WO2004050707A2 - Tumorspezifische erkennungsmoleküle - Google Patents
Tumorspezifische erkennungsmoleküle Download PDFInfo
- Publication number
- WO2004050707A2 WO2004050707A2 PCT/DE2003/003994 DE0303994W WO2004050707A2 WO 2004050707 A2 WO2004050707 A2 WO 2004050707A2 DE 0303994 W DE0303994 W DE 0303994W WO 2004050707 A2 WO2004050707 A2 WO 2004050707A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- seq
- recognition molecule
- recognition
- sequences
- core
- Prior art date
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 202
- 210000004027 cell Anatomy 0.000 claims description 120
- 238000000034 method Methods 0.000 claims description 94
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 89
- 239000000427 antigen Substances 0.000 claims description 60
- 102000036639 antigens Human genes 0.000 claims description 60
- 108091007433 antigens Proteins 0.000 claims description 60
- 239000013598 vector Substances 0.000 claims description 42
- 239000000203 mixture Substances 0.000 claims description 41
- 210000001519 tissue Anatomy 0.000 claims description 40
- 238000011282 treatment Methods 0.000 claims description 36
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 32
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 32
- 108090000623 proteins and genes Proteins 0.000 claims description 31
- 150000007523 nucleic acids Chemical class 0.000 claims description 28
- 230000014509 gene expression Effects 0.000 claims description 27
- 206010027476 Metastases Diseases 0.000 claims description 26
- 102000004169 proteins and genes Human genes 0.000 claims description 25
- 238000001514 detection method Methods 0.000 claims description 24
- 108060003951 Immunoglobulin Proteins 0.000 claims description 21
- 102000018358 immunoglobulin Human genes 0.000 claims description 21
- 239000008194 pharmaceutical composition Substances 0.000 claims description 21
- 150000001413 amino acids Chemical class 0.000 claims description 20
- 239000012636 effector Substances 0.000 claims description 20
- 108020004707 nucleic acids Proteins 0.000 claims description 18
- 102000039446 nucleic acids Human genes 0.000 claims description 18
- 238000003745 diagnosis Methods 0.000 claims description 17
- 241000700605 Viruses Species 0.000 claims description 15
- 239000002738 chelating agent Substances 0.000 claims description 15
- 238000004519 manufacturing process Methods 0.000 claims description 15
- 238000002560 therapeutic procedure Methods 0.000 claims description 15
- 210000002540 macrophage Anatomy 0.000 claims description 14
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 14
- 238000011321 prophylaxis Methods 0.000 claims description 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 13
- 238000012360 testing method Methods 0.000 claims description 13
- 206010009944 Colon cancer Diseases 0.000 claims description 12
- 238000001727 in vivo Methods 0.000 claims description 11
- 230000008878 coupling Effects 0.000 claims description 10
- 238000010168 coupling process Methods 0.000 claims description 10
- 238000005859 coupling reaction Methods 0.000 claims description 10
- 239000007850 fluorescent dye Substances 0.000 claims description 10
- 102000037865 fusion proteins Human genes 0.000 claims description 10
- 108020001507 fusion proteins Proteins 0.000 claims description 10
- 230000035772 mutation Effects 0.000 claims description 10
- 208000026310 Breast neoplasm Diseases 0.000 claims description 9
- 206010033128 Ovarian cancer Diseases 0.000 claims description 9
- 229940121354 immunomodulator Drugs 0.000 claims description 9
- 239000002502 liposome Substances 0.000 claims description 9
- 241000894007 species Species 0.000 claims description 9
- 201000009030 Carcinoma Diseases 0.000 claims description 8
- 208000029742 colonic neoplasm Diseases 0.000 claims description 8
- 210000004185 liver Anatomy 0.000 claims description 8
- 229920001184 polypeptide Polymers 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 8
- 239000003053 toxin Substances 0.000 claims description 8
- 231100000765 toxin Toxicity 0.000 claims description 8
- 108700012359 toxins Proteins 0.000 claims description 8
- 241001529936 Murinae Species 0.000 claims description 7
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 7
- 206010060862 Prostate cancer Diseases 0.000 claims description 7
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 7
- 239000012642 immune effector Substances 0.000 claims description 7
- 230000003993 interaction Effects 0.000 claims description 7
- 238000000163 radioactive labelling Methods 0.000 claims description 7
- 229910052727 yttrium Inorganic materials 0.000 claims description 7
- 206010006187 Breast cancer Diseases 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 6
- 206010027457 Metastases to liver Diseases 0.000 claims description 6
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 6
- 201000005202 lung cancer Diseases 0.000 claims description 6
- 208000020816 lung neoplasm Diseases 0.000 claims description 6
- 230000009401 metastasis Effects 0.000 claims description 6
- 238000012544 monitoring process Methods 0.000 claims description 6
- 238000004393 prognosis Methods 0.000 claims description 6
- 230000009467 reduction Effects 0.000 claims description 6
- 206010008342 Cervix carcinoma Diseases 0.000 claims description 5
- 208000002699 Digestive System Neoplasms Diseases 0.000 claims description 5
- 241000588724 Escherichia coli Species 0.000 claims description 5
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 claims description 5
- 230000001580 bacterial effect Effects 0.000 claims description 5
- 201000010881 cervical cancer Diseases 0.000 claims description 5
- 238000012217 deletion Methods 0.000 claims description 5
- 230000037430 deletion Effects 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 5
- 206010017758 gastric cancer Diseases 0.000 claims description 5
- 229960005486 vaccine Drugs 0.000 claims description 5
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical group COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 claims description 4
- 239000004472 Lysine Substances 0.000 claims description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 4
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims description 4
- 208000006265 Renal cell carcinoma Diseases 0.000 claims description 4
- 239000002671 adjuvant Substances 0.000 claims description 4
- 238000001839 endoscopy Methods 0.000 claims description 4
- 238000010166 immunofluorescence Methods 0.000 claims description 4
- 238000003780 insertion Methods 0.000 claims description 4
- 230000037431 insertion Effects 0.000 claims description 4
- 229910052740 iodine Inorganic materials 0.000 claims description 4
- 208000008443 pancreatic carcinoma Diseases 0.000 claims description 4
- 101100227726 Arabidopsis thaliana FRL3 gene Proteins 0.000 claims description 3
- 101100120609 Caenorhabditis elegans frh-1 gene Proteins 0.000 claims description 3
- 101150023991 FMNL1 gene Proteins 0.000 claims description 3
- 101150005226 FRL1 gene Proteins 0.000 claims description 3
- 101150065691 FRL2 gene Proteins 0.000 claims description 3
- 102100028930 Formin-like protein 1 Human genes 0.000 claims description 3
- 102100032789 Formin-like protein 3 Human genes 0.000 claims description 3
- 101000840258 Homo sapiens Immunoglobulin J chain Proteins 0.000 claims description 3
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims description 3
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims description 3
- 102100029571 Immunoglobulin J chain Human genes 0.000 claims description 3
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 3
- 210000003527 eukaryotic cell Anatomy 0.000 claims description 3
- 101150029401 fmnl3 gene Proteins 0.000 claims description 3
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims description 3
- 210000004962 mammalian cell Anatomy 0.000 claims description 3
- 201000002528 pancreatic cancer Diseases 0.000 claims description 3
- 210000001236 prokaryotic cell Anatomy 0.000 claims description 3
- 238000009589 serological test Methods 0.000 claims description 3
- 201000002314 small intestine cancer Diseases 0.000 claims description 3
- 201000011549 stomach cancer Diseases 0.000 claims description 3
- 241000699800 Cricetinae Species 0.000 claims description 2
- 241000238631 Hexapoda Species 0.000 claims description 2
- 108010027412 Histocompatibility Antigens Class II Proteins 0.000 claims description 2
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 claims description 2
- 241000235058 Komagataella pastoris Species 0.000 claims description 2
- 102000004856 Lectins Human genes 0.000 claims description 2
- 108090001090 Lectins Proteins 0.000 claims description 2
- 102000019298 Lipocalin Human genes 0.000 claims description 2
- 108050006654 Lipocalin Proteins 0.000 claims description 2
- 102000043129 MHC class I family Human genes 0.000 claims description 2
- 108091054437 MHC class I family Proteins 0.000 claims description 2
- 241001599018 Melanogaster Species 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 230000003197 catalytic effect Effects 0.000 claims description 2
- 239000002299 complementary DNA Substances 0.000 claims description 2
- 230000001461 cytolytic effect Effects 0.000 claims description 2
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 claims description 2
- 208000010749 gastric carcinoma Diseases 0.000 claims description 2
- 210000005260 human cell Anatomy 0.000 claims description 2
- 239000002955 immunomodulating agent Substances 0.000 claims description 2
- 239000002523 lectin Substances 0.000 claims description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 claims description 2
- 230000006641 stabilisation Effects 0.000 claims description 2
- 238000011105 stabilization Methods 0.000 claims description 2
- 201000000498 stomach carcinoma Diseases 0.000 claims description 2
- 230000009261 transgenic effect Effects 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 5
- 238000006722 reduction reaction Methods 0.000 claims 5
- 241000196324 Embryophyta Species 0.000 claims 1
- 239000004231 Riboflavin-5-Sodium Phosphate Substances 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- 208000021045 exocrine pancreatic carcinoma Diseases 0.000 claims 1
- 238000011084 recovery Methods 0.000 claims 1
- 230000027455 binding Effects 0.000 description 47
- 210000003169 central nervous system Anatomy 0.000 description 38
- 239000000126 substance Substances 0.000 description 37
- 210000004881 tumor cell Anatomy 0.000 description 34
- 239000002953 phosphate buffered saline Substances 0.000 description 26
- 239000003814 drug Substances 0.000 description 20
- 238000009472 formulation Methods 0.000 description 19
- 235000018102 proteins Nutrition 0.000 description 19
- 201000011510 cancer Diseases 0.000 description 18
- 150000001720 carbohydrates Chemical class 0.000 description 18
- 235000001014 amino acid Nutrition 0.000 description 17
- 229940024606 amino acid Drugs 0.000 description 17
- 238000006243 chemical reaction Methods 0.000 description 17
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
- 238000002965 ELISA Methods 0.000 description 15
- 238000005516 engineering process Methods 0.000 description 15
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 14
- 210000002966 serum Anatomy 0.000 description 14
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 208000001976 Endocrine Gland Neoplasms Diseases 0.000 description 11
