- Methods for humanizing non-human antibodies have been described in the art. Preferably, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as “import” residues, which are typically taken from an “import” variable domain. Humanization can be essentially performed following the method of Winter and co-workers (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting hypervariable region sequences for the corresponding sequences of a human antibody. Accordingly, such “humanized” antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567) wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some hypervariable region residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
- The choice of human variable domains, both light and heavy, to be used in making the humanized antibodies is very important to reduce antigenicity. According to the so-called “best-fit” method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable-domain sequences. The human sequence which is closest to that of the rodent is then accepted as the human framework for the humanized antibody (Sims et al., J. Immunol., 151:2296 (1993); Chothia et al., J. Mol. Biol., 196:901 (1987)). Another method uses a particular framework derived from the consensus sequence of all human antibodies of a particular subgroup of light or heavy chains. The same framework may be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623 (1993)).
- It is further important that antibodies be humanized with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, according to a preferred method, humanized antibodies are prepared by a process of analysis of the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate immunoglobulin sequence, L e., the analysis of residues that influence the ability of the candidate immunoglobulin to bind its antigen. In this way, FR residues can be selected and combined from the recipient and import sequences so that the desired antibody characteristic, such as increased affinity for the target antigen(s), is achieved. In general, the hypervariable region residues are directly and most substantially involved in influencing antigen binding.
- (v) Antibody Fragments
- Various techniques have been developed for the production of antibody fragments. Traditionally, these fragments were derived via proteolytic digestion of intact antibodies (see, e.g., Morimoto et al., Journal of Biochemical and Biophysical Methods 24:107-117 (1992); and Brennan et al., Science, 229:81 (1985)). However, these fragments can now be produced directly by recombinant host cells. For example, the antibody fragments can be isolated from the antibody phage libraries discussed above. Alternatively, Fab′-SH fragments can be directly recovered from E. coli and chemically coupled to form F(ab′)2 fragments (Carter et al., Bio/Technology 10:163-167 (1992)). According to another approach, F(ab′)2 fragments can be isolated directly from recombinant host cell culture. Other techniques for the production of antibody fragments will be apparent to the skilled practitioner. In other embodiments, the antibody of choice is a single chain Fv fragment (scFv). See WO 93/16185; U.S. Pat. No. 5,571,894; and U.S. Pat. No. 5,587,458. The antibody fragment may also be a “linear antibody”, e.g., as described in U.S. Pat. No. 5,641,870 for example. Such linear antibody fragments may be monospecific or bispecific.
- (vi) Antibody Variant Sequences
- Amino acid sequence modification(s) of the antibodies described herein are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of the antibody. Amino acid sequence variants of the antibody are prepared by introducing appropriate nucleotide changes into the antibody nucleic acid, or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into and/or substitutions of, residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics. The amino acid changes also may alter post-translational processes of the antibody, such as changing the number or position of glycosylation sites. Such alterations may be made to the parent antibody and/or multivalent antibody and/or may be introduced in the multivalent antibody amino acid sequence at the time that sequence is made.
- A useful method for identification of certain residues or regions of the antibody that are preferred locations for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells Science, 244:1081-1085 (1989). Here, a residue or group of target residues are identified (e.g., charged residues such as arg, asp, his, lys, and glu) and replaced by a neutral or negatively charged amino acid (most preferably alanine or polyalanine) to affect the interaction of the amino acids with antigen. Those amino acid locations demonstrating functional sensitivity to the substitutions then are refined by introducing further or other variants at, or for, the sites of substitution. Thus, while the site for introducing an amino acid sequence variation is predetermined, the nature of the mutation per se need not be predetermined. For example, to analyze the performance of a mutation at a given site, ala scanning or random mutagenesis is conducted at the target codon or region and the expressed multivalent antibodies are screened for the desired activity.
- Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue or the antibody fused to a cytotoxic polypeptide. Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody to an enzyme (e.g. for ADEPT) or a polypeptide which increases the serum half-life of the antibody.
- Another type of variant is an amino acid substitution variant. These variants have at least one amino acid residue in the antibody molecule replaced by a different residue. The sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but FR alterations are also contemplated. Conservative substitutions are shown in Table 1 under the heading of “preferred substitutions”. If such substitutions result in a change in biological activity, then more substantial changes, denominated “exemplary substitutions” in Table 1, or as further described below in reference to amino acid classes, may be introduced and the products screened.
-
TABLE 1 Original Exemplary Preferred Residue Substitutions Substitutions Ala (A) val; leu; ile val Arg (R) lys; gin; asn lys Asn (N) gln; his; asp, lys; arg gln Asp (D) glu; asn glu Cys (C) ser; ala ser Gln (Q) asn; glu asn Glu (E) asp; gln asp Gly (G) ala ala His (H) asn; gln; lys; arg arg Ile (I) leu; val; met; ala; leu phe; norleucine Leu (L) norleucine; ile; val; ile met; ala; phe Lys (K) arg; gln; asn arg Met (M) leu; phe; ile leu Phe (F) leu; val; ile; ala; tyr tyr Pro (P) ala ala Ser (S) thr thr Thr (T) ser ser Trp (W) tyr; phe tyr Tyr (Y) trp; phe; thr; ser phe Val (V) ile; leu; met; phe; leu ala; norleucine - Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. Naturally occurring residues are divided into groups based on common side-chain properties:
- (1) hydrophobic: norleucine, met, ala, val, leu, ile;
- (2) neutral hydrophilic: cys, ser, thr;
- (3) acidic: asp, glu;
- (4) basic: asn, gln, his, lys, arg;
- (5) residues that influence chain orientation: gly, pro; and
- (6) aromatic: trp, tyr, phe.
- Non-conservative substitutions will entail exchanging a member of one of these classes for another class.
- Any cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability.
- A particularly preferred type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human antibody). Generally, the resulting variant(s) selected for further development will have improved biological properties relative to the parent antibody from which they are generated. A convenient way for generating such substitutional variants involves affinity maturation using phage display. Briefly, several hypervariable region sites (e.g. 6-7 sites) are mutated to generate all possible amino substitutions at each site. The multivalent antibodies thus generated are displayed in a monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage-displayed variants are then screened for their biological activity (e.g. binding affinity) as herein disclosed. In order to identify candidate hypervariable region sites for modification, alanine scanning mutagenesis can be performed to identify hypervariable region residues contributing significantly to antigen binding. Alternatively, or additionally, it may be beneficial to analyze a crystal structure of the antigen-antibody complex to identify contact points between the antibody and antigen. Such contact residues and neighboring residues are candidates for substitution according to the techniques elaborated herein.
- Once such variants are generated, the panel of variants is subjected to screening as described herein and antibodies with superior properties in one or more relevant assays may be selected for further development.
- Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. By altering is meant deleting one or more carbohydrate moieties found in the antibody, and/or adding one or more glycosylation sites that are not present in the antibody.
- Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5-hydroxyproline or 5-hydroxylysine may also be used.
- Addition of glycosylation sites to the antibody is conveniently accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).
- Nucleic acid molecules encoding amino acid sequence variants of the antibody are prepared by a variety of methods known in the art. These methods include, but are not limited to, isolation from a natural source (in the case of naturally occurring amino acid sequence variants) or preparation by oligonucleotide-mediated (or site-directed) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared variant or a non-variant version of the antibody.
- It may be desirable to modify the antibody of the invention with respect to effector function, e.g. so as to enhance antigen-dependent cell-mediated cyotoxicity (ADCC) and/or complement dependent cytotoxicity (CDC) of the antibody. This may be achieved by introducing one or more amino acid modifications in an Fc region of the antibody, thereby generating a variant Fc region. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.
- In one embodiment, the variant Fc region may mediate antibody-dependent cell-mediated cytotoxicity (ADCC) in the presence of human effector cells more effectively, or bind an Fc gamma receptor (FcγR) with better affinity, than a native sequence Fc region. Such Fc region variants may comprise an amino acid modification at any one or more of
positions - The Fc region variant with reduced binding to an FcγR may comprise an amino acid modification at any one or more of amino acid positions 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 292, 293, 294, 295, 296, 298, 301, 303, 322, 324, 327, 329, 333, 335, 338, 340, 373, 376, 382, 388, 389, 414, 416, 419, 434, 435, 437, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- For example, the Fc region variant may display reduced binding to an FcγRI and comprise an amino acid modification at any one or more of amino acid positions 238, 265, 269, 270, 327 or 329 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- The Fc region variant may display reduced binding to an FcγRII and comprise an amino acid modification at any one or more of amino acid positions 238, 265, 269, 270, 292, 294, 295, 298, 303, 324, 327, 329, 333, 335, 338, 373, 376, 414, 416, 419, 435, 438 or 439 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- The Fc region variant of interest may display reduced binding to an FcγRIII and comprise an amino acid modification at one or more of amino acid positions 238, 239, 248, 249, 252, 254, 265, 268, 269, 270, 272, 278, 289, 293, 294, 295, 296, 301, 303, 322, 327, 329, 338, 340, 373, 376, 382, 388, 389, 416, 434, 435 or 437 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- In another embodiment, the Fc region variant displays improved binding to the FcγR and comprises an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 298, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 333, 334, 337, 340, 360, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- For example, the Fc region variant may display increased binding to an FcγRIII and, optionally, may further display decreased binding to an FcγRII. An exemplary such variant comprises amino acid modification(s) at position(s) 298 and/or 333 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- The Fc region variant may display increased binding to an FcγRII and comprise an amino acid modification at any one or more of amino acid positions 255, 256, 258, 267, 268, 272, 276, 280, 283, 285, 286, 290, 301, 305, 307, 309, 312, 315, 320, 322, 326, 330, 331, 337, 340, 378, 398 or 430 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat. Such Fc region variants with increased binding to an FcγRII may optionally further display decreased binding to an FcγRIII and may, for example, comprise an amino acid modification at any one or more of amino acid positions 268, 272, 298, 301, 322 or 340 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- The variant Fc region may alternatively or additionally have altered neonatal Fc receptor (FcRn) binding affinity. Such variant Fc regions may comprise an amino acid modification at any one or more of amino acid positions 238, 252, 253, 254, 255, 256, 265, 272, 286, 288, 303, 305, 307, 309, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 386, 388, 400, 413, 415, 424, 433, 434, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat. Fc region variants with reduced binding to an FcRn may comprise an amino acid modification at any one or more of amino acid positions 252, 253, 254, 255, 288, 309, 386, 388, 400, 415, 433, 435, 436, 439 or 447 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat. The above-mentioned Fc region variants may, alternatively, display increased binding to FcRn and comprise an amino acid modification at any one or more of amino acid positions 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434 of the Fc region, wherein the numbering of the residues in the Fc region is that of the EU index as in Kabat.
- Fc region variants with altered (i.e. improved or diminished) C1q binding and/or Complement Dependent Cytotoxicity (CDC) are described in WO99/51642. Such variants may comprise an amino acid substitution at one or more of amino acid positions 270, 322, 326, 327, 329, 331, 333 or 334 of the Fc region. See, also, Duncan & Winter Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821; and WO94/29351 concerning Fc region variants.
- To increase the serum half life of the antibody, one may incorporate a salvage receptor binding epitope into the antibody (especially an antibody fragment) as described in U.S. Pat. No. 5,739,277, for example. As used herein, the term “salvage receptor binding epitope” refers to an epitope of the Fc region of an IgG molecule (e.g., IgG1, IgG2, IgG3, or IgG4) that is responsible for increasing the in vivo serum half-life of the IgG molecule.
- (vii) Immunoconjugates
- The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g. a small molecule toxin or an enzymatically active toxin of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof), or a radioactive isotope (i.e., a radioconjugate).
- Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above.
- Conjugates of an antibody and one or more small molecule toxins, such as a calicheamicin, a maytansine (U.S. Pat. No. 5,208,020), a trichothene, and CC1065 are also contemplated herein.
- In one preferred embodiment of the invention, the antibody is conjugated to one or more maytansine molecules (e.g. about 1 to about 10 maytansine molecules per antibody molecule). Maytansine may, for example, be converted to May-SS-Me which may be reduced to May-SH3 and reacted with modified antibody (Chari et al. Cancer Research 52: 127-131 (1992)) to generate a maytansinoid-antibody immunoconjugate.
- Another immunoconjugate of interest comprises an antibody conjugated to one or more calicheamicin molecules. The calicheamicin family of antibiotics are capable of producing double-stranded DNA breaks at sub-picomolar concentrations. Structural analogues of calicheamicin which may be used include, but are not limited to, γ1 I, α2 I, α3 I, N-acetyl-γ1 I, PSAG and θI 1 (Hinman et al. Cancer Research 53: 3336-3342 (1993) and Lode et al. Cancer Research 58: 2925-2928 (1998)). See, also, U.S. Pat. Nos. 5,714,586; 5,712,374; 5,264,586; and 5,773,001 expressly incorporated herein by reference.
- Enzymatically active toxins and fragments thereof which can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin and the tricothecenes. See, for example, WO 93/21232 published Oct. 28, 1993.
- The present invention further contemplates an immunoconjugate formed between an antibody and a compound with nucleolytic activity (e.g. a ribonuclease or a DNA endonuclease such as a deoxyribonuclease; DNase).
- A variety of radioactive isotopes are available for the production of radioconjugated antibodies. Examples include At211, I131, I125, Y90, Re186, Re188, Sm153, Bi212, P32 and radioactive isotopes of Lu.
- Conjugates of the antibody and cytotoxic agent may be made using a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate, iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p-diazoniumbenzoylyethylenediamine), diisocyanates (such as
tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al. Science 238: 1098 (1987). Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026. The linker may be a “cleavable linker” facilitating release of the cytotoxic drug in the cell. For example, an acid-labile linker, peptidase-sensitive linker, dimethyl linker or disulfide-containing linker (Chari et al. Cancer Research 52: 127-131 (1992)) may be used. - Alternatively, a fusion protein comprising the antibody and cytotoxic agent may be made, e.g. by recombinant techniques or peptide synthesis.
- In yet another embodiment, the antibody may be conjugated to a “receptor” (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a “ligand” (e.g. avidin) which is conjugated to a cytotoxic agent (e.g. a radionucleotide).
- (viii) Antibody Dependent Enzyme Mediated Prodrug Therapy (ADEPT)
- The antibodies of the present invention may also be used in ADEPT by conjugating the antibody to a prodrug-activating enzyme which converts a prodrug (e.g. a peptidyl chemotherapeutic agent, see WO81/01145) to an active anti-cancer drug. See, for example, WO 88/07378 and U.S. Pat. No. 4,975,278.
- The enzyme component of the immunoconjugate useful for ADEPT includes any enzyme capable of acting on a prodrug in such a way so as to covert it into its more active, cytotoxic form.
- Enzymes that are useful in the method of this invention include, but are not limited to, alkaline phosphatase useful for converting phosphate-containing prodrugs into free drugs; arylsulfatase useful for converting sulfate-containing prodrugs into free drugs; cytosine deaminase useful for converting non-toxic 5-fluorocytosine into the anti-cancer drug, 5-fluorouracil; proteases, such as serratia protease, thermolysin, subtilisin, carboxypeptidases and cathepsins (such as cathepsins B and L), that are useful for converting peptide-containing prodrugs into free drugs; D-alanylcarboxypeptidases, useful for converting prodrugs that contain D-amino acid substituents; carbohydrate-cleaving enzymes such as beta-galactosidase and neuraminidase useful for converting glycosylated prodrugs into free drugs; beta-lactamase useful for converting drugs derivatized with beta-lactams into free drugs; and penicillin amidases, such as penicillin V amidase or penicillin G amidase, useful for converting drugs derivatized at their amine nitrogens with phenoxyacetyl or phenylacetyl groups, respectively, into free drugs. Alternatively, antibodies with enzymatic activity, also known in the art as “abzymes”, can be used to convert the prodrugs of the invention into free active drugs (see, e.g., Massey, Nature 328: 457-458 (1987)). Antibody-abzyme conjugates can be prepared as described herein for delivery of the abzyme to a tumor cell population.
- The enzymes of this invention can be covalently bound to the antibodies by techniques well known in the art such as the use of the heterobifunctional crosslinking reagents discussed above. Alternatively, fusion proteins comprising at least the antigen binding region of an antibody of the invention linked to at least a functionally active portion of an enzyme of the invention can be constructed using recombinant DNA techniques well known in the art (see, e.g., Neuberger et al., Nature, 312: 604-608 (1984).
- (ix) Other Antibody Modifications
- Other modifications of the antibody are contemplated herein. For example, the antibody may be linked to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol, polypropylene glycol, polyoxyalkylenes, or copolymers of polyethylene glycol and polypropylene glycol. The antibody also may be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively), in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980).
- The antibodies disclosed herein may also be formulated as immunoliposomes. Liposomes containing the antibody are prepared by methods known in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA, 82:3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA, 77:4030 (1980); U.S. Pat. Nos. 4,485,045 and 4,544,545; and WO97/38731 published Oct. 23, 1997. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556.
- Particularly useful liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter. Fab′ fragments of the antibody of the present invention can be conjugated to the liposomes as described in Martin et al. J. Biol. Chem. 257: 286-288 (1982) via a disulfide interchange reaction. A chemotherapeutic agent is optionally contained within the liposome. See Gabizon et al. J. National Cancer Inst. 81(19)1484 (1989).
- D. Vectors, Host Cells and Recombinant Methods
- The invention also provides isolated nucleic acid encoding an antibody as disclosed herein, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the antibody.
- For recombinant production of the antibody, the nucleic acid encoding it is isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression. DNA encoding the antibody is readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the antibody). Many vectors are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
- (i) Signal Sequence Component
- The multivalent antibody of this invention may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which is preferably a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide. The heterologous signal sequence selected preferably is one that is recognized and processed (i.e., cleaved by a signal peptidase) by the host cell. For prokaryotic host cells that do not recognize and process the native multivalent antibody signal sequence, the signal sequence is substituted by a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II leaders. For yeast secretion the native signal sequence may be substituted by, e.g., the yeast invertase leader, α factor leader (including Saccharomyces and Kluyveromyces α-factor leaders), or acid phosphatase leader, the C. albicans glucoamylase leader, or the signal described in WO 90/13646. In mammalian cell expression, mammalian signal sequences as well as viral secretory leaders, for example, the herpes simplex gD signal, are available.
- The DNA for such precursor region is ligated in reading frame to DNA encoding the multivalent antibody.
- (ii) Origin of Replication Component
- Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Generally, in cloning vectors this sequence is one that enables the vector to replicate independently of the host chromosomal DNA, and includes origins of replication or autonomously replicating sequences. Such sequences are well known for a variety of bacteria, yeast, and viruses. The origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2μ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells. Generally, the origin of replication component is not needed for mammalian expression vectors (the SV40 origin may typically be used only because it contains the early promoter).
- (iii) Selection Gene Component
- Expression and cloning vectors may contain a selection gene, also termed a selectable marker. Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli.
- One example of a selection scheme utilizes a drug to arrest growth of a host cell. Those cells that are successfully transformed with a heterologous gene produce a protein conferring drug resistance and thus survive the selection regimen. Examples of such dominant selection use the drugs neomycin, mycophenolic acid and hygromycin.
- Another example of suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the multivalent antibody nucleic acid, such as DHFR, thymidine kinase, metallothionein-I and -II, preferably primate metallothionein genes, adenosine deaminase, ornithine decarboxylase, etc.
- For example, cells transformed with the DHFR selection gene are first identified by culturing all of the transformants in a culture medium that contains methotrexate (Mtx), a competitive antagonist of DHFR. An appropriate host cell when wild-type DHFR is employed is the Chinese hamster ovary (CHO) cell line deficient in DHFR activity.
- Alternatively, host cells (particularly wild-type hosts that contain endogenous DHFR) transformed or co-transformed with DNA sequences encoding multivalent antibody, wild-type DHFR protein, and another selectable marker such as
aminoglycoside 3′-phosphotransferase (APH) can be selected by cell growth in medium containing a selection agent for the selectable marker such as an aminoglycosidic antibiotic, e.g., kanamycin, neomycin, or G418. See U.S. Pat. No. 4,965,199. - A suitable selection gene for use in yeast is the trp1 gene present in the yeast plasmid YRp7 (Stinchcomb et al., Nature, 282:39 (1979)). The trp1 gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1. Jones, Genetics, 85:12 (1977). The presence of the trp1 lesion in the yeast host cell genome then provides an effective environment for detecting transformation by growth in the absence of tryptophan. Similarly, Leu2-deficient yeast strains (ATCC 20,622 or 38,626) are complemented by known plasmids bearing the Leu2 gene.
- In addition, vectors derived from the 1.6 μm circular plasmid pKD1 can be used for transformation of Kluyveromyces yeasts. Alternatively, an expression system for large-scale production of recombinant calf chymosin was reported for K. lactis. Van den Berg, Bio/Technology, 8:135 (1990). Stable multi-copy expression vectors for secretion of mature recombinant human serum albumin by industrial strains of Kluyveromyces have also been disclosed. Fleer et al., Bio/Technology, 9:968-975 (1991).
- (iv) Promoter Component
- Expression and cloning vectors usually contain a promoter that is recognized by the host organism and is operably linked to the multivalent antibody nucleic acid. Promoters suitable for use with prokaryotic hosts include the phoA promoter, β-lactamase and lactose promoter systems, alkaline phosphatase, a tryptophan (trp) promoter system, and hybrid promoters such as the tac promoter. However, other known bacterial promoters are suitable. Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding the multivalent antibody.
- Promoter sequences are known for eukaryotes. Virtually all eukaryotic genes have an AT-rich region located approximately 25 to 30 bases upstream from the site where transcription is initiated. Another sequence found 70 to 80 bases upstream from the start of transcription of many genes is a CNCAAT region where N may be any nucleotide. At the 3′ end of most eukaryotic genes is an AATAAA sequence that may be the signal for addition of the poly A tail to the 3′ end of the coding sequence. All of these sequences are suitably inserted into eukaryotic expression vectors.
- Examples of suitable promoting sequences for use with yeast hosts include the promoters for 3-phosphoglycerate kinase or other glycolytic enzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase, hexokinase, pyruvate decarboxylase, phosphofructokinase, glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvate kinase, triosephosphate isomerase, phosphoglucose isomerase, and glucokinase.
- Other yeast promoters, which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for
alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization. Suitable vectors and promoters for use in yeast expression are further described in EP 73,657. Yeast enhancers also are advantageously used with yeast promoters. - Multivalent antibody transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus, adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and most preferably Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, from heat-shock promoters, provided such promoters are compatible with the host cell systems.
- The early and late promoters of the SV40 virus are conveniently obtained as an SV40 restriction fragment that also contains the SV40 viral origin of replication. The immediate early promoter of the human cytomegalovirus is conveniently obtained as a HindIII E restriction fragment. A system for expressing DNA in mammalian hosts using the bovine papilloma virus as a vector is disclosed in U.S. Pat. No. 4,419,446. A modification of this system is described in U.S. Pat. No. 4,601,978. See also Reyes et al., Nature 297:598-601 (1982) on expression of human β-interferon cDNA in mouse cells under the control of a thymidine kinase promoter from herpes simplex virus. Alternatively, the rous sarcoma virus long terminal repeat can be used as the promoter.
- (v) Enhancer Element Component
- Transcription of a DNA encoding the multivalent antibody of this invention by higher eukaryotes is often increased by inserting an enhancer sequence into the vector. Many enhancer sequences are now known from mammalian genes (globin, elastase, albumin, α-fetoprotein, and insulin). Typically, however, one will use an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. See also Yaniv, Nature 297:17-18 (1982) on enhancing elements for activation of eukaryotic promoters. The enhancer may be spliced into the vector at a
position 5′ or 3′ to the multivalent antibody-encoding sequence, but is preferably located at asite 5′ from the promoter. - (vi) Transcription Termination Component
- Expression vectors used in eukaryotic host cells (yeast, fungi, insect, plant, animal, human, or nucleated cells from other multicellular organisms) will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5′ and, occasionally 3′, untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding the multivalent antibody. One useful transcription termination component is the bovine growth hormone polyadenylation region. See WO94/11026 and the expression vector disclosed therein.
- (vii) Selection and Transformation of Host Cells
- Suitable host cells for cloning or expressing the DNA in the vectors herein are the prokaryote, yeast, or higher eukaryote cells described above. Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as B. subtilis and B. licheniformis (e.g., B. licheniformis 41P disclosed in DD 266,710 published 12 Apr. 1989), Pseudomonas such as P. aeruginosa, and Streptomyces. One preferred E. coli cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coli X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable. These examples are illustrative rather than limiting.
- In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for multivalent antibody-encoding vectors. Saccharomyces cerevisiae, or common baker's yeast, is the most commonly used among lower eukaryotic host microorganisms. However, a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pombe; Kluyveromyces hosts such as, e.g., K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans, and K. marxianus; yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora crassa; Schwanniomyces such as Schwanniomyces occidentalis; and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.
- Suitable host cells for the expression of glycosylated multivalent antibody are derived from multicellular organisms. Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruitfly), and Bombyx mori have been identified. A variety of viral strains for transfection are publicly available, e.g., the L-1 variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present invention, particularly for transfection of Spodoptera frugiperda cells.
- Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
- However, interest has been greatest in vertebrate cells, and propagation of vertebrate cells in culture (tissue culture) has become a routine procedure. Examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR(CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; a human hepatoma line (Hep G2); and myeloma or lymphoma cells (e.g. Y0, J558L, P3 and NS0 cells) (see U.S. Pat. No. 5,807,715).
- Host cells are transformed with the above-described expression or cloning vectors for multivalent antibody production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
- (viii) Culturing the Host Cells
- The host cells used to produce the multivalent antibody of this invention may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells. In addition, any of the media described in Ham et al., Meth. Enz. 58:44 (1979), Barnes et al., Anal. Biochem. 102:255 (1980), U.S. Pat. No. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO 87/00195; or U.S. Pat. Re. No. 30,985 may be used as culture media for the host cells. Any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCIN™ drug), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art. The culture conditions, such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
- (ix) Purification
- When using recombinant techniques, the multivalent antibody can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the multivalent antibody is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/Technology 10:163-167 (1992) describe a procedure for isolating antibodies which are secreted to the periplasmic space of E. coli. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min. Cell debris can be removed by centrifugation. Where the multivalent antibody is secreted into the medium, supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
- The multivalent antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc region that is present in the multivalent antibody. Protein A can be used to purify antibodies that are based on human γ1, γ2, or γ4 heavy chains (Lindmark et al., J. Immunol. Meth. 62:1-13 (1983)). Protein G is recommended for all mouse isotypes and for human γ3 (Guss et al., EMBO J. 5:15671575 (1986)). The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. Where the multivalent antibody comprises a
C H3 domain, the Bakerbond ABX™ resin (J. T. Baker, Phillipsburg, N.J.) is useful for purification. Other techniques for protein purification such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE™ chromatography on an anion or cation exchange resin (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available depending on the multivalent antibody to be recovered. - E. Pharmaceutical Formulations
- Therapeutic formulations of the multivalent antibody are prepared for storage by mixing the multivalent antibody having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of aqueous solutions, lyophilized or other dried formulations. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEENT™, PLURONICS™ or polyethylene glycol (PEG).
- The formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Examples of combinations of active compounds are provided in Section G below entitled “In Vivo Uses for the Multivalent Antibody”. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
- The active ingredients may also be entrapped in microcapsule prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsule and poly-(methylmethacylate) microcapsule, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980).
- The formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
- Sustained-release preparations may be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the multivalent antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and γ ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(−)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37° C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S—S bond formation through thio-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
- F. Non-Therapeutic Uses for the Multivalent Antibody
- The multivalent antibody of the invention may be used as an affinity purification agent. In this process, the multivalent antibody is immobilized on a solid phase such a Sephadex resin or filter paper, using methods well known in the art. The immobilized multivalent antibody is contacted with a sample containing the antigen to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the antigen to be purified, which is bound to the immobilized multivalent antibody. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, that will release the antigen from the multivalent antibody.
- The multivalent antibody may also be useful in diagnostic assays, e.g., for detecting expression of an antigen of interest in specific cells, tissues, or serum.
- For diagnostic applications, the multivalent antibody typically will be labeled with a detectable moiety. Numerous labels are available which can be generally grouped into the following categories:
- (a) Radioisotopes, such as 35S, 14C, 125I, 3H, and 131I. The multivalent antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology,
Volumes - (b) Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin and Texas Red are available. The fluorescent labels can be conjugated to the multivalent antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter.
- (c) Various enzyme-substrate labels are available and U.S. Pat. No. 4,275,149 provides a review of some of these. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described in O'Sullivan et al., Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73:147-166 (1981).
- Examples of enzyme-substrate combinations include, for example
- (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor (e.g., orthophenylene diamine (OPD) or 3,3′,5,5′-tetramethyl benzidine hydrochloride (TMB));
- (ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and
- (iii) β-D-galactosidase (β-D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) or fluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase.
- Numerous other enzyme-substrate combinations are available to those skilled in the art. For a general review of these, see U.S. Pat. Nos. 4,275,149 and 4,318,980.
- Sometimes, the label is indirectly conjugated with the multivalent antibody. The skilled artisan will be aware of various techniques for achieving this. For example, the multivalent antibody can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the multivalent antibody in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the multivalent antibody, the multivalent antibody is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten multivalent antibody (e.g., anti-digoxin antibody). Thus, indirect conjugation of the label with the multivalent antibody can be achieved.
- In another embodiment of the invention, the multivalent antibody need not be labeled, and the presence thereof can be detected using a labeled antibody which binds to the multivalent antibody.
- The multivalent antibody of the present invention may be employed in any known assay method, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987).
- The multivalent antibody may also be used for in vivo diagnostic assays. Generally, the multivalent antibody is labeled with a radionuclide (such as 111In, 99Tc, 14C, 131I, 125I, 3H, 32P or 35S) so that the antigen or cells expressing it can be localized using immunoscintiography.
- G. In Vivo Uses for the Multivalent Antibody
- It is contemplated that the multivalent antibody of the present invention may be used to treat a mammal e.g. a patient suffering from, or predisposed to, a disease or disorder who could benefit from administration of the multivalent antibody.
- Where the antibody binds an ErbB receptor, such as HER2, conditions to be treated therewith include benign or malignant tumors; leukemias and lymphoid malignancies; other disorders such as neuronal, glial, astrocytal, hypothalamic, glandular, macrophagal, epithelial, stromal, blastocoelic, inflammatory, angiogenic and immunologic disorders. Generally, the disease or disorder to be treated with the antibody that binds an ErbB receptor is cancer.
- Examples of cancer to be treated herein include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More particular examples of such cancers include squamous cell cancer, lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma as well as head and neck cancer.
- The cancer will generally comprise cells that express an antigen bound by the antibody, such that the antibody is able to bind to the cancer. In one embodiment, the cancer may be characterized by overexpression of the antigen (e.g. overexpression of an ErbB receptor). To determine expression of the antigen by the cancer, various diagnostic/prognostic assays are available. In one embodiment, antigen overexpression may be analyzed by INC, e.g. using the HERCEPTEST® (Dako) where the antigen is HER2. In the HER2 IHC test, parrafin embedded tissue sections from a tumor biopsy may be subjected to the IHC assay and accorded a HER2 protein staining intensity criteria as follows:
-
Score 0 no staining is observed or membrane staining is observed in less than 10% of tumor cells. -
Score 1+ a faint/barely perceptible membrane staining is detected in more than 10% of the tumor cells. The cells are only stained in part of their membrane. -
Score 2+ a weak to moderate complete membrane staining is observed in more than 10% of the tumor cells. -
Score 3+ a moderate to strong complete membrane staining is observed in more than 10% of the tumor cells. - Those tumors with 0 or 1+ scores for HER2 overexpression assessment may be characterized as not overexpressing HER2, whereas those tumors with 2+ or 3+ scores may be characterized as overexpressing HER2.
- Alternatively, or additionally, FISH assays such as the INFORM™ (sold by Ventana, Ariz.) or PATHVISION™ (Vysis, Ill.) may be carried out on formalin-fixed, paraffin-embedded tumor tissue to determine the extent (if any) of antigen overexpression by the tumor.
- In one embodiment, the cancer will be one which expresses (and may overexpress) an ErbB receptor selected from the group consisting of EGFR, ErbB3 and ErbB4. Examples of cancers which may express/overexpress EGFR, ErbB3 or ErbB4 include squamous cell cancer, lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma as well as head and neck cancer as well as glioblastomas.
- The cancer to be treated herein may be one characterized by excessive activation of an ErbB receptor, e.g. EGFR. Such excessive activation may be attributable to overexpression or increased production of the ErbB receptor or an ErbB ligand. In one embodiment of the invention, a diagnostic or prognostic assay will be performed to determine whether the patient's cancer is characterized by excessive activation of an ErbB receptor. For example, ErbB gene amplification and/or overexpression of an ErbB receptor in the cancer may be determined. Various assays for determining such amplification/overexpression are available in the art and include the IHC, FISH and shed antigen assays described above. Alternatively, or additionally, levels of an ErbB ligand, such as TGF-alpha, in or associated with the tumor may be determined according to known procedures. Such assays may detect protein and/or nucleic acid encoding it in the sample to be tested. In one embodiment, ErbB ligand levels in the tumor may be determined using immunohistochemistry (1HC); see, for example, Scher et al. Clin. Cancer Research 1:545-550 (1995). Alternatively, or additionally, one may evaluate levels of ErbB ligand-encoding nucleic acid in the sample to be tested; e.g. via fluorescent in situ hybridization or FISH, southern blotting, or polymerase chain reaction (PCR) techniques.
- Moreover, ErbB receptor or ErbB ligand overexpression or amplification may be evaluated using an in vivo diagnostic assay, e.g. by administering a molecule (such as an antibody) which binds the molecule to be detected and is tagged with a detectable label (e.g. a radioactive isotope) and externally scanning the patient for localization of the label.
- Where the antibody binds a B cell surface antigen, the antibody may be used to treat a B cell lymphoma (including low grade/follicular non-Hodkin's lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; and chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD).
- The antibody, e.g. the anti-B cell surface antigen antibody, may also be used to treat an autoimmune disease. Examples of autoimmune diseases or disorders include, but are not limited to, inflammatory responses such as inflammatory skin diseases including psoriasis and dermatitis (e.g. atopic dermatitis); systemic scleroderma and sclerosis; responses associated with inflammatory bowel disease (such as Crohn's disease and ulcerative colitis); respiratory distress syndrome (including adult respiratory distress syndrome; ARDS); dermatitis; meningitis; encephalitis; uveitis; colitis; glomerulonephritis; allergic conditions such as eczema and asthma and other conditions involving infiltration of T cells and chronic inflammatory responses; atherosclerosis; leukocyte adhesion deficiency; rheumatoid arthritis; systemic lupus erythematosus (SLE); diabetes mellitus (e.g. Type I diabetes mellitus or insulin dependent diabetes mellitis); multiple sclerosis; Reynaud's syndrome; autoimmune thyroiditis; allergic encephalomyelitis; Sjorgen's syndrome; juvenile onset diabetes; and immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T-lymphocytes typically found in tuberculosis, sarcoidosis, polymyositis, granulomatosis and vasculitis; pernicious anemia (Addison's disease); diseases involving leukocyte diapedesis; central nervous system (CNS) inflammatory disorder; multiple organ injury syndrome; hemolytic anemia (including, but not limited to cryoglobinemia or Coombs positive anemia); myasthenia gravis; antigen-antibody complex mediated diseases; anti-glomerular basement membrane disease; antiphospholipid syndrome; allergic neuritis; Graves' disease; Lambert-Eaton myasthenic syndrome; pemphigoid bullous; pemphigus; autoimmune polyendocrinopathies; Reiter's disease; stiff-man syndrome; Behcet disease; giant cell arteritis; immune complex nephritis; IgA nephropathy; IgM polyneuropathies; immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia etc.
- Antibodies directed against B cell surface antigens may also be used to block an immune response to a foreign antigen. By “foreign antigen” here is meant a molecule or molecules which is/are not endogenous or native to a mammal which is exposed to it. The foreign antigen may elicit an immune response, e.g. a humoral and/or T cell mediated response in the mammal. Generally, the foreign antigen will provoke the production of antibodies thereagainst. Examples of foreign antigens contemplated herein include immunogenic therapeutic agents, e.g. proteins such as antibodies, particularly antibodies comprising non-human amino acid residues (e.g. rodent, chimeric/humanized, and primatized antibodies); toxins (optionally conjugated to a targeting molecule such as an antibody, wherein the targeting molecule may also be immunogenic); gene therapy viral vectors, such as retroviruses and adenoviruses; grafts; infectious agents (e.g. bacteria and virus); alloantigens (i.e. an antigen that occurs in some, but not in other members of the same species) such as differences in blood types, human lymphocyte antigens (HLA), platelet antigens, antigens expressed on transplanted organs, blood components, pregnancy (Rh), and hemophilic factors (e.g. Factor VIII and Factor IX).
- The anti-B cell surface antigen antibody may also be used to desenzitize a mammal awaiting transplantation.
- Antibodies directed against a receptor in the TNF receptor superfamily may be employed to activate or stimulate apoptosis in cancer cells.
- In certain embodiments, an immunoconjugate comprising the antibody conjugated with a cytotoxic agent is administered to the patient. Preferably, the immunoconjugate and/or antigen to which it is bound is/are internalized by the cell, resulting in increased therapeutic efficacy of the immunoconjugate in killing the cancer cell to which it binds. In a preferred embodiment, the cytotoxic agent targets or interferes with nucleic acid in the cancer cell. Examples of such cytotoxic agents include any of the chemotherapeutic agents noted herein (such as a maytansinoid or a calicheamicin), a radioactive isotope, or a ribonuclease or a DNA endonuclease. As noted above, the multivalent antibody may also be used for ADEPT.
- The present application contemplates combining the multivalent antibody (or immunoconjugate thereof) with one or more other therapeutic agent(s), especially for treating cancer. For instance, the multivalent antibody may be co-administered with another multivalent antibody (or multivalent antibodies), a monovalent or bivalent antibody (or antibodies), chemotherapeutic agent(s) (including cocktails of chemotherapeutic agents), other cytotoxic agent(s), anti-angiogenic agent(s), cytokines, and/or growth inhibitory agent(s). Where the multivalent antibody induces apoptosis, it may be particularly desirable to combine the multivalent antibody with one or more other therapeutic agent(s) which also induce apoptosis. For instance, pro-apoptotic antibodies (e.g. bivalent or multivalent antibodies) directed against B cell surface antigens (e.g. RITUXAN®, ZEVALIN® or BEXXAR® anti-CD20 antibodies) may be combined with (1) pro-apoptotic antibodies (e.g. bivalent or multivalent antibodies directed against a receptor in the TNF receptor superfamily, such as anti-DR4 or anti-DR5 antibodies) or (2) with cytokines in the TNF family of cytokines (e.g. Apo2L). Likewise, anti-ErbB antibodies (e.g. HERCEPTIN® anti-HER2 antibody) may be combined with (1) and/or (2). Alternatively, or additionally, the patient may receive combined radiation therapy (e.g. external beam irradiation or therapy with a radioactive labelled agent, such as an antibody). Such combined therapies noted above include combined administration (where the two or more agents are included in the same or separate formulations), and separate administration, in which case, administration of the multivalent antibody can occur prior to, and/or following, administration of the adjunct therapy or therapies.
- The multivalent antibody (and adjunct therapeutic agent) is/are administered by any suitable means, including parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In addition, the multivalent antibody is suitably administered by pulse infusion, particularly with declining doses of the multivalent antibody. Preferably the dosing is given by injections, most preferably intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic.
- Aside from administration of the antibody protein to the patient, the present application contemplates administration of the antibody by gene therapy. Such administration of nucleic acid encoding the antibody is encompassed by the expression “administering a therapeutically effective amount of an antibody”. See, for example, WO96/07321 published Mar. 14, 1996 concerning the use of gene therapy to generate intracellular antibodies.
- There are two major approaches to getting the nucleic acid (optionally contained in a vector) into the patient's cells; in vivo and ex vivo. For in vivo delivery the nucleic acid is injected directly into the patient, usually at the site where the antibody is required. For ex vivo treatment, the patient's cells are removed, the nucleic acid is introduced into these isolated cells and the modified cells are administered to the patient either directly or, for example, encapsulated within porous membranes which are implanted into the patient (see, e.g. U.S. Pat. Nos. 4,892,538 and 5,283,187). There are a variety of techniques available for introducing nucleic acids into viable cells. The techniques vary depending upon whether the nucleic acid is transferred into cultured cells in vitro, or in vivo in the cells of the intended host. Techniques suitable for the transfer of nucleic acid into mammalian cells in vitro include the use of liposomes, electroporation, microinjection, cell fusion, DEAE-dextran, the calcium phosphate precipitation method, etc. A commonly used vector for ex vivo delivery of the gene is a retrovirus. The currently preferred in vivo nucleic acid transfer techniques include transfection with viral vectors (such as adenovirus, Herpes simplex I virus, or adeno-associated virus) and lipid-based systems (useful lipids for lipid-mediated transfer of the gene are DOTMA, DOPE and DC-Chol, for example). In some situations it is desirable to provide the nucleic acid source with an agent that targets the target cells, such as an antibody specific for a cell surface membrane protein or the target cell, a ligand for a receptor on the target cell, etc. Where liposomes are employed, proteins which bind to a cell surface membrane protein associated with endocytosis may be used for targeting and/or to facilitate uptake, e.g. capsid proteins or fragments thereof tropic for a particular cell type, antibodies for proteins which undergo internalization in cycling, and proteins that target intracellular localization and enhance intracellular half-life. The technique of receptor-mediated endocytosis is described, for example, by Wu et al., J. Biol. Chem. 262:4429-4432 (1987); and Wagner et al., Proc. Natl. Acad. Sci. USA 87:3410-3414 (1990). For review of the currently known gene marking and gene therapy protocols see Anderson et al., Science 256:808-813 (1992). See also WO 93/25673 and the references cited therein.
- For the prevention or treatment of disease, the appropriate dosage of multivalent antibody will depend on the type of disease to be treated, the severity and course of the disease, whether the multivalent antibody is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the multivalent antibody, and the discretion of the attending physician. The multivalent antibody is suitably administered to the patient at one time or over a series of treatments.
- Depending on the type and severity of the disease, about 1 μg/kg to 15 mg/kg (e.g., 0.1-20 mg/kg) of multivalent antibody is an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. A typical daily dosage might range from about 1 μg/kg to 100 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays.
- The multivalent antibody composition will be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The “therapeutically effective amount” of the multivalent antibody to be administered will be governed by such considerations, and is the minimum amount necessary to prevent, ameliorate, or treat a disease or disorder. The multivalent antibody need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of multivalent antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as used hereinbefore or about from 1 to 99% of the heretofore employed dosages.
- H. Articles of Manufacture
- In another embodiment of the invention, an article of manufacture containing materials useful for the treatment of the disorders described above is provided. The article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is effective for treating the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a multivalent antibody. The label or package insert indicates that the composition is used for treating the condition of choice, such as cancer. Moreover, the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises a multivalent antibody; and (b) a second container with a composition contained therein, wherein the composition comprises a further cytotoxic agent. The article of manufacture in this embodiment of the invention may further comprises a package insert indicating that the first and second antibody compositions can be used to treat cancer. Alternatively, or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.
- I. Deposit of Materials
- The following hybridoma cell lines have been deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, USA (ATCC):
-
Antibody Designation ATCC No. Deposit Date 7C2 (anti-HER2) ATCC HB-12215 Oct. 17, 1996 7F3 (anti-HER2) ATCC HB-12216 Oct. 17, 1996 4D5 (anti-HER2) ATCC CRL 10463 May 24, 1990 2C4 (anti-HER2) ATCC HB-12697 Apr. 8, 1999 3F11.39.7 (anti-DR5) HB-12456 Jan. 13, 1998 3H3.14.5 (anti-DR5) HB-12534 Jun. 2, 1998 3D5.1.10 (anti-DR5) HB-12536 Jun. 2, 1998 3H1.18.10 (anti-DR5) HB-12535 Jun. 2, 1998 4E7.24.3 (anti-DR4) HB-12454 Jan. 13, 1998 4H6.17.8 (anti-DR4) HB-12455 Jan. 13, 1998 - The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of this invention. All literature and patent citations mentioned herein are expressly incorporated by reference.
- The construct used to generate a tetravalent anti-HER2 antibody, called an “Octopus antibody” (OctHER2), is illustrated in
FIG. 5 herein. The backbone of this Octopus antibody is the recombinant, humanized monoclonal antibody 4D5 variant 8 (rhuMAb 4D5-8) (U.S. Pat. No. 5,821,337, Carter et al., expressly incorporated herein by reference). The heavy chain of rhuMAb 4D5-8 was subcloned into the pRK5 vector (EP 307,247, published Mar. 15, 1989). The VH-CH1 region of the heavy chain was removed by mutagenesis, and three unique restriction sites (BamHI; NheI; BspEI) were inserted. These sites were incorporated into PCR primers designed to amplify the VH-CH1 region from different antibodies. The resulting fragments were subcloned into the vector to create the Octopus heavy chain. Co-expression of the Octopus heavy chain with the appropriate light chain in a pRK5 vector in mammalian cell transfections results in the completed Octopus antibody (FIG. 4 ). - Octopus constructs containing flexible linkers inserted between the tandem Fd regions were are also engineered. Through mutagenesis, DNA encoding either “gly-ser” (
flex 1 linker) or “gly-ser-gly-ser” (SEQ ID NO:10) (flex 2 linker) was inserted between the DNA encoding the VH-CH1 regions of the heavy chain. - OctHER2 was expressed in transiently transfected 293 cells (Graham et al. J. Gen. Virol. 36:59-72 (1977)) and purified over a Protein A sepharose column. The complete antibody is approximately 245 kDa, as compared to the 150 kDa molecular weight of the parent antibody. The Octopus heavy chain is 75 kDa (without carbohydrate), and the light chain is 30 kDa.
- Binding of OctHER2 to antigen, HER2 extracellular domain (HER2ECD), was analyzed using a HER2ELISA assay (Sias et al. J. Immunol. Methods 132:73-80 (1990)). Ninety-six well plates were coated with the HER2 extracellular domain (ECD) (WO90/14357), and incubated with different dilutions of anti-HER2 antibodies. After washing to remove unbound antibody, a secondary peroxidase-conjugated antibody was then added to detect the anti-HER2 antibody bound to the ECD. The appropriate substrate was then added, and the wells were visualized and then quantitated on a plate reader at 562 nm.
- The ELISA results for OctHER2, bivalent human IgG1 anti-HER2 antibody rhuMAb 4D5-8 expressed by 293 cells, or bivalent anti-HER2 antibody HERCEPTIN® (commercially available from Genentech, Inc., South San Francisco, USA), are shown in
FIGS. 6A-C . OctHER2 binds the HER2ECD similar to HERCEPTIN® when analyzed in an ELISA assay. The rhuMAb 4D5-8 expressed by 293 cells binds identically to the vialed HERCEPTIN® (produced by Chinese Hamster Ovary (CHO) cells), indicating that 293 cells do not substantially alter the antigen binding capability of the antibodies. - Ultracentrifugation analysis was used to determine whether OctHER2 was capable of binding target with all four antigen binding sites. Different amounts of the HER2 extracellular domain (ECD) (WO90/14357) were titrated in with the Octopus antibody, and based upon these ratios, the average molecular weight of the complexes was calculated assuming that the Octopus antibody either had four fully functional binding sites, or three functional binding sites. These theoretical values (circles, assuming OctHER2 has four functional binding sites; and squares, assuming OctHER2 has three functional binding sites) were compared to the actual experimental values obtained (triangles). The experimental values depicted in
FIG. 7 more closely follow the curve representing four binding sites, however the drift observed is an indicator that all four sites probably do not bind with the same affinity. - Antiproliferation Assays: OctHER2 was compared to HERCEPTIN® in functional assays measuring growth inhibition of HER2 overexpressing tumor cell lines. The growth inhibition assay described in Lewis et al. Cancer Immuno. Immunother. 37:255-263 (1993) was used. Briefly, serial dilutions of OctHER2 and HERCEPTIN® were added to the media of plated cells which were then allowed to continue growing for five days. After this time, the media was removed and the cells were stained with crystal violet and quantitated by spectrophotometry. Crystal violet is a colorimetric dye that stains cells, thus allowing measurement of cell growth after treatment.
- In 3+ HER2 overexpressing cells (on which HERCEPTIN® is very effective), OctHER2 was similar to slightly better at inhibiting growth of SKBR3 cells (
FIG. 8A ), however was not as effective on BT474 cells (FIG. 8B ). Interestingly, OctHER2 inhibited more effectively than HERCEPTIN® a 2+ overexpressing cell line, MDA 361 (FIG. 8B ). - As shown in
FIG. 9 , the flexible linker Octopus constructs (OctHER2flex1, OctHER2flex2) inhibited cell growth more effectively than HERCEPTIN®. - Internalization Assays: In order to assess the application of the Octopus antibody for immunotoxin therapy, its internalization capabilities were evaluated. For antibody arming or immunotoxin therapy, a cytotoxic agent is conjugated with or fused to the antibody and the immunotoxin thus produced binds specifically to its cellular target; the thus-bound cell internalizes the antibody, and catabolizes or degrades the antibody releasing the toxin which kills the cell.
- In the internalization assays performed herein, the antibody was radioiodinated, and incubated for varying times with the cells. This was followed by measurements of the amount of intact, unbound antibody in the supernatant, the amount bound to the cell surface, the amount internalized, and finally, the amount catabolized and degraded.
- The results of internalization assays performed with respect to a 3+ overexpressing cell line (SKBR3) and a 2+ overexpressing cell line (MDA453) (the solid lines represent 2+ HER2 overexpressors, and the dashed lines, 3+overexpressors) are depicted in
FIGS. 10A-B . These results indicate that OctHER2, surprisingly, internalized and catabolized twice as fast as HERCEPTIN® in both cell lines. The rapid internalization and catabolism displayed by the Octopus antibody is ideal for an armed antibody. In comparison to unbound HERCEPTIN®, there is very little free Octopus antibody in a 2+overexpressing cell. Once again, these results suggest that the Octopus antibody would be an excellent candidate for conjugating cytotoxic agents for tumor delivery. - Electron Microscopy Autoradiography: To confirm that the Octopus antibody was being internalized and degraded in the appropriate vesicles, and not just nonspecifically, Electron Microscopy (EM) autoradiography was used. The Octopus antibody was iodinated and incubated with the cells in the same fashion as in the internalization assays. The results depicted in
FIGS. 11A-C confirm that the Octopus antibody was being internalized into the correct vesicles (early endosome,FIG. 11B ; and lysosome,FIG. 11C ). Additionally, the percentage of internalization observed with OctHER2 and HERCEPTIN® in these assays matched with the measurements in the internalization assays. - DR5 a member of the TNF receptor superfamily that binds the trimeric Apo2L/TRAIL (Apo2L). After Apo2L receptors bind Apo2L and are clustered, death domains in the cytosolic region of the receptors induce caspases to trigger cellular apoptosis. Two versions of anti-DR5 Octopus constructs were made: one from 16E2, an anti-DR5 cloned from a single-chain human Fv phage library (see WO98/51793, expressly incorporated herein by reference); the second anti-DR5 Octopus antibody was made from Mab 3H3.14.5 (the “3H3” antibody; ATCC HB-12534, WO99/64461), a murine anti-DR5MAb that induces apoptosis when it is crosslinked. Since anti-Death receptor monoclonal antibodies may require crosslinking to trigger apoptosis, they are candidates for the Octopus antibody construct. The anti-DR5 Octopus antibodies were prepared by replacing the variable domains of the OctHER2 construct described above with the VL and VH domains from 16E2 or 3H3.
- The anti-DR5 Octopus antibodies were analyzed in apoptosis assays using either crystal violet or alamarBlue staining. Briefly, serial dilutions of the Octopus antibody or Apo2L were added to the media of plated cells which were then allowed to continue growing for 24 hours. After this time, the media was either removed and the cells were stained with crystal violet, or alamarBlue was added to the media and incubated briefly with the cells. Crystal violet stains the cells, whereas alamarBlue detects metabolic activity in the culture media, thus these dyes allow for measurement of cells that survive treatment. Staining by both colorimetric dyes, crystal violet and alamarBlue, was quantitated by spectrophotometry.
- As shown in
FIGS. 12A-E , the 16E2 Octopus, surprisingly, induces apoptosis with comparable potency to Apo2L in lung (SK-MES-1; HOP 92) and colon (HCT116; COLO 205) tumor cell lines, however does not cause apoptosis on normal control cell line (HUMEC). The apoptosis induced by the 16E2 Octopus is caspase-dependent. - The anti-DR516E2 Octopus was also effective in vivo in inducing apoptosis and shrinking a colon tumor, human COLO205, in athymic nude mice. As shown in
FIG. 13A-D , histology slides of tumor tissues stained with hematoxylin and eosin from mice treated with the 16E2 Octopus or Apo2L induced similar levels of apoptotic cells. - The 16E2 Octopus-treated mice also demonstrated significant decrease in tumor volume, similar to that measured for the Apo2L and two bivalent anti-DR5 mAbs, 16E2 and 3H3, as shown in
FIG. 14 . Mice that did not receive any anti-DR5 antibodies or Apo2L (Vehicle) showed dramatic increase in their tumor volume due to uncontrolled growth. - The apoptotic activity of the material used in the mouse studies was confirmed in an in vitro apoptotic assay in
FIG. 15 . The anti-DR516E2 Octopus and the Apo2L used in the study were compared to an Apo2L standard positive control and an anti-IgE MAb (E25) negative control in an alamarBlue apoptosis assay. -
FIG. 16 demonstrates that another anti-DR5 Octopus, 3H3 Octopus, is capable of inducing apoptosis similar to the 16E2 Octopus. Additionally,FIG. 16 shows that the apoptotic activity of the Octopus antibody is not lot dependent, as several 16E2 Octopus antibodies prepared on different dates retain similar function. - In
FIGS. 17A and B, the apoptotic activity of both the 16E2 and 3H3 Octopus antibodies is better than Apo2L on a lung tumor cell line, SK-MES-1 (FIG. 17A ), and a T cell tumor line, Jurkat (FIG. 17B ). The anti-DR5 Octopus antibodies may be more effective at clustering DR5 on the tumor cell surface than Apo2L. - The 16E2 Octopus was analyzed in a 2-day and 6-day screen against the National Cancer Institute (NCI) panel of human tumor cell lines in comparison with the Apo2L.
FIGS. 18A-C depict the 2-day dose response curves showing the effects of the 16E2 Octopus and Apo2L on the growth of several human leukemia, non-small cell lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer tumor cell lines, whileFIGS. 19A-C show dose response curves from the 6 day screens. Comparable results were observed for 16E2 Octopus and Apo2L against most of the tumor cell lines, again indicating that the anti-DR5 Octopus functions similar to Apo2L. Similar inhibition of the lung and colon cancer cell lines confirmed the previous in vitro and in vivo results from apoptosis assays comparing 16E2 Octopus and Apo2L on cell lines of these cancers. The ability of 16E2 Octopus to kill certain tumor cell lines was unexpected; for example, a CNS cancer cell line, SF-295 (FIG. 19B ), as well as two renal cancer cell lines, ACHN and RXF393 (FIG. 19C ). - The results of the NCI tumor panel screens are depicted quantitatively in
FIGS. 20 A and B (2-day results) andFIGS. 21A and B (6-day results) which summarize the effect of 16E2 Octopus compared to Apo2L on growth inhibition (GI50), stasis (TGI), and toxicity (LC50) of the treated tumor cell lines. Again, these results suggest that 16E2 Octopus may be effective against more types of cancer than previously observed. - In an effort to improve the potency of the chimeric anti-CD20 antibody C2B8 (RITUXAN®; U.S. Pat. No. 5,736,137, expressly incorporated herein by reference), one approach being investigated is the ability of the antibody to trigger apoptosis of tumor cells. The apoptosis assay in Koopman et al. Blood 84:1415-1420 (1994) was performed. An Octopus anti-CD20 antibody (OctCD20) was prepared by using the C2B8 VL and VH domains in the preparation of an anti-CD20 Octopus antibody. The OctCD20 antibody was expressed in 293 cells and purified via Protein A sepharose chromatograpy as described for the previous examples.
- As shown in
FIG. 22 , RITUXAN® alone does not trigger much apoptosis of a non-Hodgkins lymphoma B cell line, Wil-2, unless it is crosslinked with anti-human IgG (RITUXAN®-IgG). OctCD20, however, is capable of inducing apoptosis in Wil-2 cells independent of crosslinking. The level of apoptosis observed with OctCD20 is lower than that of crosslinked RITUXAN®, however, suggesting that the apoptotic activity of OctCD20 could be improved, perhaps through the use of the flexible linkers. - Versions of the Octopus antibodies of Example 2 (anti-HER2), Example 3 (anti-DR5) and Example 4 (anti-CD20) with an antibody hing region dimerization domain (designated “Octopus F(ab′)2” herein) were engineered. The anti-HER2 Octopus F(ab′)2 construct was engineered by replacing the Fc region of the heavy chain cDNA with sequence encoding a leucine-zipper motif which, when expressed as protein, dimerizes to effectively join the Octopus Fab arms (
FIG. 23C ). The octopus F(ab′)2 can maintain the leucine zipper motif, or that motif can, e.g., be proteolytically removed as desired. As depicted inFIG. 24 , PCR was used to amplify the duplicate VH/CH1 domains and to insert a restriction site onto the end of the Octopus heavy chain cDNA (NotI) to permit in-frame subcloning into a vector (VG15) containing a leucine-zipper motif. PCR was again utilized to add another restriction site downstream of the heavy chain termination codon (XhoI) to allow subcloning into the pRK vector for expression in mammalian cells. The VH/CH1 domains of anti-DR5Mab16E2 and anti-CD20 Mab C2B8 were substituted into the Oct F(ab)′2 heavy chain backbone using the unique restriction sites BamHI, NheI, and BspEI. - “POPoctopus” antibodies were created by linking together Fab domains in tandem repeats to form linear Fab multimers. “POPoct-3” contains three linked Fab domains (
FIG. 23D ), while “POPoct-4” has four Fab repeats (FIG. 23E ). Anti-HER2 (rhuMab 4D5), anti-DR5 (16E2), and anti-CD20 (C2B8) POPoct-3 constructs were generated, as were anti-HER2 (rhuMab 4D5) and anti-DR5 (16E2) POPoct-4 constructs. POPoct-3 antibodies were engineered both with and withoutflex 1 linkers. -
FIG. 25 depicts the construction of the POPoct-3 heavy chain cDNA. PCR was used to amplify the VH/CH1 domain adding a 5′-BspEI site and a 3′-NotI site. This sequence was digested and along with BamHI/BspEI digested Octopus heavy chain, ligated into a pRK vector to yield an Octopus heavy chain containing sequence for three VH/CH1 domains. The BspEI site encodes for a serine and a glycine residue. - To engineer the POPoct-4 antibody (
FIG. 26 ), site-directed oligomutagenesis was used to introduce a silent mutation, resulting in the elimination of the NheI restriction site in-between the duplicate VH/CH1 domains on the Octopus heavy chain cDNA. Oligomutagenesis was again employed to add a NheI restriction site immediately downstream of the second VH/CH1 sequence. This cDNA along with the POPoct-3 construct were digested with BamHI/NheI restriction endonucleases, and ligated together with the pRK vector to produce a heavy chain cDNA containing sequence for four VH/CH1 domains. - The different Octopus heavy chains were transiently cotransfected with the appropriate light chain cDNAs into 293 mammalian cells to express antibodies containing either three Fab domains (POPoct-3 Fab) or four Fab domains (full-length Octopus; Octopus F(ab)′2; POPoct-4 Fab). While native IgG Mabs and full-length Octopus antibodies were purified over Protein A sepharose, Octopus F(ab)′2 and POPoct-3 and -4 were purified over Protein G sepharose columns.
- The Octopus F(ab)′2 is approximately 200 kDa (
FIG. 23F , lane 4), smaller than the 240 kDa of the full-length Octopus antibody (FIG. 23F , lane 3), but larger than the 150 kDa native IgG Mab (FIG. 23F ,lanes 1 and 2). At approximately 140 kDa (FIG. 23F , lane 5), POPoct-3 is slightly smaller than native IgG Mab, while POPoct-4 is slightly larger at 190 kDa. The heavy chain of the Octopus F(ab)′2 (FIG. 23G , lane 4) is approximately the same size as the native IgG Mab heavy chain (FIG. 23G ,lanes 1 and 2) at 55 kDa. The POPoct-3 heavy chain (FIG. 23G , lane 5) is similar in size to the full-length Octopus heavy chain (FIG. 23G , lane 3), while at approximately 97 kDa the POPoct-4 has the largest heavy chain - Antiproliferation Assays OctHER F(ab)′2, POPoct-3 HER2, OctHER2,
OctHER2 flex 1, and rhuMAb 4D5 (HERCEPTIN®) were added to the 3+HER2 over-expressing tumor cell line, BT474, at equimolar concentrations and evaluated for their ability to inhibit cell growth as measured by crystal violet staining. The results of these assays are shown inFIG. 27 . Although all of the antibodies induced some cytostasis of the BT474 cells, POPoct-3HER2 and rhuMAb 4D5 showed the most efficacy and inhibited growth equivalently, while OctHER2F(ab)′2 lost potency rapidly as its concentration decreased. OctHER2 flex1 demonstrated a slight but consistent improvement over OctHER2 (n=6), suggesting that improved flexibility may result in better access of the Fab to the HER2 target. - OctHER2, OctHER2 flex-1, POPoct-3HER2, POPoct-3HER2 flex-1 and rhuMAb 4D5 (HERCEPTIN®) were also evaluated at equimolar concentrations on another 3+HER2 over-expressing cell line, SKBR3, in crystal violet cytostasis assays. The results of this assay are depicted in
FIG. 28 . On this cell line, all Octopus constructs tested inhibited cell growth equivalently, and better than rhuMab 4D5 (n=4). Any improvement in efficacy due to the flexible-linker in between the Fab arms of OctHER2 or POPoct-3 was less evident on this cell line. - Internalization Assays: POPoct-3HER2 was compared to OctHER and HERCEPTIN® in internalization assays on two 3+HER2 over-expressing tumor cell lines, SKBR3 and BT474, to assess its candidacy for applications in immunotoxin therapies. Although structurally different than the full-length OctHER2 antibody, POPoct-3HER2 was internalized and catabolized identically to OctHER2 by both cell lines (
FIGS. 29A and B) and at twice the rate of HERCEPTIN®. - Apoptosis assays: Multivalent versions of the anti-DR516E2 MAb were evaluated in this example. Oct1DR5, OctDR5flex-1, OctDR5F(ab)′2, POPoct-3DR5, POPoct-3DR5flex-1 and POPoct-4 DR5 were added at equimolar concentrations to the colon tumor cell line COLO205 and analyzed in crystal violet apoptosis assays in comparison to the 16E2 MAb (n=4). The results are shown in
FIGS. 30A and B. All Octopus antibodies induced more apoptosis than the 16E2 MAb, with the order of efficacy from most potent to least: OctDR5flex-1>OctDR5=POPoct-4 DR5=POPoct-3flex-1 DR5=POPoct-3DR5 >OctDR5F(ab)′2>16E2 MAb. OctDR5flex-1 showed increased potency compared to OctDR5, especially at lower concentrations (FIG. 30A ), indicating that flexibility between the Fab arms improves efficacy. POPoct-3flex-1DR5 induced equivalent levels of apoptosis as OctHER (FIG. 30A ) and showed similar efficacy to POPoct16-3 and POPoct16-4 (FIG. 30B ). - Cell signaling: Apo2L binds to the death receptors and triggers cellular apoptosis through the caspase signaling pathway. As shown in
FIGS. 31A and B, the anti-DR5 Octopus antibodies were shown to induce apoptosis through the same signaling pathway as Apo2L. Oct16E2 triggered similar levels of apoptosis as APO2L on the lung tumor cell line SK-MES-1 (FIG. 31A , dashed lines), but after the addition of ZVAD, an inhibitor ofcaspase FIG. 31B solid lines). Further evidence that the anti-DR5 Octopus antibodies signaled through the same pathway as Apo2L was obtained by DISC (Death Induced Signaling Complex) analyses (FIG. 31B ). BJAB cells, a B-cell lymphoma line that expresses DR5, was incubated at two different concentrations of two anti-DR5 Octopus antibodies, Oct16E2 and Oct3H3, for varying times. Purification of the antibody-DR5 complexes was followed Western blot analysis to identify the signaling molecules that copurified with the complexes. As with Apo2L, the signaling molecules caspase 8 and FADD associated with DR5 after the receptor was bound by both Oct16E2 and Oct3H3 (FIG. 31B ). - Apoptosis assays: As shown in
FIG. 22 , RITUXAN® did not efficiently trigger apoptosis in vitro on the B-cell lymphoma cell line WIL-2 unless first crosslinked by anti-IgG antibody. OctCD20 was capable of inducing apoptosis of WIL-2 cells independent of crosslinking, at levels higher than RITUXAN® alone, yet slightly lower than anti-IgG-crosslinked RITUXAN®. When crosslinked with anti-IgG antibody, OctCD20 induced more apoptosis of the WIL-2 cells than crosslinked RITUXAN® (FIG. 32 ). Since one potential explanation for the efficacy of RITUXAN® in vivo is that the antibody is being crosslinked by either complement or FcγR bearing cells, this observation suggests that OctCD20 will be even more efficacious in vivo. - OctCD20 F(ab)′2, POPoct-3CD20 and POPoct-3CD20flex-1 were tested at various concentrations in apoptosis assays with WIL-2 cells, and the optimal doses are shown in the maximum response curves in
FIG. 33 . The Octopus antibodies were compared to the anti-CD20 antibody 1F5 (Clark et al. supra), which functions similar to RITUXAN® in that it does not induce apoptosis unless crosslinked with anti-IgG antibody. Both Octopus antibodies tested induced either similar (OctCD20 F(ab)′2) or higher (POPoct-3CD20, POPoct-3CD20flex-1) levels of apoptosis than crosslinked IF5 anti-CD20. Additionally, the Octopus antibodies were efficacious at considerably lower concentrations than the crosslinked anti-CD20. - When crosslinked anti-CD20 antibodies are added to the B cell lymphoma line WIL-2S, a homotypic adhesion of the cells is observed. This cell clumping is one indication that the cells have been activated through CD20. The Octopus anti-CD20 antibodies induce this same homotypic adhesion phenomenon independent of crosslinker, and as shown in
FIG. 34 with POPoct-3CD20, at much lower concentrations than crosslinked IF5 anti-CD20. - Apoptosis induction by the various anti-CD20 antibodies was further assessed using blood from a patient with chronic lymphocytic leukemia (CLL). PBL's were separated out using dextran sedimentation, washed and plated in serum-free lymphocyte medium treated overnight with no sample, 1F5 (20 μg/ml), 1F5+cross-linking mouse anti-IgG (100 μg/ml), OctCD20 F(ab′)2 at approx 0.5 or 1.0 μg/mland POPoct-3 CD20 at 0.5 μg/ml.
- An apoptosis assay was performed using annexin and PI staining. The percentage of apoptotic cells were:
-
Untreated 38.5% 1F5 37.1% 1F5 X-linked with anti-IgG 25.1% POPoct-3 CD20 (0.5 μg) 50.2% OctCD20 F(ab′)2 (0.5 μg) 37.7% OctCD20 F(ab′)2 (1.0 μg) 48.6% - The data indicate that multivalent anti-CD20 antibodies (especially POPoct-3 CD20) enhance apoptosis in a dose-dependent manner.
- Internalization Assays: OctCD20 was also evaluated as a candidate for immunotoxin therapy in internalization assays on three B-cell lymphoma lines, DB, WIL-2, and Ramos, and compared to RITUXAN®. As shown in
FIG. 35 , twice as much OctCD20 was internalized by the cells as compared to RITUXAN®, which was not internalized by the cells at appreciable levels. The higher avidity that would be expected for the multivalent antibodies due to the increased number of binding sites is evident in the fact that more OctCD20 remains bound to the cell surface of the cells over time as compared to RITUXAN®.
Claims (38)
1.-93. (canceled)
94. A binding protein comprising four polypeptide chains, wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, C is a heavy chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 is an Fc region; and two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region; and n is 0 or 1; wherein said four polypeptide chains of said binding protein form four functional antigen binding sites.
95. The binding protein according to claim 94 , wherein said binding protein is capable of binding one or more targets.
96. The binding protein according to claim 95 , wherein said one or more targets is selected from the group consisting of ABCF1; ACVR1; ACVR1B; ACVR2; ACVR2B; ACVRL1; ADORA2A; Aggrecan; AGR2; AICDA; AIF1; AIG1; AKAP1; AKAP2; AMH; AMHR2; ANGPT1; ANGPT2; ANGPTL3; ANGPTL4; ANPEP; APC; APOC1; AR; AZGP1 (zinc-a-glycoprotein); B7.1; B7.2; BAD; BAFF; BAG1; BAI1; BCL2; BCL6; BDNF; BLNK; BLR1 (MDR15); BlyS; BMP1; BMP2; BMP3B (GDF10); BMP4; BMP6; BMP8; BMPR1A; BMPR1B; BMPR2; BPAG1 (plectin); BRCA1; C19orf10 (IL27w); C3; C4A; C5; C5R1; CANT1; CASP1; CASP4; CAV1; CCBP2 (D6/JAB61); CCL1 (1-309); CCL11 (eotaxin); CCL13 (MCP-4); CCL15 (MT-1d); CCL16 (HCC-4); CCL17 (TARC); CCL18 (PARC); CCL19 (MIP-3b); CCL2 (MCP-1); MCAF; CCL20 (MIP-3a); CCL21 (MIP-2); SLC; exodus-2; CCL22 (MDC/STC-1); CCL23 (MPIF-1); CCL24 (MPIF-2/eotaxin-2); CCL25 (TECK); CCL26 (eotaxin-3); CCL27 (CTACK/ILC); CCL28; CCL3 (MT-1a); CCL4 (MIP-1b); CCL5 (RANTES); CCL7 (MCP-3); CCL8 (mcp-2); CCNA1; CCNA2; CCND1; CCNE1; CCNE2; CCR1 (CKR1/HM145); CCR2 (mcp-1RB/RA); CCR3 (CKR3/CMKBR3); CCR4; CCR5 (CMKBRS/ChemR13); CCR6 (CMKBR6/CKR-L3/STRL22/DRY6); CCR7 (CKR7/EBI1); CCR8 (CMKBR8/TER1/CKR-L1); CCR9 (GPR-9-6); CCRL1 (VSHK1); CCRL2 (L-CCR); CD164; CD19; CD1C; CD20; CD200; CD-22; CD24; CD28; CD3; CD37; CD38; CD3E; CD3G; CD3Z; CD4; CD40; CD40L; CD44; CD45RB; CD52; CD69; CD72; CD74; CD79A; CD79B; CD8; CD80; CD81; CD83; CD86; CDH1 (E-cadherin); CDH10; CDH12; CDH13; CDH18; CDH19; CDH20; CDH5; CDH7; CDH8; CDH9; CDK2; CDK3; CDK4; CDK5; CDK6; CDK7; CDK9; CDKN1A (p21Wap1/Cip1); CDKN1B (p27Kip1); CDKN1C; CDKN2A (p161NK4a); CDKN2B; CDKN2C; CDKN3; CEBPB; CER1; CHGA; CHGB; Chitinase; CHST10; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (claudin-7); CLN3; CLU (clusterin); CMKLR1; CMKOR1 (RDC1); CNR1; COL18A1; COL1A1; COL4A3; COL6A1; CR2; CRP; CSF1 (M-CSF); CSF2 (GM-CSF); CSF3 (GCSF); CTLA4; CTNNB1 (b-catenin); CTSB (cathepsin B); CX3CL1 (SCYD1); CX3CR1 (V28); CXCL1 (GRO1); CXCL10(IP-10); CXCL11 (1-TAC/IP-9); CXCL12 (SDF1); CXCL13; CXCL14; CXCL16; CXCL2 (GRO2); CXCL3 (GRO3); CXCL5 (ENA-78/LIX); CXCL6 (GCP-2); CXCL9 (MIG); CXCR3 (GPR9/CKR-L2); CXCR4; CXCR6 (TYMSTR/STRL33/Bonzo); CYB5; CYC1; CYSLTR1; DAB21P; DES; DKFZp451J0118; DNCL1; DPP4; E2F1; ECGF1; EDG1; EFNA1; EFNA3; EFNB2; EGF; EGFR; ELAC2; ENG; ENO1; ENO2; ENO3; EPHB4; EPO; ERBB2 (Her-2); EREG; ERK8; ESR1; ESR2; F3 (TF); FADD; FasL; FASN; FCER1A; FCER2; FCGR3A; FGF; FGF1 (aFGF); FGF10; FGF11; FGF12; FGF12B; FGF13; FGF14; FGF16; FGF17; FGF18; FGF19; FGF2 (bFGF); FGF20; FGF21; FGF22; FGF23; FGF3 (int-2); FGF4 (HST); FGF5; FGF6 (HST-2); FGF7 (KGF); FGF8; FGF9; FGFR3; FIGF (VEGFD); FIL1 (EPSILON); FIL1 (ZETA); FLJ12584; FLJ25530; FLRT1 (fibronectin); FLT1; FOS; FOSL1 (FRA-1); FY (DARC); GABRP (GABAa); GAGEB1; GAGEC1; GALNAC4S-6ST; GATA3; GDF5; GFI1; GGT1; GM-CSF; GNAS1; GNRH1; GPR2 (CCR10); GPR31; GPR44; GPR81 (FKSG80); GRCC10 (C10); GRP; GSN (Gelsolin); GSTP1; HAVCR2; HDAC4; HDAC5; HDAC7A; HDAC9; HGF; HIF1A; HIP1; histamine and histamine receptors; HLA-A; HLA-DRA; HM74; HMOX1; HUMCYT2A; ICEBERG; ICOSL; ID2; IFN-a; IFNA1; IFNA2; IFNA4; IFNA5; IFNA6; IFNA7; IFNB1; IFNgamma; IFNW1; IGBP1; IGF1; IGF1R; IGF2; IGFBP2; IGFBP3; IGFBP6; IL-1; IL10; IL10RA; IL10RB; IL11; IL11RA; IL-12; IL12A; IL12B; IL12RB1; IL12RB2; IL13; IL13RA1; IL13RA2; IL14; IL15; IL15RA; IL16; IL17; IL17B; IL17C; IL17R; IL18; IL18BP; IL18R1; IL18RAP; IL19; ILIA; IL1B; IL1F10; IL1F5; IL1F6; IL1F7; IL1F8; IL1F9; IL1HY1; EMU; IL1R2; IL1RAP; IL1RAPL1; IL1RAPL2; IL1RL1; IL1RL2; IL1RN; IL2; 1L20; IL20RA; IL21R; IL22; IL22R; IL22RA2; IL23; IL24; IL25; IL26; IL27; IL28A; IL28B; IL29; IL2RA; IL2RB; IL2RG; IL3; IL30; IL3RA; IL4; IL4R; IL5; IL5RA; IL6; IL6R; IL6ST (glycoprotein 130); IL7; IL7R; IL8; IL8RA; IL8RB; IL8RB; IL9; IL9R; ILK; INHA; INHBA; INSL3; INSL4; IRAK1; IRAK2; ITGA1; ITGA2; ITGA3; ITGA6 (a6 integrin); ITGAV; ITGB3; ITGB4 (b 4 integrin); JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC Box BP); KLF6; KLK10; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19 (Keratin 19); KRT2A; KRTHB6 (hair-specific type II keratin); LAMAS; LEP (leptin); Lingo-p75; Lingo-Troy; LPS; LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTBR; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK; MIB1; midkine; MIF; MIP-2; MKI67 (Ki-67); MMP2; MMP9; MS4A1; MSMB; MT3 (metallothionectin-III); MTS S1; MUC1 (mucin); MYC; MYD88; NCK2; neurocan; NFKB1; NFKB2; NGFB (NGF); NGFR; NgR-Lingo; NgR-Nogo66 (Nogo); NgR-p75; NgR-Troy; NME1 (NM23A); NOX5; NPPB; NROB1; NROB2; NR1D1; NR1D2; NR1H2; NR1H3; NR1H4; NRII2; NRII3; NR2C1; NR2C2; NR2E1; NR2E3; NR2F1; NR2F2; NR2F6; NR3C1; NR3C2; NR4A1; NR4A2; NR4A3; NR5A1; NR5A2; NR6A1; NRP1; NRP2; NT5E; NTN4; ODZ1; OPRD1; P2RX7; PAP; PART1; PATE; PAWR; PCA3; PCNA; PDGFA; PDGFB; PECAM1; PF4 (CXCL4); PGF; PGR; phosphacan; PIAS2; PIK3CG; PLAU (uPA); PLG; PLXDC1; PPBP (CXCL7); PPID; PR1; PRKCQ; PRKD1; PRL; PROC; PROK2; PSAP; PSCA; PTAFR; PTEN; PTGS2 (COX-2); PTN; RAC2 (p21Rac2); RARB; RGS1; RGS13; RGS3; RNF110 (ZNF144); ROBO2; SI00A2; SCGB1D2 (lipophilin B); SCGB2A1 (mammaglobin 2); SCGB2A2 (mammaglobin 1); SCYE1 (endothelial Monocyte-activating cytokine); SDF2; SERPINA1; SERPINA3; SERPINB5 (maspin); SERPINE1 (PAI-1); SERPINF1; SHBG; SLA2; SLC2A2; SLC33A1; SLC43A1; SLIT2; SPP1; SPRR1B (Sprl); ST6GAL1; STAB1; STATE; STEAP; STEAP2; TB4R2; TBX21; TCP10; TDGF1; TEK; TGFA; TGFB1; TGFB111; TGFB2; TGFB3; TGFBI; TGFBR1; TGFBR2; TGFBR3; TH1L; THBS1 (thrombospondin-1); THBS2; THBS4; THPO; TIE (Tie-1); TIMP3; tissue factor; TLR10; TLR2; TLR3; TLR4; TLR5; TLR6; TLR7; TLR8; TLR9; TNF; TNF-a; TNFAIP2 (B94); TNFAIP3; TNFRSF11A; TNFRSF1A; TNFRSF1B; TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF8; TNFRSF9; TNFSF10 (TRAIL); TNFSF11 (TRANCE); TNFSF12 (APO3L); TNFSF13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF15 (VEGI); TNFSF18; TNFSF4 (OX40 ligand); TNFSF5 (CD40 ligand); TNFSF6 (FasL); TNFSF7 (CD27 ligand); TNFSF8 (CD30 ligand); TNFSF9 (4-1BB ligand); TOLLIP; Toll-like receptors; TOP2A (topoisomerase Iia); TP53; TPM1; TPM2; TRADD; TRAF1; TRAF2; TRAF3; TRAF4; TRAF5; TRAF6; TREM1; TREM2; TRPC6; TSLP; TWEAK; VEGF; VEGFB; VEGFC; versican; VHL C5; VLA-4; XCL1 (lymphotactin); XCL2 (SCM-1b); XCR1 (GPR5/CCXCR1); YY1; and ZFPM2.
97. The binding protein according to claim 94 , wherein said binding protein is capable of binding a two targets, wherein the two targets are selected from the group consisting of CD138 and CD20; CD138 and CD40; CD20 and CD3; CD38 & CD138; CD38 and CD20; CD38 and CD40; CD40 and CD20; CD19 and CD20; CD-8 and IL-6; PDL-1 and CTLA-4; CTLA-4 and BTNO2; CSPGs and RGM A; IGF1 and IGF2; IGF1/2 and Erb2B; IL-12 and IL-18; IL-12 and TWEAK; IL-13 and ADAMS; IL-13 and CL25; IL-13 and IL-1beta; IL-13 and IL-25; IL-13 and IL-4; IL-13 and IL-5; IL-13 and IL-9; IL-13 and LHR agonist; IL-13 and MDC; IL-13 and MIF; IL-13 and PED2; IL-13 and SPRR2a; IL-13 and SPRR2b; IL-13 and TARC; IL-13 and TGF-β; IL-1α and IL-1β; MAG and RGM A; NgR and RGM A; NogoA and RGM A; OMGp and RGM A; RGM A and RGM B; Te38 and TNFα; TNFα and IL-12; TNFα and IL-12p40; TNFα and IL-13; TNFα and IL-15; TNFα and IL-17; TNFα and IL-18; TNFα and IL-1beta; TNFα and IL-23; TNFα and MIF; TNFα and PEG2; TNFα and PGE4; TNFα and VEGF; and VEGFR and EGFR; TNFα and RANK ligand; TNFα and Blys; TNFα and GP130; TNFα and CD-22; and TNFα and CTLA-4.
98. The binding protein according to claim 95 , wherein the binding protein is capable of modulating a biological function of one or more targets.
99. The binding protein according to claim 95 , wherein the binding protein is capable of neutralizing one or more targets.
100. The binding protein according to claim 95 , wherein said one or more targets is selected from the group consisting of cytokine, chemokine, cell surface protein, enzyme and receptor.
101. The binding protein according to claim 100 , wherein the cytokine is selected from the group consisting of lymphokines, monokines, and polypeptide hormones.
102. The binding protein according to claim 101 , wherein said cytokines are IL-1α and IL-1β.
103. The binding protein according to claim 101 , wherein said cytokines are TNF-α and IL-13.
104. The binding protein according to claim 101 , wherein said cytokines are IL-12 and IL-18.
105. The binding protein according to claim 100 , wherein the cell surface protein is an integrin.
106. The binding protein according to claim 100 , wherein the cell surface proteins are CD-20 and CD3.
107. The binding protein according to claim 100 , wherein the enzyme is selected from the group consisting of kinases and proteases.