- 201000011523 endocrine gland cancer Diseases 0.000 description 10
- 229940072221 immunoglobulins Drugs 0.000 description 10
- 208000008385 Urogenital Neoplasms Diseases 0.000 description 9
- 201000010099 disease Diseases 0.000 description 9
- 239000012634 fragment Substances 0.000 description 9
- 210000000056 organ Anatomy 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 9
- 208000037964 urogenital cancer Diseases 0.000 description 9
- 102100035360 Cerebellar degeneration-related antigen 1 Human genes 0.000 description 8
- 108090000288 Glycoproteins Proteins 0.000 description 8
- 102000003886 Glycoproteins Human genes 0.000 description 8
- 230000008901 benefit Effects 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 230000009870 specific binding Effects 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 7
- 239000008280 blood Substances 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- 238000010367 cloning Methods 0.000 description 7
- 239000012228 culture supernatant Substances 0.000 description 7
- 208000032839 leukemia Diseases 0.000 description 7
- 238000000746 purification Methods 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- 108020004705 Codon Proteins 0.000 description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 238000003384 imaging method Methods 0.000 description 6
- 230000001404 mediated effect Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 230000002285 radioactive effect Effects 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 230000001225 therapeutic effect Effects 0.000 description 6
- 239000011534 wash buffer Substances 0.000 description 6
- 102000028180 Glycophorins Human genes 0.000 description 5
- 108091005250 Glycophorins Proteins 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 102000005348 Neuraminidase Human genes 0.000 description 5
- 108010006232 Neuraminidase Proteins 0.000 description 5
- 102000003992 Peroxidases Human genes 0.000 description 5
- 238000001042 affinity chromatography Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 235000014633 carbohydrates Nutrition 0.000 description 5
- 238000004113 cell culture Methods 0.000 description 5
- 239000000824 cytostatic agent Substances 0.000 description 5
- 230000001085 cytostatic effect Effects 0.000 description 5
- 230000006378 damage Effects 0.000 description 5
- 210000004443 dendritic cell Anatomy 0.000 description 5
- 239000000032 diagnostic agent Substances 0.000 description 5
- 229940039227 diagnostic agent Drugs 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000001415 gene therapy Methods 0.000 description 5
- 210000004602 germ cell Anatomy 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 229910052738 indium Inorganic materials 0.000 description 5
- APFVFJFRJDLVQX-UHFFFAOYSA-N indium atom Chemical compound [In] APFVFJFRJDLVQX-UHFFFAOYSA-N 0.000 description 5
- PSCMQHVBLHHWTO-UHFFFAOYSA-K indium(iii) chloride Chemical compound Cl[In](Cl)Cl PSCMQHVBLHHWTO-UHFFFAOYSA-K 0.000 description 5
- 238000011068 loading method Methods 0.000 description 5
- 108040007629 peroxidase activity proteins Proteins 0.000 description 5
- 239000008363 phosphate buffer Substances 0.000 description 5
- 238000003118 sandwich ELISA Methods 0.000 description 5
- 230000004614 tumor growth Effects 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 108091035707 Consensus sequence Proteins 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 4
- 201000003741 Gastrointestinal carcinoma Diseases 0.000 description 4
- 208000017604 Hodgkin disease Diseases 0.000 description 4
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 4
- 208000009956 adenocarcinoma Diseases 0.000 description 4
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 210000001124 body fluid Anatomy 0.000 description 4
- 239000010839 body fluid Substances 0.000 description 4
- 208000003362 bronchogenic carcinoma Diseases 0.000 description 4
- 230000009920 chelation Effects 0.000 description 4
- 230000000295 complement effect Effects 0.000 description 4
- 238000009826 distribution Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003623 enhancer Substances 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000004927 fusion Effects 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 238000010348 incorporation Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 230000036210 malignancy Effects 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 230000009871 nonspecific binding Effects 0.000 description 4
- 239000008188 pellet Substances 0.000 description 4
- 230000035515 penetration Effects 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 210000002307 prostate Anatomy 0.000 description 4
- 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 4
- 238000004904 shortening Methods 0.000 description 4
- 210000003491 skin Anatomy 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- VWQVUPCCIRVNHF-UHFFFAOYSA-N yttrium atom Chemical compound [Y] VWQVUPCCIRVNHF-UHFFFAOYSA-N 0.000 description 4
- 208000030507 AIDS Diseases 0.000 description 3
- 102000014914 Carrier Proteins Human genes 0.000 description 3
- 102000019034 Chemokines Human genes 0.000 description 3
- 108010012236 Chemokines Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 208000006168 Ewing Sarcoma Diseases 0.000 description 3
- 102100026122 High affinity immunoglobulin gamma Fc receptor I Human genes 0.000 description 3
- 101000913074 Homo sapiens High affinity immunoglobulin gamma Fc receptor I Proteins 0.000 description 3
- 108010063738 Interleukins Proteins 0.000 description 3
- 102000015696 Interleukins Human genes 0.000 description 3
- 206010025323 Lymphomas Diseases 0.000 description 3
- 206010027406 Mesothelioma Diseases 0.000 description 3
- 241000699660 Mus musculus Species 0.000 description 3
- 206010029260 Neuroblastoma Diseases 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- 208000007660 Residual Neoplasm Diseases 0.000 description 3
- 206010039491 Sarcoma Diseases 0.000 description 3
- 108010090804 Streptavidin Proteins 0.000 description 3
- 229960000723 ampicillin Drugs 0.000 description 3
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 3
- 108091008324 binding proteins Proteins 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000002405 diagnostic procedure Methods 0.000 description 3
- 208000018554 digestive system carcinoma Diseases 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 238000001215 fluorescent labelling Methods 0.000 description 3
- -1 for example Substances 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 210000004907 gland Anatomy 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 238000011532 immunohistochemical staining Methods 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 229940047122 interleukins Drugs 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 210000001165 lymph node Anatomy 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 210000004379 membrane Anatomy 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 210000000822 natural killer cell Anatomy 0.000 description 3
- 210000001331 nose Anatomy 0.000 description 3
- 238000011580 nude mouse model Methods 0.000 description 3
- 230000001575 pathological effect Effects 0.000 description 3
- 210000001322 periplasm Anatomy 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 230000000069 prophylactic effect Effects 0.000 description 3
- 230000005855 radiation Effects 0.000 description 3
- 238000013391 scatchard analysis Methods 0.000 description 3
- 230000003248 secreting effect Effects 0.000 description 3
- 208000000649 small cell carcinoma Diseases 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 208000002008 AIDS-Related Lymphoma Diseases 0.000 description 2
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 2
- 206010006417 Bronchial carcinoma Diseases 0.000 description 2
- 101710132601 Capsid protein Proteins 0.000 description 2
- 101710094648 Coat protein Proteins 0.000 description 2
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 108010087819 Fc receptors Proteins 0.000 description 2
- 102000009109 Fc receptors Human genes 0.000 description 2
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 102100021181 Golgi phosphoprotein 3 Human genes 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
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 2
- 101001133056 Homo sapiens Mucin-1 Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 206010062016 Immunosuppression Diseases 0.000 description 2
- 108010065805 Interleukin-12 Proteins 0.000 description 2
- 108010002350 Interleukin-2 Proteins 0.000 description 2
- 208000007766 Kaposi sarcoma Diseases 0.000 description 2
- 101710125418 Major capsid protein Proteins 0.000 description 2
- 206010061269 Malignant peritoneal neoplasm Diseases 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 102100034256 Mucin-1 Human genes 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 208000005927 Myosarcoma Diseases 0.000 description 2
- OVRNDRQMDRJTHS-KEWYIRBNSA-N N-acetyl-D-galactosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-KEWYIRBNSA-N 0.000 description 2
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 2
- 208000009905 Neurofibromatoses Diseases 0.000 description 2
- 101710141454 Nucleoprotein Proteins 0.000 description 2
- 101710083689 Probable capsid protein Proteins 0.000 description 2
- 108010072866 Prostate-Specific Antigen Proteins 0.000 description 2
- 102100038358 Prostate-specific antigen Human genes 0.000 description 2
- 206010038389 Renal cancer Diseases 0.000 description 2
- 208000005678 Rhabdomyoma Diseases 0.000 description 2
- 208000000453 Skin Neoplasms Diseases 0.000 description 2
- 206010041067 Small cell lung cancer Diseases 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 108091008874 T cell receptors Proteins 0.000 description 2
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 description 2
- 206010043276 Teratoma Diseases 0.000 description 2
- 208000024313 Testicular Neoplasms Diseases 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000005875 antibody response Effects 0.000 description 2
- 238000009175 antibody therapy Methods 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 239000012148 binding buffer Substances 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 206010006007 bone sarcoma Diseases 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 230000004154 complement system Effects 0.000 description 2
- 238000002591 computed tomography Methods 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 210000000232 gallbladder Anatomy 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 238000003365 immunocytochemistry Methods 0.000 description 2
- 230000016784 immunoglobulin production Effects 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 230000003308 immunostimulating effect Effects 0.000 description 2
- 230000001506 immunosuppresive effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- APFVFJFRJDLVQX-AHCXROLUSA-N indium-111 Chemical compound [111In] APFVFJFRJDLVQX-AHCXROLUSA-N 0.000 description 2
- 229940055742 indium-111 Drugs 0.000 description 2
- 238000007912 intraperitoneal administration Methods 0.000 description 2
- 201000010260 leiomyoma Diseases 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 210000002751 lymph Anatomy 0.000 description 2
- 208000012804 lymphangiosarcoma Diseases 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 210000001370 mediastinum Anatomy 0.000 description 2