108. The binding protein according to claim 100 , wherein the receptor is selected from the group consisting of lymphokine receptor, monokine receptor, and polypeptide hormone receptor.
109. A binding protein conjugate comprising a binding protein described in any one of claims 94 -101 and 104, said binding protein conjugate further comprising an agent selected from the group consisting of; an immunoadhesion molecule, an imaging agent, a therapeutic agent, and a cytotoxic agent.
110. The binding protein conjugate according to claim 109 , wherein said agent is an imaging agent selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
111. The binding protein conjugate according to claim 110 , wherein said imaging agent is a radiolabel selected from the group consisting of: 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, and 153Sm.
112. The binding protein conjugate according to claim 109 , wherein said agent is a therapeutic or cytotoxic agent selected from the group consisting of; an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, toxin, and an apoptotic agent.
113. A binding protein described in claim 94 produced according to a method comprising culturing a host cell in culture medium under conditions sufficient to produce said binding protein, wherein said host cell comprises a vector, said vector comprising a nucleic acid encoding said binding protein.
114. A pharmaceutical composition comprising a binding protein of any one of claims 94 -101, 104 and 113, and a pharmaceutically acceptable carrier.
115. The pharmaceutical composition of claim 114 , further comprising at least one additional therapeutic agent.
116. The pharmaceutical composition according to claim 115 , wherein said additional agent is a therapeutic or imaging agent.
117. The pharmaceutical composition of claim 116 , wherein said additional agent is selected from the group consisting of: Therapeutic agent, imaging agent, cytotoxic agent, angiogenesis inhibitors; kinase inhibitors; co-stimulation molecule blockers; adhesion molecule blockers; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporin; rapamycin; FK506; detectable label or reporter; a TNF antagonist; an antirheumatic; a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteriod, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.
118. A pharmaceutical composition comprising a binding protein conjugate according to claim 109 and a pharmaceutically acceptable carrier.
119. The pharmaceutical composition according to claim 118 , wherein said binding protein conjugate comprises an imaging agent selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.
120. The pharmaceutical composition according to claim 119 , wherein said imaging agent is a radiolabel selected from the group consisting of: 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, and 153Sm.
121. The pharmaceutical composition according to claim 118 , wherein said binding protein conjugate comprises a therapeutic or cytotoxic agent selected from the group consisting of an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, a toxin, and an apoptotic agent.
122. The pharmaceutical composition of claim 118 further comprising a second agent.
123. The pharmaceutical composition of claim 122 , wherein said second agent is a therapeutic or imaging agent.
124. The pharmaceutical composition of claim 123 , wherein said therapeutic or imaging agent is selected from the group: cytotoxic agent, angiogenesis inhibitors, kinase inhibitors; co-stimulation molecule blockers; adhesion molecule blockers; anti-cytokine antibody or functional fragment thereof; methotrexate; cyclosporin; rapamycin; FK506; detectable label or reporter; a TNF antagonist; an antiheumatic; a muscle relaxant, a narcotic, anon-steroid anti-inflammatory dug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteriod, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, 153Sm, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, biotin, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.
125. An isolated antibody comprising four polypeptide chains, wherein two polypeptide chains comprise VD1-(X1)n-VD2-(X2)n-Fc, wherein VD1 is a first heavy chain variable domain, VD2 is a second heavy chain variable domain, X1 is a linker with the proviso that it is not CH1, and X2 is a heavy chain constant domain; and two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, wherein VD1 is a first light chain variable domain, VD2 is a second light chain variable domain, C is a light chain constant domain, X1 is a linker with the proviso that it is not CH1, and X2 does not comprise an Fc region; and n is 0 or 1; wherein said four polypeptide chains of said binding protein form four functional antigen binding sites.
126. The binding protein according to claim 125 , wherein said binding protein is capable of binding one or more targets.
127. The isolated antibody according to claim 126 , wherein said one or more targets is selected from the group consisting of BDNF; CCL3 (MIP-1a); CCL5 (RANTES); CD19; CD1c; CD20; CD22; CD24; CD28; CD3; CD37; CD38; CD4; CD40; CD44; CD52; CD69; CD72; CD74; CD79A; CD79B; C D8; CD80; CD81; CD83; CD86; CSF1 (M-CSF); CSF2 (MG-CSF); CSF3 (G-CSF); CTLA-4; EGF; EGFR; EPO; ErbB2 (Her-2); FGFI (aFGF); FGF2 (bFGF); GM-CSF; IFN-a; IFN-gamma; IGF-I; IGF-H; IL-1; IL-2; IL-3; IL-4; IL-5; IL6; IL-7; IL-8; IL-9; IL-10; NGFB; TGF-alpha; TGF-beta1; TGF-beta2; TGF-beta3; TGF-beta4; TNF; TNF-alpha; and VEGF.
128. The isolated antibody according to claim 126 , wherein said one or more targets is selected from the group consisting of ABCFI; ACVRI; ACVRIB; ACVR2; ACVR2B; ACVRL1; ADORA2A; Aggrecan; AGR2; AICDA; AIFI; AlGI; AKAPI; AKAP2; AMH; AMHR2; ANGPT1; ANGPT2; ANGPTL3; ANGPTL4; ANPEP; APC; APOCI; AR;AZGPI (zinc-a-glycoprotein); B7.1; B7.2; BAD; BAFF; BAGI; BAH; BCL2; BCL6; BLNK; BLR1 (MDR15); BlyS; BMPI; BMP2; BMP3B (GDFIO); BMP4; BMP6; BMP8; BMPRIA; BMPRIB; BMPR2; BPAGI (plectin); BRCAI; CI9orflO (IL27w); C3; C4A; C5; C5R1; CANT!; CASPI; CASP4; CAVI; CCBP2 (D6/JAB61); CCL1 (1-309); CCM (eotaxin); CCL13 (MCP-4); CCL15 (MIP-Id); CCL16 (HCC-4); CCL17 (TARC); CCL18 (PARC); CCL19 (MIP3b); CCL2 (MCP-I); MCAF; CCL20 (MIP-3a); CCL21 (MIP-2); SLC; exodus-2; CCL22 (MDC/STC-I); CCL23 (MPIF-I); CCL24 (MPIF-2/eotaxin-2); CCL25 (TECK); CCL26 (eotaxin-3); CCL27 (CTACK/ILC); CCL28; CCL4 (MIP-Ib); CCL7 (MCPS); CCL8 (mcp-2); CCNAI; CCNA2; CCND1; CCNEI; CCNE2; CCR1 (CKR1/HMI45); CCR2 (mcp-IRB/RA); CCR3 (CKR3/CMKBR3); CCR4; CCR5 (CMKBR5/ChemR13); CCR6 (CMKBR6/CKR-L3/STRL22/DRY6); CCR7 (CKR7/EB11); CCR8 (CMKBR8/TERI!CKR-L1); CCR9 (GPR-9-6); CCRL1 (VSHKI); CCRU (L-CCR); CD164; CD200; CD3E; CD3G; CD3Z; CD45RB; CDHI (E-cadherin); CDHIO; CDHI2; CDH13; CDHI8; CDHI9; CDH20; CDH5; CDH7; CDH8; CDH9; CDK2; CDK3;CDK4; CDK5; CDK6; CDK7; CDK9; CDKNIA (p21WapI/Cip1); CDKNIB (p27Kip1); CDKNIC; CDKN2A (p16INK4a); CDKN2B; CDKN2C; CDKN3; CEBPB; CERI; CHGA; CHGB; Chitinase; CHST!O; CKLFSF2; CKLFSF3; CKLFSF4; CKLFSF5; CKLFSF6; CKLFSF7; CKLFSF8; CLDN3; CLDN7 (claudin-7); CLN3; CLU (clusterin); CMKLRI; CMKORI (RDCI); CNRI; COL18AI; COL1AI; COL4A3; COL6AI; CR2; CRP; CTNNBI (b-catenin); CTSB (cathepsin B); CX3CL1 (SCYD1); CX3CRI (V28); CXCL1 (GRO1); CXCL10(IP-10); CXCL1I (1-TAC/IP-9); CXCL12 (SDFI); CXCL13; CXCL14; CXCL16; CXCL2 (GRO2); CXCL3 (GR03); CXCL5 (ENA-78/LIX); CXCL6 (GCP-2); CXCL9 (MIG); CXCR3 (GPR9/CKRL2); CXCR4; CXCR6 (TYMSTR/STRL33/Bonzo); CYB5; CYCI; CYSLTRI; DAB2IP; DES; DKFZp45IJO118; DNCL1; DPP4; E2FI; ECGFI; EDGI; EFNAI; EFNA3; EFNB2; ELAC2; ENG; ENO1; EN02; EN03; EPHB4; EREG; ERK8; ESRI; ESR2; F3 (TF); FADD; FasL; FASN; FCER1A; FCER2; FCGR3A; FGF; FGF1O; FGF11; FGF12; FGF12B; FGF13; FGF14; FGF16; FGF17; FGF18; FGF19; FGF20; FGF21; FGF22; FGF23; FGF3 (int-2); FGF4 (HST); FGF5; FGF6 (HST-2); FGF7 (KGF); FGF8; FGF9; FGFR3; FIGF (VEGFD); FILL (EPSILON); FIL1 (ZETA); FLJ12584; FLJ25530; FLRT1 (fibronectin); FLT1; FOS; FOSL1 (FRA-1); FY (DARC); GABRP (GABAa); GAGEB1; GAGEC1; GALNAC4S-6ST; GATA3; GDF5; GFI1; GGT1; GNAS1; GNRH1; GPR2 (CCR10); GPR31; GPR44; GPR81 (FKSG80); GRCC10(C10); GRP; GSN (Gelsolin); GSTP1; HAVCR2; HDAC4; HDAC5; HDAC7A; HDAC9; HGF; HIF1A; HIFI; histamine and histamine receptors; HLA-A; HLA-DRA; HM74; HMOX1; HUMCYT2A; ICEBERG; ICOSL; ID2; IFNA1; IFNA2; IFNA4; IFNA5; IFNA6; IFNA7; IFNB1; IFNW1; IGBP1; IGF1R; IGF2; IGFBP2; IGFBP3; IGFBP6; IL1 ORA; IL1 ORB; IL11; IL11RA; IL-12; IL12A; IL12B; IL12RB1; IL12RB2; IL13; IL13RA1; IL13RA2; IL14; IL15; IL15RA; IL16; IL17; IL17B; IL17C; IL17R; IL18; IL18BP; IL18R1; IL18RAP; IL19; ILIA; IL1B; IL1F10; IL1F5; IL1F6; IL1F7; IL1F8; IL1F9; IL1HY1; IL1R1; IL1R2; IL1RAP; IL1RAPL1; IL1RAPL2; IL1RL1; IL1RL2; IL1RN; IL20; IL20RA; IL21R; IL22; 1L22R; IL22RA2; IL23; IL24; IU5; IL26; 1L27; IL28A; IL28B; IL29; IURA; IURB; IL2RG; IL30; IL3RA; IL4R; IL5RA; IL6R; IL6ST (glycoprotein 130); IL7R; ILSRA; ILSRB; ILSRB; IL9R; ILK; INHA; INHBA; INSL3; INSL4; IRAK1; IRAK2; ITGA1; ITGA2; ITGA3; ITGA6 (a6 integrin); ITGAV; ITGB3; ITGB4 (b 4 integrin); JAG1; JAK1; JAK3; JUN; K6HF; KAI1; KDR; KITLG; KLF5 (GC Box BP); KLF6; KLKIO; KLK12; KLK13; KLK14; KLK15; KLK3; KLK4; KLK5; KLK6; KLK9; KRT1; KRT19 (Keratin 19); KRT2A; KRTHB6 (hair-specific type II keratin); LAMAS; LEP (leptin); Lingo-p75; Lingo-Troy; LPS; LTA (TNF-b); LTB; LTB4R (GPR16); LTB4R2; LTBR; MACMARCKS; MAG or Omgp; MAP2K7 (c-Jun); MDK; MEM; midkine; MIF; MIP-2; MKI67 (Ki-67); MMP2; MMP9; MS4A1; MSMB; MT3 (metallothionectin-III); MTSS1; MUCI (mucin); MYC; MYD88; NCK2; neurocan; NFKB1; NFKB2; NGFR; NgR-Lingo; NgR-Nog066 (Nogo); NgR-p75; NgR-Troy; NMEI (NM23A); NOX5; NPPB; NROB1; NROB2; NRID1; NRID2; NRIH2; NRIH3; NRIH4; NRII2; NRII3; NR2C1; NR2C2; NR2E1; NR2E3; NR2F1; NR2F2; NR2F6; NR3C1; NR3C2; NR4A1; NR4A2; NR4A3; NR5A1; NR5A2; NR6A1; NRP1; NRP2; NT5E; NTN4; ODZ1; OPRD1; P2RX7; PAP; PART1; PATE; PAWR; PCA3; PCNA; PDGFA; PDGFB; PECAM1; PF4 (CXCL4); PGF; PGR; phosphacan; PIAS2; PIK3CG; PLAU (uPA); PLG; PLXDC1; PPBP(CXCL7); PPID; PR1; PRKCQ; PRKD1; PRL; PROC; PROK2; PSAP; PSCA; PTAFR; PTEN; PTGS2 (COX-2); PTN; RAC2 (p21Rac2); RARB; RGS1; RGS13; RGS3; RNF11O (ZNF144); ROB02; SIOOA2; SCGBID2 (lipophilin B); SCGB2A1 (mammaglobin 2); SCGB2A2 (mammaglobin 1); SCYEI (endothelial Monocyte-activating cytokine); SDF2; SERPINA1; SERPINA3; SERPINB5 (maspin); SERPINEI (PA1-I); SERPINF1; SHBG; SLA2; SLC2A2; SLC33A1; SLC43A1; SLIT2; SPP1; SPRRIB (Spr1); ST6GAL1; STAB1; STAT6; STEAP; STEAP2; TB4R2; TBX21; TCPIO; TDGF1; TEK; TGFBR1; TGFBR2; TGFBR3; TH1L; THBSI (thrombospondin-1); THBS2; THBS4; THPO; TIE (Tie-I); TIMP3; tissue factor; TLR10; TLR2; TLR3; TLR4; TLR5; TLR6; TLR7; TLR8; TLR9; TNF; TNF-ALPHAIP2 (B94); TNF-ALPHAIP3; TNFRSF11A; TNFRSF1A; TNFRSF1B; TNFRSF21; TNFRSF5; TNFRSF6 (Fas); TNFRSF7; TNFRSF8; TNFRSF9; TNFSF1O (TRAIL); TNFSF11 (TRANCE); TNFSF12 (AP03L); TNFSF13 (April); TNFSF13B; TNFSF14 (HVEM-L); TNFSF15 (VEGI); TNFSF18; TNFSF4 (OX40 ligand); TNFSF5 (CD40 ligand); TNFSF6 (FasL); TNFSF7 (C027 ligand); TNFSF8 (CmO ligand); TNFSF9 (4-1BB ligand); TOLLIP; Toll-like receptors; TOP2A (topoisomerase Iia); TP53; TPM1; TPM2; TRADD; TRAF1; TRAF2; TRAF3; TRAF4; TRAF5; TRAF6; TREM1; TREM2; TRPC6; TSLP; TWEAK; VEGF; VEGFB; VEGFC; versican; VHL C5; VLA-4; XCL1 (lymphotactin); XCL2 (SCM-1b); XCR1 (GPR5/CCXCR1); YY1; and ZFPM2.
129. The binding protein according to claim 125 , wherein said binding protein is capable of binding a two targets, wherein the two targets are selected from the group consisting of CD20 and CD3; CD38 and CD20; CD38 and CD40; CD40 and CD20; CD19 and CD20; CD8 and IL-6; IGF-I and IGF-II; IGF-I/II and Erb2B; TNF-alpha and IL-1beta; TNF-alpha and MIF; TNF-alpha and VEGF; TNF-alpha and CTLA-4.
130. The binding protein according to claim 125 , wherein said binding protein is capable of binding a two targets, wherein the two targets are selected from the group consisting of CD138 and CD20; CD138 and CD40; CD38 and CD138; PDL-1 and CTLA4;CTLA-4 and BTN02; CSPGs and RGM A; IL-12 and IL-18; IL-12 and TWEAK; IL-13 and ADAM8; IL-13 and CL25; IL-13 and IL-1beta; IL-13 and IL-25; IL-13 and IL-4; IL-13 and IL-5; IL-13 and IL-9; IL-13 and LHR agonist; IL-13 and MDC; IL-13 and MIF; IL-13 and PED2; IL-13 and SPRR2a; IL-13 and SPRR2b; IL-13 and TARC; IL-13 and TGF-beta; IL-1 alpha and IL-1 beta; MAG and RGM A; NgR and RGM A; NogoA and RGMA; OMGp and RGMA; RGMA and RGM B; Te38 and TNF-alpha; TNF-alpha and IL-12; TNF-alpha and IL-12p40; TNF-alpha and IL-13; TNF-alpha and IL-15; TNF-alpha and IL-17; TNF-alpha and IL-18; TNF-alpha and IL-23; TNF-alpha and MIF; TNF-alpha and PEG2; TNF-alpha and PGE4; and VEGFR and EGFR; TNF-alpha and RANK ligand; TNF-alpha and Blys; TNF-alpha and GP130; and TNF-alpha and CD-22.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/939,117 US20110110852A1 (en) | 2000-04-11 | 2010-11-03 | Multivalent Antibodies and Uses Therefor |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US19581900P | 2000-04-11 | 2000-04-11 | |
US09/813,341 US20020004587A1 (en) | 2000-04-11 | 2001-03-20 | Multivalent antibodies and uses therefor |
US11/218,821 US20060025576A1 (en) | 2000-04-11 | 2005-09-02 | Multivalent antibodies and uses therefor |
US11/535,031 US20080299120A1 (en) | 2000-04-11 | 2006-09-25 | Multivalent antibodies and uses therefor |
US12/939,117 US20110110852A1 (en) | 2000-04-11 | 2010-11-03 | Multivalent Antibodies and Uses Therefor |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/535,031 Continuation US20080299120A1 (en) | 2000-04-11 | 2006-09-25 | Multivalent antibodies and uses therefor |
Publications (1)
Publication Number | Publication Date |
---|---|
US20110110852A1 true US20110110852A1 (en) | 2011-05-12 |
Family
ID=22722944
Family Applications (6)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/813,341 Abandoned US20020004587A1 (en) | 2000-04-11 | 2001-03-20 | Multivalent antibodies and uses therefor |
US11/218,821 Abandoned US20060025576A1 (en) | 2000-04-11 | 2005-09-02 | Multivalent antibodies and uses therefor |
US11/535,031 Abandoned US20080299120A1 (en) | 2000-04-11 | 2006-09-25 | Multivalent antibodies and uses therefor |
US12/939,117 Abandoned US20110110852A1 (en) | 2000-04-11 | 2010-11-03 | Multivalent Antibodies and Uses Therefor |
US13/470,189 Expired - Fee Related US8722859B2 (en) | 2000-04-11 | 2012-05-11 | Multivalent antibodies and uses therefor |
US14/229,683 Expired - Lifetime US9493579B2 (en) | 2000-04-11 | 2014-03-28 | Multivalent antibodies and uses therefor |
Family Applications Before (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/813,341 Abandoned US20020004587A1 (en) | 2000-04-11 | 2001-03-20 | Multivalent antibodies and uses therefor |
US11/218,821 Abandoned US20060025576A1 (en) | 2000-04-11 | 2005-09-02 | Multivalent antibodies and uses therefor |
US11/535,031 Abandoned US20080299120A1 (en) | 2000-04-11 | 2006-09-25 | Multivalent antibodies and uses therefor |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/470,189 Expired - Fee Related US8722859B2 (en) | 2000-04-11 | 2012-05-11 | Multivalent antibodies and uses therefor |
US14/229,683 Expired - Lifetime US9493579B2 (en) | 2000-04-11 | 2014-03-28 | Multivalent antibodies and uses therefor |
Country Status (22)
Country | Link |
---|---|
US (6) | US20020004587A1 (en) |
EP (2) | EP2857516B1 (en) |
JP (1) | JP2003531588A (en) |
KR (1) | KR20020093029A (en) |
CN (2) | CN100390288C (en) |
AU (2) | AU2001247616B2 (en) |
BR (1) | BR0110610A (en) |
CA (1) | CA2403425C (en) |
CY (1) | CY1119291T1 (en) |
DK (2) | DK1272647T3 (en) |
ES (2) | ES2637801T3 (en) |
HK (1) | HK1208705A1 (en) |
HU (1) | HUP0300369A2 (en) |
IL (3) | IL151853A0 (en) |
LT (1) | LT2857516T (en) |
MX (1) | MXPA02010011A (en) |
NZ (1) | NZ521540A (en) |
PL (1) | PL357939A1 (en) |
PT (1) | PT2857516T (en) |
SI (1) | SI2857516T1 (en) |
WO (1) | WO2001077342A1 (en) |
ZA (1) | ZA200207589B (en) |
Cited By (55)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080226659A1 (en) * | 2000-03-16 | 2008-09-18 | Sharon Erickson | Methods of treatment using anti-erbb antibody-maytansinoid conjugates |
US20100028340A1 (en) * | 2008-02-29 | 2010-02-04 | Abbott Gmbh & Co. Kg | Antibodies against the rgm a protein and uses thereof |
US20100047239A1 (en) * | 2005-08-19 | 2010-02-25 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
US20100291113A1 (en) * | 2007-10-03 | 2010-11-18 | Cornell University | Treatment of Proliferative Disorders Using Antibodies to PSMA |
US20100322948A1 (en) * | 2007-09-06 | 2010-12-23 | Abbott Gmbh & Co. Kg | Bone morphogenetic protein (BMP)-binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and use of same |
US20110059852A1 (en) * | 2008-03-26 | 2011-03-10 | Cellerant Therapeutics, Inc. | Compositions and methods for treating haematological proliferative disorders of meyloid origin |
US20110135664A1 (en) * | 2009-12-08 | 2011-06-09 | Abbott Gmbh & Co. Kg | Monoclonal antibodies against the rgm a protein for use in the treatment of retinal nerve fiber layer degeneration |
US20120009596A1 (en) * | 2010-07-07 | 2012-01-12 | Sen-Yung Hsieh | Protein markers for detecting liver cancer and method for identifying the markers thereof |
WO2013036543A2 (en) * | 2011-09-10 | 2013-03-14 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Molecular imaging of cancer cells in vivo |
WO2013177264A1 (en) * | 2012-05-22 | 2013-11-28 | Shire Human Genetic Therapies, Inc. | Anti-ccl2 antibodies for treatment of scleroderma |
US8680239B2 (en) | 2000-12-22 | 2014-03-25 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V. | Use of RGM and its modulators |
US8772459B2 (en) | 2009-12-02 | 2014-07-08 | Imaginab, Inc. | J591 minibodies and Cys-diabodies for targeting human prostate specific membrane antigen (PSMA) and methods for their use |
US8906864B2 (en) | 2005-09-30 | 2014-12-09 | AbbVie Deutschland GmbH & Co. KG | Binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and their use |
US8911734B2 (en) | 2010-12-01 | 2014-12-16 | Alderbio Holdings Llc | Methods of preventing or treating pain using anti-NGF antibodies that selectively inhibit the association of NGF with TrkA, without affecting the association of NGF with p75 |
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US8951737B2 (en) | 1996-05-06 | 2015-02-10 | Cornell Research Foundation, Inc. | Treatment and diagnosis of cancer |
US9067988B2 (en) | 2010-12-01 | 2015-06-30 | Alderbio Holdings Llc | Methods of preventing or treating pain using anti-NGF antibodies |
US9078878B2 (en) | 2010-12-01 | 2015-07-14 | Alderbio Holdings Llc | Anti-NGF antibodies that selectively inhibit the association of NGF with TrkA, without affecting the association of NGF with p75 |
US9085622B2 (en) | 2010-09-03 | 2015-07-21 | Glaxosmithkline Intellectual Property Development Limited | Antigen binding proteins |
US20150203584A1 (en) * | 2011-11-16 | 2015-07-23 | Boehringer Ingelheim International Gmbh | Anti il-36r antibodies |
US9102722B2 (en) | 2012-01-27 | 2015-08-11 | AbbVie Deutschland GmbH & Co. KG | Composition and method for the diagnosis and treatment of diseases associated with neurite degeneration |
US9212225B1 (en) | 2014-07-01 | 2015-12-15 | Amphivena Therapeutics, Inc. | Bispecific CD33 and CD3 binding proteins |
US9539324B2 (en) | 2010-12-01 | 2017-01-10 | Alderbio Holdings, Llc | Methods of preventing inflammation and treating pain using anti-NGF compositions |
US9605070B2 (en) | 2014-01-31 | 2017-03-28 | Novartis Ag | Antibody molecules to TIM-3 and uses thereof |
US9610360B2 (en) | 2007-01-24 | 2017-04-04 | Ceruliean Pharma Inc. | Polymer drug conjugates with tether groups for controlled drug delivery |
US9718884B2 (en) | 2014-11-05 | 2017-08-01 | Eli Lilly And Company | Anti-TNF-/anti-IL-23 bispecific antibodies |
US9771417B2 (en) | 2014-08-07 | 2017-09-26 | Novartis Ag | Angiopoietin-like 4 antibodies and methods of use |
US9884909B2 (en) | 2010-12-01 | 2018-02-06 | Alderbio Holdings Llc | Anti-NGF compositions and use thereof |
US9988443B2 (en) | 2014-08-07 | 2018-06-05 | Novartis Ag | Angiopoetin-like 4 (ANGPTL4) antibodies and methods of use |
US10059763B2 (en) | 2014-09-03 | 2018-08-28 | Boehringer Ingelheim International Gmbh | Compound targeting IL-23A and TNF-alpha and uses thereof |
US10308712B2 (en) | 2014-03-27 | 2019-06-04 | Bird Rock Bio, Inc. | Antibodies that bind human cannabinoid 1 (CB1) receptor |
US10400038B2 (en) | 2014-04-03 | 2019-09-03 | Igm Biosciences, Inc. | Modified J-chain |
US10517969B2 (en) | 2009-02-17 | 2019-12-31 | Cornell University | Methods and kits for diagnosis of cancer and prediction of therapeutic value |
US10570204B2 (en) | 2013-09-26 | 2020-02-25 | The Medical College Of Wisconsin, Inc. | Methods for treating hematologic cancers |
US10618978B2 (en) | 2015-09-30 | 2020-04-14 | Igm Biosciences, Inc. | Binding molecules with modified J-chain |
US10689449B2 (en) | 2015-01-20 | 2020-06-23 | Igm Biosciences, Inc. | Multimeric death domain-containing receptor-5 (DR5) antibodies and uses thereof |
US10738118B2 (en) | 2015-05-29 | 2020-08-11 | Amphivena Therapeutics, Inc. | Methods of using bispecific CD33 and CD3 binding proteins |
US10738095B2 (en) | 2015-06-03 | 2020-08-11 | The Medical College Of Wisconsin, Inc. | Engineered CCL20 locked dimer polypeptide |
US10752687B2 (en) | 2014-01-24 | 2020-08-25 | Novartis Ag | Antibody molecules to PD-1 and uses thereof |
US10787520B2 (en) | 2015-03-04 | 2020-09-29 | Igm Biosciences, Inc. | Multimeric bispecific binding molecules specific for CD20 and CD3 |
US10844135B2 (en) | 2003-10-10 | 2020-11-24 | Immunogen, Inc. | Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates and methods of making said |
US10927185B2 (en) | 2014-11-21 | 2021-02-23 | Astellas Pharma Inc. | Bispecific antibody format |
US11214610B2 (en) | 2010-12-01 | 2022-01-04 | H. Lundbeck A/S | High-purity production of multi-subunit proteins such as antibodies in transformed microbes such as Pichia pastoris |
US11254744B2 (en) | 2015-08-07 | 2022-02-22 | Imaginab, Inc. | Antigen binding constructs to target molecules |
US11266745B2 (en) | 2017-02-08 | 2022-03-08 | Imaginab, Inc. | Extension sequences for diabodies |
US11344620B2 (en) | 2014-09-13 | 2022-05-31 | Novartis Ag | Combination therapies |
US11421026B2 (en) | 2015-09-30 | 2022-08-23 | Bird Rock Bio, Inc. | Antibodies that bind human cannabinoid 1 (CB1) receptor |
US11464871B2 (en) | 2012-10-02 | 2022-10-11 | Novartis Ag | Methods and systems for polymer precipitation and generation of particles |
US11571462B2 (en) | 2015-06-03 | 2023-02-07 | The Medical College Of Wisconsin, Inc. | Engineered CCL20 locked dimer polypeptide |
US11634489B2 (en) | 2017-08-03 | 2023-04-25 | Alector Llc | Anti-TREM2 antibodies and methods of use thereof |
US11639389B2 (en) | 2015-09-30 | 2023-05-02 | Igm Biosciences, Inc. | Binding molecules with modified J-chain |
US11692042B2 (en) | 2018-03-09 | 2023-07-04 | Agenus Inc. | Anti-CD73 antibodies and methods of use thereof |
US11730812B2 (en) | 2019-03-08 | 2023-08-22 | Boehringer Ingelheim International Gmbh | Anti-IL-36R antibody formulations |
US11884719B2 (en) | 2018-12-21 | 2024-01-30 | 23Andme, Inc. | Anti-IL-36 antibodies and methods of use thereof |
Families Citing this family (1023)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7833529B1 (en) | 1999-01-07 | 2010-11-16 | Zymogenetics, Inc. | Methods for inhibiting B lymphocyte proliferation with soluble ztnf4 receptor |
US20050100548A1 (en) * | 2001-07-24 | 2005-05-12 | Biogen Idec Ma Inc. | BAFF, inhibitors thereof and their use in the modulation of B-cell response |
US20030095967A1 (en) * | 1999-01-25 | 2003-05-22 | Mackay Fabienne | BAFF, inhibitors thereof and their use in the modulation of B-cell response and treatment of autoimmune disorders |
US7696325B2 (en) | 1999-03-10 | 2010-04-13 | Chugai Seiyaku Kabushiki Kaisha | Polypeptide inducing apoptosis |
EP1642596A3 (en) * | 1999-05-07 | 2006-04-12 | Genentech, Inc. | Treatment of autoimmune diseases with antagonists which bind to B cell surface markers |
MXPA02000419A (en) * | 1999-07-12 | 2004-09-10 | Genentech Inc | Blocking immune response to a foreign antigen using an antagonist which binds to cd20. |
DK1255558T3 (en) * | 2000-02-16 | 2006-10-23 | Genentech Inc | Anti-April antibodies and hybridoma cells |
LT2857516T (en) | 2000-04-11 | 2017-09-11 | Genentech, Inc. | Multivalent antibodies and uses therefor |
TWI318983B (en) | 2000-05-02 | 2010-01-01 | Uab Research Foundation | An antibody selective for a tumor necrosis factor-related apoptosis-inducing ligand receptor and uses thereof |
US7476383B2 (en) | 2000-05-02 | 2009-01-13 | The Uab Research Foundation | Antibody selective for DR4 and uses thereof |
US7279160B2 (en) | 2000-05-02 | 2007-10-09 | The Uab Research Foundation | Combinations of DR5 antibodies and other therapeutic agents |
KR20040023565A (en) * | 2000-09-18 | 2004-03-18 | 아이덱 파마슈티칼즈 코포레이션 | Combination therapy for treatment of autoimmune diseases using b cell depleting/immunoregulatory antibody combination |
CN1469735A (en) * | 2000-10-16 | 2004-01-21 | ����˹��ҩ�﹫˾ | Liposomal formulation of mitoxantrone |
CN1308448C (en) | 2000-10-20 | 2007-04-04 | 中外制药株式会社 | Degraded TPO agonist antibody |
US20030228309A1 (en) * | 2000-11-08 | 2003-12-11 | Theodora Salcedo | Antibodies that immunospecifically bind to TRAIL receptors |
US20060062786A1 (en) * | 2000-11-08 | 2006-03-23 | Human Genome Sciences, Inc. | Antibodies that immunospecifically bind to TRAIL receptors |
US7754208B2 (en) * | 2001-01-17 | 2010-07-13 | Trubion Pharmaceuticals, Inc. | Binding domain-immunoglobulin fusion proteins |
US20030133939A1 (en) * | 2001-01-17 | 2003-07-17 | Genecraft, Inc. | Binding domain-immunoglobulin fusion proteins |
US7829084B2 (en) * | 2001-01-17 | 2010-11-09 | Trubion Pharmaceuticals, Inc. | Binding constructs and methods for use thereof |
EP2301971A1 (en) | 2001-02-20 | 2011-03-30 | ZymoGenetics, L.L.C. | Antibodies that bind both BCMA and TACI |
US7589180B2 (en) | 2001-05-11 | 2009-09-15 | Abbott Laboratories Inc. | Specific binding proteins and uses thereof |
US20100056762A1 (en) | 2001-05-11 | 2010-03-04 | Old Lloyd J | Specific binding proteins and uses thereof |
EP1436003B3 (en) | 2001-05-24 | 2012-03-14 | ZymoGenetics, Inc. | Taci-immunoglobulin fusion proteins |
US7348003B2 (en) | 2001-05-25 | 2008-03-25 | Human Genome Sciences, Inc. | Methods of treating cancer using antibodies that immunospecifically bind to TRAIL receptors |
US20050129616A1 (en) * | 2001-05-25 | 2005-06-16 | Human Genome Sciences, Inc. | Antibodies that immunospecifically bind to TRAIL receptors |
US20090226429A1 (en) * | 2001-05-25 | 2009-09-10 | Human Genome Sciences, Inc. | Antibodies That Immunospecifically Bind to TRAIL Receptors |
US20050214209A1 (en) * | 2001-05-25 | 2005-09-29 | Human Genome Sciences, Inc. | Antibodies that immunospecifically bind to TRAIL receptors |
US7361341B2 (en) * | 2001-05-25 | 2008-04-22 | Human Genome Sciences, Inc. | Methods of treating cancer using antibodies that immunospecifically bind to trail receptors |
US7595378B2 (en) * | 2001-06-13 | 2009-09-29 | Genmab A/S | Human monoclonal antibodies to epidermal growth factor receptor (EGFR) |
RU2335507C2 (en) * | 2001-06-13 | 2008-10-10 | Генмаб А/С | Human monoclonal antibodies to receptor of epidermal growth factor (egfr), method of their preparation and usage, hybridome, transfectome, transgene animal, expression vector |
ATE477280T1 (en) * | 2001-06-28 | 2010-08-15 | Domantis Ltd | DOUBLE-SPECIFIC LIGAND AND USE THEREOF |
US20050271663A1 (en) * | 2001-06-28 | 2005-12-08 | Domantis Limited | Compositions and methods for treating inflammatory disorders |
WO2003011909A1 (en) * | 2001-07-31 | 2003-02-13 | University Of Southampton | Binding agents with differential activity |
JP2005510208A (en) * | 2001-08-03 | 2005-04-21 | ジェネンテック・インコーポレーテッド | TACIs and BR3 polypeptides and uses thereof |
BRPI0213846B8 (en) * | 2001-11-01 | 2021-05-25 | Uab Research Foundation | composition comprising an antibody that specifically binds a trail receptor dr5 and one or more therapeutic agents |
JP2005516958A (en) * | 2001-12-20 | 2005-06-09 | ヒューマン ジノーム サイエンシーズ, インコーポレイテッド | Antibodies that immunospecifically bind to TRAIL receptors |
EP2075256A2 (en) | 2002-01-14 | 2009-07-01 | William Herman | Multispecific binding molecules |
US20030149246A1 (en) * | 2002-02-01 | 2003-08-07 | Russell John C. | Macromolecular conjugates and processes for preparing the same |
US8287864B2 (en) * | 2002-02-14 | 2012-10-16 | Immunomedics, Inc. | Structural variants of antibodies for improved therapeutic characteristics |
CN101914158A (en) * | 2002-02-14 | 2010-12-15 | 免疫医疗公司 | Anti-cd20 antibodies and fusion proteins thereof and methods of use |
US20040002587A1 (en) * | 2002-02-20 | 2004-01-01 | Watkins Jeffry D. | Fc region variants |
US20080267965A1 (en) * | 2002-02-21 | 2008-10-30 | Kalled Susan L | Use of Bcma as an Immunoregulatory Agent |
US20080254027A1 (en) * | 2002-03-01 | 2008-10-16 | Bernett Matthew J | Optimized CD5 antibodies and methods of using the same |
US20080260731A1 (en) * | 2002-03-01 | 2008-10-23 | Bernett Matthew J | Optimized antibodies that target cd19 |
US8188231B2 (en) | 2002-09-27 | 2012-05-29 | Xencor, Inc. | Optimized FC variants |
US20040132101A1 (en) | 2002-09-27 | 2004-07-08 | Xencor | Optimized Fc variants and methods for their generation |
US20070148171A1 (en) * | 2002-09-27 | 2007-06-28 | Xencor, Inc. | Optimized anti-CD30 antibodies |
US7317091B2 (en) * | 2002-03-01 | 2008-01-08 | Xencor, Inc. | Optimized Fc variants |
US8093357B2 (en) * | 2002-03-01 | 2012-01-10 | Xencor, Inc. | Optimized Fc variants and methods for their generation |
US9321832B2 (en) * | 2002-06-28 | 2016-04-26 | Domantis Limited | Ligand |
US9028822B2 (en) | 2002-06-28 | 2015-05-12 | Domantis Limited | Antagonists against TNFR1 and methods of use therefor |
SI1517921T1 (en) * | 2002-06-28 | 2006-10-31 | Domantis Ltd | Dual specific ligands with increased serum half-life |
US7696320B2 (en) | 2004-08-24 | 2010-04-13 | Domantis Limited | Ligands that have binding specificity for VEGF and/or EGFR and methods of use therefor |
WO2004011611A2 (en) * | 2002-07-25 | 2004-02-05 | Genentech, Inc. | Taci antibodies and uses thereof |
ATE516818T1 (en) | 2002-07-31 | 2011-08-15 | Seattle Genetics Inc | AURISTATIN CONJUGATES AND THEIR USE IN THE TREATMENT OF CANCER, AN AUTOIMMUNE DISEASE OR AN INFECTIOUS DISEASE |
US8946387B2 (en) * | 2002-08-14 | 2015-02-03 | Macrogenics, Inc. | FcγRIIB specific antibodies and methods of use thereof |
US8044180B2 (en) * | 2002-08-14 | 2011-10-25 | Macrogenics, Inc. | FcγRIIB specific antibodies and methods of use thereof |
US20090017023A1 (en) * | 2002-08-14 | 2009-01-15 | Macrogenics, Inc. | FcGammaRIIB Specific Antibodies and Methods of Use Thereof |
US8968730B2 (en) * | 2002-08-14 | 2015-03-03 | Macrogenics Inc. | FcγRIIB specific antibodies and methods of use thereof |
US8530627B2 (en) | 2002-08-14 | 2013-09-10 | Macrogenics, Inc. | FcγRIIB specific antibodies and methods of use thereof |
US8193318B2 (en) * | 2002-08-14 | 2012-06-05 | Macrogenics, Inc. | FcγRIIB specific antibodies and methods of use thereof |
US20060165700A1 (en) * | 2002-09-04 | 2006-07-27 | Ostroff Gary R | Cancer therapy using whole glucan particles and antibodies |
CN1705491B (en) * | 2002-09-27 | 2013-05-29 | 赞科股份有限公司 | Optimized Fc variants and methods for their generation |
US20060235208A1 (en) * | 2002-09-27 | 2006-10-19 | Xencor, Inc. | Fc variants with optimized properties |
WO2004039329A2 (en) * | 2002-10-31 | 2004-05-13 | Biogen Idec Ma Inc. | Treatment of immunological renal disorders by lymphotoxin pathway inhibitors |
US8505468B2 (en) * | 2002-11-19 | 2013-08-13 | Sharp Kabushiki Kaisha | Substrate accommodating tray |
JP4351674B2 (en) * | 2002-12-16 | 2009-10-28 | ジェネンテック・インコーポレーテッド | Immunoglobulin variants and their use and use |
JP2006515750A (en) * | 2002-12-20 | 2006-06-08 | バイオジェン・アイデック・エムエイ・インコーポレイテッド | Multivalent lymphotoxin β receptor agonist and treatment using the same |
CA2511910A1 (en) * | 2002-12-27 | 2004-07-15 | Domantis Limited | Dual specific single domain antibodies specific for a ligand and for the receptor of the ligand |
US7960512B2 (en) * | 2003-01-09 | 2011-06-14 | Macrogenics, Inc. | Identification and engineering of antibodies with variant Fc regions and methods of using same |
EP2368578A1 (en) | 2003-01-09 | 2011-09-28 | Macrogenics, Inc. | Identification and engineering of antibodies with variant Fc regions and methods of using same |
US20070275460A1 (en) * | 2003-03-03 | 2007-11-29 | Xencor.Inc. | Fc Variants With Optimized Fc Receptor Binding Properties |
US20090010920A1 (en) * | 2003-03-03 | 2009-01-08 | Xencor, Inc. | Fc Variants Having Decreased Affinity for FcyRIIb |
US8388955B2 (en) | 2003-03-03 | 2013-03-05 | Xencor, Inc. | Fc variants |
US8084582B2 (en) | 2003-03-03 | 2011-12-27 | Xencor, Inc. | Optimized anti-CD20 monoclonal antibodies having Fc variants |
JP2004279086A (en) | 2003-03-13 | 2004-10-07 | Konica Minolta Holdings Inc | Radiation image conversion panel and method for manufacturing it |
ES2537015T3 (en) * | 2003-03-19 | 2015-06-01 | Biogen Ma Inc. | NOGO receptor binding protein |
RS51686B (en) * | 2003-04-09 | 2011-10-31 | Genentech Inc. | Therapy of autoimmune disease in a patient with an inadequate response to a tnf-alpha inhibitor |
US9051373B2 (en) | 2003-05-02 | 2015-06-09 | Xencor, Inc. | Optimized Fc variants |
US7619068B2 (en) * | 2003-05-09 | 2009-11-17 | Diadexus, Inc. | Ovr110 antibody compositions and methods of use |
PT1631313E (en) * | 2003-06-05 | 2015-07-02 | Genentech Inc | Combination therapy for b cell disorders |
US20050163775A1 (en) * | 2003-06-05 | 2005-07-28 | Genentech, Inc. | Combination therapy for B cell disorders |
US7172751B2 (en) * | 2003-06-13 | 2007-02-06 | Immunomedics, Inc. | D-amino acid peptides |
AU2004260936B2 (en) | 2003-06-27 | 2010-06-10 | Amgen Fremont Inc. | Antibodies directed to the deletion mutants of epidermal growth factor receptor and uses thereof |
ATE416190T1 (en) | 2003-07-04 | 2008-12-15 | Affibody Ab | POLYPEPTIDES WITH BINDING AFFINITY FOR HER2 |
TW200510532A (en) | 2003-07-15 | 2005-03-16 | Chugai Pharmaceutical Co Ltd | IgM production by transformed cell and method of quantifying the same |
US7696322B2 (en) * | 2003-07-28 | 2010-04-13 | Catalent Pharma Solutions, Inc. | Fusion antibodies |
WO2005017529A1 (en) * | 2003-07-29 | 2005-02-24 | Genentech, Inc. | Assay for human anti cd20 antibodies and uses therefor |
ZA200601218B (en) * | 2003-08-29 | 2007-05-30 | Genentech Inc | Anti-CD20 therapy of ocular disorders |
US9714282B2 (en) | 2003-09-26 | 2017-07-25 | Xencor, Inc. | Optimized Fc variants and methods for their generation |
US8101720B2 (en) * | 2004-10-21 | 2012-01-24 | Xencor, Inc. | Immunoglobulin insertions, deletions and substitutions |
US20060127404A1 (en) * | 2003-09-30 | 2006-06-15 | Chichi Huang | Hinge core mimetibodies, compositions, methods and uses |
US20070212346A1 (en) | 2003-10-09 | 2007-09-13 | Tomoyuki Igawa | Highly Concentrated Stabilized Igm Solution |
EP1710255A4 (en) * | 2003-12-12 | 2008-09-24 | Chugai Pharmaceutical Co Ltd | Modified antibodies recognising receptor trimers or higher multimers |
AU2004303848A1 (en) * | 2003-12-19 | 2005-07-07 | Genentech, Inc. | Detection of CD20 in transplant rejection |
AU2004305560A1 (en) * | 2003-12-19 | 2005-07-07 | Genentech, Inc. | Detection of CD20 in therapy of autoimmune diseases |
EP1697520A2 (en) * | 2003-12-22 | 2006-09-06 | Xencor, Inc. | Fc polypeptides with novel fc ligand binding sites |
CN1925871A (en) * | 2004-01-27 | 2007-03-07 | 南加州大学 | Polymer-bound antibody concer therapeutic agent |
DK1716181T3 (en) | 2004-02-19 | 2010-03-01 | Genentech Inc | CDR repaired antibodies |
KR20060132006A (en) * | 2004-03-23 | 2006-12-20 | 비오겐 아이덱 엠에이 아이엔씨. | Receptor coupling agents and therapeutic uses thereof |
EP2053062A1 (en) * | 2004-03-24 | 2009-04-29 | Xencor, Inc. | Immunoglobin variants outside the Fc region |
JP2007532681A (en) * | 2004-04-16 | 2007-11-15 | ジェネンテック・インコーポレーテッド | Methods for increasing B cell depletion |
MXPA06012674A (en) * | 2004-05-05 | 2007-03-26 | Genentech Inc | Preventing autoimmune disease. |
AU2005244058B2 (en) | 2004-05-10 | 2011-07-28 | Macrogenics, Inc. | Humanized FcgammaRIIB specific antibodies and methods of use thereof |
KR20070039911A (en) | 2004-06-01 | 2007-04-13 | 도만티스 리미티드 | Bispecific fusion antibodies with enhanced serum half-life |
BRPI0510883B8 (en) | 2004-06-01 | 2021-05-25 | Genentech Inc | drug-antibody conjugate compound, pharmaceutical composition, method of manufacturing a drug-antibody conjugate compound, and uses of a formulation, a drug-antibody conjugate and a chemotherapeutic agent, and a combination |
BRPI0510885A (en) * | 2004-06-04 | 2007-12-26 | Genentech Inc | lupus treatment method and industrialized article |
TW201422238A (en) | 2004-06-04 | 2014-06-16 | Genentech Inc | Use of CD20 antibody in treatment of multiple sclerosis and an article for the use |
US8486893B2 (en) | 2004-06-24 | 2013-07-16 | Biogen Idec Ma Inc. | Treatment of conditions involving demyelination |
AU2005272993B2 (en) * | 2004-07-15 | 2010-02-11 | Xencor, Inc | Optimized Fc variants |
US20150010550A1 (en) | 2004-07-15 | 2015-01-08 | Xencor, Inc. | OPTIMIZED Fc VARIANTS |
WO2006012508A2 (en) * | 2004-07-22 | 2006-02-02 | Genentech, Inc. | Method of treating sjögren's syndrome |
US20070286855A1 (en) * | 2004-08-03 | 2007-12-13 | Mayo Foundation For Medical Education And Research | Improving treatments |
WO2006031994A2 (en) * | 2004-09-14 | 2006-03-23 | Xencor, Inc. | Monomeric immunoglobulin fc domains |
SG165344A1 (en) * | 2004-10-05 | 2010-10-28 | Genentech Inc | Method for treating vasculitis |
JO3000B1 (en) | 2004-10-20 | 2016-09-05 | Genentech Inc | Antibody Formulations. |
US20060121042A1 (en) * | 2004-10-27 | 2006-06-08 | Medimmune, Inc. | Modulation of antibody specificity by tailoring the affinity to cognate antigens |
US7632497B2 (en) * | 2004-11-10 | 2009-12-15 | Macrogenics, Inc. | Engineering Fc Antibody regions to confer effector function |
US8367805B2 (en) | 2004-11-12 | 2013-02-05 | Xencor, Inc. | Fc variants with altered binding to FcRn |
US8802820B2 (en) * | 2004-11-12 | 2014-08-12 | Xencor, Inc. | Fc variants with altered binding to FcRn |
US8546543B2 (en) | 2004-11-12 | 2013-10-01 | Xencor, Inc. | Fc variants that extend antibody half-life |
WO2006053301A2 (en) * | 2004-11-12 | 2006-05-18 | Xencor, Inc. | Fc variants with altered binding to fcrn |
US20070135620A1 (en) * | 2004-11-12 | 2007-06-14 | Xencor, Inc. | Fc variants with altered binding to FcRn |
WO2006076594A2 (en) * | 2005-01-12 | 2006-07-20 | Xencor, Inc. | Antibodies and fc fusion proteins with altered immunogenicity |
ZA200705459B (en) * | 2005-01-13 | 2008-09-25 | Genentech Inc | Treatment method |
KR20190110637A (en) | 2005-01-21 | 2019-09-30 | 제넨테크, 인크. | Fixed dosing of her antibodies |
EP1846034A4 (en) | 2005-02-02 | 2010-11-10 | Uab Research Foundation | Agents and methods related to reducing resistance to apoptosis-inducing death receptor agonists |
DOP2006000029A (en) * | 2005-02-07 | 2006-08-15 | Genentech Inc | ANTIBODY VARIANTS AND USES THEREOF. (VARIATIONS OF AN ANTIBODY AND USES OF THE SAME) |
RU2404806C2 (en) | 2005-02-23 | 2010-11-27 | Дженентек, Инк. | Extension of time to progression of disease or lifetime of oncologic patients with application of her dimerisation inhibitors |
TW200714289A (en) * | 2005-02-28 | 2007-04-16 | Genentech Inc | Treatment of bone disorders |
US20060204505A1 (en) * | 2005-03-08 | 2006-09-14 | Sliwkowski Mark X | Methods for identifying tumors responsive to treatment with HER dimerization inhibitors (HDIs) |
TW200722518A (en) | 2005-03-31 | 2007-06-16 | Chugai Pharmaceutical Co Ltd | Sc(fv)2 structural isomers |
CA2602663A1 (en) * | 2005-03-31 | 2006-10-05 | Xencor, Inc. | Fc variants with optimized properties |
US8349332B2 (en) | 2005-04-06 | 2013-01-08 | Ibc Pharmaceuticals, Inc. | Multiple signaling pathways induced by hexavalent, monospecific and bispecific antibodies for enhanced toxicity to B-cell lymphomas and other diseases |
US8475794B2 (en) | 2005-04-06 | 2013-07-02 | Ibc Pharmaceuticals, Inc. | Combination therapy with anti-CD74 antibodies provides enhanced toxicity to malignancies, Autoimmune disease and other diseases |
AR053579A1 (en) * | 2005-04-15 | 2007-05-09 | Genentech Inc | TREATMENT OF INTESTINAL INFLAMMATORY DISEASE (IBD) |
AU2006251647A1 (en) * | 2005-05-20 | 2006-11-30 | Genentech, Inc. | Pretreatment of a biological sample from an autoimmune disease subject |
AU2006250068A1 (en) * | 2005-05-26 | 2006-11-30 | Schering Corporation | Interferon-IGG fusion |
JP5068167B2 (en) | 2005-06-10 | 2012-11-07 | 中外製薬株式会社 | Stabilizer for protein preparation containing meglumine and use thereof |
JP5085322B2 (en) | 2005-06-10 | 2012-11-28 | 中外製薬株式会社 | Pharmaceutical composition containing sc (Fv) 2 |
KR101245462B1 (en) | 2005-07-08 | 2013-03-20 | 바이오겐 아이덱 엠에이 인코포레이티드 | Sp35 antibodies and uses thereof |
JP2009502936A (en) | 2005-07-25 | 2009-01-29 | トルビオン ファーマシューティカルズ, インコーポレイテッド | Single dose of CD20 specific binding molecule |
US20080279850A1 (en) * | 2005-07-25 | 2008-11-13 | Trubion Pharmaceuticals, Inc. | B-Cell Reduction Using CD37-Specific and CD20-Specific Binding Molecules |
CN105012953B (en) | 2005-07-25 | 2018-06-22 | 阿普泰沃研发有限责任公司 | B- cells are reduced with CD37- specificity and CD20- specific binding molecules |
KR20080056714A (en) | 2005-08-09 | 2008-06-23 | 지모제넥틱스, 인코포레이티드 | Methods for treating b-cell malignancies using taci-ig fusion molecule |
WO2007019573A2 (en) | 2005-08-09 | 2007-02-15 | Zymogenetics, Inc. | Methods for the treatment and prevention of abnormal cell proliferation using taci-fusion molecules |
CA2618681C (en) | 2005-08-10 | 2015-10-27 | Macrogenics, Inc. | Identification and engineering of antibodies with variant fc regions and methods of using same |
WO2007024715A2 (en) | 2005-08-19 | 2007-03-01 | Abbott Laboratories | Dual variable domain immunoglobin and uses thereof |
AU2014203217B2 (en) * | 2005-08-19 | 2017-03-30 | Abbvie Inc. | Dual variable domain immunoglobin and uses thereof |
AU2012205249B2 (en) * | 2005-08-19 | 2014-03-20 | Abbvie Inc. | Dual variable domain immunoglobin and uses thereof |
EP2500358A3 (en) | 2005-08-19 | 2012-10-17 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
JP2012228248A (en) * | 2005-08-19 | 2012-11-22 | Abbott Lab | Dual variable domain immunoglobulin and use thereof |
US20090215992A1 (en) * | 2005-08-19 | 2009-08-27 | Chengbin Wu | Dual variable domain immunoglobulin and uses thereof |
US8129114B2 (en) * | 2005-08-24 | 2012-03-06 | Bristol-Myers Squibb Company | Biomarkers and methods for determining sensitivity to epidermal growth factor receptor modulators |
US8124738B2 (en) * | 2005-09-26 | 2012-02-28 | Medarex, Inc. | Human monoclonal antibodies to CD70 |
DK1931709T3 (en) * | 2005-10-03 | 2017-03-13 | Xencor Inc | FC VARIETIES WITH OPTIMIZED FC RECEPTOR BINDING PROPERTIES |
AU2006302254B2 (en) | 2005-10-06 | 2011-05-26 | Xencor, Inc. | Optimized anti-CD30 antibodies |
MY149159A (en) | 2005-11-15 | 2013-07-31 | Hoffmann La Roche | Method for treating joint damage |
AU2006318539B2 (en) | 2005-11-23 | 2012-09-13 | Genentech, Inc. | Methods and compositions related to B cell assays |
US20090155283A1 (en) * | 2005-12-01 | 2009-06-18 | Drew Philip D | Noncompetitive Domain Antibody Formats That Bind Interleukin 1 Receptor Type 1 |
US20090175872A1 (en) | 2005-12-02 | 2009-07-09 | Biogen Idec Ma Inc. | Treatment of Conditions Involving Demyelination |
EP1994141B1 (en) * | 2006-02-23 | 2017-11-15 | ViaCyte, Inc. | Compositions and methods useful for culturing differentiable cells |
WO2007106707A2 (en) * | 2006-03-10 | 2007-09-20 | Macrogenics, Inc. | Identification and engineering of antibodies with variant heavy chains and methods of using same |
EP2035028A2 (en) | 2006-05-15 | 2009-03-18 | Ares Trading S.A. | Methods for treating autoimmune diseases using a taci-ig fusion molecule |
US7786270B2 (en) | 2006-05-26 | 2010-08-31 | Macrogenics, Inc. | Humanized FcγRIIB-specific antibodies and methods of use thereof |
CA2654317A1 (en) * | 2006-06-12 | 2007-12-21 | Trubion Pharmaceuticals, Inc. | Single-chain multivalent binding proteins with effector function |
SI2029173T1 (en) | 2006-06-26 | 2016-12-30 | Macrogenics, Inc. | Fc riib-specific antibodies and methods of use thereof |
CA2656224C (en) | 2006-06-26 | 2018-01-09 | Macrogenics, Inc. | Combination of fc.gamma.riib antibodies and cd20-specific antibodies and methods of use thereof |
MX2009000696A (en) | 2006-07-19 | 2009-01-30 | Univ Pennsylvania | Wsx-1/p28 as a target for anti-inflammatory responses. |
RS53263B (en) | 2006-08-14 | 2014-08-29 | Xencor Inc. | Optimized antibodies that target cd19 |
AU2007299843B2 (en) * | 2006-09-18 | 2012-03-08 | Xencor, Inc | Optimized antibodies that target HM1.24 |
US20080112961A1 (en) * | 2006-10-09 | 2008-05-15 | Macrogenics, Inc. | Identification and Engineering of Antibodies with Variant Fc Regions and Methods of Using Same |
WO2008063776A2 (en) * | 2006-10-12 | 2008-05-29 | Genentech, Inc. | Antibodies to lymphotoxin-alpha |
US8652466B2 (en) | 2006-12-08 | 2014-02-18 | Macrogenics, Inc. | Methods for the treatment of disease using immunoglobulins having Fc regions with altered affinities for FcγRactivating and FcγRinhibiting |
KR20090088946A (en) * | 2006-12-14 | 2009-08-20 | 메다렉스, 인코포레이티드 | Human antibodies that bind cd70 and uses thereof |
US20090186034A1 (en) * | 2006-12-19 | 2009-07-23 | Genetech, Inc. | Gene expression markers for inflammatory bowel disease |
US8128926B2 (en) | 2007-01-09 | 2012-03-06 | Biogen Idec Ma Inc. | Sp35 antibodies and uses thereof |
WO2008091701A2 (en) * | 2007-01-25 | 2008-07-31 | Dana-Farber Cancer Institute | Use of anti-egfr antibodies in treatment of egfr mutant mediated disease |
PE20090681A1 (en) | 2007-03-02 | 2009-06-10 | Genentech Inc | PREDICTION OF RESPONSE TO A HER INHIBITOR |
WO2008115404A1 (en) * | 2007-03-15 | 2008-09-25 | Ludwing Institute For Cancer Research | Treatment method using egfr antibodies and src inhibitors and related formulations |
US20100233689A1 (en) * | 2007-03-22 | 2010-09-16 | The General Hospital Corporation | Pyrazoloanthrone and derivatives thereof for the treatment of cancer expressing 'mullerian inhibiting substance' type ii receptor (misrii) and of excess androgen states |
AU2008259513B2 (en) * | 2007-06-06 | 2014-07-24 | Domantis Limited | Polypeptides, antibody variable domains and antagonists |
WO2008154249A2 (en) | 2007-06-08 | 2008-12-18 | Genentech, Inc. | Gene expression markers of tumor resistance to her2 inhibitor treatment |
US7580304B2 (en) * | 2007-06-15 | 2009-08-25 | United Memories, Inc. | Multiple bus charge sharing |
US20100322939A1 (en) * | 2007-06-21 | 2010-12-23 | Genmab A/S | Novel methods for treating egfr-associated tumors |
AU2008275229B2 (en) | 2007-07-09 | 2014-11-06 | Genentech, Inc. | Prevention of disulfide bond reduction during recombinant production of polypeptides |
JP2010535032A (en) | 2007-07-31 | 2010-11-18 | メディミューン,エルエルシー | Multispecific epitope binding proteins and uses thereof |
CA2696360C (en) | 2007-08-14 | 2018-11-20 | Ludwig Institute For Cancer Research | Monoclonal antibody targeting the egfr receptor and uses thereof |
EP3789400A1 (en) | 2007-09-26 | 2021-03-10 | Chugai Seiyaku Kabushiki Kaisha | Modified antibody constant region |
CN101842387B (en) | 2007-09-26 | 2014-05-07 | Ucb医药有限公司 | Dual specificity antibody fusions |
US20090155255A1 (en) * | 2007-09-27 | 2009-06-18 | Biogen Idec Ma Inc. | Cd23 binding molecules and methods of use thereof |
EP2205071B1 (en) * | 2007-10-11 | 2015-07-22 | Biogen MA Inc. | Lingo-1 antagonists and trkb agonists for use in the treatment of glaucoma |
EP2050764A1 (en) | 2007-10-15 | 2009-04-22 | sanofi-aventis | Novel polyvalent bispecific antibody format and uses thereof |
AU2008312406B2 (en) | 2007-10-16 | 2014-03-06 | Ares Trading S.A. | Combination of BLyS inhibition and anti-CD 20 agents for treatment of autoimmune disease |
JP5754046B2 (en) * | 2007-11-08 | 2015-07-22 | バイオジェン・エムエイ・インコーポレイテッドBiogen MA Inc. | Use of a LINGO-4 antagonist in the treatment of conditions involving demyelination |
AU2008328785A1 (en) | 2007-11-27 | 2009-06-04 | Ablynx N.V. | Method for obtaining polypeptide constructs comprising two or more single domain antibodies |
CN101970689A (en) | 2007-11-29 | 2011-02-09 | 健泰科生物技术公司 | Gene expression markers for inflammatory bowel disease |
EP2615115A3 (en) * | 2007-11-30 | 2014-01-08 | Glaxo Group Limited | Antigen-binding constructs |
US9266967B2 (en) | 2007-12-21 | 2016-02-23 | Hoffmann-La Roche, Inc. | Bivalent, bispecific antibodies |
US20090162359A1 (en) | 2007-12-21 | 2009-06-25 | Christian Klein | Bivalent, bispecific antibodies |
SI2808343T1 (en) | 2007-12-26 | 2019-10-30 | Xencor Inc | Fc variants with altered binding to FcRn |
EP2077281A1 (en) | 2008-01-02 | 2009-07-08 | Bergen Teknologioverforing AS | Anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome |
US7914785B2 (en) | 2008-01-02 | 2011-03-29 | Bergen Teknologieverforing As | B-cell depleting agents, like anti-CD20 antibodies or fragments thereof for the treatment of chronic fatigue syndrome |
US8557242B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | ERBB2 antibodies comprising modular recognition domains |
US8454960B2 (en) | 2008-01-03 | 2013-06-04 | The Scripps Research Institute | Multispecific antibody targeting and multivalency through modular recognition domains |
US20110189206A1 (en) * | 2008-01-03 | 2011-08-04 | Barbas Iii Carlos F | Antibody Targeting Through a Modular Recognition Domain |
US8557243B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | EFGR antibodies comprising modular recognition domains |
US8574577B2 (en) | 2008-01-03 | 2013-11-05 | The Scripps Research Institute | VEGF antibodies comprising modular recognition domains |
WO2009092383A2 (en) * | 2008-01-22 | 2009-07-30 | Multimerics Aps | Products and methods to prevent infection |
TWI472339B (en) | 2008-01-30 | 2015-02-11 | Genentech Inc | Composition comprising antibody that binds to domain ii of her2 and acidic variants thereof |
TW200940064A (en) * | 2008-03-06 | 2009-10-01 | Genentech Inc | Combination therapy with C-MET and EGFR antagonists |
KR102070761B1 (en) * | 2008-03-31 | 2020-01-29 | 제넨테크, 인크. | Compositions and methods for treating and diagnosing asthma |
CN102046195A (en) | 2008-04-02 | 2011-05-04 | 宏观基因有限公司 | HER2/neu-specific antibodies and methods of using same |
EP2365003A1 (en) * | 2008-04-11 | 2011-09-14 | Emergent Product Development Seattle, LLC | CD37 immunotherapeutic and combination with bifunctional chemotherapeutic thereof |
JP5522405B2 (en) * | 2008-04-25 | 2014-06-18 | 協和発酵キリン株式会社 | Stable multivalent antibody |
US20100260668A1 (en) * | 2008-04-29 | 2010-10-14 | Abbott Laboratories | Dual Variable Domain Immunoglobulins and Uses Thereof |
NZ588554A (en) | 2008-04-29 | 2013-03-28 | Abbott Lab | Dual variable domain immunoglobulins and uses thereof |
CN102112494A (en) | 2008-06-03 | 2011-06-29 | 雅培制药有限公司 | Dual variable domain immunoglobulins and uses thereof |
AR072001A1 (en) | 2008-06-03 | 2010-07-28 | Abbott Lab | IMMUNOGLOBULIN WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
WO2010006060A2 (en) * | 2008-07-08 | 2010-01-14 | Abbott Laboratories | Prostaglandin e2 dual variable domain immunoglobulins and uses thereof |
CA2729961C (en) | 2008-07-09 | 2018-05-01 | Biogen Idec Ma Inc. | Li113, li62 variant co2, anti-lingo antibodies |
EA201100228A1 (en) * | 2008-07-21 | 2011-08-30 | Иммьюномедикс, Инк. | STRUCTURAL VARIANTS OF ANTIBODIES TO IMPROVE THE THERAPEUTIC CHARACTERISTICS |
TW201438738A (en) | 2008-09-16 | 2014-10-16 | Genentech Inc | Methods for treating progressive multiple sclerosis |
DK2334705T3 (en) * | 2008-09-26 | 2017-03-27 | Ucb Biopharma Sprl | BIOLOGICAL PRODUCTS |
US8268314B2 (en) | 2008-10-08 | 2012-09-18 | Hoffmann-La Roche Inc. | Bispecific anti-VEGF/anti-ANG-2 antibodies |
WO2010051502A2 (en) | 2008-10-31 | 2010-05-06 | Biogen Idec Ma Inc. | Light targeting molecules and uses thereof |
JP6041489B2 (en) | 2008-11-22 | 2016-12-07 | ジェネンテック, インコーポレイテッド | Use of anti-VEGF antibodies in combination with chemotherapy for the treatment of breast cancer |
RU2011127198A (en) * | 2008-12-04 | 2013-01-10 | Эбботт Лэборетриз | IMMUNOGLOBULINS WITH DOUBLE VARIABLE DOMAINS AND THEIR APPLICATION |
WO2010075249A2 (en) | 2008-12-22 | 2010-07-01 | Genentech, Inc. | A method for treating rheumatoid arthritis with b-cell antagonists |
US20110142836A1 (en) * | 2009-01-02 | 2011-06-16 | Olav Mella | B-cell depleting agents for the treatment of chronic fatigue syndrome |
JP5836807B2 (en) | 2009-03-05 | 2015-12-24 | アッヴィ・インコーポレイテッド | IL-17 binding protein |
CN101830986A (en) * | 2009-03-13 | 2010-09-15 | 北京表源生物技术有限公司 | Fusion protein polymer |
TW201544123A (en) | 2009-03-20 | 2015-12-01 | Genentech Inc | Anti-HER antibodies |
UA108199C2 (en) | 2009-03-25 | 2015-04-10 | ANTIBODY AGAINST α5β1 AND ITS APPLICATION | |
WO2010112194A1 (en) | 2009-04-02 | 2010-10-07 | F. Hoffmann-La Roche Ag | Antigen-binding polypeptides and multispecific antibodies comprising them |
RU2598248C2 (en) | 2009-04-02 | 2016-09-20 | Роше Гликарт Аг | Polyspecific antibodies containing antibody of full length and one-chain fragments fab |
KR20110124368A (en) | 2009-04-07 | 2011-11-16 | 로슈 글리카트 아게 | Bispecific anti-erbb-2/anti-c-met antibodies |
AU2010234031B2 (en) | 2009-04-07 | 2015-10-01 | Roche Glycart Ag | Trivalent, bispecific antibodies |
CN102378768A (en) | 2009-04-07 | 2012-03-14 | 罗氏格黎卡特股份公司 | Bispecific anti-erbb-3/anti-c-met antibodies |
BRPI1007602A2 (en) | 2009-05-27 | 2016-02-16 | Hoffmann La Roche | "tri or tetraspecific antibody, method for preparing a trispecific or tetraspecific antibody, host cell, composition, pharmaceutical composition and method for treating a patient in need of therapy" |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
US8703132B2 (en) | 2009-06-18 | 2014-04-22 | Hoffmann-La Roche, Inc. | Bispecific, tetravalent antigen binding proteins |
JP5758888B2 (en) | 2009-07-06 | 2015-08-05 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Bispecific digoxigenin binding antibody |
EP2456890A1 (en) | 2009-07-20 | 2012-05-30 | Genentech, Inc. | Gene expression markers for crohn's disease |
UY32808A (en) * | 2009-07-29 | 2011-02-28 | Abbott Lab | IMMUNOGLOBULINS AS A DUAL VARIABLE DOMAIN AND USES OF THE SAME |
RU2639288C2 (en) | 2009-08-11 | 2017-12-20 | Дженентек, Инк. | Proteins production in cultural media without glutamine |
WO2011022264A1 (en) | 2009-08-15 | 2011-02-24 | Genentech, Inc. | Anti-angiogenesis therapy for the treatment of previously treated breast cancer |
CN105131112A (en) | 2009-08-29 | 2015-12-09 | Abbvie公司 | Therapeutic dll4 binding proteins |
EP2473524A4 (en) | 2009-09-01 | 2013-05-22 | Abbott Lab | Dual variable domain immunoglobulins and uses thereof |
WO2011028952A1 (en) | 2009-09-02 | 2011-03-10 | Xencor, Inc. | Compositions and methods for simultaneous bivalent and monovalent co-engagement of antigens |
US20120283415A1 (en) * | 2009-09-10 | 2012-11-08 | Ucb Pharma S.A. | Multivalent Antibodies |
SG179196A1 (en) | 2009-09-16 | 2012-04-27 | Genentech Inc | Coiled coil and/or tether containing protein complexes and uses thereof |
US20110076232A1 (en) * | 2009-09-29 | 2011-03-31 | Ludwig Institute For Cancer Research | Specific binding proteins and uses thereof |
EP2488200A4 (en) * | 2009-10-14 | 2013-06-12 | Merck Sharp & Dohme | April antagonists and methods of use |
EP2488658A4 (en) | 2009-10-15 | 2013-06-19 | Abbvie Inc | Dual variable domain immunoglobulins and uses thereof |
UY32979A (en) | 2009-10-28 | 2011-02-28 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
WO2011091078A2 (en) | 2010-01-19 | 2011-07-28 | Xencor, Inc. | Antibody fc variants with enhanced complement activity |
RU2573994C2 (en) | 2010-02-10 | 2016-01-27 | Иммьюноджен, Инк | Anti-cd20 antibodies and thereof application |
US9120855B2 (en) | 2010-02-10 | 2015-09-01 | Novartis Ag | Biologic compounds directed against death receptor 5 |
WO2011102342A1 (en) | 2010-02-16 | 2011-08-25 | 国立大学法人京都工芸繊維大学 | Antibody-immobilized carrier, method of producing antibody-immobilized carrier, and use of said antibody-immobilized carrier |
US9556249B2 (en) | 2010-02-18 | 2017-01-31 | Genentech, Inc. | Neuregulin antagonists and use thereof in treating cancer |
EP3696194A1 (en) | 2010-02-23 | 2020-08-19 | F. Hoffmann-La Roche AG | Anti-angiogenesis therapy for the treatment of ovarian cancer |
MY160628A (en) | 2010-03-02 | 2017-03-15 | Abbvie Inc | Therapeutic DLL4 Binding Proteins |
KR101899835B1 (en) | 2010-03-24 | 2018-09-19 | 제넨테크, 인크. | Anti-lrp6 antibodies |
EP3153521B1 (en) | 2010-03-26 | 2019-09-04 | Trustees of Dartmouth College | Vista regulatory t cell mediator protein, vista binding agents and use thereof |
AR080793A1 (en) | 2010-03-26 | 2012-05-09 | Roche Glycart Ag | BISPECIFIC ANTIBODIES |
AR080794A1 (en) | 2010-03-26 | 2012-05-09 | Hoffmann La Roche | BIVING SPECIFIC ANTIBODIES ANTI-VEGF / ANTI-ANG-2 |
US10745467B2 (en) | 2010-03-26 | 2020-08-18 | The Trustees Of Dartmouth College | VISTA-Ig for treatment of autoimmune, allergic and inflammatory disorders |
US20150231215A1 (en) | 2012-06-22 | 2015-08-20 | Randolph J. Noelle | VISTA Antagonist and Methods of Use |
CA2796633C (en) | 2010-04-23 | 2020-10-27 | Genentech, Inc. | Production of heteromultimeric proteins |
JP2013533211A (en) | 2010-05-04 | 2013-08-22 | メリマック ファーマシューティカルズ インコーポレーティッド | Antibodies against epidermal growth factor receptor (EGFR) and uses thereof |
PE20130205A1 (en) | 2010-05-14 | 2013-03-24 | Abbvie Inc | IL-1 BINDING PROTEINS |
WO2011146568A1 (en) | 2010-05-19 | 2011-11-24 | Genentech, Inc. | Predicting response to a her inhibitor |
WO2011147834A1 (en) | 2010-05-26 | 2011-12-01 | Roche Glycart Ag | Antibodies against cd19 and uses thereof |
WO2011153243A2 (en) | 2010-06-02 | 2011-12-08 | Genentech, Inc. | Anti-angiogenesis therapy for treating gastric cancer |
EP2582729A4 (en) | 2010-06-18 | 2014-05-28 | Hoffmann La Roche | Anti-axl antibodies and methods of use |
WO2011161119A1 (en) | 2010-06-22 | 2011-12-29 | F. Hoffmann-La Roche Ag | Antibodies against insulin-like growth factor i receptor and uses thereof |
WO2011161189A1 (en) | 2010-06-24 | 2011-12-29 | F. Hoffmann-La Roche Ag | Anti-hepsin antibodies and methods of use |
WO2012006500A2 (en) | 2010-07-08 | 2012-01-12 | Abbott Laboratories | Monoclonal antibodies against hepatitis c virus core protein |
UY33492A (en) | 2010-07-09 | 2012-01-31 | Abbott Lab | IMMUNOGLOBULINS WITH DUAL VARIABLE DOMAIN AND USES OF THE SAME |
EP2591004A1 (en) | 2010-07-09 | 2013-05-15 | F.Hoffmann-La Roche Ag | Anti-neuropilin antibodies and methods of use |
WO2012009705A1 (en) | 2010-07-15 | 2012-01-19 | Zyngenia, Inc. | Ang-2 binding complexes and uses thereof |
WO2012010549A1 (en) | 2010-07-19 | 2012-01-26 | F. Hoffmann-La Roche Ag | Method to identify a patient with an increased likelihood of responding to an anti-cancer therapy |
SG187120A1 (en) | 2010-07-19 | 2013-02-28 | Hoffmann La Roche | Method to identify a patient with an increased likelihood of responding to an anti-cancer therapy |
WO2012010582A1 (en) | 2010-07-21 | 2012-01-26 | Roche Glycart Ag | Anti-cxcr5 antibodies and methods of use |
US20130177500A1 (en) | 2010-07-23 | 2013-07-11 | Trustee Of Boston University | Anti-despr inhibitors as therapeutics for inhibition of pathological angiogenesis and tumor cell invasiveness and for molecular imaging and targeted delivery |
EP3252072A3 (en) | 2010-08-03 | 2018-03-14 | AbbVie Inc. | Dual variable domain immunoglobulins and uses thereof |
RU2013106216A (en) | 2010-08-03 | 2014-09-10 | Ф. Хоффманн-Ля Рош Аг | BIOMARKERS OF CHRONIC Lymphocytic Leukemia |
KR20130049196A (en) | 2010-08-05 | 2013-05-13 | 에프. 호프만-라 로슈 아게 | Anti-mhc antibody anti-viral cytokine fusion protein |
WO2012020038A1 (en) | 2010-08-13 | 2012-02-16 | Roche Glycart Ag | Anti-tenascin-c a2 antibodies and methods of use |
CN104474546A (en) | 2010-08-13 | 2015-04-01 | 弗·哈夫曼-拉罗切有限公司 | Antibodies to il-1beta and il-18, for treatment of disease |
CR20180142A (en) | 2010-08-13 | 2018-04-05 | Roche Glycart Ag | ANTI-FAP ANTIBODIES AND METHODS OF USE (Divisional Exp: 2013-0038) |
WO2012025530A1 (en) | 2010-08-24 | 2012-03-01 | F. Hoffmann-La Roche Ag | Bispecific antibodies comprising a disulfide stabilized - fv fragment |
CA2807269A1 (en) | 2010-08-24 | 2012-03-01 | Roche Glycart Ag | Activatable bispecific antibodies |
CN103080132B (en) | 2010-08-25 | 2016-06-08 | 弗·哈夫曼-拉罗切有限公司 | The antibody of anti-IL-18R1 and purposes thereof |
KR20130139884A (en) | 2010-08-26 | 2013-12-23 | 애브비 인코포레이티드 | Dual variable domain immunoglobulins and uses thereof |
DK2612151T3 (en) | 2010-08-31 | 2017-10-02 | Genentech Inc | BIOMARKETS AND METHODS OF TREATMENT |
AU2011324870B2 (en) | 2010-11-01 | 2015-01-29 | Symphogen A/S | pan-HER antibody composition |
KR20190120439A (en) | 2010-11-08 | 2019-10-23 | 제넨테크, 인크. | Subcutaneously administered anti-il-6 receptor antibody |
EP2638070B1 (en) | 2010-11-10 | 2016-10-19 | F.Hoffmann-La Roche Ag | Methods and compositions for neural disease immunotherapy |
WO2012071436A1 (en) | 2010-11-24 | 2012-05-31 | Genentech, Inc. | Method of treating autoimmune inflammatory disorders using il-23r loss-of-function mutants |
CN103443125B (en) | 2010-11-30 | 2019-05-03 | 霍夫曼-拉罗奇有限公司 | Low-affinity blood-brain barrier receptor antibody and application thereof |
SG191124A1 (en) | 2010-12-15 | 2013-07-31 | Kek High Energy Accelerator | Protein production method |
AR084342A1 (en) | 2010-12-16 | 2013-05-08 | Genentech Inc | DIAGNOSIS AND TREATMENTS RELATED TO THE INHIBITION OF TH2 |
CA3167037A1 (en) * | 2010-12-20 | 2012-06-28 | The Rockefeller University | Modulating agonistic tnfr antibodies |
JP6253987B2 (en) | 2010-12-20 | 2017-12-27 | ジェネンテック, インコーポレイテッド | Anti-mesothelin antibodies and immunoconjugates |
SG191312A1 (en) | 2010-12-21 | 2013-07-31 | Abbvie Inc | Il-1 -alpha and -beta bispecific dual variable domain immunoglobulins and their use |
US20120195910A1 (en) | 2010-12-22 | 2012-08-02 | Genentech, Inc. | Anti-pcsk9 antibodies and methods of use |
JP5766296B2 (en) | 2010-12-23 | 2015-08-19 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Polypeptide-polynucleotide complexes and their use in targeted delivery of effector components |
US10689447B2 (en) | 2011-02-04 | 2020-06-23 | Genentech, Inc. | Fc variants and methods for their production |
KR101913448B1 (en) | 2011-02-04 | 2018-10-30 | 제넨테크, 인크. | Fc VARIANTS AND METHODS FOR THEIR PRODUCTION |
BR112013019975A2 (en) | 2011-02-28 | 2017-08-01 | Hoffmann La Roche | "Antigen binding proteins, pharmaceutical composition, use of an antigen binding protein, method for treating a patient and method for preparing an antigen binding protein, nucleic acid, vector and host cell" |
EP2681240B1 (en) | 2011-02-28 | 2017-08-16 | F. Hoffmann-La Roche AG | Monovalent antigen binding proteins |
CN103649118A (en) | 2011-03-01 | 2014-03-19 | 安进公司 | Bispecific binding agents |
WO2012119989A2 (en) | 2011-03-04 | 2012-09-13 | Oryzon Genomics, S.A. | Methods and antibodies for the diagnosis and treatment of cancer |
AR085911A1 (en) | 2011-03-16 | 2013-11-06 | Sanofi Sa | SAFE THERAPEUTIC DOSE OF A SIMILAR PROTEIN TO AN ANTIBODY WITH VUAL REGION |
TWI671315B (en) * | 2011-03-28 | 2019-09-11 | 法商賽諾菲公司 | Dual variable region antibody-like binding proteins having cross-over binding region orientation |
WO2012130831A1 (en) | 2011-03-29 | 2012-10-04 | Roche Glycart Ag | Antibody fc variants |
MX342240B (en) | 2011-04-07 | 2016-09-21 | Genentech Inc | Anti-fgfr4 antibodies and methods of use. |
GB201106395D0 (en) | 2011-04-14 | 2011-06-01 | Hubrecht Inst | Compounds |
CN103502273A (en) | 2011-04-20 | 2014-01-08 | 罗氏格黎卡特股份公司 | Method and constructs for the pH dependent passage of the blood-brain-barrier |
JP5987053B2 (en) | 2011-05-12 | 2016-09-06 | ジェネンテック, インコーポレイテッド | Multiple reaction monitoring LC-MS / MS method for detecting therapeutic antibodies in animal samples using framework signature peptides |
MX370233B (en) | 2011-05-16 | 2019-12-06 | Genentech Inc | Fgfr1 agonists and methods of use. |
WO2012162561A2 (en) | 2011-05-24 | 2012-11-29 | Zyngenia, Inc. | Multivalent and monovalent multispecific complexes and their uses |
US8691231B2 (en) | 2011-06-03 | 2014-04-08 | Merrimack Pharmaceuticals, Inc. | Methods of treatment of tumors expressing predominantly high affinity EGFR ligands or tumors expressing predominantly low affinity EGFR ligands with monoclonal and oligoclonal anti-EGFR antibodies |
BR112013032235A2 (en) | 2011-06-15 | 2016-11-22 | Hoffmann La Roche | anti-human epo receptor antibodies and methods of use |
KR20140045440A (en) | 2011-06-30 | 2014-04-16 | 제넨테크, 인크. | Anti-c-met antibody formulations |
EP2543680A1 (en) * | 2011-07-07 | 2013-01-09 | Centre National de la Recherche Scientifique | Multispecific mutated antibody Fab fragments |
US20130022551A1 (en) | 2011-07-22 | 2013-01-24 | Trustees Of Boston University | DEspR ANTAGONISTS AND AGONISTS AS THERAPEUTICS |
CN103890007A (en) | 2011-08-17 | 2014-06-25 | 霍夫曼-拉罗奇有限公司 | Neuregulin antibodies and uses thereof |
PT2748202T (en) * | 2011-08-23 | 2018-10-08 | Roche Glycart Ag | Bispecific antigen binding molecules |
HUE038225T2 (en) * | 2011-08-23 | 2018-10-29 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
RU2014109038A (en) | 2011-08-23 | 2015-09-27 | Рош Гликарт Аг | ANTIBODIES TO CHONDROITINSULFATE PROTEOGLYCAN MELANOMA |
EP2748200B1 (en) * | 2011-08-23 | 2018-04-11 | Roche Glycart AG | Fc-free antibodies comprising two fab fragments and methods of use |
BR112014004168A2 (en) * | 2011-08-23 | 2017-12-12 | Roche Glycart Ag | bispecific antibody, pharmaceutical composition, use of bispecific antibody, prokaryotic or eukaryotic host cell, antibody production method and invention |
US20130058947A1 (en) | 2011-09-02 | 2013-03-07 | Stem Centrx, Inc | Novel Modulators and Methods of Use |
MX2014003094A (en) | 2011-09-15 | 2014-04-25 | Genentech Inc | Methods of promoting differentiation. |
BR112014006419A2 (en) | 2011-09-19 | 2018-08-07 | Genentech Inc | Methods to Treat a Cancer Patient, Kit and Article |