- 229940126601 medicinal product Drugs 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 201000002077 muscle cancer Diseases 0.000 description 2
- 208000001611 myxosarcoma Diseases 0.000 description 2
- 201000004931 neurofibromatosis Diseases 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 208000003154 papilloma Diseases 0.000 description 2
- 208000007312 paraganglioma Diseases 0.000 description 2
- 208000012111 paraneoplastic syndrome Diseases 0.000 description 2
- 201000002524 peritoneal carcinoma Diseases 0.000 description 2
- 208000010626 plasma cell neoplasm Diseases 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229950010131 puromycin Drugs 0.000 description 2
- 238000001959 radiotherapy Methods 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000000405 serological effect Effects 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 125000006850 spacer group Chemical group 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 208000011580 syndromic disease Diseases 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 210000001685 thyroid gland Anatomy 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 206010069754 Acquired gene mutation Diseases 0.000 description 1
- 208000007876 Acrospiroma Diseases 0.000 description 1
- 206010000830 Acute leukaemia Diseases 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 1
- 208000000583 Adenolymphoma Diseases 0.000 description 1
- 208000003200 Adenoma Diseases 0.000 description 1
- 206010001233 Adenoma benign Diseases 0.000 description 1
- 208000005034 Angiolymphoid Hyperplasia with Eosinophilia Diseases 0.000 description 1
- 201000003076 Angiosarcoma Diseases 0.000 description 1
- 108010032595 Antibody Binding Sites Proteins 0.000 description 1
- 102000005427 Asialoglycoprotein Receptor Human genes 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 101100136076 Aspergillus oryzae (strain ATCC 42149 / RIB 40) pel1 gene Proteins 0.000 description 1
- 208000004736 B-Cell Leukemia Diseases 0.000 description 1
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 1
- 206010004146 Basal cell carcinoma Diseases 0.000 description 1
- 208000003609 Bile Duct Adenoma Diseases 0.000 description 1
- 208000035462 Biphenotypic Acute Leukemia Diseases 0.000 description 1
- 206010004950 Birth mark Diseases 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 208000003170 Bronchiolo-Alveolar Adenocarcinoma Diseases 0.000 description 1
- 206010058354 Bronchioloalveolar carcinoma Diseases 0.000 description 1
- 208000002908 Brown-Pearce carcinoma Diseases 0.000 description 1
- 206010007270 Carcinoid syndrome Diseases 0.000 description 1
- 201000000274 Carcinosarcoma Diseases 0.000 description 1
- 208000007389 Cementoma Diseases 0.000 description 1
- 206010008642 Cholesteatoma Diseases 0.000 description 1
- 201000005262 Chondroma Diseases 0.000 description 1
- 208000005243 Chondrosarcoma Diseases 0.000 description 1
- 201000009047 Chordoma Diseases 0.000 description 1
- 208000006332 Choriocarcinoma Diseases 0.000 description 1
- 101100007328 Cocos nucifera COS-1 gene Proteins 0.000 description 1
- 102100035324 Complement factor H-related protein 4 Human genes 0.000 description 1
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 1
- 208000009798 Craniopharyngioma Diseases 0.000 description 1
- 201000005171 Cystadenoma Diseases 0.000 description 1
- YVGGHNCTFXOJCH-UHFFFAOYSA-N DDT Chemical compound C1=CC(Cl)=CC=C1C(C(Cl)(Cl)Cl)C1=CC=C(Cl)C=C1 YVGGHNCTFXOJCH-UHFFFAOYSA-N 0.000 description 1
- 208000006402 Ductal Carcinoma Diseases 0.000 description 1
- 208000007033 Dysgerminoma Diseases 0.000 description 1
- 206010058314 Dysplasia Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 208000003468 Ehrlich Tumor Carcinoma Diseases 0.000 description 1
- 208000005163 Extra-Adrenal Paraganglioma Diseases 0.000 description 1
- 208000009849 Female Genital Neoplasms Diseases 0.000 description 1
- 201000008808 Fibrosarcoma Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010053717 Fibrous histiocytoma Diseases 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 208000007569 Giant Cell Tumors Diseases 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 206010018338 Glioma Diseases 0.000 description 1
- 206010018381 Glomus tumour Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 108010015899 Glycopeptides Proteins 0.000 description 1
- 102000002068 Glycopeptides Human genes 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 208000005234 Granulosa Cell Tumor Diseases 0.000 description 1
- 206010053759 Growth retardation Diseases 0.000 description 1
- 208000002927 Hamartoma Diseases 0.000 description 1
- 208000006050 Hemangiopericytoma Diseases 0.000 description 1
- 208000001258 Hemangiosarcoma Diseases 0.000 description 1
- 208000002291 Histiocytic Sarcoma Diseases 0.000 description 1
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000878133 Homo sapiens Complement factor H-related protein 4 Proteins 0.000 description 1
- 101000766306 Homo sapiens Serotransferrin Proteins 0.000 description 1
- 241000714259 Human T-lymphotropic virus 2 Species 0.000 description 1
- 241000701024 Human betaherpesvirus 5 Species 0.000 description 1
- 102000012745 Immunoglobulin Subunits Human genes 0.000 description 1
- 108010079585 Immunoglobulin Subunits Proteins 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 229910021617 Indium monochloride Inorganic materials 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 108090000172 Interleukin-15 Proteins 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002586 Interleukin-7 Proteins 0.000 description 1
- 208000032177 Intestinal Polyps Diseases 0.000 description 1
- 206010061252 Intraocular melanoma Diseases 0.000 description 1
- 208000009164 Islet Cell Adenoma Diseases 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 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
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-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
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 208000018142 Leiomyosarcoma Diseases 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 201000004462 Leydig Cell Tumor Diseases 0.000 description 1
- 206010024612 Lipoma Diseases 0.000 description 1
- 206010025219 Lymphangioma Diseases 0.000 description 1
- 208000004138 Lymphangiomyoma Diseases 0.000 description 1
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 1
- 208000028018 Lymphocytic leukaemia Diseases 0.000 description 1
- 208000008095 Malignant Carcinoid Syndrome Diseases 0.000 description 1
- 206010025538 Malignant ascites Diseases 0.000 description 1
- 208000000172 Medulloblastoma Diseases 0.000 description 1
- 208000002030 Merkel cell carcinoma Diseases 0.000 description 1
- 208000010153 Mesonephroma Diseases 0.000 description 1
- 206010027452 Metastases to bone Diseases 0.000 description 1
- 206010059282 Metastases to central nervous system Diseases 0.000 description 1
- 206010027458 Metastases to lung Diseases 0.000 description 1
- 108010063954 Mucins Proteins 0.000 description 1
- 208000007727 Muscle Tissue Neoplasms Diseases 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 1
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 1
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 1
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 1
- JOCBASBOOFNAJA-UHFFFAOYSA-N N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid Chemical compound OCC(CO)(CO)NCCS(O)(=O)=O JOCBASBOOFNAJA-UHFFFAOYSA-N 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 206010029098 Neoplasm skin Diseases 0.000 description 1
- 206010029266 Neuroendocrine carcinoma of the skin Diseases 0.000 description 1
- 201000004404 Neurofibroma Diseases 0.000 description 1
- 208000005890 Neuroma Diseases 0.000 description 1
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 1
- 230000004989 O-glycosylation Effects 0.000 description 1
- 108010079246 OMPA outer membrane proteins Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 208000009095 Orbital Neoplasms Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 206010061332 Paraganglion neoplasm Diseases 0.000 description 1
- 102000012288 Phosphopyruvate Hydratase Human genes 0.000 description 1
- 108010022181 Phosphopyruvate Hydratase Proteins 0.000 description 1
- 208000002163 Phyllodes Tumor Diseases 0.000 description 1
- 206010071776 Phyllodes tumour Diseases 0.000 description 1
- 208000007641 Pinealoma Diseases 0.000 description 1
- 208000033014 Plasma cell tumor Diseases 0.000 description 1
- 208000007452 Plasmacytoma Diseases 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 1
- 102100022019 Pregnancy-specific beta-1-glycoprotein 2 Human genes 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 208000033759 Prolymphocytic T-Cell Leukemia Diseases 0.000 description 1
- 208000034541 Rare lymphatic malformation Diseases 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 206010038802 Reticuloendothelial system stimulated Diseases 0.000 description 1
- 201000000582 Retinoblastoma Diseases 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- 208000003274 Sertoli cell tumor Diseases 0.000 description 1
- 102220497176 Small vasohibin-binding protein_T47D_mutation Human genes 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 206010042658 Sweat gland tumour Diseases 0.000 description 1
- 208000000389 T-cell leukemia Diseases 0.000 description 1
- 208000028530 T-cell lymphoblastic leukemia/lymphoma Diseases 0.000 description 1
- 208000026651 T-cell prolymphocytic leukemia Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 239000007994 TES buffer Substances 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- WDLRUFUQRNWCPK-UHFFFAOYSA-N Tetraxetan Chemical compound OC(=O)CN1CCN(CC(O)=O)CCN(CC(O)=O)CCN(CC(O)=O)CC1 WDLRUFUQRNWCPK-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
- 208000033781 Thyroid carcinoma Diseases 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 101710120037 Toxin CcdB Proteins 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 102000007238 Transferrin Receptors Human genes 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
- 206010054094 Tumour necrosis Diseases 0.000 description 1
- 206010046458 Urethral neoplasms Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 201000005969 Uveal melanoma Diseases 0.000 description 1
- 241000700618 Vaccinia virus Species 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 208000008383 Wilms tumor Diseases 0.000 description 1
- NKVLDFAVEWLOCX-GUSKIFEASA-N [(2s,3r,4s,5r,6r)-3-[(2s,3r,4s,5r,6s)-5-[(2s,3r,4s,5r)-4-[(2s,3r,4r)-3,4-dihydroxy-4-(hydroxymethyl)oxolan-2-yl]oxy-3,5-dihydroxyoxan-2-yl]oxy-3,4-dihydroxy-6-methyloxan-2-yl]oxy-4,5-dihydroxy-6-methyloxan-2-yl] (4ar,5r,6as,6br,9s,10s,12ar)-10-[(2r,3r,4s, Chemical compound O([C@H]1[C@H](O)CO[C@H]([C@@H]1O)O[C@H]1[C@H](C)O[C@H]([C@@H]([C@@H]1O)O)O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](C)O[C@H]1OC(=O)[C@]12CCC(C)(C)CC1C1=CCC3[C@@]([C@@]1(C[C@H]2O)C)(C)CCC1[C@]3(C)CC[C@@H]([C@@]1(C)C=O)O[C@@H]1O[C@@H]([C@H]([C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)CO2)O)[C@H]1O[C@H]1[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O1)O)O)C(=O)NCCCCCCCCCCCC)[C@@H]1OC[C@](O)(CO)[C@H]1O NKVLDFAVEWLOCX-GUSKIFEASA-N 0.000 description 1
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 208000018234 adnexal spiradenoma/cylindroma of a sweat gland Diseases 0.000 description 1