AU2012319150B2 (en) | 2011-10-05 | 2017-08-17 | Genentech, Inc. | Methods of treating liver conditions using Notch2 antagonists |
US9175308B2 (en) | 2011-10-07 | 2015-11-03 | Mie University | Chimeric antigen receptor |
PL2766397T3 (en) | 2011-10-11 | 2018-10-31 | F.Hoffmann-La Roche Ag | Improved assembly of bispecific antibodies |
PT2766393T (en) | 2011-10-14 | 2018-10-15 | Hoffmann La Roche | Anti-htra1 antibodies and methods of use |
RU2014119426A (en) | 2011-10-15 | 2015-11-20 | Дженентек, Инк. | WAYS OF APPLICATION OF SCD1 ANTAGONISTS |
WO2013059531A1 (en) | 2011-10-20 | 2013-04-25 | Genentech, Inc. | Anti-gcgr antibodies and uses thereof |
AR088514A1 (en) | 2011-10-24 | 2014-06-18 | Abbvie Inc | BISPECIFIC IMMUNOLIGANTS DIRECTED AGAINST TNF |
UY34411A (en) | 2011-10-24 | 2013-05-31 | Abbvie Inc | IMMUNO LINKERS AGAINST SCLEROSTINE |
AR088509A1 (en) | 2011-10-28 | 2014-06-11 | Genentech Inc | THERAPEUTIC COMBINATIONS AND METHODS TO TREAT MELANOMA |
CN103906533A (en) * | 2011-11-07 | 2014-07-02 | 米迪缪尼有限公司 | Multispecific and multivalent binding proteins and uses thereof |
MX2014005885A (en) | 2011-11-21 | 2014-09-04 | Genentech Inc | Purification of anti-c-met antibodies. |
US20140335084A1 (en) | 2011-12-06 | 2014-11-13 | Hoffmann-La Roche Inc. | Antibody formulation |
WO2013092720A1 (en) | 2011-12-22 | 2013-06-27 | F. Hoffmann-La Roche Ag | Full length antibody display system for eukaryotic cells and its use |
SG10201700169PA (en) | 2011-12-22 | 2017-02-27 | Hoffmann La Roche | Expression vector element combinations, novel production cell generation methods and their use for the recombinant production of polypeptides |
SG11201403223PA (en) | 2011-12-22 | 2014-07-30 | Hoffmann La Roche | Expression vector organization, novel production cell generation methods and their use for the recombinant production of polypeptides |
WO2013096346A1 (en) * | 2011-12-22 | 2013-06-27 | Development Center For Biotechnology | Bispecific t-cell activator antibody |
AR089434A1 (en) | 2011-12-23 | 2014-08-20 | Genentech Inc | PROCEDURE TO PREPARE FORMULATIONS WITH HIGH CONCENTRATION OF PROTEINS |
WO2013101771A2 (en) | 2011-12-30 | 2013-07-04 | Genentech, Inc. | Compositions and method for treating autoimmune diseases |
CN104159920A (en) | 2011-12-30 | 2014-11-19 | 艾伯维公司 | Dual specific binding proteins directed against il-13 and/or il-17 |
EP2915818A3 (en) | 2011-12-30 | 2015-11-11 | AbbVie Inc. | Dual variable domain immunoglobulins and uses thereof |
CN104066449B (en) | 2012-01-18 | 2018-04-27 | 霍夫曼-拉罗奇有限公司 | Anti- LRP5 antibody and application method |
CA2862424A1 (en) | 2012-01-18 | 2013-07-25 | Genentech, Inc. | Methods of using fgf19 modulators |
MX366965B (en) | 2012-02-03 | 2019-07-31 | Hoffmann La Roche | Bispecific antibody molecules with antigen-transfected t-cells and their use in medicine. |
KR20140127854A (en) | 2012-02-10 | 2014-11-04 | 제넨테크, 인크. | Single-chain antibodies and other heteromultimers |
RU2014133069A (en) | 2012-02-11 | 2016-04-10 | Дженентек, Инк. | R-SPONDIN TRANSLOCATIONS AND WAYS WITH THEIR USE |
JP6152120B2 (en) | 2012-02-15 | 2017-06-21 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Affinity chromatography based on Fc receptors |
PT2825559T (en) * | 2012-03-13 | 2019-06-07 | Novimmune Sa | Readily isolated bispecific antibodies with native immunoglobulin format |
WO2013139956A1 (en) | 2012-03-22 | 2013-09-26 | Thrombogenics Nv | Antibodies abrogating cell binding to lactadherin |
RU2014136886A (en) | 2012-03-27 | 2016-05-20 | Дженентек, Инк. | DIAGNOSTIC AND TREATMENT TYPES RELATED TO HER3 INHIBITORS |
AR090549A1 (en) | 2012-03-30 | 2014-11-19 | Genentech Inc | ANTI-LGR5 AND IMMUNOCATE PLAYERS |
KR101674784B1 (en) | 2012-04-05 | 2016-11-09 | 에프. 호프만-라 로슈 아게 | Bispecific antibodies against human tweak and human il17 and uses thereof |
MX2014012843A (en) | 2012-04-24 | 2017-01-23 | Thrombogenics Nv | Anti-pdgf-c antibodies. |
BR112014027166A2 (en) | 2012-05-01 | 2017-06-27 | Genentech Inc | antibody, nucleic acid, host cell, method for producing an antibody, immunoconjugate, pharmaceutical formulation, treatment method, proliferation inhibiting method and detection methods. |
WO2013170191A1 (en) | 2012-05-11 | 2013-11-14 | Genentech, Inc. | Methods of using antagonists of nad biosynthesis from nicotinamide |
EP2849787A4 (en) | 2012-05-14 | 2016-06-15 | Biogen Ma Inc | Lingo-2 antagonists for treatment of conditions involving motor neurons |
EP2852618A2 (en) | 2012-05-21 | 2015-04-01 | F. Hoffmann-La Roche AG | Methods for improving safety of blood-brain barrier transport |
SI2852615T1 (en) | 2012-05-22 | 2019-02-28 | Bristol-Myers Squibb Company | Il-17a/f il-23 bispecific antibodies and their uses |
RU2625771C2 (en) | 2012-05-23 | 2017-07-18 | Дженентек, Инк. | Therapeutics selection method |
KR20150013188A (en) | 2012-05-24 | 2015-02-04 | 에프. 호프만-라 로슈 아게 | Multispecific antibodies |
MX2014014830A (en) | 2012-06-15 | 2015-05-11 | Genentech Inc | Anti-pcsk9 antibodies, formulations, dosing, and methods of use. |
US9890215B2 (en) | 2012-06-22 | 2018-02-13 | King's College London | Vista modulators for diagnosis and treatment of cancer |
WO2014039983A1 (en) | 2012-09-07 | 2014-03-13 | The Trustees Of Dartmouth College | Vista modulators for diagnosis and treatment of cancer |
BR112014028368A2 (en) | 2012-06-27 | 2017-11-14 | Hoffmann La Roche | method of producing antibody fc region conjugate, antibody fc region conjugate and pharmaceutical formulation |
CA2871880A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
BR112014030843A2 (en) | 2012-07-04 | 2019-10-15 | Hoffmann La Roche | anti-theophylline antibody, pharmaceutical formulation and use of the antibody |
CA2872192A1 (en) | 2012-07-04 | 2014-01-09 | F. Hoffmann-La Roche Ag | Anti-biotin antibodies and methods of use |
CN107973856B (en) | 2012-07-04 | 2021-11-23 | 弗·哈夫曼-拉罗切有限公司 | Covalently linked antigen-antibody conjugates |
CN110042114A (en) | 2012-07-05 | 2019-07-23 | 弗·哈夫曼-拉罗切有限公司 | Expression and excretory system |
BR112015000441A2 (en) | 2012-07-09 | 2017-12-19 | Genentech Inc | Immunoconjugates, Pharmaceutical Formulation and Treatment Method and Method for Inhibiting Proliferation of a CD22 Positive Cell |
IN2014DN10652A (en) | 2012-07-09 | 2015-09-11 | Genentech Inc | |
EA201590173A1 (en) | 2012-07-09 | 2015-09-30 | Дженентек, Инк. | IMMUNOCONGUTS CONTAINING ANTIBODIES TO CD22 |
CN104411337A (en) | 2012-07-09 | 2015-03-11 | 基因泰克公司 | Immunoconjugates comprising anti-cd79b antibodies |
WO2014011955A2 (en) | 2012-07-12 | 2014-01-16 | Abbvie, Inc. | Il-1 binding proteins |
MA37794B1 (en) | 2012-07-13 | 2017-07-31 | Roche Glycart Ag | Anti-vegf / anti-ang-2 bispecific antibodies and their use in the treatment of ocular vascular pathologies |
ES2786263T3 (en) | 2012-07-13 | 2020-10-09 | Univ Pennsylvania | Enhancement of T-lymphocyte CAR activity by co-introduction of a bispecific antibody |
US20150203591A1 (en) * | 2012-08-02 | 2015-07-23 | Regeneron Pharmaceuticals, Inc. | Mutivalent antigen-binding proteins |
EP3315514A1 (en) | 2012-08-29 | 2018-05-02 | F. Hoffmann-La Roche AG | Blood brain barrier shuttle |
WO2014056783A1 (en) | 2012-10-08 | 2014-04-17 | Roche Glycart Ag | Fc-free antibodies comprising two fab-fragments and methods of use |
KR20180008921A (en) | 2012-11-01 | 2018-01-24 | 애브비 인코포레이티드 | Anti-vegf/dll4 dual variable domain immunoglobulins and uses thereof |
CA2890207A1 (en) | 2012-11-05 | 2014-05-08 | Foundation Medicine, Inc. | Novel ntrk1 fusion molecules and uses thereof |
US9725512B2 (en) | 2012-11-08 | 2017-08-08 | Hoffmann-La Roche Inc. | HER3 antibodies binding to the beta-hairpin of HER3 |
MA38176A1 (en) | 2012-11-13 | 2017-06-30 | Genentech Inc | Novel anti-haemagglutinin antibody, useful for the treatment, inhibition or prevention of viral influenza infection |
EP2928919A2 (en) | 2012-12-04 | 2015-10-14 | AbbVie Inc. | Blood-brain barrier (bbb) penetrating dual specific binding proteins |
US9458244B2 (en) | 2012-12-28 | 2016-10-04 | Abbvie Inc. | Single chain multivalent binding protein compositions and methods |
EP2938637A2 (en) | 2012-12-28 | 2015-11-04 | AbbVie Inc. | Multivalent binding protein compositions |
WO2014106001A2 (en) | 2012-12-28 | 2014-07-03 | Abbvie, Inc. | Dual specific binding proteins having a receptor sequence |
US10980804B2 (en) | 2013-01-18 | 2021-04-20 | Foundation Medicine, Inc. | Methods of treating cholangiocarcinoma |
EP2948177A1 (en) | 2013-01-22 | 2015-12-02 | AbbVie Inc. | Methods for optimizing domain stability of binding proteins |
WO2014116749A1 (en) | 2013-01-23 | 2014-07-31 | Genentech, Inc. | Anti-hcv antibodies and methods of using thereof |
US10047163B2 (en) | 2013-02-08 | 2018-08-14 | Abbvie Stemcentrx Llc | Multispecific constructs |
CN104994879A (en) | 2013-02-22 | 2015-10-21 | 霍夫曼-拉罗奇有限公司 | Methods of treating cancer and preventing drug resistance |
RU2015140921A (en) | 2013-02-26 | 2017-04-03 | Роше Гликарт Аг | ANTIBODIES TO MCSP |
SG11201504497TA (en) * | 2013-02-26 | 2015-09-29 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
KR20150122761A (en) * | 2013-02-26 | 2015-11-02 | 로슈 글리카트 아게 | Bispecific t cell activating antigen binding molecules |
JP2016510751A (en) | 2013-03-06 | 2016-04-11 | ジェネンテック, インコーポレイテッド | Methods of treating and preventing anticancer drug resistance |
DK2970487T3 (en) | 2013-03-12 | 2020-05-11 | Molecular Templates Inc | Cytotoxic proteins comprising cell-targeted binding regions and Shiga toxin A subunit regions to selectively kill specific cell types |
CA2903480A1 (en) | 2013-03-14 | 2014-09-25 | Genentech, Inc. | Combinations of a mek inhibitor compound with an her3/egfr inhibitor compound and methods of use |
BR112015022604A2 (en) | 2013-03-14 | 2017-10-24 | Genentech Inc | Uses of a Chromatin Modifier Modulator and an Egfr Antagonist |
US9562099B2 (en) | 2013-03-14 | 2017-02-07 | Genentech, Inc. | Anti-B7-H4 antibodies and immunoconjugates |
MX2015010777A (en) | 2013-03-14 | 2016-04-25 | Genentech Inc | Anti-b7-h4 antibodies and immunoconjugates. |
CN105451767B (en) | 2013-03-15 | 2019-10-18 | 泽恩格尼亚股份有限公司 | Multivalence and monovalent polyspecific compound and application thereof |
CN105324396A (en) | 2013-03-15 | 2016-02-10 | 艾伯维公司 | Dual specific binding proteins directed against il-1 beta and il-17 |
TW201446800A (en) | 2013-03-15 | 2014-12-16 | Abbvie Inc | Dual specific binding proteins directed against TNF α |
BR112015023262B8 (en) | 2013-03-15 | 2024-02-06 | Ac Immune Sa | Isolated antibody, immunoconjugate, pharmaceutical formulation and uses of antibody |
CN105143265A (en) | 2013-03-15 | 2015-12-09 | 豪夫迈·罗氏有限公司 | Anti-crth2 antibodies and their use |
EP2968537A1 (en) | 2013-03-15 | 2016-01-20 | Genentech, Inc. | Methods of treating cancer and preventing cancer drug resistance |
EP2970484B2 (en) | 2013-03-15 | 2022-09-21 | Amgen Inc. | Heterodimeric bispecific antibodies |
SG11201507427QA (en) | 2013-03-15 | 2015-10-29 | Genentech Inc | Compositions and methods for diagnosis and treatment of hepatic cancers |
KR102158924B1 (en) | 2013-03-15 | 2020-09-22 | 제넨테크, 인크. | Biomarkers and methods of treating pd-1 and pd-l1 related conditions |
UA118028C2 (en) | 2013-04-03 | 2018-11-12 | Рош Глікарт Аг | Bispecific antibodies specific for fap and dr5, antibodies specific for dr5 and methods of use |
WO2014177459A2 (en) | 2013-04-29 | 2014-11-06 | F. Hoffmann-La Roche Ag | Fc-receptor binding modified asymmetric antibodies and methods of use |
SG10201810481UA (en) | 2013-04-29 | 2018-12-28 | Hoffmann La Roche | Fcrn-binding abolished anti-igf-1r antibodies and their use in the treatment of vascular eye diseases |
KR102282134B1 (en) | 2013-04-29 | 2021-07-27 | 에프. 호프만-라 로슈 아게 | Human fcrn-binding modified antibodies and methods of use |
EP2999716A2 (en) | 2013-05-20 | 2016-03-30 | F. Hoffmann-La Roche AG | Anti-transferrin receptor antibodies and methods of use |
CN104341504B (en) | 2013-08-06 | 2017-10-24 | 百奥泰生物科技(广州)有限公司 | Bispecific antibody |
US10093978B2 (en) | 2013-08-12 | 2018-10-09 | Genentech, Inc. | Compositions for detecting complement factor H (CFH) and complement factor I (CFI) polymorphisms |
WO2015025054A1 (en) | 2013-08-22 | 2015-02-26 | Medizinische Universität Wien | Dye-specific antibodies for prestained molecular weight markers and methods producing the same |
CN105518027A (en) | 2013-09-17 | 2016-04-20 | 豪夫迈·罗氏有限公司 | Methods of using anti-LGR5 antibodies |
US20150125397A1 (en) | 2013-10-06 | 2015-05-07 | Abbvie Inc. | Dual specific binding proteins directed against immune cell receptors and autoantigens |
WO2015052230A1 (en) | 2013-10-11 | 2015-04-16 | F. Hoffmann-La Roche Ag | Multispecific domain exchanged common variable light chain antibodies |
WO2015054670A1 (en) | 2013-10-11 | 2015-04-16 | Genentech, Inc. | Nsp4 inhibitors and methods of use |
KR20160070136A (en) | 2013-10-18 | 2016-06-17 | 제넨테크, 인크. | Anti-rsp02 and/or anti-rsp03 antibodies and their uses |
RU2016119425A (en) | 2013-10-23 | 2017-11-28 | Дженентек, Инк. | METHODS FOR DIAGNOSIS AND TREATMENT OF EOSINOPHILIC DISEASES |
WO2015066543A1 (en) | 2013-11-01 | 2015-05-07 | Board Of Regents, The University Of Texas System | Targeting her2 and her3 with bispecific antibodies in cancerous cells |
EP3071597B1 (en) | 2013-11-21 | 2020-07-29 | F.Hoffmann-La Roche Ag | Anti-alpha-synuclein antibodies and methods of use |
TW201536320A (en) | 2013-12-02 | 2015-10-01 | Abbvie Inc | Compositions and methods for treating osteoarthritis |
WO2015089344A1 (en) | 2013-12-13 | 2015-06-18 | Genentech, Inc. | Anti-cd33 antibodies and immunoconjugates |
MX2016007958A (en) | 2013-12-17 | 2016-08-03 | Genentech Inc | Anti-cd3 antibodies and methods of use. |
PL3083680T3 (en) | 2013-12-20 | 2020-06-29 | F. Hoffmann-La Roche Ag | Humanized anti-tau(ps422) antibodies and methods of use |
TWI670283B (en) | 2013-12-23 | 2019-09-01 | 美商建南德克公司 | Antibodies and methods of use |
TW201609805A (en) | 2013-12-23 | 2016-03-16 | 美國禮來大藥廠 | Multifunctional antibodies binding to EGFR and MET |
CN106661107B (en) | 2013-12-24 | 2021-12-24 | 杨森制药公司 | anti-VISTA antibodies and fragments |
US11014987B2 (en) | 2013-12-24 | 2021-05-25 | Janssen Pharmaceutics Nv | Anti-vista antibodies and fragments, uses thereof, and methods of identifying same |
RU2769133C2 (en) * | 2013-12-30 | 2022-03-28 | Эпимаб Биотерапьютикс Инк. | Immunoglobulin with tandem arrangement of fab fragments and application thereof |
CN105873615B (en) | 2014-01-03 | 2020-12-25 | 豪夫迈·罗氏有限公司 | Covalently linked helicar-anti-helicar antibody conjugates and uses thereof |
CN105873616B (en) | 2014-01-03 | 2020-06-05 | 豪夫迈·罗氏有限公司 | Covalently linked polypeptide toxin-antibody conjugates |
WO2015101586A1 (en) | 2014-01-03 | 2015-07-09 | F. Hoffmann-La Roche Ag | Bispecific anti-hapten/anti-blood brain barrier receptor antibodies, complexes thereof and their use as blood brain barrier shuttles |
PL3092251T3 (en) | 2014-01-06 | 2021-08-02 | F. Hoffmann-La Roche Ag | Monovalent blood brain barrier shuttle modules |
UA118278C2 (en) | 2014-01-10 | 2018-12-26 | Глаксосмітклайн Інтеллектьюел Проперті (№ 2) Лімітед | Hydroxy formamide derivatives and their use |
CA2931986A1 (en) | 2014-01-15 | 2015-07-23 | F. Hoffmann-La Roche Ag | Fc-region variants with modified fcrn- and maintained protein a-binding properties |
BR112016015693A2 (en) | 2014-01-24 | 2017-10-24 | Genentech Inc | Method To Treat Prostate Cancer And Antibody |
US20160347798A1 (en) | 2014-01-27 | 2016-12-01 | Molecular Templates, Inc. | Mhc class i epitope delivering polypeptides and cell-targeted molecules for direct cell killing and immune stimulation via mhc class i presentation and methods regarding the same |
PL3102230T3 (en) | 2014-02-08 | 2021-11-15 | F.Hoffmann-La Roche Ag | Methods of treating alzheimer's disease |
AU2015214058B2 (en) | 2014-02-08 | 2020-07-09 | Genentech, Inc. | Methods of treating Alzheimer's Disease |
AR099465A1 (en) | 2014-02-12 | 2016-07-27 | Genentech Inc | ANTI-JAGGED1 ANTIBODIES AND METHODS OF USE |
US10675352B2 (en) | 2014-02-14 | 2020-06-09 | Centrose, Llc | Extracellular targeted drug conjugates |
MX2016010729A (en) | 2014-02-21 | 2016-10-26 | Genentech Inc | Anti-il-13/il-17 bispecific antibodies and uses thereof. |
US11142584B2 (en) | 2014-03-11 | 2021-10-12 | Molecular Templates, Inc. | CD20-binding proteins comprising Shiga toxin A subunit effector regions for inducing cellular internalization and methods using same |
PT3116909T (en) | 2014-03-14 | 2020-01-30 | Novartis Ag | Antibody molecules to lag-3 and uses thereof |
PL3116999T3 (en) | 2014-03-14 | 2021-12-27 | F.Hoffmann-La Roche Ag | Methods and compositions for secretion of heterologous polypeptides |
EP3593812A3 (en) | 2014-03-15 | 2020-05-27 | Novartis AG | Treatment of cancer using chimeric antigen receptor |
US20170107294A1 (en) | 2014-03-21 | 2017-04-20 | Nordlandssykehuset Hf | Anti-cd14 antibodies and uses thereof |
MA39776A (en) | 2014-03-24 | 2017-02-01 | Hoffmann La Roche | Cancer treatment with c-met antagonists and correlation of the latter with hgf expression |
RU2016142476A (en) | 2014-03-31 | 2018-05-07 | Дженентек, Инк. | COMBINED THERAPY, INCLUDING ANTI-ANGIOGENESIS AGENTS AND AGONISTS BINDING OX40 |
MA51552A (en) | 2014-03-31 | 2021-03-24 | Hoffmann La Roche | ANTI-OX40 ANTIBODIES AND METHODS OF USE |
UA117289C2 (en) | 2014-04-02 | 2018-07-10 | Ф. Хоффманн-Ля Рош Аг | Multispecific antibodies |
SG11201608054YA (en) | 2014-04-02 | 2016-10-28 | Hoffmann La Roche | Method for detecting multispecific antibody light chain mispairing |
AU2015244814B2 (en) | 2014-04-07 | 2020-12-24 | Chugai Seiyaku Kabushiki Kaisha | Immunoactivating antigen-binding molecule |
US20150361163A1 (en) | 2014-04-18 | 2015-12-17 | Acceleron Pharma, Inc. | Methods for increasing red blood cell levels and treating sickle-cell disease |
WO2015164615A1 (en) | 2014-04-24 | 2015-10-29 | University Of Oslo | Anti-gluten antibodies and uses thereof |
WO2015171822A1 (en) | 2014-05-06 | 2015-11-12 | Genentech, Inc. | Production of heteromultimeric proteins using mammalian cells |
WO2015175375A1 (en) | 2014-05-13 | 2015-11-19 | Short Jay M | Conditionally active biological proteins |
MA39599A (en) | 2014-05-14 | 2016-10-05 | Merrimack Pharmaceuticals Inc | Dosage and administration anti-egfr therapeutics |
RU2016150370A (en) | 2014-05-22 | 2018-06-26 | Дженентек, Инк. | Antibodies and immunoconjugates against GPC3 |
KR20170005016A (en) | 2014-05-23 | 2017-01-11 | 제넨테크, 인크. | Mit biomarkers and methods using the same |
LT3148579T (en) | 2014-05-28 | 2021-05-25 | Agenus Inc. | Anti-gitr antibodies and methods of use thereof |
CA2949982A1 (en) | 2014-06-11 | 2015-12-17 | Genentech, Inc. | Anti-lgr5 antibodies and uses thereof |
JP6997619B2 (en) | 2014-06-11 | 2022-01-17 | キャシー・エイ・グリーン | Use of VISTA agonists and VISTA antagonists for suppression or enhancement of humoral immunity |
AU2015274647C1 (en) | 2014-06-11 | 2020-01-30 | Molecular Templates, Inc. | Protease-cleavage resistant, Shiga toxin a subunit effector polypeptides and cell-targeted molecules comprising the same |
EP3154566B1 (en) | 2014-06-13 | 2022-08-03 | Acceleron Pharma Inc. | Actrii antagonist for the treatment or prevention of a cutaneous ulcer in a subject that has anemia |
EP3154589A1 (en) | 2014-06-13 | 2017-04-19 | Genentech, Inc. | Methods of treating and preventing cancer drug resistance |
GB201411320D0 (en) | 2014-06-25 | 2014-08-06 | Ucb Biopharma Sprl | Antibody construct |
AR100978A1 (en) | 2014-06-26 | 2016-11-16 | Hoffmann La Roche | ANTI-Tau HUMANIZED ANTIBODY BRAIN LAUNCHERS (pS422) AND USES OF THE SAME |
CA2947504A1 (en) | 2014-06-26 | 2015-12-30 | F. Hoffmann-La Roche Ag | Anti-brdu antibodies and methods of use |
TW201623329A (en) | 2014-06-30 | 2016-07-01 | 亞佛瑞司股份有限公司 | Vaccines and monoclonal antibodies targeting truncated variants of osteopontin and uses thereof |
DK3160513T3 (en) | 2014-06-30 | 2020-04-06 | Glykos Finland Oy | SACCHARIDE DERIVATIVES OF A TOXIC PAYLOAD AND ANTIBODY CONJUGATES THEREOF |
EP3164417A1 (en) | 2014-07-01 | 2017-05-10 | Pfizer Inc. | Bispecific heterodimeric diabodies and uses thereof |
EP3680336A1 (en) | 2014-07-03 | 2020-07-15 | F. Hoffmann-La Roche AG | Polypeptide expression systems |
NZ728041A (en) | 2014-07-10 | 2023-01-27 | Affiris Ag | Substances and methods for the use in prevention and/or treatment in huntington’s disease |
CN106488775A (en) | 2014-07-11 | 2017-03-08 | 基因泰克公司 | NOTCH approach suppresses |
JP7032929B2 (en) | 2014-07-11 | 2022-03-09 | ヴェンタナ メディカル システムズ, インク. | Anti-PD-L1 antibody and its diagnostic use |
TWI685502B (en) * | 2014-07-15 | 2020-02-21 | 日商安斯泰來製藥股份有限公司 | Novel Anti-Human Tie2 Antibody |
TWI750110B (en) | 2014-07-21 | 2021-12-21 | 瑞士商諾華公司 | Treatment of cancer using humanized anti- bcma chimeric antigen receptor |
US11542488B2 (en) | 2014-07-21 | 2023-01-03 | Novartis Ag | Sortase synthesized chimeric antigen receptors |
WO2016014530A1 (en) | 2014-07-21 | 2016-01-28 | Novartis Ag | Combinations of low, immune enhancing. doses of mtor inhibitors and cars |
KR102594343B1 (en) | 2014-07-21 | 2023-10-26 | 노파르티스 아게 | Treatment of cancer using a cd33 chimeric antigen receptor |
ES2781175T3 (en) | 2014-07-31 | 2020-08-31 | Novartis Ag | Optimized subset of T cells containing a chimeric antigen receptor |
PE20170263A1 (en) | 2014-08-04 | 2017-03-30 | Hoffmann La Roche | T-CELL ACTIVATING ANTIGEN-BINDING BI-SPECIFIC MOLECULES |
CA2958200A1 (en) | 2014-08-14 | 2016-02-18 | Novartis Ag | Treatment of cancer using a gfr alpha-4 chimeric antigen receptor |
PT3183268T (en) | 2014-08-19 | 2020-05-15 | Novartis Ag | Anti-cd123 chimeric antigen receptor (car) for use in cancer treatment |
JP7286267B2 (en) | 2014-08-28 | 2023-06-05 | バイオアトラ インコーポレイテッド | Conditionally active chimeric antigen receptor for modified T cells |
US11111288B2 (en) | 2014-08-28 | 2021-09-07 | Bioatla, Inc. | Conditionally active chimeric antigen receptors for modified t-cells |
JP7085837B2 (en) | 2014-09-12 | 2022-06-17 | ジェネンテック, インコーポレイテッド | Anti-HER2 antibody and immune complex |
PE20170670A1 (en) | 2014-09-12 | 2017-06-06 | Genentech Inc | ANTI-CLL-1 ANTIBODIES AND IMMUNOCONJUGATES |
SG11201701128YA (en) | 2014-09-12 | 2017-03-30 | Genentech Inc | Cysteine engineered antibodies and conjugates |
TW201625689A (en) | 2014-09-12 | 2016-07-16 | 建南德克公司 | Anti-B7-H4 antibodies and immunoconjugates |
KR20170055521A (en) | 2014-09-17 | 2017-05-19 | 제넨테크, 인크. | Immunoconjugates comprising anti-her2 antibodies and pyrrolobenzodiazepines |
AU2015317608B2 (en) | 2014-09-17 | 2021-03-11 | Novartis Ag | Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy |
LT3262071T (en) | 2014-09-23 | 2020-06-25 | F. Hoffmann-La Roche Ag | Method of using anti-cd79b immunoconjugates |
MX2017004810A (en) | 2014-10-14 | 2017-10-16 | Novartis Ag | Antibody molecules to pd-l1 and uses thereof. |
US9732148B2 (en) | 2014-10-16 | 2017-08-15 | Genentech, Inc. | Anti-α-synuclein antibodies and methods of use |
CN106796235B (en) | 2014-11-03 | 2021-01-29 | 豪夫迈·罗氏有限公司 | Assays for detecting T cell immune subsets and methods of use thereof |
CN107109484B (en) | 2014-11-03 | 2021-12-14 | 豪夫迈·罗氏有限公司 | Methods and biomarkers for efficacy prediction and assessment of OX40 agonist treatment |
WO2016071376A2 (en) | 2014-11-06 | 2016-05-12 | F. Hoffmann-La Roche Ag | Fc-region variants with modified fcrn-binding and methods of use |
WO2016073157A1 (en) | 2014-11-06 | 2016-05-12 | Genentech, Inc. | Anti-ang2 antibodies and methods of use thereof |
JP6576456B2 (en) | 2014-11-06 | 2019-09-18 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Fc region variants with modified FcRn binding properties and protein A binding properties |
EP3218403B1 (en) | 2014-11-10 | 2020-05-13 | F.Hoffmann-La Roche Ag | Anti-interleukin-33 antibodies and uses thereof |
EP3552488A1 (en) | 2014-11-10 | 2019-10-16 | F. Hoffmann-La Roche AG | Animal model for nephropathy and agents for treating the same |
HUE049982T2 (en) * | 2014-11-14 | 2020-11-30 | Hoffmann La Roche | Antigen binding molecules comprising a tnf family ligand trimer |
US10508151B2 (en) | 2014-11-19 | 2019-12-17 | Genentech, Inc. | Anti-transferrin receptor antibodies and methods of use |
CN107108745B (en) | 2014-11-19 | 2021-01-12 | 基因泰克公司 | Antibodies against BACE1 and their use for immunotherapy of neurological diseases |
US11008403B2 (en) | 2014-11-19 | 2021-05-18 | Genentech, Inc. | Anti-transferrin receptor / anti-BACE1 multispecific antibodies and methods of use |
SG11201704056XA (en) | 2014-11-20 | 2017-06-29 | Hoffmann La Roche | Combination therapy of t cell activating bispecific antigen binding molecules cd3 abd folate receptor 1 (folr1) and pd-1 axis binding antagonists |
EP3227341A1 (en) | 2014-12-02 | 2017-10-11 | CeMM - Forschungszentrum für Molekulare Medizin GmbH | Anti-mutant calreticulin antibodies and their use in the diagnosis and therapy of myeloid malignancies |
PL3227332T3 (en) | 2014-12-03 | 2020-06-15 | F. Hoffmann-La Roche Ag | Multispecific antibodies |
US20180334490A1 (en) | 2014-12-03 | 2018-11-22 | Qilong H. Wu | Methods for b cell preconditioning in car therapy |
MA41119A (en) | 2014-12-03 | 2017-10-10 | Acceleron Pharma Inc | METHODS OF TREATMENT OF MYELODYSPLASIC SYNDROMES AND SIDEROBLASTIC ANEMIA |
AU2015357463B2 (en) | 2014-12-05 | 2021-10-07 | Immunext, Inc. | Identification of VSIG8 as the putative vista receptor and its use thereof to produce vista/VSIG8 modulators |
US9975949B2 (en) | 2014-12-05 | 2018-05-22 | Genentech, Inc. | Anti-CD79b antibodies and methods of use |
WO2016094566A2 (en) | 2014-12-10 | 2016-06-16 | Genentech, Inc. | Blood brain barrier receptor antibodies and methods of use |
US10093733B2 (en) | 2014-12-11 | 2018-10-09 | Abbvie Inc. | LRP-8 binding dual variable domain immunoglobulin proteins |
US10358502B2 (en) | 2014-12-18 | 2019-07-23 | Siwa Corporation | Product and method for treating sarcopenia |
RU2698205C2 (en) | 2014-12-18 | 2019-08-23 | Ф. Хоффманн-Ля Рош Аг | Method for determining antibodies that induce complement-dependent cytotoxicity |
US9993535B2 (en) | 2014-12-18 | 2018-06-12 | Siwa Corporation | Method and composition for treating sarcopenia |
PT3233921T (en) | 2014-12-19 | 2021-12-09 | Chugai Pharmaceutical Co Ltd | Anti-c5 antibodies and methods of use |
JP2018504400A (en) | 2015-01-08 | 2018-02-15 | バイオジェン・エムエイ・インコーポレイテッドBiogen MA Inc. | LINGO-1 antagonist and use for treatment of demyelinating disorders |
WO2016115274A1 (en) * | 2015-01-14 | 2016-07-21 | Compass Therapeutics Llc | Multispecific immunomodulatory antigen-binding constructs |
CN113956354A (en) | 2015-01-22 | 2022-01-21 | 中外制药株式会社 | Combinations and methods of use of two or more anti-C5 antibodies |
CN107250161A (en) | 2015-01-26 | 2017-10-13 | 宏观基因有限公司 | Include the multivalent molecule of DR5 binding structural domains |
JP2018506275A (en) | 2015-01-28 | 2018-03-08 | ジェネンテック, インコーポレイテッド | Gene expression markers and treatment of multiple sclerosis |
US10828353B2 (en) | 2015-01-31 | 2020-11-10 | The Trustees Of The University Of Pennsylvania | Compositions and methods for T cell delivery of therapeutic molecules |
WO2016126950A1 (en) | 2015-02-05 | 2016-08-11 | Molecular Templates, Inc. | Multivalent cd20-binding molecules comprising shiga toxin a subunit effector regions and enriched compositions thereof |
US9969800B2 (en) | 2015-02-05 | 2018-05-15 | Chugai Seiyaku Kabushiki Kaisha | IL-8 antibodies |
EP3262064A1 (en) | 2015-02-23 | 2018-01-03 | Seagull Therapeutics SAS | Non-natural semaphorins 3 and their medical use |
CA2977687C (en) | 2015-02-24 | 2024-02-13 | Hwai Wen Chang | Conditionally active proteins |
EP3265557B1 (en) | 2015-03-06 | 2019-10-16 | F. Hoffmann-La Roche AG | Ultrapurified dsba and dsbc and methods of making and using the same |
CN107430117A (en) | 2015-03-16 | 2017-12-01 | 豪夫迈·罗氏有限公司 | Detection and quantitative IL 13 method and the purposes in diagnosing and treating Th2 relevant diseases |
WO2016146833A1 (en) | 2015-03-19 | 2016-09-22 | F. Hoffmann-La Roche Ag | Biomarkers for nad(+)-diphthamide adp ribosyltransferase resistance |
MA41919A (en) | 2015-04-06 | 2018-02-13 | Acceleron Pharma Inc | ALK4 HETEROMULTIMERS: ACTRIIB AND THEIR USES |
WO2016164089A2 (en) | 2015-04-06 | 2016-10-13 | Acceleron Pharma Inc. | Tgf-beta superfamily type i and type ii receptor heteromultimers and uses thereof |
MX2017012805A (en) | 2015-04-07 | 2018-04-11 | Genentech Inc | Antigen binding complex having agonistic activity and methods of use. |
DK3280441T3 (en) | 2015-04-07 | 2021-11-15 | Alector Llc | ANTI-SORTILINE ANTIBODIES AND PROCEDURES FOR USE |
IL303972A (en) | 2015-04-08 | 2023-08-01 | Novartis Ag | Cd20 therapies, cd22 therapies, and combination therapies with a cd19 chimeric antigen receptor (car) - expressing cell |
CN108064245A (en) | 2015-04-17 | 2018-05-22 | 埃尔萨里斯生物技术公司 | Anti- Tyro3 antibody and application thereof |
EP3286211A1 (en) | 2015-04-23 | 2018-02-28 | Novartis AG | Treatment of cancer using chimeric antigen receptor and protein kinase a blocker |
SI3286315T1 (en) | 2015-04-24 | 2021-09-30 | F. Hoffmann-La Roche Ag | Methods of identifying bacteria comprising binding polypeptides |
MY189692A (en) | 2015-05-07 | 2022-02-26 | Memorial Sloan Kettering Cancer Center | Anti-ox40 antibodies and methods of use thereof |
JP6963508B2 (en) | 2015-05-11 | 2021-11-10 | ジェネンテック, インコーポレイテッド | Compositions and Methods for Treating Lupus Nephritis |
US10570185B2 (en) | 2015-05-11 | 2020-02-25 | Northwestern University | Method to detect autoantibody reactivity for deamidated insulin autoantigen in diabetes |
JP7048319B2 (en) | 2015-05-12 | 2022-04-05 | ジェネンテック, インコーポレイテッド | Treatment and diagnosis methods for cancer |
KR20180013881A (en) | 2015-05-29 | 2018-02-07 | 제넨테크, 인크. | PD-L1 promoter methylation in cancer |
JP7144935B2 (en) | 2015-05-29 | 2022-09-30 | ジェネンテック, インコーポレイテッド | Therapeutic and diagnostic methods for cancer |
CN107771182A (en) | 2015-05-29 | 2018-03-06 | 豪夫迈·罗氏有限公司 | The anti-Ebola virus glycoproteins antibody of humanization and application method |
HUE051246T2 (en) | 2015-05-30 | 2021-03-01 | Molecular Templates Inc | De-immunized, shiga toxin a subunit scaffolds and cell-targeting molecules comprising the same |
CN107810012A (en) | 2015-06-02 | 2018-03-16 | 豪夫迈·罗氏有限公司 | Use the composition and method of the anti-Antybody therapy sacred diseases of IL 34 |
US10112990B2 (en) | 2015-06-05 | 2018-10-30 | Genentech, Inc. | Anti-Tau antibodies and methods of use |
MX2017015937A (en) | 2015-06-08 | 2018-12-11 | Genentech Inc | Methods of treating cancer using anti-ox40 antibodies and pd-1 axis binding antagonists. |
MX2017014740A (en) | 2015-06-08 | 2018-08-15 | Genentech Inc | Methods of treating cancer using anti-ox40 antibodies. |
TW201710286A (en) | 2015-06-15 | 2017-03-16 | 艾伯維有限公司 | Binding proteins against VEGF, PDGF, and/or their receptors |
EP3307780A1 (en) | 2015-06-15 | 2018-04-18 | Genentech, Inc. | Antibodies and immunoconjugates |
JP6996983B2 (en) | 2015-06-16 | 2022-02-21 | ジェネンテック, インコーポレイテッド | Anti-CLL-1 antibody and how to use |
JP2018526972A (en) | 2015-06-16 | 2018-09-20 | ジェネンテック, インコーポレイテッド | Anti-CD3 antibody and method of use |
WO2016205531A2 (en) | 2015-06-17 | 2016-12-22 | Genentech, Inc. | Anti-her2 antibodies and methods of use |
LT3313879T (en) | 2015-06-24 | 2022-03-25 | F. Hoffmann-La Roche Ag | Anti-transferrin receptor antibodies with tailored affinity |
US9862763B2 (en) | 2015-06-24 | 2018-01-09 | Hoffmann-La Roche Inc. | Humanized anti-tau(pS422) antibodies and methods of use |
CN107922497B (en) | 2015-06-24 | 2022-04-12 | 詹森药业有限公司 | anti-VISTA antibodies and fragments |
EP3314255B1 (en) | 2015-06-25 | 2019-07-31 | H. Hoffnabb-La Roche Ag | Cell based assay for determining antibody or ligand binding and function |
CA2989936A1 (en) | 2015-06-29 | 2017-01-05 | Genentech, Inc. | Type ii anti-cd20 antibody for use in organ transplantation |
WO2017001350A1 (en) | 2015-06-29 | 2017-01-05 | Ventana Medical Systems, Inc. | Materials and methods for performing histochemical assays for human pro-epiregulin and amphiregulin |
EP3112381A1 (en) * | 2015-07-01 | 2017-01-04 | FONDAZIONE IRCCS Istituto Nazionale dei Tumori | Bispecific antibodies for use in cancer immunotherapy |
SG10201913807QA (en) * | 2015-07-23 | 2020-03-30 | Inhibrx Inc | Multivalent and multispecific gitr-binding fusion proteins |
US20180207273A1 (en) | 2015-07-29 | 2018-07-26 | Novartis Ag | Combination therapies comprising antibody molecules to tim-3 |
DK3317301T3 (en) | 2015-07-29 | 2021-06-28 | Immutep Sas | COMBINATION THERAPIES INCLUDING ANTIBODY MOLECULES AGAINST LAYER-3 |
EP3331550B1 (en) | 2015-08-04 | 2022-11-02 | Acceleron Pharma Inc. | Composition for treating myeloproliferative disorders |
CN105384825B (en) | 2015-08-11 | 2018-06-01 | 南京传奇生物科技有限公司 | A kind of bispecific chimeric antigen receptor and its application based on single domain antibody |
CN108135932A (en) | 2015-08-28 | 2018-06-08 | 宾夕法尼亚大学董事会 | Express the method and composition of the cell of signal transduction molecule in chimeric cell |
WO2017040195A1 (en) | 2015-08-28 | 2017-03-09 | The Trustees Of The University Of Pennsylvania | Methods and compositions for cells expressing a chimeric intracellular signaling molecule |
JP6266164B2 (en) | 2015-09-18 | 2018-01-31 | 中外製薬株式会社 | Antibodies that bind to IL-8 and uses thereof |
EA201890613A1 (en) | 2015-09-21 | 2018-10-31 | Аптево Рисёрч Энд Девелопмент Ллс | POLYPEPTIDES CONNECTING CD3 |
JP6904947B2 (en) | 2015-09-22 | 2021-07-21 | スプリング バイオサイエンス コーポレーション | Anti-OX40 antibody and its diagnostic applications |
CN116987187A (en) | 2015-09-23 | 2023-11-03 | 豪夫迈·罗氏有限公司 | Optimized variants of anti-VEGF antibodies |
BR112018005931A2 (en) | 2015-09-24 | 2018-10-09 | Abvitro Llc | hiv antibody compositions and methods of use |
AR106188A1 (en) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | ANTI-CD19 HUMANIZED HUMAN ANTIBODIES AND METHODS OF USE |
AR106189A1 (en) | 2015-10-02 | 2017-12-20 | Hoffmann La Roche | BIESPECTIFIC ANTIBODIES AGAINST HUMAN A-b AND THE HUMAN TRANSFERRINE RECEIVER AND METHODS OF USE |
CN107849137B (en) | 2015-10-02 | 2021-11-26 | 豪夫迈·罗氏有限公司 | Bispecific anti-CEAXCD 3T cell activating antigen binding molecules |
WO2017055399A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Cellular based fret assay for the determination of simultaneous binding |
EP3150636A1 (en) | 2015-10-02 | 2017-04-05 | F. Hoffmann-La Roche AG | Tetravalent multispecific antibodies |
PE20240096A1 (en) | 2015-10-02 | 2024-01-18 | Hoffmann La Roche | BISPECIFIC ANTIBODIES SPECIFIC FOR A CO-STIMULATORY TNF RECEPTOR |
NZ741067A (en) | 2015-10-02 | 2023-07-28 | Hoffmann La Roche | Bispecific anti-human cd20/human transferrin receptor antibodies and methods of use |
EP3150637A1 (en) | 2015-10-02 | 2017-04-05 | F. Hoffmann-La Roche AG | Multispecific antibodies |
KR102072317B1 (en) | 2015-10-02 | 2020-01-31 | 에프. 호프만-라 로슈 아게 | Anti-PD1 Antibodies and Methods of Use |
MA43345A (en) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | PYRROLOBENZODIAZEPINE ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
EP3359572A2 (en) | 2015-10-06 | 2018-08-15 | H. Hoffnabb-La Roche Ag | Method for treating multiple sclerosis |
JP7074665B2 (en) * | 2015-10-07 | 2022-05-24 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Field of Invention of Tetravalent Bispecific Antibodies to Co-Stimulated TNF Receptors |
MA43354A (en) | 2015-10-16 | 2018-08-22 | Genentech Inc | CONJUGATE DRUG CONJUGATES WITH CLOUDY DISULPHIDE |
MA45326A (en) | 2015-10-20 | 2018-08-29 | Genentech Inc | CALICHEAMICIN-ANTIBODY-DRUG CONJUGATES AND METHODS OF USE |
SG10201913247XA (en) | 2015-10-23 | 2020-02-27 | Eureka Therapeutics Inc | Antibody/t-cell receptor chimeric constructs and uses thereof |
EP3184547A1 (en) | 2015-10-29 | 2017-06-28 | F. Hoffmann-La Roche AG | Anti-tpbg antibodies and methods of use |
WO2017072210A1 (en) | 2015-10-29 | 2017-05-04 | F. Hoffmann-La Roche Ag | Anti-variant fc-region antibodies and methods of use |
MX2018004509A (en) | 2015-10-30 | 2018-08-01 | Genentech Inc | Anti-htra1 antibodies and methods of use thereof. |
WO2017075173A2 (en) | 2015-10-30 | 2017-05-04 | Genentech, Inc. | Anti-factor d antibodies and conjugates |
CA3003399A1 (en) | 2015-11-02 | 2017-05-11 | Bioatla, Llc | Conditionally active polypeptides |
WO2017079768A1 (en) | 2015-11-08 | 2017-05-11 | Genentech, Inc. | Methods of screening for multispecific antibodies |
WO2017085693A1 (en) | 2015-11-19 | 2017-05-26 | AbbVie Deutschland GmbH & Co. KG | Reporter gene assay methods for identifying and analyzing multi-specific binding proteins |
CN106729743B (en) | 2015-11-23 | 2021-09-21 | 四川科伦博泰生物医药股份有限公司 | anti-ErbB 2 antibody-drug conjugate, and composition, preparation method and application thereof |
EP3380121B1 (en) | 2015-11-23 | 2023-12-20 | Acceleron Pharma Inc. | Actrii antagonist for use in treating eye disorders |
KR20180083944A (en) | 2015-12-02 | 2018-07-23 | 아게누스 인코포레이티드 | Antibodies and methods for their use |
WO2017096189A1 (en) * | 2015-12-02 | 2017-06-08 | Agenus Inc. | Anti-gitr antibodies and methods of use thereof |
WO2017097723A2 (en) | 2015-12-09 | 2017-06-15 | F. Hoffmann-La Roche Ag | Treatment method |
EP3178848A1 (en) | 2015-12-09 | 2017-06-14 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies |
US20200261573A1 (en) | 2015-12-17 | 2020-08-20 | Novartis Ag | Combination of c-met inhibitor with antibody molecule to pd-1 and uses thereof |
EP3389712B1 (en) | 2015-12-17 | 2024-04-10 | Novartis AG | Antibody molecules to pd-1 and uses thereof |
CA3005592C (en) | 2015-12-18 | 2024-01-23 | Chugai Seiyaku Kabushiki Kaisha | Anti-c5 antibodies and methods of use |
ES2837428T3 (en) | 2016-01-08 | 2021-06-30 | Hoffmann La Roche | CEA-Positive Cancer Treatment Procedures Using PD-1 Axis Binding Antagonists and Anti-CEA / Anti-CD3 Bispecific Antibodies |
CN108780094B (en) | 2016-01-12 | 2022-09-13 | 生物蛋白有限公司 | Diagnostic methods using conditionally active antibodies |
US10697972B2 (en) | 2016-01-12 | 2020-06-30 | Bioatla, Llc | Diagnostics using conditionally active antibodies |
US20190031785A1 (en) * | 2016-01-13 | 2019-01-31 | Compass Therapeutics Llc | Multispecific immunomodulatory antigen-binding constructs |
CN114113625A (en) | 2016-01-20 | 2022-03-01 | 基因泰克公司 | High dose treatment for alzheimer's disease |
ES2847155T3 (en) | 2016-01-21 | 2021-08-02 | Novartis Ag | Multispecific molecules targeting CLL-1 |
CU24613B1 (en) | 2016-02-06 | 2022-07-08 | Epimab Biotherapeutics Inc | FABS TANDEM IMMUNOGLOBULIN BINDING PROTEINS (FIT-IG) BSPECIFIC BINDING TO CMET AND EGFR |
EP3413910A1 (en) | 2016-02-12 | 2018-12-19 | Janssen Pharmaceutica NV | Anti-vista (b7h5) antibodies |
ES2912064T3 (en) | 2016-02-19 | 2022-05-24 | Siwa Corp | Method and composition for treating cancer, killing metastatic cancer cells, and preventing cancer metastasis using antibody to advanced glycation end products (AGEs) |
JP6821693B2 (en) | 2016-02-29 | 2021-01-27 | ジェネンテック, インコーポレイテッド | Treatment and diagnosis for cancer |
US20200281973A1 (en) | 2016-03-04 | 2020-09-10 | Novartis Ag | Cells expressing multiple chimeric antigen receptor (car) molecules and uses therefore |
EP3216458A1 (en) | 2016-03-07 | 2017-09-13 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Modified vascular endothelial growth factor a (vegf-a) and its medical use |
CA3016563A1 (en) | 2016-03-21 | 2017-09-28 | Elstar Therapeutics, Inc. | Multispecific and multifunctional molecules and uses thereof |
WO2017165683A1 (en) | 2016-03-23 | 2017-09-28 | Novartis Ag | Cell secreted minibodies and uses thereof |
CN108700598A (en) | 2016-03-25 | 2018-10-23 | 豪夫迈·罗氏有限公司 | The drug of the total antibody of multichannel and antibody conjugate quantifies measuring method |
DK3433273T3 (en) * | 2016-03-25 | 2022-02-14 | Biomunex Pharmaceuticals | BINDING OF MOLECULES TO CD38 AND PD-L1 |
US20170319688A1 (en) | 2016-04-14 | 2017-11-09 | Genentech, Inc. | Anti-rspo3 antibodies and methods of use |
UA125382C2 (en) | 2016-04-15 | 2022-03-02 | Імьюнекст Інк. | Anti-human vista antibodies and use thereof |
CA3021027A1 (en) | 2016-04-15 | 2017-10-19 | Novartis Ag | Compositions and methods for selective expression of chimeric antigen receptors |
SG11201808994YA (en) | 2016-04-15 | 2018-11-29 | Bioatla Llc | Anti-axl antibodies, antibody fragments and their immunoconjugates and uses thereof |
KR20190003958A (en) | 2016-04-15 | 2019-01-10 | 제넨테크, 인크. | Treatment and monitoring of cancer |
WO2017181079A2 (en) | 2016-04-15 | 2017-10-19 | Genentech, Inc. | Methods for monitoring and treating cancer |
CN109311975A (en) | 2016-04-15 | 2019-02-05 | Siwa有限公司 | For treating the anti-AGE antibody of neurodegenerative disorder |
CN109071640B (en) | 2016-05-11 | 2022-10-18 | 豪夫迈·罗氏有限公司 | Modified anti-tenascin antibodies and methods of use |
ES2930255T3 (en) | 2016-05-13 | 2022-12-09 | Bioatla Inc | Anti-Ror2 antibodies, antibody fragments, their immunoconjugates and uses thereof |
AU2017268234A1 (en) | 2016-05-17 | 2018-12-13 | Genentech, Inc. | Stromal gene signatures for diagnosis and use in immunotherapy |
JP2019522633A (en) | 2016-05-20 | 2019-08-15 | ジェネンテック, インコーポレイテッド | PROTAC antibody conjugates and methods of use |
WO2017205741A1 (en) | 2016-05-27 | 2017-11-30 | Genentech, Inc. | Bioanalytical method for the characterization of site-specific antibody-drug conjugates |
TWI781934B (en) | 2016-05-27 | 2022-11-01 | 美商艾吉納斯公司 | Anti-tim-3 antibodies and methods of use thereof |
WO2017210617A2 (en) | 2016-06-02 | 2017-12-07 | Porter, David, L. | Therapeutic regimens for chimeric antigen receptor (car)- expressing cells |
CA3059010A1 (en) | 2016-06-02 | 2018-12-06 | F. Hoffmann-La Roche Ag | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
EP3252078A1 (en) | 2016-06-02 | 2017-12-06 | F. Hoffmann-La Roche AG | Type ii anti-cd20 antibody and anti-cd20/cd3 bispecific antibody for treatment of cancer |
EP3464280B1 (en) | 2016-06-06 | 2021-10-06 | F. Hoffmann-La Roche AG | Silvestrol antibody-drug conjugates and methods of use |
AU2017286676A1 (en) | 2016-06-17 | 2018-12-13 | F. Hoffmann La-Roche Ag | Purification of multispecific antibodies |
US11613572B2 (en) | 2016-06-21 | 2023-03-28 | Teneobio, Inc. | CD3 binding antibodies |
US11213585B2 (en) | 2016-06-23 | 2022-01-04 | Siwa Corporation | Vaccines for use in treating various diseases and disorders |
JP7133477B2 (en) | 2016-06-24 | 2022-09-08 | ジェネンテック, インコーポレイテッド | Anti-polyubiquitin multispecific antibody |
US10722558B2 (en) | 2016-07-15 | 2020-07-28 | Acceleron Pharma Inc. | Compositions and methods for treating pulmonary hypertension |
WO2018013918A2 (en) | 2016-07-15 | 2018-01-18 | Novartis Ag | Treatment and prevention of cytokine release syndrome using a chimeric antigen receptor in combination with a kinase inhibitor |
WO2018014260A1 (en) | 2016-07-20 | 2018-01-25 | Nanjing Legend Biotech Co., Ltd. | Multispecific antigen binding proteins and methods of use thereof |
BR112019001615A2 (en) | 2016-07-27 | 2019-04-30 | Acceleron Pharma Inc. | methods and compositions for treating myelofibrosis |
EP3491016A1 (en) | 2016-07-28 | 2019-06-05 | Novartis AG | Combination therapies of chimeric antigen receptors and pd-1 inhibitors |
KR20190036551A (en) | 2016-08-01 | 2019-04-04 | 노파르티스 아게 | Treatment of Cancer Using Chimeric Antigen Receptors in Combination with Inhibitors of PRO-M2 Macrophage Molecules |
WO2018027042A1 (en) | 2016-08-03 | 2018-02-08 | Bio-Techne Corporation | Identification of vsig3/vista as a novel immune checkpoint and use thereof for immunotherapy |
SG11201801024XA (en) | 2016-08-05 | 2018-05-30 | Chugai Pharmaceutical Co Ltd | Therapeutic or preventive compositions for il-8-related diseases |
JP7198752B2 (en) | 2016-08-09 | 2023-01-04 | カイマブ・リミテッド | Anti-ICOS antibody |
WO2018031662A1 (en) | 2016-08-11 | 2018-02-15 | Genentech, Inc. | Pyrrolobenzodiazepine prodrugs and antibody conjugates thereof |
BR112019002579A2 (en) * | 2016-08-16 | 2019-05-21 | Epimab Biotherapeutics, Inc. | monovalent, asymmetric and tandem fab bispecific antibodies |
WO2018038046A1 (en) | 2016-08-22 | 2018-03-01 | 中外製薬株式会社 | Gene-modified non-human animal expressing human gpc3 polypeptide |
PE20190976A1 (en) | 2016-09-14 | 2019-07-09 | Teneobio Inc | ANTIBODIES OF UNION TO CD3 |
SG10201607778XA (en) | 2016-09-16 | 2018-04-27 | Chugai Pharmaceutical Co Ltd | Anti-Dengue Virus Antibodies, Polypeptides Containing Variant Fc Regions, And Methods Of Use |
EP3515932B1 (en) | 2016-09-19 | 2023-11-22 | F. Hoffmann-La Roche AG | Complement factor based affinity chromatography |
EP4268845A3 (en) | 2016-09-23 | 2024-02-28 | F. Hoffmann-La Roche AG | Uses of il-13 antagonists for treating atopic dermatitis |
JP7050770B2 (en) | 2016-10-05 | 2022-04-08 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Method for preparing antibody drug conjugate |
CN116650622A (en) | 2016-10-05 | 2023-08-29 | 艾科赛扬制药股份有限公司 | Compositions and methods for treating kidney disease |
EP3523451A1 (en) | 2016-10-06 | 2019-08-14 | Genentech, Inc. | Therapeutic and diagnostic methods for cancer |
CN110225927B (en) | 2016-10-07 | 2024-01-12 | 诺华股份有限公司 | Chimeric antigen receptor for the treatment of cancer |
WO2018068201A1 (en) | 2016-10-11 | 2018-04-19 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against ctla-4 |
KR102583190B1 (en) | 2016-10-13 | 2023-09-26 | 치아타이 티안큉 파마수티컬 그룹 주식회사 | Anti-LAG-3 antibodies and compositions |
JP2019535250A (en) | 2016-10-29 | 2019-12-12 | ジェネンテック, インコーポレイテッド | Anti-MIC antibody and method of use |
WO2018089628A1 (en) | 2016-11-09 | 2018-05-17 | Agenus Inc. | Anti-ox40 antibodies, anti-gitr antibodies, and methods of use thereof |
EP3541843A1 (en) | 2016-11-15 | 2019-09-25 | Genentech, Inc. | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibodies |
TW201829463A (en) | 2016-11-18 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Anti-hla-g antibodies and use thereof |
JOP20190100A1 (en) | 2016-11-19 | 2019-05-01 | Potenza Therapeutics Inc | Anti-gitr antigen-binding proteins and methods of use thereof |
WO2018098363A2 (en) | 2016-11-23 | 2018-05-31 | Bioverativ Therapeutics Inc. | Bispecific antibodies binding to coagulation factor ix and coagulation factor x |
CN117820467A (en) | 2016-12-07 | 2024-04-05 | 基因泰克公司 | anti-TAU antibodies and methods of use |
KR102580647B1 (en) | 2016-12-07 | 2023-09-20 | 몰레큘러 템플레이츠, 인코퍼레이션. | Shiga toxin A subunit effector polypeptides, Shiga toxin effector scaffolds, and cell-targeting molecules for site-specific conjugation |
KR20230146126A (en) | 2016-12-07 | 2023-10-18 | 제넨테크, 인크. | Anti-tau antibodies and methods of use |
CN110114674B (en) | 2016-12-13 | 2023-05-09 | 豪夫迈·罗氏有限公司 | Method for determining the presence of a target antigen in a tumor sample |
EP3555120A1 (en) | 2016-12-19 | 2019-10-23 | Abcam Plc | Monovalent and divalent binding proteins |
TWI829628B (en) | 2016-12-19 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Combination therapy with targeted 4-1bb (cd137) agonists |
WO2018114748A1 (en) | 2016-12-20 | 2018-06-28 | F. Hoffmann-La Roche Ag | Combination therapy of anti-cd20/anti-cd3 bispecific antibodies and 4-1bb (cd137) agonists |
BR112019010294A2 (en) | 2016-12-21 | 2019-09-03 | Hoffmann La Roche | method for the enzymatic production of an antibody, antibody and pharmaceutical formulation |
AU2017381656B2 (en) | 2016-12-21 | 2020-07-02 | F. Hoffmann-La Roche Ag | Re-use of enzymes in in vitro glycoengineering of antibodies |
CA3044920C (en) | 2016-12-21 | 2022-06-28 | Roberto Falkenstein | In vitro glycoengineering of antibodies |
JP7215999B2 (en) | 2016-12-21 | 2023-01-31 | エフ.ホフマン-ラ ロシュ アーゲー | Assays for Determining Antibody or Ligand Binding and Function |
IL311136A (en) | 2016-12-22 | 2024-04-01 | Daiichi Sankyo Co Ltd | Anti-cd3 antibody for use in the treatment or prophylaxis of cancer and molecules containing said antibody |
TW201829469A (en) | 2017-01-03 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | Bispecific antigen binding molecules comprising anti-4-1bb clone 20h4.9 |
CN110520444A (en) * | 2017-01-09 | 2019-11-29 | 拜奥穆尼克斯制药 | It is used to prepare the peptide linker of multi-specificity antibody |
EP3573648B1 (en) | 2017-01-25 | 2023-11-22 | Molecular Templates, Inc. | Cell-targeting molecules comprising de-immunized, shiga toxin a subunit effectors and cd8+ t-cell epitopes |
EP3574005B1 (en) | 2017-01-26 | 2021-12-15 | Novartis AG | Cd28 compositions and methods for chimeric antigen receptor therapy |
KR102613874B1 (en) | 2017-01-30 | 2023-12-15 | 알렉시온 파마슈티칼스, 인코포레이티드 | Monovalent anti-properdin antibodies and antibody fragments |
MY197534A (en) | 2017-02-10 | 2023-06-21 | Genentech Inc | Anti-tryptase antibodies, compositions thereof, and uses thereof |
US20200291089A1 (en) | 2017-02-16 | 2020-09-17 | Elstar Therapeutics, Inc. | Multifunctional molecules comprising a trimeric ligand and uses thereof |
EP3589647A1 (en) | 2017-02-28 | 2020-01-08 | Novartis AG | Shp inhibitor compositions and uses for chimeric antigen receptor therapy |
MX2019010295A (en) | 2017-03-01 | 2019-11-21 | Genentech Inc | Diagnostic and therapeutic methods for cancer. |
NZ756132A (en) | 2017-03-10 | 2022-02-25 | Hoffmann La Roche | Method for producing multispecific antibodies |
CN110382544B (en) * | 2017-03-16 | 2023-12-22 | 先天制药公司 | Compositions and methods for treating cancer |
KR20200007776A (en) | 2017-03-22 | 2020-01-22 | 제넨테크, 인크. | Hydrogel Crosslinked Hyaluronic Acid Prodrug Compositions and Methods |
EP3600442A1 (en) | 2017-03-22 | 2020-02-05 | Genentech, Inc. | Optimized antibody compositions for treatment of ocular disorders |
BR112019019998A2 (en) | 2017-03-27 | 2020-04-28 | Biomunex Pharmaceuticals | stable multispecific antibodies |
SG10201911225WA (en) | 2017-03-28 | 2020-01-30 | Genentech Inc | Methods of treating neurodegenerative diseases |
ES2955852T3 (en) | 2017-04-03 | 2023-12-07 | Hoffmann La Roche | STEAP-1 binding antibodies |
MX2019011770A (en) | 2017-04-03 | 2020-01-09 | Hoffmann La Roche | Immunoconjugates of an anti-pd-1 antibody with a mutant il-2 or with il-15. |
MA49042A (en) | 2017-04-05 | 2020-02-12 | Symphogen As | TARGETING POLYTHERAPIES PD-1, TIM-3 AND LAG-3 |
CN116375876A (en) | 2017-04-05 | 2023-07-04 | 豪夫迈·罗氏有限公司 | Bispecific antibodies that specifically bind PD1 and LAG3 |
SI3606954T1 (en) | 2017-04-05 | 2022-10-28 | F. Hoffmann - La Roche Ag | Anti-lag3 antibodies |
CN110770255A (en) | 2017-04-11 | 2020-02-07 | 印希彼有限公司 | Multispecific polypeptide constructs with restricted CD3 binding and methods of use thereof |
US10919957B2 (en) | 2017-04-13 | 2021-02-16 | Siwa Corporation | Humanized monoclonal advanced glycation end-product antibody |
SG11201909048TA (en) | 2017-04-21 | 2019-11-28 | Genentech Inc | Use of klk5 antagonists for treatment of a disease |
JOP20190248A1 (en) | 2017-04-21 | 2019-10-20 | Amgen Inc | Trem2 antigen binding proteins and uses thereof |
CN110741016A (en) | 2017-04-26 | 2020-01-31 | 优瑞科生物技术公司 | Chimeric antibody/T-cell receptor constructs and uses thereof |
WO2018201051A1 (en) | 2017-04-28 | 2018-11-01 | Novartis Ag | Bcma-targeting agent, and combination therapy with a gamma secretase inhibitor |
WO2018197533A1 (en) | 2017-04-28 | 2018-11-01 | F. Hoffmann-La Roche Ag | Antibody selection method |
WO2018201056A1 (en) | 2017-04-28 | 2018-11-01 | Novartis Ag | Cells expressing a bcma-targeting chimeric antigen receptor, and combination therapy with a gamma secretase inhibitor |
EP3630836A1 (en) | 2017-05-31 | 2020-04-08 | Elstar Therapeutics, Inc. | Multispecific molecules that bind to myeloproliferative leukemia (mpl) protein and uses thereof |
CN110785187B (en) | 2017-06-22 | 2024-04-05 | 诺华股份有限公司 | Antibody molecules against CD73 and uses thereof |
JP2020525483A (en) | 2017-06-27 | 2020-08-27 | ノバルティス アーゲー | Dosing regimens for anti-TIM-3 antibodies and uses thereof |
AU2018301393A1 (en) | 2017-07-11 | 2020-02-06 | Compass Therapeutics Llc | Agonist antibodies that bind human CD137 and uses thereof |
BR112020001180A2 (en) | 2017-07-20 | 2020-09-08 | Aptevo Research And Development Llc | antigen-binding proteins that bind to 5t4 and 4-1bb and related compositions and methods |
AU2018302283A1 (en) | 2017-07-20 | 2020-02-06 | Novartis Ag | Dosage regimens of anti-LAG-3 antibodies and uses thereof |
AU2018304458B2 (en) | 2017-07-21 | 2021-12-09 | Foundation Medicine, Inc. | Therapeutic and diagnostic methods for cancer |
WO2019035938A1 (en) | 2017-08-16 | 2019-02-21 | Elstar Therapeutics, Inc. | Multispecific molecules that bind to bcma and uses thereof |
BR112020005519A2 (en) | 2017-09-20 | 2020-10-27 | The University Of British Columbia | new anti-hla-a2 antibodies and their uses |
EP3684801A1 (en) | 2017-09-22 | 2020-07-29 | H. Hoffnabb-La Roche Ag | Multivalent mono- or bispecific recombinant antibodies for analytic purpose |
AU2018338859A1 (en) | 2017-09-29 | 2020-02-06 | Chugai Seiyaku Kabushiki Kaisha | Multispecific antigen-binding molecule having blood coagulation factor VIII (FVIII) cofactor function-substituting activity, and pharmaceutical formulation containing said molecule as active ingredient |
CA3078974A1 (en) | 2017-10-12 | 2019-04-18 | Immunowake Inc. | Vegfr-antibody light chain fusion protein |
WO2019077092A1 (en) | 2017-10-20 | 2019-04-25 | F. Hoffmann-La Roche Ag | Method for generating multispecific antibodies from monospecific antibodies |
JP7438942B2 (en) | 2017-10-30 | 2024-02-27 | エフ. ホフマン-ラ ロシュ アーゲー | Methods for in vivo generation of multispecific antibodies from monospecific antibodies |
WO2019089753A2 (en) | 2017-10-31 | 2019-05-09 | Compass Therapeutics Llc | Cd137 antibodies and pd-1 antagonists and uses thereof |
CA3079036A1 (en) | 2017-11-01 | 2019-05-09 | F. Hoffmann-La Roche Ag | Combination therapy with targeted ox40 agonists |
MX2020004567A (en) | 2017-11-06 | 2020-08-13 | Genentech Inc | Diagnostic and therapeutic methods for cancer. |
CA3081854A1 (en) | 2017-11-08 | 2019-05-16 | Kyowa Kirin Co., Ltd. | Bispecific antibody which binds to cd40 and epcam |
MX2020004756A (en) | 2017-11-16 | 2020-08-20 | Novartis Ag | Combination therapies. |
US11851497B2 (en) | 2017-11-20 | 2023-12-26 | Compass Therapeutics Llc | CD137 antibodies and tumor antigen-targeting antibodies and uses thereof |
WO2019111871A1 (en) | 2017-12-05 | 2019-06-13 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule comprising altered antibody variable region binding cd3 and cd137 |
US11795226B2 (en) | 2017-12-12 | 2023-10-24 | Macrogenics, Inc. | Bispecific CD16-binding molecules and their use in the treatment of disease |
WO2019122882A1 (en) | 2017-12-19 | 2019-06-27 | Kymab Limited | Bispecific antibody for icos and pd-l1 |
AU2018390881A1 (en) | 2017-12-21 | 2020-07-02 | F. Hoffmann-La Roche Ag | Antibodies binding to HLA-A2/WT1 |
US11518801B1 (en) | 2017-12-22 | 2022-12-06 | Siwa Corporation | Methods and compositions for treating diabetes and diabetic complications |
US11873336B2 (en) | 2017-12-22 | 2024-01-16 | Teneobio, Inc. | Heavy chain antibodies binding to CD22 |
TW201929907A (en) | 2017-12-22 | 2019-08-01 | 美商建南德克公司 | Use of PILRA binding agents for treatment of a Disease |
JP7074859B2 (en) | 2017-12-22 | 2022-05-24 | エフ.ホフマン-ラ ロシュ アーゲー | Method of depletion of light chain mismatched antibody variants by hydrophobic interaction chromatography |
KR20200104886A (en) | 2017-12-28 | 2020-09-04 | 난징 레전드 바이오테크 씨오., 엘티디. | Antibodies and variants against PD-L1 |
EP3732202A4 (en) | 2017-12-28 | 2022-06-15 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against tigit |
JP7436365B2 (en) | 2017-12-29 | 2024-02-21 | エフ. ホフマン-ラ ロシュ アーゲー | Anti-VEGF antibodies and methods of use |
EP3732193A1 (en) | 2017-12-29 | 2020-11-04 | Alector LLC | Anti-tmem106b antibodies and methods of use thereof |
WO2019137541A1 (en) | 2018-01-15 | 2019-07-18 | Nanjing Legend Biotech Co., Ltd. | Single-domain antibodies and variants thereof against pd-1 |
WO2019149716A1 (en) | 2018-01-31 | 2019-08-08 | F. Hoffmann-La Roche Ag | Bispecific antibodies comprising an antigen-binding site binding to lag3 |
JP2021511782A (en) | 2018-01-31 | 2021-05-13 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | Stabilized immunoglobulin domain |
WO2019152660A1 (en) | 2018-01-31 | 2019-08-08 | Novartis Ag | Combination therapy using a chimeric antigen receptor |
EP3746476A1 (en) | 2018-01-31 | 2020-12-09 | Alector LLC | Anti-ms4a4a antibodies and methods of use thereof |
CN111868082A (en) | 2018-02-02 | 2020-10-30 | 博奥泰克尼公司 | Compounds that modulate the interaction of VISTA and VSIG3 and methods of making and using the same |
PE20211116A1 (en) | 2018-02-08 | 2021-06-23 | Genentech Inc | BISPECIFIC ANTIGEN BINDING MOLECULES AND METHODS OF USE |
CA3226165A1 (en) | 2018-02-09 | 2019-08-15 | Genentech, Inc. | Therapeutic and diagnostic methods for mast cell-mediated inflammatory diseases |
TWI829667B (en) | 2018-02-09 | 2024-01-21 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to gprc5d |
AU2019236372A1 (en) | 2018-03-13 | 2020-07-30 | F. Hoffmann-La Roche Ag | Therapeutic combination of 4-1 BB agonists with anti-CD20 antibodies |
TW202003561A (en) | 2018-03-13 | 2020-01-16 | 瑞士商赫孚孟拉羅股份公司 | Combination therapy with targeted 4-1BB (CD137) agonists |
US20210238280A1 (en) | 2018-03-14 | 2021-08-05 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to calreticulin and uses thereof |
EP3765516A2 (en) | 2018-03-14 | 2021-01-20 | Elstar Therapeutics, Inc. | Multifunctional molecules and uses thereof |
US20200040103A1 (en) | 2018-03-14 | 2020-02-06 | Genentech, Inc. | Anti-klk5 antibodies and methods of use |
CN116327926A (en) | 2018-03-15 | 2023-06-27 | 中外制药株式会社 | Anti-dengue virus antibodies with cross-reactivity to Zika virus and methods of use |
KR20200135510A (en) | 2018-03-29 | 2020-12-02 | 제넨테크, 인크. | Regulation of lactation stimulating activity in mammalian cells |
CN111936514A (en) * | 2018-03-30 | 2020-11-13 | 美勒斯公司 | Multivalent antibodies |
CN111886254B (en) | 2018-03-30 | 2023-12-08 | 南京传奇生物科技有限公司 | Single domain antibodies against LAG-3 and uses thereof |
TW202011029A (en) | 2018-04-04 | 2020-03-16 | 美商建南德克公司 | Methods for detecting and quantifying FGF21 |
US20210147547A1 (en) | 2018-04-13 | 2021-05-20 | Novartis Ag | Dosage Regimens For Anti-Pd-L1 Antibodies And Uses Thereof |
WO2019204272A1 (en) | 2018-04-17 | 2019-10-24 | Molecular Templates, Inc. | Her2-targeting molecules comprising de-immunized, shiga toxin a subunit scaffolds |
AR114789A1 (en) | 2018-04-18 | 2020-10-14 | Hoffmann La Roche | ANTI-HLA-G ANTIBODIES AND THE USE OF THEM |
AR115052A1 (en) | 2018-04-18 | 2020-11-25 | Hoffmann La Roche | MULTI-SPECIFIC ANTIBODIES AND THE USE OF THEM |
WO2019210153A1 (en) | 2018-04-27 | 2019-10-31 | Novartis Ag | Car t cell therapies with enhanced efficacy |
CN112166196A (en) | 2018-05-18 | 2021-01-01 | 豪夫迈·罗氏有限公司 | Targeted intracellular delivery of large nucleic acids |
JP2021525243A (en) | 2018-05-21 | 2021-09-24 | コンパス セラピューティクス リミテッド ライアビリティ カンパニー | Compositions and Methods for Promoting Killing of Target Cells by NK Cells |
WO2019226658A1 (en) | 2018-05-21 | 2019-11-28 | Compass Therapeutics Llc | Multispecific antigen-binding compositions and methods of use |
EP3801769A1 (en) | 2018-05-25 | 2021-04-14 | Novartis AG | Combination therapy with chimeric antigen receptor (car) therapies |
UA128113C2 (en) | 2018-05-25 | 2024-04-10 | ЕЛЕКТОР ЕлЕлСі | Anti-sirpa antibodies and methods of use thereof |
CN110551216B (en) * | 2018-05-31 | 2022-11-18 | 信达生物制药(苏州)有限公司 | Multivalent anti-OX 40 antibodies and uses thereof |
WO2019232244A2 (en) | 2018-05-31 | 2019-12-05 | Novartis Ag | Antibody molecules to cd73 and uses thereof |
EP3805400A4 (en) | 2018-06-04 | 2022-06-29 | Chugai Seiyaku Kabushiki Kaisha | Antigen-binding molecule showing changed half-life in cytoplasm |
CN112203725A (en) | 2018-06-13 | 2021-01-08 | 诺华股份有限公司 | BCMA chimeric antigen receptors and uses thereof |
CA3104295A1 (en) | 2018-06-19 | 2019-12-26 | Atarga, Llc | Antibody molecules to complement component 5 and uses thereof |
TW202012445A (en) | 2018-06-26 | 2020-04-01 | 日商協和麒麟股份有限公司 | Antibody binding to chondroitin sulfate proteoglycan-5 |
JP7397445B2 (en) | 2018-06-26 | 2023-12-13 | 協和キリン株式会社 | Antibody that binds to Cell Adhesion Molecule 3 |
CA3099176A1 (en) | 2018-06-29 | 2020-01-02 | Alector Llc | Anti-sirp-beta1 antibodies and methods of use thereof |
EP3820905A1 (en) * | 2018-07-03 | 2021-05-19 | NGM Biopharmaceuticals, Inc. | Bispecific antibodies |
EP3818083A2 (en) | 2018-07-03 | 2021-05-12 | Elstar Therapeutics, Inc. | Anti-tcr antibody molecules and uses thereof |
AR116109A1 (en) | 2018-07-10 | 2021-03-31 | Novartis Ag | DERIVATIVES OF 3- (5-AMINO-1-OXOISOINDOLIN-2-IL) PIPERIDINE-2,6-DIONA AND USES OF THE SAME |
LT3618928T (en) | 2018-07-13 | 2023-04-11 | Alector Llc | Anti-sortilin antibodies and methods of use thereof |
BR112021000727A2 (en) | 2018-07-20 | 2021-04-13 | Surface Oncology, Inc. | ANTI-CD112R COMPOSITIONS AND METHODS |
WO2020021465A1 (en) | 2018-07-25 | 2020-01-30 | Advanced Accelerator Applications (Italy) S.R.L. | Method of treatment of neuroendocrine tumors |
JPWO2020027330A1 (en) | 2018-08-03 | 2021-08-19 | 中外製薬株式会社 | Antigen-binding molecule containing two antigen-binding domains linked to each other |
EP3608674A1 (en) | 2018-08-09 | 2020-02-12 | Regeneron Pharmaceuticals, Inc. | Methods for assessing binding affinity of an antibody variant to the neonatal fc receptor |
SG10202106830VA (en) | 2018-08-10 | 2021-08-30 | Chugai Pharmaceutical Co Ltd | Anti-cd137 antigen-binding molecule and utilization thereof |
GB201814281D0 (en) | 2018-09-03 | 2018-10-17 | Femtogenix Ltd | Cytotoxic agents |
WO2020054979A1 (en) * | 2018-09-12 | 2020-03-19 | 아주대학교산학협력단 | Composition comprising cd83 inhibitor as effective ingredient for preventing or treating behcet's disease |
WO2020061060A1 (en) | 2018-09-19 | 2020-03-26 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
EP3867646A1 (en) | 2018-10-18 | 2021-08-25 | F. Hoffmann-La Roche AG | Diagnostic and therapeutic methods for sarcomatoid kidney cancer |
CN113056287A (en) | 2018-10-24 | 2021-06-29 | 豪夫迈·罗氏有限公司 | Conjugated chemical degradation inducers and methods of use |
JP2022512798A (en) | 2018-10-25 | 2022-02-07 | エフ.ホフマン-ラ ロシュ アーゲー | Modification of antibody FcRn binding |
WO2020084034A1 (en) | 2018-10-26 | 2020-04-30 | F. Hoffmann-La Roche Ag | Multispecific antibody screening method using recombinase mediated cassette exchange |
WO2020092455A2 (en) | 2018-10-29 | 2020-05-07 | The Broad Institute, Inc. | Car t cell transcriptional atlas |
SG11202104864QA (en) | 2018-11-13 | 2021-06-29 | Compass Therapeutics Llc | Multispecific binding constructs against checkpoint molecules and uses thereof |
WO2020117257A1 (en) | 2018-12-06 | 2020-06-11 | Genentech, Inc. | Combination therapy of diffuse large b-cell lymphoma comprising an anti-cd79b immunoconjugates, an alkylating agent and an anti-cd20 antibody |
WO2020123275A1 (en) | 2018-12-10 | 2020-06-18 | Genentech, Inc. | Photocrosslinking peptides for site specific conjugation to fc-containing proteins |
MX2021007392A (en) | 2018-12-20 | 2021-08-24 | Novartis Ag | Dosing regimen and pharmaceutical combination comprising 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives. |
AR117453A1 (en) | 2018-12-20 | 2021-08-04 | Genentech Inc | CF OF MODIFIED ANTIBODIES AND METHODS TO USE THEM |
TW202038959A (en) | 2018-12-20 | 2020-11-01 | 瑞士商諾華公司 | Novel dosing regimens for mdm2 inhibitors |
CR20210325A (en) | 2018-12-21 | 2021-07-20 | Hoffmann La Roche | Antibodies binding to cd3 |
EP3674319A1 (en) * | 2018-12-24 | 2020-07-01 | Sanofi | Pseudofab-based multispecific binding proteins |
WO2020136564A1 (en) | 2018-12-24 | 2020-07-02 | Sanofi | Pseudofab-based multispecific binding proteins |
CN113412123A (en) | 2018-12-28 | 2021-09-17 | 豪夫迈·罗氏有限公司 | peptide-MHC-I-antibody fusion proteins for therapeutic use in patients with enhanced immune response |
CN111378044B (en) | 2018-12-28 | 2022-07-15 | 长春金赛药业有限责任公司 | Antibody fusion protein, preparation method and application thereof |
CN111378045B (en) | 2018-12-28 | 2022-08-02 | 长春金赛药业有限责任公司 | Bivalent and bispecific antibody, preparation method thereof, encoding gene, host cell and composition |
TW202039581A (en) | 2018-12-28 | 2020-11-01 | 日商協和麒麟股份有限公司 | Bispecific antibody binding to TfR |
SG11202105926QA (en) * | 2018-12-31 | 2021-07-29 | Merus Nv | Truncated multivalent multimers |
CN113329770A (en) | 2019-01-24 | 2021-08-31 | 中外制药株式会社 | Novel cancer antigen and antibody against said antigen |
GB201901197D0 (en) | 2019-01-29 | 2019-03-20 | Femtogenix Ltd | G-A Crosslinking cytotoxic agents |
JP2022520811A (en) | 2019-02-15 | 2022-04-01 | ノバルティス アーゲー | 3- (1-oxo-5- (piperidine-4-yl) isoindoline-2-yl) piperidine-2,6-dione derivative and its use |
US10871640B2 (en) | 2019-02-15 | 2020-12-22 | Perkinelmer Cellular Technologies Germany Gmbh | Methods and systems for automated imaging of three-dimensional objects |
KR20210129672A (en) | 2019-02-15 | 2021-10-28 | 노파르티스 아게 | Substituted 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof |
SG11202109056TA (en) | 2019-02-21 | 2021-09-29 | Marengo Therapeutics Inc | Multifunctional molecules that bind to calreticulin and uses thereof |
CA3130508A1 (en) | 2019-02-21 | 2020-08-27 | Marengo Therapeutics, Inc. | Antibody molecules that bind to nkp30 and uses thereof |
CA3131014A1 (en) | 2019-02-21 | 2020-08-27 | Andreas Loew | Anti-tcr antibody molecules and uses thereof |
GB2599228B (en) | 2019-02-21 | 2024-02-07 | Marengo Therapeutics Inc | Multifunctional molecules that bind to T cell related cancer cells and uses thereof |
WO2020172598A1 (en) | 2019-02-21 | 2020-08-27 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to t cells and uses thereof to treat autoimmune disorders |
WO2020172553A1 (en) | 2019-02-22 | 2020-08-27 | Novartis Ag | Combination therapies of egfrviii chimeric antigen receptors and pd-1 inhibitors |
KR20210138588A (en) | 2019-03-08 | 2021-11-19 | 제넨테크, 인크. | Methods for detecting and quantifying membrane-associated proteins on extracellular vesicles |
MA55519A (en) | 2019-03-29 | 2022-02-09 | Atarga Llc | ANTI-FGF23 ANTIBODIES |
WO2020214995A1 (en) | 2019-04-19 | 2020-10-22 | Genentech, Inc. | Anti-mertk antibodies and their methods of use |