- 210000004404 adrenal cortex Anatomy 0.000 description 1
- 210000004100 adrenal gland Anatomy 0.000 description 1
- 210000001943 adrenal medulla Anatomy 0.000 description 1
- 238000001261 affinity purification Methods 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 1
- 208000010029 ameloblastoma Diseases 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004873 anchoring Methods 0.000 description 1
- 208000000252 angiomatosis Diseases 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 108010044715 asialofetuin Proteins 0.000 description 1
- 108010028495 asialoglycophorin Proteins 0.000 description 1
- 108010006523 asialoglycoprotein receptor Proteins 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
- 238000007845 assembly PCR Methods 0.000 description 1
- 229960000190 bacillus calmette–guérin vaccine Drugs 0.000 description 1
- 229960001212 bacterial vaccine Drugs 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 208000003373 basosquamous carcinoma Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 208000021592 benign granular cell tumor Diseases 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
- 230000001588 bifunctional effect Effects 0.000 description 1
- 210000000013 bile duct Anatomy 0.000 description 1
- 210000003445 biliary tract Anatomy 0.000 description 1
- 201000009036 biliary tract cancer Diseases 0.000 description 1
- 208000020790 biliary tract neoplasm Diseases 0.000 description 1
- 230000004791 biological behavior Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 238000007413 biotinylation Methods 0.000 description 1
- 230000006287 biotinylation Effects 0.000 description 1
- 229910052797 bismuth Inorganic materials 0.000 description 1
- JCXGWMGPZLAOME-UHFFFAOYSA-N bismuth atom Chemical compound [Bi] JCXGWMGPZLAOME-UHFFFAOYSA-N 0.000 description 1
- 201000000053 blastoma Diseases 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 201000006491 bone marrow cancer Diseases 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 201000008274 breast adenocarcinoma Diseases 0.000 description 1
- 210000000621 bronchi Anatomy 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 208000005761 carcinoid heart disease Diseases 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 238000010370 cell cloning Methods 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 230000006041 cell recruitment Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 238000012412 chemical coupling Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 208000011654 childhood malignant neoplasm Diseases 0.000 description 1
- 208000012191 childhood neoplasm Diseases 0.000 description 1
- 201000005217 chondroblastoma Diseases 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 208000009060 clear cell adenocarcinoma Diseases 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000002872 contrast media Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 208000030381 cutaneous melanoma Diseases 0.000 description 1
- 208000017763 cutaneous neuroendocrine carcinoma Diseases 0.000 description 1
- 208000002445 cystadenocarcinoma Diseases 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000013024 dilution buffer Substances 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 201000008184 embryoma Diseases 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- 230000012202 endocytosis Effects 0.000 description 1
- 210000003038 endothelium Anatomy 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 210000001808 exosome Anatomy 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 206010016629 fibroma Diseases 0.000 description 1
- 201000008825 fibrosarcoma of bone Diseases 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 201000008361 ganglioneuroma Diseases 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000001 growth retardation Toxicity 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 201000011066 hemangioma Diseases 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 201000005133 hidradenoma Diseases 0.000 description 1
- 201000009379 histiocytoid hemangioma Diseases 0.000 description 1
- 201000000284 histiocytoma Diseases 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 210000003026 hypopharynx Anatomy 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 201000004933 in situ carcinoma Diseases 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- APHGZSBLRQFRCA-UHFFFAOYSA-M indium(1+);chloride Chemical compound [In]Cl APHGZSBLRQFRCA-UHFFFAOYSA-M 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 201000002313 intestinal cancer Diseases 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 201000002529 islet cell tumor Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- ZBKFYXZXZJPWNQ-UHFFFAOYSA-N isothiocyanate group Chemical group [N-]=C=S ZBKFYXZXZJPWNQ-UHFFFAOYSA-N 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 210000001865 kupffer cell Anatomy 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 101150066555 lacZ gene Proteins 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 210000000867 larynx Anatomy 0.000 description 1
- 210000000088 lip Anatomy 0.000 description 1
- 206010024627 liposarcoma Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 208000016992 lung adenocarcinoma in situ Diseases 0.000 description 1
- 201000000966 lung oat cell carcinoma Diseases 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 108010026228 mRNA guanylyltransferase Proteins 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 208000029559 malignant endocrine neoplasm Diseases 0.000 description 1
- 201000006812 malignant histiocytosis Diseases 0.000 description 1
- 201000008749 mast-cell sarcoma Diseases 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 206010027191 meningioma Diseases 0.000 description 1
- 208000011831 mesonephric neoplasm Diseases 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 208000024191 minimally invasive lung adenocarcinoma Diseases 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 102000035118 modified proteins Human genes 0.000 description 1
- 108091005573 modified proteins Proteins 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 206010051747 multiple endocrine neoplasia Diseases 0.000 description 1
- 208000025113 myeloid leukemia Diseases 0.000 description 1
- 201000004130 myoblastoma Diseases 0.000 description 1
- 208000009091 myxoma Diseases 0.000 description 1
- UUORTJUPDJJXST-UHFFFAOYSA-N n-(2-hydroxyethyl)prop-2-enamide Chemical compound OCCNC(=O)C=C UUORTJUPDJJXST-UHFFFAOYSA-N 0.000 description 1
- 210000001989 nasopharynx Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 230000009826 neoplastic cell growth Effects 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- 208000029986 neuroepithelioma Diseases 0.000 description 1
- 231100001223 noncarcinogenic Toxicity 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 201000002575 ocular melanoma Diseases 0.000 description 1
- 208000004128 odontoma Diseases 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 208000025303 orbit neoplasm Diseases 0.000 description 1
- 210000003300 oropharynx Anatomy 0.000 description 1
- 208000008798 osteoma Diseases 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 210000003101 oviduct Anatomy 0.000 description 1
- 208000022102 pancreatic neuroendocrine neoplasm Diseases 0.000 description 1
- 201000010198 papillary carcinoma Diseases 0.000 description 1
- 210000003695 paranasal sinus Anatomy 0.000 description 1
- 210000002990 parathyroid gland Anatomy 0.000 description 1
- 101150040383 pel2 gene Proteins 0.000 description 1
- 101150050446 pelB gene Proteins 0.000 description 1
- 210000004197 pelvis Anatomy 0.000 description 1
- 210000003899 penis Anatomy 0.000 description 1
- 229960003330 pentetic acid Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 238000002823 phage display Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 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
- 230000010399 physical interaction Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 208000024724 pineal body neoplasm Diseases 0.000 description 1
- 201000004123 pineal gland cancer Diseases 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920000724 poly(L-arginine) polymer Polymers 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 108010011110 polyarginine Proteins 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 201000005825 prostate adenocarcinoma Diseases 0.000 description 1
- 108020001580 protein domains Proteins 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- XNSAINXGIQZQOO-SRVKXCTJSA-N protirelin Chemical compound NC(=O)[C@@H]1CCCN1C(=O)[C@@H](NC(=O)[C@H]1NC(=O)CC1)CC1=CN=CN1 XNSAINXGIQZQOO-SRVKXCTJSA-N 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000003608 radiolysis reaction Methods 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 201000010174 renal carcinoma Diseases 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 201000009410 rhabdomyosarcoma Diseases 0.000 description 1
- 210000003079 salivary gland Anatomy 0.000 description 1
- 201000007416 salivary gland adenoid cystic carcinoma Diseases 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 208000004259 scirrhous adenocarcinoma Diseases 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 201000003708 skin melanoma Diseases 0.000 description 1
- 201000004477 skin sarcoma Diseases 0.000 description 1
- 208000000587 small cell lung carcinoma Diseases 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 230000037439 somatic mutation Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 206010042863 synovial sarcoma Diseases 0.000 description 1
- 201000003120 testicular cancer Diseases 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 208000001644 thecoma Diseases 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 208000008732 thymoma Diseases 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 208000013077 thyroid gland carcinoma Diseases 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- 210000002993 trophoblast Anatomy 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 210000003708 urethra Anatomy 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 210000002229 urogenital system Anatomy 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 210000003905 vulva Anatomy 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
- A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
- A61K51/1045—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants
-
- 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
- A61K47/6851—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 targeting a determinant of a tumour cell
-
- 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
- A61K47/69—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6903—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 conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being semi-solid, e.g. an ointment, a gel, a hydrogel or a solidifying gel
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
- C07K16/3076—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells against structure-related tumour-associated moieties
-
- 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
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/626—Diabody or triabody
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Definitions
- the invention relates to recognition molecules which are directed against tumors and can be used for the diagnosis and therapy of tumor diseases.
- Tumor or cancer diseases are tumors that describe a localized increase in tissue volume.
- any localized swelling due to edema, acute and / or chronic inflammation, an aneurysmal enlargement or an inflammation-related swelling of the organ is a tumor.
- tissue neoplasms such as growths, blastomas and / or neoplasias in the form of spontaneous, differently disinhibited, autonomous and irreversible excess growth of the body's own tissue, which is usually associated with different levels of loss of specific cells and tissue functions, are understood as tumor diseases . It is possible to systematize tumors according to their biological behavior, but also according to a histogenetic system or according to clinical or pathological findings.
- cancer diagnosis using ultrasound and the use of radioactively labeled antibodies is possible, the antigens typical of the tumor binding to the organs to be examined and thus making the tumors recognizable within the imaging process.
- laboratory tests are another important means of early cancer detection. Samples of urine, blood or even tissue samples are examined for abnormalities. For example, this can be changed Composition of these samples,. but also the occurrence of substances that do not normally occur or only occur in small quantities. These substances are commonly referred to as tumor markers. They are either produced by the tumor tissue itself or are formed as a reaction of the body to the tumor.
- tumor markers also refer to cellular changes, the qualitative or quantitative analysis of which enables a statement to be made about the presence, the course or a prognosis of malignant diseases.
- Tumor markers are usually physiologically occurring or modified substances that can be detected in an increased or decreased concentration or in or on tumor cells compared to physiological conditions or the normal genotypic / phenotypic expression in urine, serum or other body fluids, these substances being synthesized and / or by the tumor tissue secreted and subsequently released by tumor decay or formed as a reaction of the organism to a tumor.
- a large number of tumor markers * have been described, the use of which is particularly useful for colon cancer, breast cancer, ovarian cancer, prostate and testicular cancer and for small cell lung cancer.
- These cancer markers include, for example, the CEA, CA 15-3, CA 125, alpha-fetoprotein, HCG, the prostate-specific antigen, the neuron-specific enolase, CA 19-9 and SCC.
- the markers mentioned show an increase in serum or tissues or their presence as modified proteins, lipids and / or carbohydrates on the one hand for example (i) inflammatory diseases, intestinal polyps, viral infections but in particular also (ii) cirrhosis, degeneration, tumors and metastases on.
- a large part of these markers consists of molecules that contain protein and carbohydrate structures, possibly lipids. The lower the Protein content and accordingly the greater the carbohydrate or lipid content of these markers, the more difficult it is for them to be detected, for example, with recognition molecules, such as antibodies. So far, various antibodies against carbohydrate structures have been produced by immunizing mice with the help of hybridoma technology.
- Cancer diagnosis with recognition molecules has several disadvantages. Certain tumor markers can thus also occur in the case of non-carcinogenic diseases, as a result of which the recognition molecules used indicate a positive reaction; furthermore, non-interaction of the recognition molecules does not mean that there is no tumor disease. Another disadvantage is that the known recognition substances are generally non-specific. This means that positive evidence only rarely points to a specific type of tumor. Another, very decisive disadvantage of the known recognition molecules is that they can only be used to a limited extent to monitor the course of the development of tumors, for example after an operation. This means that the known tumor markers can generally not be used for early detection or post-treatment, especially not for prophylaxis.
- the antibody JAA / F1 which is directed against Galßl-3GalNAc, recognizes not only this antigen itself, but also GlcNAcßl ⁇ 6Galßl- 3- (GlcNAcßl- »6) GalNAc and, albeit with less avidity, Galßl -3GlcNAc.
- the primary IgM antibodies most often obtained by immunization are too large for therapeutic use.
- Another disadvantage of the known recognition molecules against tumor markers is that they only make the tumor recognizable when it has already reached a critical size. This means that early stages of tumor growth cannot be determined using the known recognition molecules that are directed against tumor markers.
- Another disadvantage of the known recognition substances is that they cannot be used functionally *. Functional means that the recognition molecules don't just do that bind to the tumor markers that they are detected, but that they thus interact with the tumor cell via markers, that the growth of the tumor cell is impaired. It is possible for the recognition molecules to interact with specific tumor markers, which are immobilized, for example, on tumor cell surfaces, in such a specific manner that the tumor characterized by the tumor markers is treated therapeutically. On the one hand, these functionally active recognition molecules are able to detect tumor markers associated with tumor cells and at the same time, by binding to this tumor-specific structure, to prevent the tumor cell from further growth or from the formation of metastases.
- the known recognition molecules are disadvantageously able to influence tumor growth in only a few cases. As a rule, therefore, additional substances that restrict or inhibit tumor growth must be coupled to the antibody so that it is the “ferry” of this substance, but not the agent of the treatment.
- the object of the invention is therefore to provide recognition molecules with which tumors can be detected simply, safely and efficiently and which can also be used in the prophylaxis, therapy and / or aftertreatment of tumors.
- the invention solves this technical problem by providing recognition molecules comprising an amino acid sequence which contains the amino acid sequence SEQ ID No. 1 and the amino acid sequence SEQ ID No. 2 or 3 and the amino acid sequence SEQ ID No. 4, 5 or 6, wherein the recognition molecules specifically bind the core-1 antigen.
- recognition molecules comprising an amino acid sequence which contains the amino acid sequence SEQ ID No. 1 and the amino acid sequence SEQ ID No. 2 or 3 and the amino acid sequence SEQ ID No. 4, 5 or 6, wherein the recognition molecules specifically bind the core-1 antigen.
- recognition molecule is understood to mean a molecule which, in particular under stringent conditions, specifically binds the carbohydrate structure Core-1.
- Core-1 is understood according to the invention to mean the carbohydrate structure Galßl-3GalNAc, which can be present as an ⁇ -anomer (Galßl-3GalNAc ⁇ ) or ⁇ -anomer (Galßl-3GalNAcß).
- the ⁇ -anomeric variant is preferred here.
- the recognition molecules according to the invention can also bind only the alpha anomer Galßl-3GalNAc ⁇ or both anomers Galßl-3GalNAc ⁇ and Galßl-3GalNAcß in the same way.
- a specific binding against core-1 means a binding which only recognizes core-1, preferably the ⁇ -anomer, or which recognizes core-1 and core-2 (Galßl-3 (GlcNAcßl- 6) GalNAc ⁇ ).
- the recognition molecules show no cross-reactivity with other derivatives and anomers of these carbohydrate structures as listed in Example 7.
- a recognition molecule according to the invention which specifically binds the core-1 antigen comprises: a) a first amino acid sequence, the amino acid sequence SEQ ID No. 1 and the amino acid sequence SEQ ID No. 2 or
- the first and second amino acid sequences can occur on one or more, preferably two, polypeptides there.
- the core-1 binding recognition molecules according to the invention are characterized in that they contain a defined set of individual amino acid sequences.
- the amino acid sequence of these recognition molecules contains one or two triplets of defined sequences. These sequences represent the binding domains and define the specificity of the recognition molecule.
- the 1-triplet recognition molecule contains the amino acid sequence SEQ ID NO. 1, the amino acid sequence SEQ ID NO. 2 or 3 and the amino acid sequence SEQ ID NO. 4 or 5 or 6.
- Core 1-specific recognition molecules, which are defined by two triplets contain the amino acid sequence SEQ ID NO for the first triplet. 1, the amino acid sequence SEQ ID NO. 2 or 3 and the amino acid sequence SEQ ID NO. 4 or 5 or 6 and for the second triplet the amino acid sequence SEQ ID NO.
- the first and second triplet can occur either on one or more polypeptide chains, which together form the binding recognition molecule in the latter case.
- these triplets are referred to as triplet sequence 1 for the first amino acid sequence included and as triplet sequence 2 for the second amino acid sequence included, see definition a) and b) of the above description.
- the recognition molecule can be an antibody, in particular a murine, chimeric or human IgG or IgM, an scFv structure or another.
- a further embodiment of the invention relates to recognition molecules in which at least one amino acid sequence of SEQ ID NO. 1 . to 13 is changed by mutation, deletion and / or insertion, but the property of binding specificity against Core-1 still exists. This advantageously serves to improve the recognition molecules, for example in terms of affinity, solubility and / or producibility.
- the recognition molecule is modified by one or more mutations in one or more amino acid sequences selected from SEQ ID NO:
- amino acids with analogous physicochemical properties which advantageously do not fundamentally change the 3-dimensional structure of the binding domain of the recognition molecule, so that the core-1 specificity of the recognition molecule is retained.
- At least one amino acid sequence is the amino acid sequences SEQ ID NO. 1, 2, 3, 7, 8 and / or 9 through canonical structure variants or equivalent structures with the amino acid sequences SEQ ID NO. 14 to 45 replaced, the SEQ ID NO. 1 by a sequence of the sequences SEQ ID NO. 14 to 17 (CDRH1), SEQ ID NO. 2 or 3 by a sequence of the sequences SEQ ID NO. 18 to 27 (CDRH2) and SEQ ID NO. 7 or 8 or 9 by a sequence of the sequences SEQ ID NO. 28 to 45 (CDRL1) is replaced.
- amino acid sequences SEQ ID NO. 1 to 13 or their modifications in a core-1-specific recognition molecule in the sense of the invention form spatial structures, for example so-called loops, which are characterized in that they have a definable tertiary structure and / or quaternary structure.
- the binding region of a recognition molecule with the Core-1 antigen is formed by amino acid residues, which are provided by up to six variable loops on the surface of the molecule and which interact specifically with Core-1.
- recognition molecules that specifically bind Core-1 are provided, in which at least one sequence of the sequences of the triplet that is not directly involved in the interaction with the Core-1 antigen is omitted.
- the recognition molecules comprise at least one of the amino acid sequences of SEQ ID NO. 1 to 13 or the variants described above twice or more, whereby these doublings * can also occur as variants of the same amino acid sequence. All of the recognition molecules described in this section advantageously recognize the antigen Core-1 specifically. In the following, these recognition molecules, which strictly speaking do not carry triplet sequences due to the omission or duplication of sequences, are nevertheless referred to as triplet sequence 1 or triplet sequence 2 in order to simplify the illustration.
- the recognition molecules according to the invention which specifically bind the Core-1 antigen comprise amino acid sequences which have a homology of at least 60%, preferably 70%, preferably 80%, very particularly preferably 90% compared to the sequences SEQ ID NO. 1 to 13.
- the recognition molecules in the sense of the invention can further comprise framework sequences which comprise the comprehensive amino acid sequences amino acid sequence SEQ ID NO. 1 and the amino acid sequence SEQ ID NO. 2 or 3 and the amino acid sequence SEQ ID No. 4 or 5 or 6, or their variants described above, separate from one another, and framework sequences, the amino acid sequences SEQ .ID No. 7 or 8 or 9 and the amino acid sequence SEQ ID No. 10 or 11 and the amino acid sequence SEQ ID No. 12 or 13, or their variants described above, separate.
- the first and second amino acid sequences can occur on one or more, preferably two, polypeptide chains.
- these scaffold sequences are referred to as spacers or framework sequences and can have different lengths and sequences.
- recognition molecules are also expressly included, in which not all amino acid sequences of SEQ ID No. 1 to 13 or their variants described above are separated by spacers.
- the recognition molecules preferably have further flanking amino acid sequences which are in the For the purposes of the invention, they are also referred to as framework sequences.
- the scaffold sequences have the particular task of arranging the amino acid sequences described, which are responsible or involved for the core-1-specific binding of the recognition molecules, in a suitable arrangement and spatial structure so that the binding to the core-1 can take place. It can be provided that the amino acid sequences SEQ ID NO. 1 to NO. 13 can not specifically bind the core-1 antigen in the sense of the invention without at least one additional amino acid sequence as a framework sequence.
- the scaffold sequences can e.g. provide the necessary biological and chemical stability so that the spatial structure can be effectively built up and maintained for the function and application in a suitable functional form that includes the Core-1 binding.
- the triplet sequences are inserted into existing proteins by exchanging amino acid sequences and / or by addition, the existing protein sequences serving as framework sequences in the sense of the invention, or framework sequences being taken from suitable proteins. These framework sequences can be changed, for example, by mutations, deletions or insertions.
- the person skilled in the art uses known methods of molecular biology, biochemistry and protein engineering.
- Preferred proteins for this are proteins of the immunoglobulin superfamily, protease inhibitors, lectins, helix bundle proteins and lipocalins, as are disclosed, for example, in: Nygren and Uhlen, 1997; Nuttall SD et al. , 1999 and Skerra, 2000.
- the framework sequences are antibody framework sequences from one or different species or amino acid sequences which mimic the consensus sequence of the framework sequences of murine, human and / or antibodies from other mammals.
- a consensus sequence is an idealized sequence in which the most frequently occurring amino acid is represented at each position when many existing sequences, for example from antibody databases, are compared with one another.
- the preferred recognition molecules here are characterized in that the framework sequences for the first triplet sequence 1 comprise the amino acid sequence SEQ ID NO. 1, the amino acid sequence SEQ ID NO. 2 or 3 and the amino acid sequence SEQ ID NO.
- variable heavy chain VH which are also referred to in the literature as framework sequences
- framework sequences for the triplet sequence 2 comprising the amino acid sequence SEQ ID NO. 7 or 8 or 9, the amino acid sequence SEQ ID NO. 10 or 11 and the amino acid sequence SEQ ID NO. 12 or 13, or their variants described above, are antibody framework sequences of the variable light chain VL.
- Antibody framework sequences of antibodies from mammals are furthermore preferred; antibody framework sequences of human and / or murine origin are particularly preferred.
- the framework sequences can be combined from antibody framework sequences of different species.
- These antibody framework sequences are known to the person skilled in the art and are available in various databases, such as the Kabat database (immuno.bme.nwu.edu) or the database of the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov).
- These antibody scaffold structures can also be extended by further amino acids and / or by an or several mutations, for example deletions and / or insertions, are changed, the specific binding to Core-1 being retained.
- the recognition molecule represents a variable chain of an antibody or a structure derived therefrom.
- antibody backbone sequences as backbone sequences in the sense of the invention are the amino acid sequences corresponding to FRH1, FRH2, FRH3 and FHR4 in Table 2 for the variable heavy chain and the amino acid sequences corresponding to FRL1, FRL2, FRL3 and FRL4 in Table 2 for the variable light chain, the Amino acid sequences of triplet sequences 1 and 2 with SEQ ID NO. 1 to 13 correspond to the corresponding CDR regions of the antibodies.
- variable heavy (VH) or light (VL) antibody chain is composed as follows: the VH: FRH1-CDRH1-FRH2- CDRH2-FRH3-CDRH3-FRH4 and the VL: FRL1-CDRL1-FRL2-CDRL2-FRL3- CDRL3-FRL4.
- Table 2 explains the positions in detail. The positions of the individual amino acids or amino acid sequences correspond to the numbering of amino acids in antibody molecules according to Kabat.
- amino acid sequences SEQ ID NO. 46 to 79 correspond to amino acid sequences with preferred framework sequences for the variable heavy chain.
- amino acid sequences SEQ ID NO. 80 to 94 correspond to amino acid sequences with preferred framework sequences for the variable light chain.
- the core-1-specific recognition molecules can be present in different formats.
- the basic structure of the recognition molecule is one or more polypeptide chains, as described above triplet sequence 1 or triplet sequences 1 and 2 and framework sequences according to the invention.
- the amino acid sequence of the variable heavy chain with the skeleton sequences and the triplet sequences 1 and the amino acid sequence of the variable light chain with the skeleton sequences and the triplet sequences 2 are linked non-covalently or covalently to one another and can lie on one or more polypeptide chains.
- Several polypeptide chains can be covalently, for example by disulfide bridges, or non-covalently linked as a recognition molecule.
- recognition molecules according to the invention include, in particular, the linking of the triplet sequences with amino acid sequences that go beyond the framework sequences described above.
- recognition molecules according to the invention therefore comprise, in addition to the triplet sequences and the framework sequences, further additional sequences. Additional sequences are, in particular, amino acid sequences which primarily do not serve the spatial arrangement of the triplet sequences as in the form of the framework sequences, but which can advantageously influence them by secondary or tertiary interactions.
- additional sequences in the form of constant domains of an antibody stabilize the antibody and bring about dimerization, which leads to improved binding of the antibody, or, for example, fusion of an scFv with a domain of a bacteriophage coat protein causes an increase in the activity of the scFv binding, as described e.g. in Jensen KB et al. , 2002 is disclosed.
- the recognition molecules comprise amino acid sequences with framework sequences based on antibodies and, in addition to the triplet sequences, further ones Additional sequences.
- the additional sequences have at least one of the following tasks:
- Scaffold sequences for example to create or improve a binding ability
- tags for example multimerization sequences - for example ⁇ -tail sequence from IgM or association domain from p53 or MBL - to multimerize the core-1-binding portions for multivalent binding or to purify the recognition molecules, for example His-Tag or for detection, for example myc tag or for labeling or chelating recognition molecules, for example by lysine-rich sequences.
- tags for example multimerization sequences - for example ⁇ -tail sequence from IgM or association domain from p53 or MBL - to multimerize the core-1-binding portions for multivalent binding or to purify the recognition molecules, for example His-Tag or for detection, for example myc tag or for labeling or chelating recognition molecules, for example by lysine-rich sequences.
- Suitable structures are known to the person skilled in the art or can be derived from the prior art by logical conclusion.
- Further preferred embodiments are recognition molecules according to the invention, which comprise the following formats: single chain antibody fragment (scFv), Fv fragment, Fab fragment, F (ab) 2 fragment, multibody (slide, tria, tetrabody), immunoglobulin of the isotypes IgG, IgM, IgA, IgE, IgD or their subclasses, for example IgGl, or recognition molecules derived from immunoglobulins, which comprise at least one constant domain.
- scFv single chain antibody fragment
- Fv fragment fragment
- Fab fragment fragment
- F (ab) 2 fragment multibody (slide, tria, tetrabody)
- immunoglobulin of the isotypes IgG, IgM, IgA, IgE, IgD or their subclasses for example IgGl
- recognition molecules derived from immunoglobulins
- the recognition molecules according to the invention are composed of a heavy and a light polypeptide chain, the amino acid sequences of the heavy and light chain each comprising one of the triplet structures described above which represent the CDR regions of the antibody, the corresponding antibody framework sequences which form the framework sequences of the Represent antibodies, and additional sequences that comprise at least one of the constant domains of the antibody isotype.
- the two chains can form covalent bonds with one another.
- the constant regions and variable regions can contain sequences of antibodies from one or different species. Parts of constant domains or entire constant domains can be deleted or mutated, for example in order to change the effector function of the additional sequences, for example to prevent or improve the binding to Fc receptors.
- the recognition molecule is a murine, chimerized, humanized, partially human or human antibody or antibody fragment.
- the chimarization takes place, for example, by linking the variable antibody domains to constant antibody domains or fragments of the constant ones Domain of antibodies of different species. Sequences of the constant domains of human antibodies are preferred.
- the antibody framework sequences can be chosen so that the sequences are largely homologous to human antibody sequences.
- the choice for the species origin of the framework sequences also depends on the application. For therapeutic use in certain areas, the largest possible proportion of human framework sequences is preferred, especially when a human anti-mouse antibody response (HAMA) is to be avoided.
- HAMA human anti-mouse antibody response
- a xeno component is advantageous because it stimulates the immune system in an additional way.
- a combination of the two is particularly suitable in some cases, especially if a xeno component is advantageous in a primary immunization and a species-compatible and thus human component is advantageous in later applications.
- HuHI being preferred for the variable heavy chain
- HuKII being preferred for the variable light chain
- Homology to human germline sequences which are known to the person skilled in the art and are, for example, accessible via the V BASE database (www.mrc-cpe.cam.ac.uk) is particularly preferred.
- the triplet sequences which generally correspond to the binding loops (CDR regions) and which preferably have strong homologies, are added the corresponding sequence areas in the human germline sequence have been gradually adapted by simple mutations without impairing the specific binding to Core-1.
- Detection molecules with these sequences are referred to here as partially human antibodies or antibody fragments.
- Preferred humanized sequences are, for example, the sequences SEQ ID NO. 56 to 79 or SEQ ID NO. 85 to 94.
- certain amino acids of the antibody framework sequences of one species are exchanged for another in order to normally generate less immunogenic regions.
- immunoglobulins are composed of the heavy chain and the light chain of an antibody, 2 light chains and 2 heavy chains preferably being one unit. Immunoglobulins of the IgM type usually consist of 5 such units, which are linked to one another in addition to disulfide bridges by the J chain.
- the J chain is not present, which also leads to multimerization of the subunits, where hexa- and pentameric structures can be present.
- the recognition molecules there are single chain antibody fragments comprising a triplet structure 1 with corresponding antibody framework sequences described above, which represent the CDR regions of the antibody and framework sequences of the variable domain of the heavy chain of antibodies, and a triplet structure 2 with the corresponding antibodies described above
- Antibody framework sequences that represent the CDR regions of the antibody and framework sequences of the variable domain of the light chain of antibodies that are covalently linked to one another in the form of a fusion protein. The sequences are linked to one another directly or by a linker.
- scFv formats without a linker or with a linker of 1 to 9 amino acids in length.
- These scFv antibodies form multimeric structures (for example slide, tria, tetrabodies), which are also referred to as multibodies in the sense of the invention and, owing to the multivalence, show higher avidity to the core 1 antigen.
- Core-1 specific recognition molecules in scFv format with different linker lengths were constructed (SEQ ID NO. 95 to 106) and their binding characteristics were examined in an ELISA. A gradual link shortening led to an increase in binding to asialoglycophorin, a core-1-bearing glycoprotein, as shown in Figure 3. The variants with the SEQ ID NO showed the best binding properties.
- 104 and 105 These multivalent constructs in the slide / triabody format are particularly preferred embodiments of the invention and are advantageous for tumor therapy due to improved pharmacokinetic properties.
- the recognition molecules are fused, chemically coupled, covalently or non-covalently associated with (i) immunoglobulin domains of different species, (ii) enzyme molecules, (iii) Interaction domains, (iv) signal sequences, (v)
- Fluorescent dyes (vi) toxins, (vii) catalytic enzymes
- Antibodies (viii) one or more antibodies or
- Antibody fragments with different specificity (ix) cytolytic components, (x) immunomodulators, (xi)
- the recognition molecules can, in particular, be fused to a tag that enable the recognition molecule to be detected and purified, such as a Myc tag or a His tag. Technologies for producing these constructs are known to the person skilled in the art, and the person skilled in the art is also able to select suitable sequences and components and to connect them in a suitable manner with the recognition molecules according to the invention.
- the described recognition molecules based on antibodies or antibody fragments are fused with peptides or proteins that are not derived from immunoglobulins.
- the multimerization domain of a non-immunoglobulin molecule is fused to an scFv, in particular the C-terminal end of the alpha chain of the C4 binding protein, as described by Tonye Libyh M. et al. , 1997, and thus constructed a multivalent recognition molecule.
- an scFv is fused to a transmembrane domain of a non-immunoglobulin molecule, for example to the transmembrane domain of c-erb B2, h-PDGFR, the human transferrin receptor or the human asialoglycoprotein receptor (Liäo et al., 2000), and Consequently enables the expression of binding molecules on the surface of cells.
- a further preferred embodiment of the invention comprises recognition molecules according to the invention which further comprise amino acid sequences which bind specifically to macrophages or other immune effector cells.
- the recognition molecules according to the invention further comprise an antibody binding site against CD64, which causes macrophages to bind to core-1 positive tumor cells in the form of a bispecific antibody or antibody fragment (diabodies), which leads to their control and / or destruction.
- a preferred embodiment of the invention relates to radioactively labeled core-1-specific recognition molecules.
- a preferred form are recognition molecules based on antibodies or antibody fragments.
- Another preferred embodiment is radioactive labeled recognition molecules according to the invention in single chain format (including as slide, tria, tetrabodies).
- Further preferred forms are radioactively labeled single chain antibody fragments and whole immunoglobulins, for example chimeric or humanized IgG or IgM antibodies according to the invention or humanized antibody fragments.
- the invention is of course not restricted to these antibodies, the radioactive labeling and these formats of the antibodies.
- Antibody fragments such as the preferred multivalent scFv fragments, in particular without or with a very short linker, offer an advantage over intact monoclonal antibodies for targeting solid tumors. If the antibodies are intact and show a specific accumulation in the tumor area in biodistribution studies, the Tumor has an inhomogeneous antibody distribution with predominant enrichment in the marginal area. Due to tumor necrosis, inhomogeneous antigen distribution and an increased interstitial tissue pressure, these tumor constructs do not reach centrally located tumor parts. Smaller antibody fragments, on the other hand, show rapid tumor marking, penetrate deeper into the tumor and are removed from the bloodstream relatively quickly.
- multivalent antibody constructs such as multibodies (diabodies, tria / tetrabodies), F (ab ”) 2 and other minibodies (multivalent antibody constructs consisting of the binding domain and a multimerization sequence, for example scFv and CH3 domain of an IgG) offer many advantages in tumor therapy.
- Multivalent constructs in slide / triabody format are preferred embodiments of the invention and, because of their improved pharmacokinetic properties, are of advantage for tumor therapy and have been further developed for use in tumor therapy and can be used as vehicles for the specific enrichment of, for example, cytotoxic substances such as chemotherapeutic agents or radionuclides in the tumor
- cytotoxic substances such as chemotherapeutic agents or radionuclides in the tumor
- tumor cells can be killed over a distance of several cell diameters, as a result of which antigen-negative tumor cells are also detected in a tumor area and the poor penetration of the antibodies into solid tumors ren can be at least partially offset.
- a particularly preferred embodiment of the invention are radio-labeled multibodies - in particular as detailed in Example 9 - which combine particularly advantageous pharmacokinetic properties with one in the Combination compared to whole immunoglobulins and - scFv improved tumor retention, tumor penetration, serum half-life and serum to tumor distribution ratio. Further advantages are the high avidity and the bacterial expression, which makes it possible to produce these recognition molecules inexpensively.
- This particular format of the recognition molecules according to the invention is thus advantageously suitable preferably for the treatment of small primary tumors, metastases and minimally residual diseases.
- a preferred embodiment of the invention is non-radioactively labeled recognition molecules.
- a preferred form here are recognition molecules based on antibodies or antibody fragments.
- a particularly preferred embodiment is chimeras and humanized immunoglobulins based on IgM molecules for inhibiting liver metastasis and for fighting residual tumor cells.
- chimerized or humanized IgG and IgM-based recognition molecules coupled to toxins or cytostatics, and in particular multibodies (diatria, tetrabodies) with particularly advantageous pharmacokinetic properties as stated above.
- liposomes which are loaded, for example, with toxins or cytostatics and which carry recognition molecules according to the invention on their surface.
- toxins or cytostatics are loaded, for example, with toxins or cytostatics and which carry recognition molecules according to the invention on their surface.
- the person skilled in the art is able to select suitable radioisotopes, toxins and cytostatics. Suitable techniques, processes, dosages and formulations are known to the person skilled in the art.
- a further preferred embodiment of the invention are effector cells of the immune system, on the surface of which recognition molecules according to the invention are bound, which direct / address the effector cells to tumor cells carrying Core-1 and thereby mediate their fight and / or destruction.
- Preferred effector cells are macrophages, dendritic cells and NK cells, which are obtained from the patient and are coupled ex vivo to the recognition molecules. Cell lines of these cell types are further preferred. The coupling is carried out, for example, by bispecific recognition molecules which, in addition to the core-1-specific components, also comprise amino acids which mediate binding to the effector cells.
- these are bispecific antibodies, complement components or constant domains of antibodies.
- a further preferred embodiment here are macrophages from the patient which, after being obtained with a bispecific antibody, for example in the form of whole antibodies, preferably chemically coupled Fab fragments or more preferably diabodies, which on the one hand recognize CD64 and on the other hand are core-1 specific according to the invention.
- a bispecific antibody for example in the form of whole antibodies, preferably chemically coupled Fab fragments or more preferably diabodies, which on the one hand recognize CD64 and on the other hand are core-1 specific according to the invention.
- These macrophages, which carry the bispecific recognition molecules via the CD64 specificity are fed back to the patient in a suitable formulation in order to combat the Core-1 positive tumor.
- the techniques used for this and the suitable methods, dosages and formulations are known to the person skilled in the art.
- a further preferred embodiment are macrophages from the patient which, after being obtained with a Core-1 specific according to the invention Antibodies or antibody fragments which comprise the constant part of an antibody which binds to macrophages via the Fc receptors known per se.
- the recognition molecules can bind to the macrophages either as whole antibodies, preferably chimeras or humanized IgG or IgM, or as antibody fragments, for example scFv, Fab or multibodies in the form of a fusion protein or chemically coupled with the part of the constant domain of antibodies known to the person skilled in the art ,
- These macrophages carrying the recognition molecules are fed back to the patient in a suitable formulation in order to combat the Core-1 positive tumor.
- the techniques used for this and the suitable methods, dosages and formulations are known to the person skilled in the art.
- a further preferred embodiment are cell lines or cells from the body, such as the effector cells described above, which are transfected with molecules, the core-1-specific recognition molecules according to the invention and further comprise elements which bring about expression and anchoring in the membrane, for example a transmembrane domain, and mediate the activation of the effector cells upon contact with a core-1-bearing tumor cell.
- the corresponding elements are known to the person skilled in the art.
- a dendritic cell line is transfected with a vector which comprises a recognition molecule which comprises an scFv or multibody according to the invention and a transmembrane domain and an activating domain.
- macrophages are transfected virally.
- the invention also relates to nucleic acid molecules which comprise one or more genetic sequences which encode at least one of the recognition molecules and / or constructs according to the invention described above. Due to the degenerate genetic code, these nucleic acid molecules can have very different sequences. The choice of the codons also depends on the cell used for the production of the recognition molecule, since different codons are often preferred in different cells from different organisms and the expression rate can be strongly influenced, for example the codons used in eukaryotic genes AGA and AGG for arginine in bacteria rarely represented. Here the codons CGC and CGU occur much more frequently.
- the nucleic acid molecule according to the invention is a genomic DNA, a cDNA and / or an RNA. The criteria for the selection of suitable codons and the production of a suitable nucleic acid molecule are known to the person skilled in the art.
- the invention further relates to vectors for expressing the recognition molecules, in particular in cells.
- a vector is understood to mean a nucleic acid molecule according to the invention which is used for expression of the recognition molecule and which comprises a nucleic acid sequence which comprises one or more genetic sequences which encode at least one of the recognition molecules described above, and in particular comprises at least one promoter which Expression of the recognition molecule causes.
- Vectors can of course include further elements which are known to the person skilled in the art and which, for example, the multiplication of vectors for production in suitable cells and for Serve cloning.
- nucleic acid sequences can be present on one or more vectors, for example in a preferred embodiment the heavy chain of an immunoglobulin according to the invention is encoded by one and the light chain by another vector.
- the variable domain of the light chain and the variable domain of the heavy chain are encoded on the same vector under a promoter as a fusion protein.
- nucleic acid sequences which code parts of a recognition molecule can be expressed by different promoters known to the person skilled in the art.
- the different nucleic acid sequences can lie on a common vector.
- Each sequence can be expressed by its own, identical or different, promoter or the sequences can be present in a bicistronic vector under a promoter.
- Different expression rates of the parts of the recognition molecules are preferably achieved by the different promoters, which improves the formation of the entire recognition molecule compared to an identical expression rate of the different parts.
- promoters are preferably used which are inducible in order to improve expression of the recognition molecule.
- the vectors particularly preferably further comprise other regulatory elements known to the person skilled in the art, for example enhancers, which enhance the expression of the recognition molecule or parts thereof, for example the CMV enhancer or immunoglobulin enhancer sequences.
- the nucleic acid molecules and vectors preferably additionally comprise nucleic acid sequences which serve as signal sequences for secretion of the recognition molecule or parts thereof which are known per se to the person skilled in the art, for example PelB, OmpA or MalE for prokaryotic cell systems or the signal peptide of the T cell receptor, the Immunoglobulin chains, t-PA or EPO for eukaryotic cell systems [Boel et al., 2000; Herrera et al. , 2000].
- This advantageously facilitates cleaning and / or improves the yield of the recognition molecules.
- the methods for producing the nucleic acids and vectors described above, suitable promoters, enhancers and vector constructs and the criteria for their selection are known to the person skilled in the art and are explained in more detail in the examples.
- the vector according to the invention further comprises nucleic acid sequences which code for viral proteins.
- the virus itself is designated as a special form of a vector, the genetic material of which comprises a nucleic acid sequence which codes for a recognition molecule according to the invention.
- the recognition molecule is a fusion protein with a virus envelope protein or parts thereof, which enables not only the genetic material to comprise the nucleic acid sequence of the recognition molecule, but also the recognition molecule itself to be present in a binding manner on the surface of the virus, for example an scFv recognition molecule according to the invention as a fusion protein with a coat protein of adenoviruses, poxviruses or vaccinia viruses suitable for gene therapy applications.
- Known systems are preferably used, which use a helper virus for replication, in order to ensure, for example, the safety of a gene therapy method comprising this vector.
- the procedures for making the described Viral vectors for infection and expression of the recognition molecules are known to the person skilled in the art.
- the vector according to the invention comprises a fusion protein from a recognition molecule according to the invention and a protein or peptide that specifically binds to a virus.
- the recognition molecules obtained can thus advantageously be used to address the virus to a cell expressing Core-1.
- the transfer of the genetic material can be mediated via infections, thereby allowing expression of specific molecules which are encoded by the genetic material of the virus in the cells in vivo in the organism in the form of a gene therapy 'or in vitro in cell culture express.
- the invention relates to a method for obtaining the recognition molecules comprising the introduction of one or more vectors according to the invention, which contain one or more nucleic acid molecules according to the invention, into a suitable host cell, the cultivation of this host cell under suitable conditions and the provision of one or more recognition molecules from the Cells or the culture medium.
- introduction of vectors is understood to mean technologies known per se to the person skilled in the art with which the vector is brought into a host cell, for example electroporation, transfection using cationic lipids or infection and remains there transiently or stably.
- Provision of one or more recognition molecules is understood in the context of the invention to mean technologies known per se to those skilled in the art with which the recognition molecules expressed during the cultivation process are obtained from the culture supernatant * and / or cells, for example differently protein chemical purification steps such as fractionation, concentration, precipitation, and / or chromatography.
- the techniques and methods to be used in the method are known to the person skilled in the art, and the person skilled in the art is also able to select suitable host cells and cultivation conditions and methods for providing them from the cells and / or the culture supernatant.
- the person skilled in the art selects nucleic acid sequences with suitable codons and promoter sequences matched to the host cell in order to obtain the greatest possible expression of active recognition molecules.
- the person skilled in the art uses, for example, affinity chromatography steps, for example chromatography on protein A or protein G or protein L or, for example, metal ion affinity chromatography via an additionally inserted His tag. This is explained in more detail in the examples.
- extraction also includes additional steps, such as pretreatments of the starting material or further treatments of