AU2020275415A1 (en) | 2019-05-14 | 2021-11-25 | Genentech, Inc. | Methods of using anti-CD79B immunoconjugates to treat follicular lymphoma |
EP3981429A4 (en) | 2019-06-10 | 2023-07-26 | Chugai Seiyaku Kabushiki Kaisha | Anti-t cell antigen-binding molecule to be used in combination with cytokine inhibitor |
KR20220031616A (en) | 2019-06-11 | 2022-03-11 | 알렉터 엘엘씨 | Anti-Sortilin Antibodies for Use in Therapy |
AR119746A1 (en) | 2019-06-14 | 2022-01-05 | Teneobio Inc | MULTISPECIFIC HEAVY CHAIN ANTIBODIES THAT BIND CD22 AND CD3 |
KR20220010549A (en) | 2019-06-19 | 2022-01-25 | 에프. 호프만-라 로슈 아게 | Methods of Generating Multivalent Bispecific Antibody Expression Cells by Targeted Integration of Multiple Expression Cassettes in Defined Tissues |
CA3140318A1 (en) | 2019-06-19 | 2020-12-24 | Johannes Auer | Method for the generation of a bivalent, bispecific antibody expressing cell by targeted integration of multiple expression cassettes in a defined organization |
EP3986925A1 (en) | 2019-06-19 | 2022-04-27 | F. Hoffmann-La Roche AG | Method for the generation of a trivalent antibody expressing cell by targeted integration of multiple expression cassettes in a defined organization |
KR20220024636A (en) | 2019-06-19 | 2022-03-03 | 에프. 호프만-라 로슈 아게 | Methods of Generation of Multivalent, Multispecific Antibody Expression Cells by Targeted Integration of Multiple Expression Cassettes of Defined Tissues |
MX2021015536A (en) | 2019-06-19 | 2022-02-10 | Hoffmann La Roche | Method for the generation of a protein expressing cell by targeted integration using cre mrna. |
BR112021026286A2 (en) | 2019-06-26 | 2022-03-03 | Hoffmann La Roche | Method for increasing the heterologous polypeptide titer and/or reducing lactate production of a recombinant mammalian cell and methods for producing a heterologous polypeptide and a recombinant mammalian cell |
AR119393A1 (en) | 2019-07-15 | 2021-12-15 | Hoffmann La Roche | ANTIBODIES THAT BIND NKG2D |
CN114174338A (en) | 2019-07-31 | 2022-03-11 | 豪夫迈·罗氏有限公司 | Antibodies that bind to GPRC5D |
SG11202112491WA (en) | 2019-07-31 | 2021-12-30 | Hoffmann La Roche | Antibodies binding to gprc5d |
KR20220058540A (en) | 2019-07-31 | 2022-05-09 | 알렉터 엘엘씨 | Anti-MS4A4A antibodies and methods of use thereof |
TW202118512A (en) | 2019-09-12 | 2021-05-16 | 美商建南德克公司 | Compositions and methods of treating lupus nephritis |
BR112022004972A2 (en) | 2019-09-18 | 2022-06-28 | Genentech Inc | ANTIBODIES, ISOLATED NUCLEIC ACIDS, ISOLATED HOST CELLS, METHODS OF PRODUCTION OF AN ANTIBODY, OF PRODUCTION OF A BIESPECIFIC ANTIBODY, AND OF TREATMENT OF AN INDIVIDUAL, BIESPECIFIC ANTIBODIES, PHARMACEUTICAL COMPOSITION, ANTIBODY, BIESPECIFIC ANTIBODY OR PHARMACEUTICAL COMPOSITION OF THE ANTIBODY, USE OF A COMBINATION OF THE ANTIBODY AND METHODS TO REDUCE INFLAMMATION AND TO IMPROVE SCALING AND/OR SKIN ERUPTION AND COMBINATION FOR USE |
US20220348687A1 (en) | 2019-09-20 | 2022-11-03 | Genentech, Inc. | Dosing for anti-tryptase antibodies |
EP4036116A4 (en) | 2019-09-27 | 2024-01-24 | Nanjing Genscript Biotech Co Ltd | Anti-vhh domain antibodies and use thereof |
TW202122114A (en) | 2019-10-18 | 2021-06-16 | 美商建南德克公司 | Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma |
TW202128191A (en) | 2019-10-21 | 2021-08-01 | 瑞士商諾華公司 | Tim-3 inhibitors and uses thereof |
CN114786679A (en) | 2019-10-21 | 2022-07-22 | 诺华股份有限公司 | Combination therapy with Vernetork and TIM-3 inhibitors |
AU2020384917A1 (en) | 2019-11-15 | 2022-03-31 | F. Hoffmann-La Roche Ag | Prevention of visible particle formation in aqueous protein solutions |
CN114761037A (en) | 2019-11-26 | 2022-07-15 | 诺华股份有限公司 | Chimeric antigen receptor binding to BCMA and CD19 and uses thereof |
AU2020403145A1 (en) | 2019-12-13 | 2022-07-07 | Genentech, Inc. | Anti-Ly6G6D antibodies and methods of use |
JP7296467B2 (en) | 2019-12-18 | 2023-06-22 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Antibodies that bind to HLA-A2/MAGE-A4 |
CN114867532A (en) | 2019-12-18 | 2022-08-05 | 豪夫迈·罗氏有限公司 | Bispecific anti-CCL 2 antibodies |
WO2021123902A1 (en) | 2019-12-20 | 2021-06-24 | Novartis Ag | Combination of anti tim-3 antibody mbg453 and anti tgf-beta antibody nis793, with or without decitabine or the anti pd-1 antibody spartalizumab, for treating myelofibrosis and myelodysplastic syndrome |
MX2022007746A (en) | 2019-12-23 | 2022-10-07 | Genentech Inc | Apolipoprotein l1-specific antibodies and methods of use. |
CR20220357A (en) | 2019-12-27 | 2022-08-24 | Chugai Pharmaceutical Co Ltd | Anti-ctla-4 antibody and use thereof |
WO2021138407A2 (en) | 2020-01-03 | 2021-07-08 | Marengo Therapeutics, Inc. | Multifunctional molecules that bind to cd33 and uses thereof |
CN110818795B (en) | 2020-01-10 | 2020-04-24 | 上海复宏汉霖生物技术股份有限公司 | anti-TIGIT antibodies and methods of use |
JP2023510382A (en) | 2020-01-15 | 2023-03-13 | エフ. ホフマン-ラ ロシュ アーゲー | Methods for reducing impurities from recombinant protein manufacturing processes |
US20210222244A1 (en) | 2020-01-17 | 2021-07-22 | Becton, Dickinson And Company | Methods and compositions for single cell secretomics |
IL293752A (en) | 2020-01-17 | 2022-08-01 | Novartis Ag | Combination comprising a tim-3 inhibitor and a hypomethylating agent for use in treating myelodysplastic syndrome or chronic myelomonocytic leukemia |
TW202144395A (en) | 2020-02-12 | 2021-12-01 | 日商中外製藥股份有限公司 | Anti-CD137 antigen-binding molecule for use in cancer treatment |
CA3173737A1 (en) | 2020-02-27 | 2021-09-02 | Novartis Ag | Methods of making chimeric antigen receptor-expressing cells |
CR20220461A (en) | 2020-03-13 | 2022-10-21 | Genentech Inc | Anti-interleukin-33 antibodies and uses thereof |
KR20220156575A (en) | 2020-03-19 | 2022-11-25 | 제넨테크, 인크. | Isotype Selective Anti-TGF-beta Antibodies and Methods of Use |
EP4107184A1 (en) | 2020-03-23 | 2022-12-28 | Genentech, Inc. | Method for treating pneumonia, including covid-19 pneumonia, with an il6 antagonist |
CN115867577A (en) | 2020-03-23 | 2023-03-28 | 基因泰克公司 | Biomarkers for predicting response to IL-6 antagonists in COVID-19 pneumonia |
WO2021194860A1 (en) | 2020-03-23 | 2021-09-30 | Genentech, Inc. | Tocilizumab and remdesivir combination therapy for covid-19 pneumonia |
CA3169967A1 (en) | 2020-03-24 | 2021-09-30 | Genentech, Inc. | Tie2-binding agents and methods of use |
WO2021194839A1 (en) | 2020-03-25 | 2021-09-30 | Eli Lilly And Company | Multispecific binding proteins and methods of developing the same |
CN115315512A (en) | 2020-03-26 | 2022-11-08 | 基因泰克公司 | Modified mammalian cells with reduced host cell proteins |
CN115397866A (en) | 2020-03-31 | 2022-11-25 | 中外制药株式会社 | DLL 3-targeted multispecific antigen-binding molecules and uses thereof |
CN115698717A (en) | 2020-04-03 | 2023-02-03 | 基因泰克公司 | Methods of treatment and diagnosis of cancer |
CN115380217A (en) | 2020-04-08 | 2022-11-22 | 豪夫迈·罗氏有限公司 | Macromolecular nonspecific clearance assay |
BR112022017215A2 (en) | 2020-04-09 | 2022-10-18 | Univ Muenchen Tech | Targeted delivery of a MIR-21 inhibitor to macrophages for the treatment of pulmonary fibrosis |
US10994021B1 (en) * | 2020-04-11 | 2021-05-04 | Bliss Biopharmaceutical (Hangzhou) Co., Ltd. | Tetravalent antibody-drug conjugates and use thereof |
JP2023523011A (en) | 2020-04-24 | 2023-06-01 | マレンゴ・セラピューティクス,インコーポレーテッド | Multifunctional molecules that bind to T cell-associated cancer cells and uses thereof |
WO2021217051A1 (en) | 2020-04-24 | 2021-10-28 | Genentech, Inc. | Methods of using anti-cd79b immunoconjugates |
WO2021222167A1 (en) | 2020-04-28 | 2021-11-04 | Genentech, Inc. | Methods and compositions for non-small cell lung cancer immunotherapy |
CN113563473A (en) * | 2020-04-29 | 2021-10-29 | 三生国健药业(上海)股份有限公司 | Tetravalent bispecific antibody, preparation method and application thereof |
EP4146283A1 (en) | 2020-05-03 | 2023-03-15 | Levena (Suzhou) Biopharma Co., Ltd. | Antibody-drug conjugates (adcs) comprising an anti-trop-2 antibody, compositions comprising such adcs, as well as methods of making and using the same |
JP2023520249A (en) | 2020-05-15 | 2023-05-16 | エフ. ホフマン-ラ ロシュ アーゲー | Method for preventing visible particle formation in parenteral protein solutions |
CN115605185A (en) | 2020-05-19 | 2023-01-13 | 豪夫迈·罗氏有限公司(Ch) | Use of a chelating agent to prevent the formation of visible particles in parenteral protein solutions |
CR20220659A (en) | 2020-06-08 | 2023-08-21 | Hoffmann La Roche | Anti-hbv antibodies and methods of use |
CN115698719A (en) | 2020-06-12 | 2023-02-03 | 基因泰克公司 | Methods and compositions for cancer immunotherapy |
EP4168456A1 (en) | 2020-06-19 | 2023-04-26 | F. Hoffmann-La Roche AG | Antibodies binding to cd3 |
EP4168446A1 (en) | 2020-06-19 | 2023-04-26 | F. Hoffmann-La Roche AG | Antibodies binding to cd3 and folr1 |
WO2021255146A1 (en) | 2020-06-19 | 2021-12-23 | F. Hoffmann-La Roche Ag | Antibodies binding to cd3 and cea |
TWI811703B (en) | 2020-06-19 | 2023-08-11 | 瑞士商赫孚孟拉羅股份公司 | Antibodies binding to cd3 and cd19 |
PE20230435A1 (en) | 2020-06-19 | 2023-03-08 | Chugai Pharmaceutical Co Ltd | ANTI-T CELL ANTIGEN BINDING MOLECULES TO BE USED IN COMBINATION WITH AN ANGIOGENESIS INHIBITOR |
JP2023531676A (en) | 2020-06-23 | 2023-07-25 | ノバルティス アーゲー | Dosing Regimens Containing 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione Derivatives |
CA3184747A1 (en) | 2020-06-24 | 2021-12-30 | Genentech, Inc. | Apoptosis resistant cell lines |
JP2023532764A (en) | 2020-07-07 | 2023-07-31 | エフ. ホフマン-ラ ロシュ アーゲー | Alternative surfactants as stabilizers for therapeutic protein formulations |
US20230181471A1 (en) | 2020-07-09 | 2023-06-15 | Hoffmann-La Roche Inc. | Concentrated compositions of proteins, their preparation and use thereof |
CA3188000A1 (en) | 2020-07-13 | 2022-01-20 | Genentech, Inc. | Cell-based methods for predicting polypeptide immunogenicity |
EP4182025A1 (en) | 2020-07-16 | 2023-05-24 | Novartis AG | Anti-betacellulin antibodies, fragments thereof, and multi-specific binding molecules |
WO2022016119A1 (en) | 2020-07-17 | 2022-01-20 | Simurx, Inc. | Chimeric myd88 receptors for redirecting immunosuppressive signaling and related compositions and methods |
US20230303682A1 (en) | 2020-07-17 | 2023-09-28 | Genentech, Inc. | Anti-Notch2 Antibodies and Methods of Use |
TW202216215A (en) | 2020-07-21 | 2022-05-01 | 美商建南德克公司 | Antibody-conjugated chemical inducers of degradation of brm and methods thereof |
WO2022026592A2 (en) | 2020-07-28 | 2022-02-03 | Celltas Bio, Inc. | Antibody molecules to coronavirus and uses thereof |
GB2597532A (en) | 2020-07-28 | 2022-02-02 | Femtogenix Ltd | Cytotoxic compounds |
CA3190649A1 (en) | 2020-07-31 | 2022-02-03 | Chugai Seiyaku Kabushiki Kaisha | Pharmaceutical composition comprising cell expressing chimeric receptor |
JP2023536164A (en) | 2020-08-03 | 2023-08-23 | ノバルティス アーゲー | Heteroaryl-substituted 3-(1-oxoisoindolin-2-yl)piperidine-2,6-dione derivatives and uses thereof |
WO2022031948A1 (en) | 2020-08-07 | 2022-02-10 | Genentech, Inc. | T cell-based methods for predicting polypeptide immunogenicity |
GB2616128A (en) | 2020-08-26 | 2023-08-30 | Marengo Therapeutics Inc | Antibody molecules that bind to NKp30 and uses thereof |
CN116761818A (en) | 2020-08-26 | 2023-09-15 | 马伦戈治疗公司 | Method for detecting TRBC1 or TRBC2 |
CA3190755A1 (en) | 2020-08-26 | 2022-03-03 | Andreas Loew | Multifunctional molecules that bind to calreticulin and uses thereof |
TW202227625A (en) | 2020-08-28 | 2022-07-16 | 美商建南德克公司 | Crispr/cas9 multiplex knockout of host cell proteins |
EP4204020A1 (en) | 2020-08-31 | 2023-07-05 | Advanced Accelerator Applications International S.A. | Method of treating psma-expressing cancers |
WO2022043557A1 (en) | 2020-08-31 | 2022-03-03 | Advanced Accelerator Applications International Sa | Method of treating psma-expressing cancers |
EP4213877A1 (en) | 2020-09-17 | 2023-07-26 | Genentech, Inc. | Results of empacta: a randomized, double-blind, placebo-controlled, multicenter study to evaluate the efficacy and safety of tocilizumab in hospitalized patients with covid-19 pneumonia |
AU2021342566A1 (en) | 2020-09-21 | 2023-03-02 | Genentech, Inc. | Purification of multispecific antibodies |
CA3195257A1 (en) | 2020-09-24 | 2022-03-31 | F. Hoffmann-La Roche Ag | Mammalian cell lines with gene knockout |
CA3193952A1 (en) | 2020-10-05 | 2022-04-14 | Bernard Martin Fine | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
AR123855A1 (en) | 2020-10-20 | 2023-01-18 | Genentech Inc | PEG-CONJUGATED ANTI-MERTK ANTIBODIES AND METHODS OF USE |
JP2023545566A (en) | 2020-10-20 | 2023-10-30 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Combination therapy with PD-1 axis binding antagonist and LRRK2 inhibitor |
JP2023547499A (en) | 2020-11-06 | 2023-11-10 | ノバルティス アーゲー | Antibody Fc variant |
US20240033358A1 (en) | 2020-11-13 | 2024-02-01 | Novartis Ag | Combination therapies with chimeric antigen receptor (car)-expressing cells |
US20240101681A1 (en) | 2020-12-02 | 2024-03-28 | Alector Llc | Methods of use of anti-sortilin antibodies |
JP2023553157A (en) | 2020-12-10 | 2023-12-20 | ユーティレックス カンパニー リミテッド | Anti-PD-1 antibody and its uses |
AU2021399841A1 (en) | 2020-12-17 | 2023-07-06 | F. Hoffmann-La Roche Ag | Anti-hla-g antibodies and use thereof |
WO2022136140A1 (en) | 2020-12-22 | 2022-06-30 | F. Hoffmann-La Roche Ag | Oligonucleotides targeting xbp1 |
CN114716548A (en) | 2021-01-05 | 2022-07-08 | (株)爱恩德生物 | anti-FGFR 3 antibodies and uses thereof |
WO2022162587A1 (en) | 2021-01-27 | 2022-08-04 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
EP4284510A1 (en) | 2021-01-29 | 2023-12-06 | Novartis AG | Dosage regimes for anti-cd73 and anti-entpd2 antibodies and uses thereof |
JP2024505600A (en) | 2021-02-03 | 2024-02-06 | モーツァルト セラピューティクス, インコーポレイテッド | Binders and how to use them |
JP2024509695A (en) | 2021-02-03 | 2024-03-05 | ジェネンテック, インコーポレイテッド | Multispecific binding proteolysis platform and methods of use |
JP2024509169A (en) | 2021-03-03 | 2024-02-29 | ソレント・セラピューティクス・インコーポレイテッド | Antibody-drug conjugates including anti-BCMA antibodies |
WO2022192647A1 (en) | 2021-03-12 | 2022-09-15 | Genentech, Inc. | Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use |
WO2022198192A1 (en) | 2021-03-15 | 2022-09-22 | Genentech, Inc. | Compositions and methods of treating lupus nephritis |
JP2024512002A (en) | 2021-03-18 | 2024-03-18 | アレクトル エルエルシー | Anti-TMEM106B antibody and method of use thereof |
WO2022197877A1 (en) | 2021-03-19 | 2022-09-22 | Genentech, Inc. | Methods and compositions for time delayed bio-orthogonal release of cytotoxic agents |
WO2022204274A1 (en) | 2021-03-23 | 2022-09-29 | Alector Llc | Anti-tmem106b antibodies for treating and preventing coronavirus infections |
KR20230162793A (en) | 2021-03-26 | 2023-11-28 | 얀센 바이오테크 인코포레이티드 | Humanized antibodies against paired helical filament tau and uses thereof |
TW202304979A (en) | 2021-04-07 | 2023-02-01 | 瑞士商諾華公司 | USES OF ANTI-TGFβ ANTIBODIES AND OTHER THERAPEUTIC AGENTS FOR THE TREATMENT OF PROLIFERATIVE DISEASES |
KR20240004462A (en) | 2021-04-08 | 2024-01-11 | 마렝고 테라퓨틱스, 인크. | Multifunctional molecules that bind to TCR and their uses |
JP2024513474A (en) | 2021-04-09 | 2024-03-25 | エフ. ホフマン-ラ ロシュ アーゲー | Method for selecting cell clones expressing heterologous polypeptides |
EP4319820A1 (en) | 2021-04-10 | 2024-02-14 | Profoundbio Us Co. | Folr1 binding agents, conjugates thereof and methods of using the same |
AR125344A1 (en) | 2021-04-15 | 2023-07-05 | Chugai Pharmaceutical Co Ltd | ANTI-C1S ANTIBODY |
EP4326855A1 (en) | 2021-04-19 | 2024-02-28 | Genentech, Inc. | Modified mammalian cells |
WO2022226317A1 (en) | 2021-04-23 | 2022-10-27 | Profoundbio Us Co. | Anti-cd70 antibodies, conjugates thereof and methods of using the same |
IL307821A (en) | 2021-04-30 | 2023-12-01 | Hoffmann La Roche | Dosing for treatment with anti-cd20/anti-cd3 bispecific antibody |
CN117321078A (en) | 2021-04-30 | 2023-12-29 | 豪夫迈·罗氏有限公司 | Administration for combination therapy with anti-CD 20/anti-CD 3 bispecific antibody and anti-CD 79B antibody drug conjugates |
WO2022241446A1 (en) | 2021-05-12 | 2022-11-17 | Genentech, Inc. | Methods of using anti-cd79b immunoconjugates to treat diffuse large b-cell lymphoma |
AU2022273303A1 (en) | 2021-05-14 | 2023-11-02 | Genentech, Inc. | Agonists of trem2 |
AR125874A1 (en) | 2021-05-18 | 2023-08-23 | Novartis Ag | COMBINATION THERAPIES |
WO2022246259A1 (en) | 2021-05-21 | 2022-11-24 | Genentech, Inc. | Modified cells for the production of a recombinant product of interest |
CN113214393B (en) * | 2021-05-25 | 2022-11-18 | 深圳市新产业生物医学工程股份有限公司 | IL-6 antibody or antigen-binding fragment thereof and detection kit comprising same |
CN113278071B (en) | 2021-05-27 | 2021-12-21 | 江苏荃信生物医药股份有限公司 | Anti-human interferon alpha receptor1 monoclonal antibody and application thereof |
WO2022255440A1 (en) | 2021-06-04 | 2022-12-08 | Chugai Seiyaku Kabushiki Kaisha | Anti-ddr2 antibodies and uses thereof |
KR20240019109A (en) | 2021-06-09 | 2024-02-14 | 에프. 호프만-라 로슈 아게 | Combination of a specific BRAF inhibitor (Paradox Break) and a PD-1 axis binding antagonist for use in the treatment of cancer |
CA3221924A1 (en) | 2021-06-11 | 2022-12-15 | Genentech, Inc. | Method for treating chronic obstructive pulmonary disease with an st2 antagonist |
WO2022263638A1 (en) | 2021-06-17 | 2022-12-22 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
WO2022263501A1 (en) | 2021-06-18 | 2022-12-22 | F. Hoffmann-La Roche Ag | Bispecific anti-ccl2 antibodies |
CN117616123A (en) | 2021-06-25 | 2024-02-27 | 中外制药株式会社 | anti-CTLA-4 antibodies |
CA3220353A1 (en) | 2021-06-25 | 2022-12-29 | Chugai Seiyaku Kabushiki Kaisha | Use of anti-ctla-4 antibody |
TW202320857A (en) | 2021-07-06 | 2023-06-01 | 美商普方生物製藥美國公司 | Linkers, drug linkers and conjugates thereof and methods of using the same |
WO2023287663A1 (en) | 2021-07-13 | 2023-01-19 | Genentech, Inc. | Multi-variate model for predicting cytokine release syndrome |
CA3219606A1 (en) | 2021-07-22 | 2023-01-26 | F. Hoffmann-La Roche Ag | Heterodimeric fc domain antibodies |
WO2023004386A1 (en) | 2021-07-22 | 2023-01-26 | Genentech, Inc. | Brain targeting compositions and methods of use thereof |
WO2023012147A1 (en) | 2021-08-03 | 2023-02-09 | F. Hoffmann-La Roche Ag | Bispecific antibodies and methods of use |
WO2023019239A1 (en) | 2021-08-13 | 2023-02-16 | Genentech, Inc. | Dosing for anti-tryptase antibodies |
AU2022332303A1 (en) | 2021-08-27 | 2024-02-01 | Genentech, Inc. | Methods of treating tau pathologies |
WO2023034750A1 (en) | 2021-08-30 | 2023-03-09 | Genentech, Inc. | Anti-polyubiquitin multispecific antibodies |
CN113603775B (en) | 2021-09-03 | 2022-05-20 | 江苏荃信生物医药股份有限公司 | Anti-human interleukin-33 monoclonal antibody and application thereof |
CN113683694B (en) | 2021-09-03 | 2022-05-13 | 江苏荃信生物医药股份有限公司 | Anti-human TSLP monoclonal antibody and application thereof |
WO2023044483A2 (en) | 2021-09-20 | 2023-03-23 | Voyager Therapeutics, Inc. | Compositions and methods for the treatment of her2 positive cancer |
TW202333781A (en) | 2021-10-08 | 2023-09-01 | 日商中外製藥股份有限公司 | Anti-hla-dq2.5 antibody formulation |
CA3234162A1 (en) | 2021-10-15 | 2023-04-20 | Michele Fiscella | Antibodies and methods of using thereof |
WO2023076876A1 (en) | 2021-10-26 | 2023-05-04 | Mozart Therapeutics, Inc. | Modulation of immune responses to viral vectors |
WO2023086807A1 (en) | 2021-11-10 | 2023-05-19 | Genentech, Inc. | Anti-interleukin-33 antibodies and uses thereof |
WO2023092004A1 (en) | 2021-11-17 | 2023-05-25 | Voyager Therapeutics, Inc. | Compositions and methods for the treatment of tau-related disorders |
AR127887A1 (en) | 2021-12-10 | 2024-03-06 | Hoffmann La Roche | ANTIBODIES THAT BIND CD3 AND PLAP |
WO2023117325A1 (en) | 2021-12-21 | 2023-06-29 | F. Hoffmann-La Roche Ag | Method for the determination of hydrolytic activity |
WO2023129974A1 (en) | 2021-12-29 | 2023-07-06 | Bristol-Myers Squibb Company | Generation of landing pad cell lines |
WO2023141445A1 (en) | 2022-01-19 | 2023-07-27 | Genentech, Inc. | Anti-notch2 antibodies and conjugates and methods of use |
WO2023147329A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation and methods thereof |
WO2023147328A1 (en) | 2022-01-26 | 2023-08-03 | Genentech, Inc. | Antibody-conjugated chemical inducers of degradation with hydolysable maleimide linkers and methods thereof |
TW202342548A (en) | 2022-02-07 | 2023-11-01 | 美商威特拉公司 | Anti-idiotype antibody molecules and uses thereof |
WO2023173026A1 (en) | 2022-03-10 | 2023-09-14 | Sorrento Therapeutics, Inc. | Antibody-drug conjugates and uses thereof |
TW202346355A (en) | 2022-03-11 | 2023-12-01 | 比利時商健生藥品公司 | Multispecific antibodies and uses thereof |
WO2023170295A1 (en) | 2022-03-11 | 2023-09-14 | Janssen Pharmaceutica Nv | Multispecific antibodies and uses thereof |
WO2023170291A1 (en) | 2022-03-11 | 2023-09-14 | Janssen Pharmaceutica Nv | Multispecific antibodies and uses thereof |
US20230414750A1 (en) | 2022-03-23 | 2023-12-28 | Hoffmann-La Roche Inc. | Combination treatment of an anti-cd20/anti-cd3 bispecific antibody and chemotherapy |
WO2023191816A1 (en) | 2022-04-01 | 2023-10-05 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023201291A1 (en) | 2022-04-13 | 2023-10-19 | Genentech, Inc. | Pharmaceutical compositions of mosunetuzumab and methods of use |
WO2023198727A1 (en) | 2022-04-13 | 2023-10-19 | F. Hoffmann-La Roche Ag | Pharmaceutical compositions of anti-cd20/anti-cd3 bispecific antibodies and methods of use |
WO2023202967A1 (en) | 2022-04-19 | 2023-10-26 | F. Hoffmann-La Roche Ag | Improved production cells |
WO2023215737A1 (en) | 2022-05-03 | 2023-11-09 | Genentech, Inc. | Anti-ly6e antibodies, immunoconjugates, and uses thereof |
WO2023219613A1 (en) | 2022-05-11 | 2023-11-16 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2023220695A2 (en) | 2022-05-13 | 2023-11-16 | Voyager Therapeutics, Inc. | Compositions and methods for the treatment of her2 positive cancer |
WO2023232961A1 (en) | 2022-06-03 | 2023-12-07 | F. Hoffmann-La Roche Ag | Improved production cells |
WO2024015897A1 (en) | 2022-07-13 | 2024-01-18 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024020432A1 (en) | 2022-07-19 | 2024-01-25 | Genentech, Inc. | Dosing for treatment with anti-fcrh5/anti-cd3 bispecific antibodies |
WO2024020564A1 (en) | 2022-07-22 | 2024-01-25 | Genentech, Inc. | Anti-steap1 antigen-binding molecules and uses thereof |
WO2024020579A1 (en) | 2022-07-22 | 2024-01-25 | Bristol-Myers Squibb Company | Antibodies binding to human pad4 and uses thereof |
WO2024030956A2 (en) | 2022-08-03 | 2024-02-08 | Mozart Therapeutics, Inc. | Cd39-specific binding agents and methods of using the same |
WO2024030976A2 (en) | 2022-08-03 | 2024-02-08 | Voyager Therapeutics, Inc. | Compositions and methods for crossing the blood brain barrier |
WO2024049949A1 (en) | 2022-09-01 | 2024-03-07 | Genentech, Inc. | Therapeutic and diagnostic methods for bladder cancer |
WO2024068996A1 (en) | 2022-09-30 | 2024-04-04 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Anti-sars-cov-2 antibodies and use thereof in the treatment of sars-cov-2 infection |
Citations (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816467A (en) * | 1987-01-09 | 1989-03-28 | Farmitalia Carlo Erba S.R.L | Heteroaryl 3-oxo-propanenitrile derivatives, pharmaceutical compositions and use |
US4968603A (en) * | 1986-12-31 | 1990-11-06 | The Regents Of The University Of California | Determination of status in neoplastic disease |
US5183884A (en) * | 1989-12-01 | 1993-02-02 | United States Of America | Dna segment encoding a gene for a receptor related to the epidermal growth factor receptor |
US5434340A (en) * | 1988-12-05 | 1995-07-18 | Genpharm International, Inc. | Transgenic mice depleted in mature T-cells and methods for making transgenic mice |
US5530101A (en) * | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5545806A (en) * | 1990-08-29 | 1996-08-13 | Genpharm International, Inc. | Ransgenic non-human animals for producing heterologous antibodies |
US5545807A (en) * | 1988-10-12 | 1996-08-13 | The Babraham Institute | Production of antibodies from transgenic animals |
US5569825A (en) * | 1990-08-29 | 1996-10-29 | Genpharm International | Transgenic non-human animals capable of producing heterologous antibodies of various isotypes |
US5591699A (en) * | 1993-02-24 | 1997-01-07 | E. I. Du Pont De Nemours And Company | Particle transport fluids thickened with acetylate free xanthan heteropolysaccharide biopolymer plus guar gum |
US5591828A (en) * | 1989-06-22 | 1997-01-07 | Behringwerke Aktiengesellschaft | Bispecific and oligospecific mono-and oligovalent receptors, the preparation and use thereof |
US5595721A (en) * | 1993-09-16 | 1997-01-21 | Coulter Pharmaceutical, Inc. | Radioimmunotherapy of lymphoma using anti-CD20 |
US5641870A (en) * | 1995-04-20 | 1997-06-24 | Genentech, Inc. | Low pH hydrophobic interaction chromatography for antibody purification |
US5641869A (en) * | 1991-05-24 | 1997-06-24 | Genentech, Inc. | Method for purifying heregulin |
US5677171A (en) * | 1988-01-12 | 1997-10-14 | Genentech, Inc. | Monoclonal antibodies directed to the HER2 receptor |
US5736137A (en) * | 1992-11-13 | 1998-04-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
US5783186A (en) * | 1995-12-05 | 1998-07-21 | Amgen Inc. | Antibody-induced apoptosis |
US5821337A (en) * | 1991-06-14 | 1998-10-13 | Genentech, Inc. | Immunoglobulin variants |
US5824311A (en) * | 1987-11-30 | 1998-10-20 | Trustees Of The University Of Pennsylvania | Treatment of tumors with monoclonal antibodies against oncogene antigens |
US5959083A (en) * | 1991-06-03 | 1999-09-28 | Behringwerke Aktiengellschaft | Tetravalent bispecific receptors, the preparation and use thereof |
US5989830A (en) * | 1995-10-16 | 1999-11-23 | Unilever Patent Holdings Bv | Bifunctional or bivalent antibody fragment analogue |
US6027725A (en) * | 1991-11-25 | 2000-02-22 | Enzon, Inc. | Multivalent antigen-binding proteins |
US6368596B1 (en) * | 1997-07-08 | 2002-04-09 | Board Of Regents, The University Of Texas System | Compositions and methods for homoconjugates of antibodies which induce growth arrest or apoptosis of tumor cells |
US6476198B1 (en) * | 1993-07-13 | 2002-11-05 | The Scripps Research Institute | Multispecific and multivalent antigen-binding polypeptide molecules |
US6670453B2 (en) * | 1997-10-27 | 2003-12-30 | Unilever Patent Holdings B.V. | Multivalent antigen-binding proteins |
Family Cites Families (140)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3773919A (en) | 1969-10-23 | 1973-11-20 | Du Pont | Polylactide-drug mixtures |
USRE30985E (en) | 1978-01-01 | 1982-06-29 | Serum-free cell culture media | |
US4275149A (en) | 1978-11-24 | 1981-06-23 | Syva Company | Macromolecular environment control in specific receptor assays |
US4318980A (en) | 1978-04-10 | 1982-03-09 | Miles Laboratories, Inc. | Heterogenous specific binding assay employing a cycling reactant as label |
WO1981001145A1 (en) | 1979-10-18 | 1981-04-30 | Univ Illinois | Hydrolytic enzyme-activatible pro-drugs |
US4419446A (en) | 1980-12-31 | 1983-12-06 | The United States Of America As Represented By The Department Of Health And Human Services | Recombinant DNA process utilizing a papilloma virus DNA as a vector |
US4485045A (en) | 1981-07-06 | 1984-11-27 | Research Corporation | Synthetic phosphatidyl cholines useful in forming liposomes |
NZ201705A (en) | 1981-08-31 | 1986-03-14 | Genentech Inc | Recombinant dna method for production of hepatitis b surface antigen in yeast |
US4601978A (en) | 1982-11-24 | 1986-07-22 | The Regents Of The University Of California | Mammalian metallothionein promoter system |
US4560655A (en) | 1982-12-16 | 1985-12-24 | Immunex Corporation | Serum-free cell culture medium and process for making same |
US4657866A (en) | 1982-12-21 | 1987-04-14 | Sudhir Kumar | Serum-free, synthetic, completely chemically defined tissue culture media |
US4816567A (en) | 1983-04-08 | 1989-03-28 | Genentech, Inc. | Recombinant immunoglobin preparations |
DD266710A3 (en) | 1983-06-06 | 1989-04-12 | Ve Forschungszentrum Biotechnologie | Process for the biotechnical production of alkaline phosphatase |
US4544545A (en) | 1983-06-20 | 1985-10-01 | Trustees University Of Massachusetts | Liposomes containing modified cholesterol for organ targeting |
US4767704A (en) | 1983-10-07 | 1988-08-30 | Columbia University In The City Of New York | Protein-free culture medium |
US4943533A (en) | 1984-03-01 | 1990-07-24 | The Regents Of The University Of California | Hybrid cell lines that produce monoclonal antibodies to epidermal growth factor receptor |
US4965199A (en) | 1984-04-20 | 1990-10-23 | Genentech, Inc. | Preparation of functional human factor VIII in mammalian cells using methotrexate based selection |
US5807715A (en) | 1984-08-27 | 1998-09-15 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and transformed mammalian lymphocyte cells for producing functional antigen-binding protein including chimeric immunoglobulin |
US4879231A (en) | 1984-10-30 | 1989-11-07 | Phillips Petroleum Company | Transformation of yeasts of the genus pichia |
US4737456A (en) | 1985-05-09 | 1988-04-12 | Syntex (U.S.A.) Inc. | Reducing interference in ligand-receptor binding assays |
GB8516415D0 (en) | 1985-06-28 | 1985-07-31 | Celltech Ltd | Culture of animal cells |
US5091178A (en) | 1986-02-21 | 1992-02-25 | Oncogen | Tumor therapy with biologically active anti-tumor antibodies |
US4927762A (en) | 1986-04-01 | 1990-05-22 | Cell Enterprises, Inc. | Cell culture medium with antioxidant |
GB8610600D0 (en) | 1986-04-30 | 1986-06-04 | Novo Industri As | Transformation of trichoderma |
US5182368A (en) | 1986-06-13 | 1993-01-26 | Ledbetter Jeffrey A | Ligands and methods for augmenting B-cell proliferation |
US5567610A (en) | 1986-09-04 | 1996-10-22 | Bioinvent International Ab | Method of producing human monoclonal antibodies and kit therefor |
IL85035A0 (en) | 1987-01-08 | 1988-06-30 | Int Genetic Eng | Polynucleotide molecule,a chimeric antibody with specificity for human b cell surface antigen,a process for the preparation and methods utilizing the same |
GB8705477D0 (en) | 1987-03-09 | 1987-04-15 | Carlton Med Prod | Drug delivery systems |
EP0307434B2 (en) | 1987-03-18 | 1998-07-29 | Scotgen Biopharmaceuticals, Inc. | Altered antibodies |
US4975278A (en) | 1988-02-26 | 1990-12-04 | Bristol-Myers Company | Antibody-enzyme conjugates in combination with prodrugs for the delivery of cytotoxic agents to tumor cells |
IL87737A (en) | 1987-09-11 | 1993-08-18 | Genentech Inc | Method for culturing polypeptide factor dependent vertebrate recombinant cells |
US4892538A (en) | 1987-11-17 | 1990-01-09 | Brown University Research Foundation | In vivo delivery of neurotransmitters by implanted, encapsulated cells |
US5283187A (en) | 1987-11-17 | 1994-02-01 | Brown University Research Foundation | Cell culture-containing tubular capsule produced by co-extrusion |
US5223409A (en) | 1988-09-02 | 1993-06-29 | Protein Engineering Corp. | Directed evolution of novel binding proteins |
EP0435911B1 (en) | 1988-09-23 | 1996-03-13 | Cetus Oncology Corporation | Cell culture medium for enhanced cell growth, culture longevity and product expression |
FR2646437B1 (en) | 1989-04-28 | 1991-08-30 | Transgene Sa | NOVEL DNA SEQUENCES, THEIR APPLICATION AS A SEQUENCE ENCODING A SIGNAL PEPTIDE FOR THE SECRETION OF MATURE PROTEINS BY RECOMBINANT YEASTS, EXPRESSION CASSETTES, PROCESSED YEASTS AND PROCESS FOR PREPARING THE SAME |
DK0474727T3 (en) | 1989-05-19 | 1998-01-12 | Genentech Inc | HER2 extracellular domain |
EP0402226A1 (en) | 1989-06-06 | 1990-12-12 | Institut National De La Recherche Agronomique | Transformation vectors for yeast yarrowia |
DE10399023I2 (en) | 1989-09-12 | 2006-11-23 | Ahp Mfg B V | TFN-binding proteins |
WO1991005264A1 (en) * | 1989-09-29 | 1991-04-18 | Oncogenetics Partners | Detection and quantification of neu related proteins in the biological fluids of humans |
US5013556A (en) | 1989-10-20 | 1991-05-07 | Liposome Technology, Inc. | Liposomes with enhanced circulation time |
US5208020A (en) | 1989-10-25 | 1993-05-04 | Immunogen Inc. | Cytotoxic agents comprising maytansinoids and their therapeutic use |
US5229275A (en) | 1990-04-26 | 1993-07-20 | Akzo N.V. | In-vitro method for producing antigen-specific human monoclonal antibodies |
US5427908A (en) | 1990-05-01 | 1995-06-27 | Affymax Technologies N.V. | Recombinant library screening methods |
US5723286A (en) | 1990-06-20 | 1998-03-03 | Affymax Technologies N.V. | Peptide library and screening systems |
WO1991019515A1 (en) | 1990-06-21 | 1991-12-26 | The Board Of Trustees Of The Leland Stanford Junior University | Oligomeric immunoglobulin constant domain mutant with enhanced complement-mediated cytolytic activity |
US5625126A (en) | 1990-08-29 | 1997-04-29 | Genpharm International, Inc. | Transgenic non-human animals for producing heterologous antibodies |
US5661016A (en) | 1990-08-29 | 1997-08-26 | Genpharm International Inc. | Transgenic non-human animals capable of producing heterologous antibodies of various isotypes |
US5633425A (en) | 1990-08-29 | 1997-05-27 | Genpharm International, Inc. | Transgenic non-human animals capable of producing heterologous antibodies |
IL99553A0 (en) | 1990-09-28 | 1992-08-18 | Ixsys Inc | Compositions containing oligonucleotides linked to expression elements,a kit for the preparation of vectors useful for the expression of a diverse population of random peptides and methods utilizing the same |
US5122469A (en) | 1990-10-03 | 1992-06-16 | Genentech, Inc. | Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins |
EP0564531B1 (en) | 1990-12-03 | 1998-03-25 | Genentech, Inc. | Enrichment method for variant proteins with altered binding properties |
US5571894A (en) | 1991-02-05 | 1996-11-05 | Ciba-Geigy Corporation | Recombinant antibodies specific for a growth factor receptor |
GB9112536D0 (en) | 1991-06-11 | 1991-07-31 | Celltech Ltd | Chemical compounds |
US5264586A (en) | 1991-07-17 | 1993-11-23 | The Scripps Research Institute | Analogs of calicheamicin gamma1I, method of making and using the same |
US5587458A (en) | 1991-10-07 | 1996-12-24 | Aronex Pharmaceuticals, Inc. | Anti-erbB-2 antibodies, combinations thereof, and therapeutic and diagnostic uses thereof |
US5270170A (en) | 1991-10-16 | 1993-12-14 | Affymax Technologies N.V. | Peptide library and screening method |
CA2122732C (en) | 1991-11-25 | 2008-04-08 | Marc D. Whitlow | Multivalent antigen-binding proteins |
ATE503496T1 (en) | 1992-02-06 | 2011-04-15 | Novartis Vaccines & Diagnostic | BIOSYNTHETIC BINDING PROTEIN FOR TUMOR MARKERS |
CA2128862C (en) | 1992-02-11 | 2008-05-20 | Jonathan G. Seidman | Homogenotization of gene-targeting events |
AU4025193A (en) | 1992-04-08 | 1993-11-18 | Cetus Oncology Corporation | Humanized C-erbB-2 specific antibodies |
ZA932522B (en) | 1992-04-10 | 1993-12-20 | Res Dev Foundation | Immunotoxins directed against c-erbB-2(HER/neu) related surface antigens |
WO1993024640A2 (en) | 1992-06-04 | 1993-12-09 | The Regents Of The University Of California | Methods and compositions for in vivo gene therapy |
AU687346B2 (en) | 1992-06-30 | 1998-02-26 | Oncologix, Inc. | A combination of anti-erbB-2 monoclonal antibodies and method of using |
US5397703A (en) | 1992-07-09 | 1995-03-14 | Cetus Oncology Corporation | Method for generation of antibodies to cell surface molecules |
ES2091684T3 (en) | 1992-11-13 | 1996-11-01 | Idec Pharma Corp | THERAPEUTIC APPLICATION OF CHEMICAL AND RADIO-MARKED ANTIBODIES AGAINST THE RESTRICTED DIFFERENTIATION ANTIGEN OF HUMAN B-LYMPHOCYTES FOR THE TREATMENT OF B-CELL LYMPHOMA. |
CA2103323A1 (en) | 1992-11-24 | 1994-05-25 | Gregory D. Plowman | Her4 human receptor tyrosine kinase |
JP3720353B2 (en) | 1992-12-04 | 2005-11-24 | メディカル リサーチ カウンシル | Multivalent and multispecific binding proteins, their production and use |
ATE187494T1 (en) | 1992-12-11 | 1999-12-15 | Dow Chemical Co | MULTIVALENT SINGLE CHAIN ANTIBODIES |
US5861156A (en) | 1993-01-08 | 1999-01-19 | Creative Biomolecules | Methods of delivering agents to target cells |
DE614989T1 (en) | 1993-02-17 | 1995-09-28 | Morphosys Proteinoptimierung | Method for in vivo selection of ligand binding proteins. |
WO1994022478A1 (en) | 1993-03-30 | 1994-10-13 | The Trustees Of The University Of Pennsylvania | PREVENTION OF TUMORS WITH MONOCLONAL ANTIBODIES AGAINST $i(NEU) |
AU691811B2 (en) | 1993-06-16 | 1998-05-28 | Celltech Therapeutics Limited | Antibodies |
WO1995009917A1 (en) | 1993-10-07 | 1995-04-13 | The Regents Of The University Of California | Genetically engineered bispecific tetravalent antibodies |
SE9304060D0 (en) | 1993-12-06 | 1993-12-06 | Bioinvent Int Ab | Methods to select specific bacteriophages |
JPH09507074A (en) | 1993-12-23 | 1997-07-15 | イミュネックス・コーポレーション | Method for preventing or treating a disease characterized by tumor cells expressing CD40 |
US5773001A (en) | 1994-06-03 | 1998-06-30 | American Cyanamid Company | Conjugates of methyltrithio antitumor agents and intermediates for their synthesis |
US5516637A (en) | 1994-06-10 | 1996-05-14 | Dade International Inc. | Method involving display of protein binding pairs on the surface of bacterial pili and bacteriophage |
US5910486A (en) | 1994-09-06 | 1999-06-08 | Uab Research Foundation | Methods for modulating protein function in cells using, intracellular antibody homologues |
US6214388B1 (en) | 1994-11-09 | 2001-04-10 | The Regents Of The University Of California | Immunoliposomes that optimize internalization into target cells |
WO1996016673A1 (en) | 1994-12-02 | 1996-06-06 | Chiron Corporation | Method of promoting an immune response with a bispecific antibody |
US5731168A (en) | 1995-03-01 | 1998-03-24 | Genentech, Inc. | Method for making heteromultimeric polypeptides |
US5739277A (en) | 1995-04-14 | 1998-04-14 | Genentech Inc. | Altered polypeptides with increased half-life |
AU700592B2 (en) | 1995-04-17 | 1999-01-07 | University Of Utah Research Foundation | Conotoxin peptides |
US5702892A (en) | 1995-05-09 | 1997-12-30 | The United States Of America As Represented By The Department Of Health And Human Services | Phage-display of immunoglobulin heavy chain libraries |
JPH11507535A (en) | 1995-06-07 | 1999-07-06 | イムクローン システムズ インコーポレイテッド | Antibodies and antibody fragments that suppress tumor growth |
US5874409A (en) * | 1995-06-07 | 1999-02-23 | La Jolla Pharmaceutical Company | APL immunoreactive peptides, conjugates thereof and methods of treatment for APL antibody-mediated pathologies |
US5837234A (en) | 1995-06-07 | 1998-11-17 | Cytotherapeutics, Inc. | Bioartificial organ containing cells encapsulated in a permselective polyether suflfone membrane |
US5712374A (en) | 1995-06-07 | 1998-01-27 | American Cyanamid Company | Method for the preparation of substantiallly monomeric calicheamicin derivative/carrier conjugates |
US5714586A (en) | 1995-06-07 | 1998-02-03 | American Cyanamid Company | Methods for the preparation of monomeric calicheamicin derivative/carrier conjugates |
AU6113396A (en) | 1995-06-14 | 1997-01-15 | Regents Of The University Of California, The | Novel high affinity human antibodies to tumor antigens |
DK1666591T3 (en) | 1995-06-29 | 2011-05-23 | Immunex Corp | Cytokine that induces apoptosis |
ATE219517T1 (en) | 1995-08-18 | 2002-07-15 | Morphosys Ag | PROTEIN/(POLY)PEPTIDE LIBRARIES |
US6030945A (en) | 1996-01-09 | 2000-02-29 | Genentech, Inc. | Apo-2 ligand |
WO1997035196A1 (en) | 1996-03-20 | 1997-09-25 | Dyax Corp. | Engineering affinity ligands for macromolecules |
US5968511A (en) | 1996-03-27 | 1999-10-19 | Genentech, Inc. | ErbB3 antibodies |
US6239259B1 (en) * | 1996-04-04 | 2001-05-29 | Unilever Patent Holdings B.V. | Multivalent and multispecific antigen-binding protein |
US6190856B1 (en) | 1996-05-22 | 2001-02-20 | The Johns Hopkins University | Methods of detection utilizing modified bacteriophage |
US5708156A (en) | 1996-05-31 | 1998-01-13 | Ilekis; John V. | Epidermal growth factor receptor-like gene product and its uses |
CA2257873A1 (en) | 1996-06-10 | 1997-12-18 | The Scripps Research Institute | Use of substrate subtraction libraries to distinguish enzyme specificities |
US5922845A (en) | 1996-07-11 | 1999-07-13 | Medarex, Inc. | Therapeutic multispecific compounds comprised of anti-Fcα receptor antibodies |
AU3737297A (en) | 1996-08-05 | 1998-02-25 | Brigham And Women's Hospital | Bacteriophage-mediated gene therapy |
ATE230850T1 (en) | 1996-10-08 | 2003-01-15 | Bisys B V U | METHOD AND MEANS FOR SELECTING PEPTIDES AND PROTEINS WITH SPECIFIC AFFINITY FOR A TARGET MOLECULE |
NZ509480A (en) | 1996-10-18 | 2005-05-27 | Genentech Inc | Use of anti-ErbB2 antibodies which bind to domain1 of ErbB2 for treating a tumour or cancer |
KR20050004269A (en) | 1996-10-25 | 2005-01-12 | 휴먼 게놈 사이언시즈, 인코포레이티드 | Neutrokine alpha |
DE69738841D1 (en) | 1996-12-23 | 2008-08-28 | Immunex Corp | LIGAND FOR RECEPTOR ACTIVATOR OF NF-KAPPA B, LIGAND IS A MEMBER OF THE TNF SUPERFAMILY |
DE69837806T3 (en) | 1997-01-28 | 2012-01-05 | Human Genome Sciences, Inc. | "DEATH-DOMAIN" -INTERDENTING RECEPTOR 4 (DR4), A MEMBER OF THE TNF-RECEPTOR SUPERFAMILY, BINDING ON TRAIL (APO-2L) |
US6072047A (en) | 1997-02-13 | 2000-06-06 | Immunex Corporation | Receptor that binds trail |
US5969102A (en) | 1997-03-03 | 1999-10-19 | St. Jude Children's Research Hospital | Lymphocyte surface receptor that binds CAML, nucleic acids encoding the same and methods of use thereof |
IL130864A0 (en) * | 1997-03-03 | 2001-01-28 | Bristol Myers Squibb Co | Monoclonal antibodies to human cd6 |
US20010010924A1 (en) | 1997-03-14 | 2001-08-02 | Keith Charles Deen | Tumor necrosis factor related receptor, tr6 polynecleotides |
DE69836956T2 (en) | 1997-03-17 | 2007-10-31 | Human Genome Sciences, Inc. | RECEPTOR 5 CONTAINING A DOMAIN RESPONSIBLE FOR CELL DEATH |
US6268411B1 (en) * | 1997-09-11 | 2001-07-31 | The Johns Hopkins University | Use of multivalent chimeric peptide-loaded, MHC/ig molecules to detect, activate or suppress antigen-specific T cell-dependent immune responses |
EA003636B1 (en) | 1997-04-16 | 2003-08-28 | Амген Инк. | Osteoprotegerin binding proteins and receptors |
AU7126498A (en) | 1997-04-16 | 1998-11-11 | Millennium Pharmaceuticals, Inc. | Tumor necrosis factor receptor related proteins tango-63d and tango-63e |
JP2001511653A (en) | 1997-05-15 | 2001-08-14 | ジェネンテク,インコーポレイテッド | Apo-2 receptor |
JP2002503963A (en) | 1997-06-11 | 2002-02-05 | ヒューマン・ジェノム・サイエンシズ・インコーポレイテッド | Human tumor necrosis factor receptor TR9 |
EP0988321A2 (en) | 1997-06-20 | 2000-03-29 | Innogenetics N.V. | B7-binding molecules for treating immune diseases |
WO1999002653A1 (en) | 1997-07-11 | 1999-01-21 | Trustees Of The University Of Pennsylvania | Nucleic acid encoding a novel chemotherapy-induced protein, and methods of use |
WO1999004001A1 (en) | 1997-07-21 | 1999-01-28 | Zymogenetics, Inc. | Tumor necrosis factor receptor ztnfr-5 |
JP2002501721A (en) | 1997-08-01 | 2002-01-22 | モルフォシス・アクチェンゲゼルシャフト | Novel method and phage for identifying nucleic acid sequences encoding members of multimeric (poly) peptide complexes |
EP1019502A2 (en) | 1997-08-06 | 2000-07-19 | Regeneron Pharmaceuticals, Inc. | Human orphan receptor ntr-1 |
EP1003864A1 (en) | 1997-08-15 | 2000-05-31 | Idun Pharmaceuticals, Inc. | Trail receptors, nucleic acids encoding the same, and methods of use thereof |
AU9376498A (en) | 1997-09-05 | 1999-03-22 | University Of Washington | Tumor necrosis factor family receptors and ligands, encoding nucleic acids and related binding agents |
WO1999014330A1 (en) | 1997-09-18 | 1999-03-25 | Genentech, Inc. | DcR3 POLYPEPTIDE, A TNFR HOMOLOG |
WO1999019488A1 (en) | 1997-10-15 | 1999-04-22 | Children's Medical Center Corporation | Novel human egf receptors and use thereof |
WO1999033980A2 (en) | 1997-12-30 | 1999-07-08 | Chiron Corporation | Members of tnf and tnfr families |
CA2317727C (en) | 1998-01-23 | 2013-01-08 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Multipurpose antibody derivatives |
PT1053256E (en) | 1998-01-26 | 2011-09-28 | Genentech Inc | Antibodies to death receptor 4 (dr4) and uses thereof |
PT1068241E (en) | 1998-04-02 | 2007-11-19 | Genentech Inc | Antibody variants and fragments thereof |
DE19819846B4 (en) | 1998-05-05 | 2016-11-24 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Multivalent antibody constructs |
AU751647B2 (en) * | 1998-05-26 | 2002-08-22 | Innogenetics N.V. | Method for expanding primate B cells selectively in immunocompromised mice and producing large numbers of antigen-specific B lymphocytes for the production of primate monoclonal antibodies |
ES2395693T3 (en) | 1998-06-12 | 2013-02-14 | Genentech, Inc. | Monoclonal antibodies, cross-reactive antibodies and process for manufacturing them |
DK1100830T3 (en) | 1998-07-28 | 2004-01-19 | Micromet Ag | Straight Mini Antibodies |
US6824780B1 (en) * | 1999-10-29 | 2004-11-30 | Genentech, Inc. | Anti-tumor antibody compositions and methods of use |
LT2857516T (en) * | 2000-04-11 | 2017-09-11 | Genentech, Inc. | Multivalent antibodies and uses therefor |
US20020076406A1 (en) | 2000-07-25 | 2002-06-20 | Leung Shui-On | Multivalent target binding protein |
SI1517921T1 (en) | 2002-06-28 | 2006-10-31 | Domantis Ltd | Dual specific ligands with increased serum half-life |
US20060263367A1 (en) * | 2005-05-23 | 2006-11-23 | Fey Georg H | Bispecific antibody devoid of Fc region and method of treatment using same |
US20130149308A1 (en) * | 2011-08-12 | 2013-06-13 | Genentech, Inc. | Antibodies to il-1beta and il-18, for treatment of disease |
-
2001
- 2001-03-20 LT LTEP14189442.8T patent/LT2857516T/en unknown
- 2001-03-20 JP JP2001575196A patent/JP2003531588A/en not_active Withdrawn
- 2001-03-20 MX MXPA02010011A patent/MXPA02010011A/en active IP Right Grant
- 2001-03-20 KR KR1020027013596A patent/KR20020093029A/en active IP Right Grant
- 2001-03-20 CN CNB018103723A patent/CN100390288C/en not_active Expired - Lifetime
- 2001-03-20 BR BR0110610-4A patent/BR0110610A/en not_active IP Right Cessation
- 2001-03-20 WO PCT/US2001/008928 patent/WO2001077342A1/en active Application Filing
- 2001-03-20 SI SI200131064T patent/SI2857516T1/en unknown
- 2001-03-20 AU AU2001247616A patent/AU2001247616B2/en not_active Ceased
- 2001-03-20 AU AU4761601A patent/AU4761601A/en active Pending
- 2001-03-20 CN CNA2008100909964A patent/CN101289511A/en active Pending
- 2001-03-20 PL PL01357939A patent/PL357939A1/en unknown
- 2001-03-20 US US09/813,341 patent/US20020004587A1/en not_active Abandoned
- 2001-03-20 CA CA2403425A patent/CA2403425C/en not_active Expired - Lifetime
- 2001-03-20 EP EP14189442.8A patent/EP2857516B1/en not_active Expired - Lifetime
- 2001-03-20 EP EP01920580.6A patent/EP1272647B1/en not_active Expired - Lifetime
- 2001-03-20 DK DK01920580.6T patent/DK1272647T3/en active
- 2001-03-20 ES ES14189442.8T patent/ES2637801T3/en not_active Expired - Lifetime
- 2001-03-20 IL IL15185301A patent/IL151853A0/en unknown
- 2001-03-20 NZ NZ521540A patent/NZ521540A/en unknown
- 2001-03-20 PT PT141894428T patent/PT2857516T/en unknown
- 2001-03-20 DK DK14189442.8T patent/DK2857516T3/en active
- 2001-03-20 HU HU0300369A patent/HUP0300369A2/en unknown
- 2001-03-20 ES ES01920580.6T patent/ES2528794T3/en not_active Expired - Lifetime
-
2002
- 2002-09-19 IL IL151853A patent/IL151853A/en not_active IP Right Cessation
- 2002-09-20 ZA ZA200207589A patent/ZA200207589B/en unknown
-
2005
- 2005-09-02 US US11/218,821 patent/US20060025576A1/en not_active Abandoned
-
2006
- 2006-09-25 US US11/535,031 patent/US20080299120A1/en not_active Abandoned
-
2008
- 2008-04-09 IL IL190741A patent/IL190741A0/en unknown
-
2010
- 2010-11-03 US US12/939,117 patent/US20110110852A1/en not_active Abandoned
-
2012
- 2012-05-11 US US13/470,189 patent/US8722859B2/en not_active Expired - Fee Related
-
2014
- 2014-03-28 US US14/229,683 patent/US9493579B2/en not_active Expired - Lifetime
-
2015
- 2015-09-23 HK HK15109350.8A patent/HK1208705A1/en not_active IP Right Cessation
-
2017
- 2017-09-05 CY CY20171100936T patent/CY1119291T1/en unknown
Patent Citations (29)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4968603A (en) * | 1986-12-31 | 1990-11-06 | The Regents Of The University Of California | Determination of status in neoplastic disease |
US4816467A (en) * | 1987-01-09 | 1989-03-28 | Farmitalia Carlo Erba S.R.L | Heteroaryl 3-oxo-propanenitrile derivatives, pharmaceutical compositions and use |
US5824311A (en) * | 1987-11-30 | 1998-10-20 | Trustees Of The University Of Pennsylvania | Treatment of tumors with monoclonal antibodies against oncogene antigens |
US5677171A (en) * | 1988-01-12 | 1997-10-14 | Genentech, Inc. | Monoclonal antibodies directed to the HER2 receptor |
US5545807A (en) * | 1988-10-12 | 1996-08-13 | The Babraham Institute | Production of antibodies from transgenic animals |
US5434340A (en) * | 1988-12-05 | 1995-07-18 | Genpharm International, Inc. | Transgenic mice depleted in mature T-cells and methods for making transgenic mice |
US5530101A (en) * | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
US5591828A (en) * | 1989-06-22 | 1997-01-07 | Behringwerke Aktiengesellschaft | Bispecific and oligospecific mono-and oligovalent receptors, the preparation and use thereof |
US5480968A (en) * | 1989-12-01 | 1996-01-02 | The United States Of America As Represented By The Department Of Health And Human Services | Isolated polypeptide erbB-3, related to the epidermal growth factor receptor and antibody thereto |
US5183884A (en) * | 1989-12-01 | 1993-02-02 | United States Of America | Dna segment encoding a gene for a receptor related to the epidermal growth factor receptor |
US5569825A (en) * | 1990-08-29 | 1996-10-29 | Genpharm International | Transgenic non-human animals capable of producing heterologous antibodies of various isotypes |
US5545806A (en) * | 1990-08-29 | 1996-08-13 | Genpharm International, Inc. | Ransgenic non-human animals for producing heterologous antibodies |
US5641869A (en) * | 1991-05-24 | 1997-06-24 | Genentech, Inc. | Method for purifying heregulin |
US5959083A (en) * | 1991-06-03 | 1999-09-28 | Behringwerke Aktiengellschaft | Tetravalent bispecific receptors, the preparation and use thereof |
US5821337A (en) * | 1991-06-14 | 1998-10-13 | Genentech, Inc. | Immunoglobulin variants |
US6515110B1 (en) * | 1991-11-25 | 2003-02-04 | Enzon, Inc. | Multivalent antigen-binding proteins |
US6027725A (en) * | 1991-11-25 | 2000-02-22 | Enzon, Inc. | Multivalent antigen-binding proteins |
US5736137A (en) * | 1992-11-13 | 1998-04-07 | Idec Pharmaceuticals Corporation | Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma |
US5591699A (en) * | 1993-02-24 | 1997-01-07 | E. I. Du Pont De Nemours And Company | Particle transport fluids thickened with acetylate free xanthan heteropolysaccharide biopolymer plus guar gum |
US6476198B1 (en) * | 1993-07-13 | 2002-11-05 | The Scripps Research Institute | Multispecific and multivalent antigen-binding polypeptide molecules |
US5595721A (en) * | 1993-09-16 | 1997-01-21 | Coulter Pharmaceutical, Inc. | Radioimmunotherapy of lymphoma using anti-CD20 |
US6066719A (en) * | 1995-04-20 | 2000-05-23 | Genetech, Inc. | Antibody fragments |
US6214984B1 (en) * | 1995-04-20 | 2001-04-10 | Genentech, Inc. | Isolated nucleic acid encoding, and methods for preparing, antibody fragments |
US20020002271A1 (en) * | 1995-04-20 | 2002-01-03 | Genentech, Inc. | Antibody purification |
US5641870A (en) * | 1995-04-20 | 1997-06-24 | Genentech, Inc. | Low pH hydrophobic interaction chromatography for antibody purification |
US5989830A (en) * | 1995-10-16 | 1999-11-23 | Unilever Patent Holdings Bv | Bifunctional or bivalent antibody fragment analogue |
US5783186A (en) * | 1995-12-05 | 1998-07-21 | Amgen Inc. | Antibody-induced apoptosis |
US6368596B1 (en) * | 1997-07-08 | 2002-04-09 | Board Of Regents, The University Of Texas System | Compositions and methods for homoconjugates of antibodies which induce growth arrest or apoptosis of tumor cells |
US6670453B2 (en) * | 1997-10-27 | 2003-12-30 | Unilever Patent Holdings B.V. | Multivalent antigen-binding proteins |
Cited By (97)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8951737B2 (en) | 1996-05-06 | 2015-02-10 | Cornell Research Foundation, Inc. | Treatment and diagnosis of cancer |
US20080226659A1 (en) * | 2000-03-16 | 2008-09-18 | Sharon Erickson | Methods of treatment using anti-erbb antibody-maytansinoid conjugates |
US8337856B2 (en) | 2000-03-16 | 2012-12-25 | Immunogen, Inc. | Methods of treatment using anti-ERBB antibody-maytansinoid conjugates |
US8680239B2 (en) | 2000-12-22 | 2014-03-25 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V. | Use of RGM and its modulators |
US10844135B2 (en) | 2003-10-10 | 2020-11-24 | Immunogen, Inc. | Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates and methods of making said |
US20100047239A1 (en) * | 2005-08-19 | 2010-02-25 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
US8258268B2 (en) | 2005-08-19 | 2012-09-04 | Abbott Laboratories | Dual variable domain immunoglobulin and uses thereof |
US8906864B2 (en) | 2005-09-30 | 2014-12-09 | AbbVie Deutschland GmbH & Co. KG | Binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and their use |
US8940871B2 (en) | 2006-03-20 | 2015-01-27 | The Regents Of The University Of California | Engineered anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting |
US9610360B2 (en) | 2007-01-24 | 2017-04-04 | Ceruliean Pharma Inc. | Polymer drug conjugates with tether groups for controlled drug delivery |
US9527919B2 (en) | 2007-09-04 | 2016-12-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US8940298B2 (en) | 2007-09-04 | 2015-01-27 | The Regents Of The University Of California | High affinity anti-prostate stem cell antigen (PSCA) antibodies for cancer targeting and detection |
US20100322948A1 (en) * | 2007-09-06 | 2010-12-23 | Abbott Gmbh & Co. Kg | Bone morphogenetic protein (BMP)-binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and use of same |
US20100291113A1 (en) * | 2007-10-03 | 2010-11-18 | Cornell University | Treatment of Proliferative Disorders Using Antibodies to PSMA |
US9605069B2 (en) | 2008-02-29 | 2017-03-28 | AbbVie Deutschland GmbH & Co. KG | Antibodies against the RGM a protein and uses thereof |
US8962803B2 (en) | 2008-02-29 | 2015-02-24 | AbbVie Deutschland GmbH & Co. KG | Antibodies against the RGM A protein and uses thereof |
US20100028340A1 (en) * | 2008-02-29 | 2010-02-04 | Abbott Gmbh & Co. Kg | Antibodies against the rgm a protein and uses thereof |
US8715619B2 (en) | 2008-03-26 | 2014-05-06 | Cellerant Therapeutics, Inc. | Compositions and methods for treating haematological proliferative disorders of myeloid origin |
US9371390B2 (en) | 2008-03-26 | 2016-06-21 | Cellerant Therapeutics, Inc. | Cytokine receptors associated with myelogenous haematological proliferative disorders and uses thereof |
US8709715B2 (en) | 2008-03-26 | 2014-04-29 | Cellerant Therapeutics, Inc. | Cytokine receptors associated with myelogenous haematological proliferative disorders and uses thereof |
US20110059852A1 (en) * | 2008-03-26 | 2011-03-10 | Cellerant Therapeutics, Inc. | Compositions and methods for treating haematological proliferative disorders of meyloid origin |
US10517969B2 (en) | 2009-02-17 | 2019-12-31 | Cornell University | Methods and kits for diagnosis of cancer and prediction of therapeutic value |
US8772459B2 (en) | 2009-12-02 | 2014-07-08 | Imaginab, Inc. | J591 minibodies and Cys-diabodies for targeting human prostate specific membrane antigen (PSMA) and methods for their use |
US11180570B2 (en) | 2009-12-02 | 2021-11-23 | Imaginab, Inc. | J591 minibodies and cys-diabodies for targeting human prostate specific membrane antigen (PSMA) and methods for their use |
US9175075B2 (en) | 2009-12-08 | 2015-11-03 | AbbVie Deutschland GmbH & Co. KG | Methods of treating retinal nerve fiber layer degeneration with monoclonal antibodies against a retinal guidance molecule (RGM) protein |
US20110135664A1 (en) * | 2009-12-08 | 2011-06-09 | Abbott Gmbh & Co. Kg | Monoclonal antibodies against the rgm a protein for use in the treatment of retinal nerve fiber layer degeneration |
US20120009596A1 (en) * | 2010-07-07 | 2012-01-12 | Sen-Yung Hsieh | Protein markers for detecting liver cancer and method for identifying the markers thereof |
US8741288B2 (en) * | 2010-07-07 | 2014-06-03 | Chang Gung Medical Foundation, Linkou Branch | Protein markers for detecting liver cancer and method for identifying the markers thereof |
US9085622B2 (en) | 2010-09-03 | 2015-07-21 | Glaxosmithkline Intellectual Property Development Limited | Antigen binding proteins |
US9783602B2 (en) | 2010-12-01 | 2017-10-10 | Alderbio Holdings Llc | Anti-NGF compositions and use thereof |
US9884909B2 (en) | 2010-12-01 | 2018-02-06 | Alderbio Holdings Llc | Anti-NGF compositions and use thereof |
US9067988B2 (en) | 2010-12-01 | 2015-06-30 | Alderbio Holdings Llc | Methods of preventing or treating pain using anti-NGF antibodies |
US9078878B2 (en) | 2010-12-01 | 2015-07-14 | Alderbio Holdings Llc | Anti-NGF antibodies that selectively inhibit the association of NGF with TrkA, without affecting the association of NGF with p75 |
US10457727B2 (en) | 2010-12-01 | 2019-10-29 | Alderbio Holdings Llc | Methods of preventing inflammation and treating pain using anti-NGF compositions |
US9539324B2 (en) | 2010-12-01 | 2017-01-10 | Alderbio Holdings, Llc | Methods of preventing inflammation and treating pain using anti-NGF compositions |
US10344083B2 (en) | 2010-12-01 | 2019-07-09 | Alderbio Holdings Llc | Anti-NGF compositions and use thereof |
US10227402B2 (en) | 2010-12-01 | 2019-03-12 | Alderbio Holdings Llc | Anti-NGF antibodies and anti-NGF antibody fragments |
US10221236B2 (en) | 2010-12-01 | 2019-03-05 | Alderbio Holdings Llc | Anti-NGF antibodies that selectively inhibit the association of NGF with TRKA without affecting the association of NGF with P75 |
US9718882B2 (en) | 2010-12-01 | 2017-08-01 | Alderbio Holdings Llc | Anti-NGF antibodies that selectively inhibit the association of NGF with TrkA, without affecting the association of NGF with P75 |
US11214610B2 (en) | 2010-12-01 | 2022-01-04 | H. Lundbeck A/S | High-purity production of multi-subunit proteins such as antibodies in transformed microbes such as Pichia pastoris |
US9738713B2 (en) | 2010-12-01 | 2017-08-22 | Alderbio Holdings Llc | Methods of preventing or treating pain using anti-NGF antibodies |
US8911734B2 (en) | 2010-12-01 | 2014-12-16 | Alderbio Holdings Llc | Methods of preventing or treating pain using anti-NGF antibodies that selectively inhibit the association of NGF with TrkA, without affecting the association of NGF with p75 |
US9783601B2 (en) | 2010-12-01 | 2017-10-10 | Alderbio Holdings Llc | Methods of preventing inflammation and treating pain using anti-NGF compositions |
WO2013036543A2 (en) * | 2011-09-10 | 2013-03-14 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Molecular imaging of cancer cells in vivo |
US11458212B2 (en) | 2011-09-10 | 2022-10-04 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Molecular imaging of cancer cells in vivo |
WO2013036543A3 (en) * | 2011-09-10 | 2013-05-02 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Molecular imaging of cancer cells in vivo |
US10406248B2 (en) | 2011-09-10 | 2019-09-10 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Molecular imaging of cancer cells in vivo |
US20150203584A1 (en) * | 2011-11-16 | 2015-07-23 | Boehringer Ingelheim International Gmbh | Anti il-36r antibodies |
US10550189B2 (en) * | 2011-11-16 | 2020-02-04 | Boehringer Ingelheim International Gmbh | Anti IL-36R antibodies |
US9365643B2 (en) | 2012-01-27 | 2016-06-14 | AbbVie Deutschland GmbH & Co. KG | Antibodies that bind to repulsive guidance molecule A (RGMA) |
US9102722B2 (en) | 2012-01-27 | 2015-08-11 | AbbVie Deutschland GmbH & Co. KG | Composition and method for the diagnosis and treatment of diseases associated with neurite degeneration |
US10106602B2 (en) | 2012-01-27 | 2018-10-23 | AbbVie Deutschland GmbH & Co. KG | Isolated monoclonal anti-repulsive guidance molecule A antibodies and uses thereof |
WO2013177264A1 (en) * | 2012-05-22 | 2013-11-28 | Shire Human Genetic Therapies, Inc. | Anti-ccl2 antibodies for treatment of scleroderma |
US11464871B2 (en) | 2012-10-02 | 2022-10-11 | Novartis Ag | Methods and systems for polymer precipitation and generation of particles |
US11708412B2 (en) | 2013-09-26 | 2023-07-25 | Novartis Ag | Methods for treating hematologic cancers |
US10570204B2 (en) | 2013-09-26 | 2020-02-25 | The Medical College Of Wisconsin, Inc. | Methods for treating hematologic cancers |
US11827704B2 (en) | 2014-01-24 | 2023-11-28 | Novartis Ag | Antibody molecules to PD-1 and uses thereof |
US10752687B2 (en) | 2014-01-24 | 2020-08-25 | Novartis Ag | Antibody molecules to PD-1 and uses thereof |
US9605070B2 (en) | 2014-01-31 | 2017-03-28 | Novartis Ag | Antibody molecules to TIM-3 and uses thereof |
US10472419B2 (en) | 2014-01-31 | 2019-11-12 | Novartis Ag | Antibody molecules to TIM-3 and uses thereof |
US9884913B2 (en) | 2014-01-31 | 2018-02-06 | Novartis Ag | Antibody molecules to TIM-3 and uses thereof |
US11155620B2 (en) | 2014-01-31 | 2021-10-26 | Novartis Ag | Method of detecting TIM-3 using antibody molecules to TIM-3 |
US10981990B2 (en) | 2014-01-31 | 2021-04-20 | Novartis Ag | Antibody molecules to TIM-3 and uses thereof |
US11566069B2 (en) | 2014-03-27 | 2023-01-31 | Bird Rock Bio, Inc. | Treatment of disease responsive to modulation of cannabanoid 1(CB1) receptor signaling |
US10308712B2 (en) | 2014-03-27 | 2019-06-04 | Bird Rock Bio, Inc. | Antibodies that bind human cannabinoid 1 (CB1) receptor |
US10975147B2 (en) | 2014-04-03 | 2021-04-13 | Igm Biosciences, Inc. | Modified J-chain |
US11555075B2 (en) | 2014-04-03 | 2023-01-17 | Igm Biosciences, Inc. | Modified J-chain |
US10400038B2 (en) | 2014-04-03 | 2019-09-03 | Igm Biosciences, Inc. | Modified J-chain |
US9803029B2 (en) | 2014-07-01 | 2017-10-31 | Amphivena Therapeutics, Inc. | Bispecific CD33 and CD3 binding proteins |
US9212225B1 (en) | 2014-07-01 | 2015-12-15 | Amphivena Therapeutics, Inc. | Bispecific CD33 and CD3 binding proteins |
US10626190B2 (en) | 2014-07-01 | 2020-04-21 | Amphivena Therapeutics, Inc. | Bispecific CD33 and CD3 binding proteins |
US10577411B2 (en) | 2014-08-07 | 2020-03-03 | Novartis Ag | Angiopoietin-like 4 antibodies and methods of use |
US9771417B2 (en) | 2014-08-07 | 2017-09-26 | Novartis Ag | Angiopoietin-like 4 antibodies and methods of use |
US9988443B2 (en) | 2014-08-07 | 2018-06-05 | Novartis Ag | Angiopoetin-like 4 (ANGPTL4) antibodies and methods of use |
US10059763B2 (en) | 2014-09-03 | 2018-08-28 | Boehringer Ingelheim International Gmbh | Compound targeting IL-23A and TNF-alpha and uses thereof |
US10793629B2 (en) | 2014-09-03 | 2020-10-06 | Boehringer Ingelheim International Gmbh | Compound targeting IL-23A and TNF-alpha and uses thereof |
US11680096B2 (en) | 2014-09-03 | 2023-06-20 | Boehringer Ingelheim International Gmbh | Compound targeting IL-23A and TNF-alpha and uses thereof |
US11344620B2 (en) | 2014-09-13 | 2022-05-31 | Novartis Ag | Combination therapies |
US9718884B2 (en) | 2014-11-05 | 2017-08-01 | Eli Lilly And Company | Anti-TNF-/anti-IL-23 bispecific antibodies |
US10927185B2 (en) | 2014-11-21 | 2021-02-23 | Astellas Pharma Inc. | Bispecific antibody format |
US10689449B2 (en) | 2015-01-20 | 2020-06-23 | Igm Biosciences, Inc. | Multimeric death domain-containing receptor-5 (DR5) antibodies and uses thereof |
US11578131B2 (en) | 2015-01-20 | 2023-02-14 | Igm Biosciences, Inc. | Polynucleotides encoding death domain-containing receptor-5 (DR5) binding molecules |
US10787520B2 (en) | 2015-03-04 | 2020-09-29 | Igm Biosciences, Inc. | Multimeric bispecific binding molecules specific for CD20 and CD3 |
US11753469B2 (en) | 2015-05-29 | 2023-09-12 | Anji Bruno, Llc | Methods of using bispecific CD33 and CD3 binding proteins |
US10738118B2 (en) | 2015-05-29 | 2020-08-11 | Amphivena Therapeutics, Inc. | Methods of using bispecific CD33 and CD3 binding proteins |
US10738095B2 (en) | 2015-06-03 | 2020-08-11 | The Medical College Of Wisconsin, Inc. | Engineered CCL20 locked dimer polypeptide |
US11571462B2 (en) | 2015-06-03 | 2023-02-07 | The Medical College Of Wisconsin, Inc. | Engineered CCL20 locked dimer polypeptide |
US11254744B2 (en) | 2015-08-07 | 2022-02-22 | Imaginab, Inc. | Antigen binding constructs to target molecules |
US10618978B2 (en) | 2015-09-30 | 2020-04-14 | Igm Biosciences, Inc. | Binding molecules with modified J-chain |
US11639389B2 (en) | 2015-09-30 | 2023-05-02 | Igm Biosciences, Inc. | Binding molecules with modified J-chain |
US11542342B2 (en) | 2015-09-30 | 2023-01-03 | Igm Biosciences, Inc. | Binding molecules with modified J-chain |
US11421026B2 (en) | 2015-09-30 | 2022-08-23 | Bird Rock Bio, Inc. | Antibodies that bind human cannabinoid 1 (CB1) receptor |
US11266745B2 (en) | 2017-02-08 | 2022-03-08 | Imaginab, Inc. | Extension sequences for diabodies |
US11634489B2 (en) | 2017-08-03 | 2023-04-25 | Alector Llc | Anti-TREM2 antibodies and methods of use thereof |
US11692042B2 (en) | 2018-03-09 | 2023-07-04 | Agenus Inc. | Anti-CD73 antibodies and methods of use thereof |
US11884719B2 (en) | 2018-12-21 | 2024-01-30 | 23Andme, Inc. | Anti-IL-36 antibodies and methods of use thereof |
US11730812B2 (en) | 2019-03-08 | 2023-08-22 | Boehringer Ingelheim International Gmbh | Anti-IL-36R antibody formulations |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20110110852A1 (en) | Multivalent Antibodies and Uses Therefor | |
US11912773B2 (en) | Fc variants and methods for their production | |
US9994646B2 (en) | Coiled coil and/or tether containing protein complexes and uses thereof | |
US10584181B2 (en) | Methods of making and using multispecific antibody panels and antibody analog panels | |
CN103649117B (en) | Fc variant and the method for generation thereof | |
US11459405B2 (en) | Bispecific antibodies having constant region mutations and uses therefor | |
US20220002386A1 (en) | Production of heteromultimeric proteins using mammalian cells | |
JP2019503172A5 (en) |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |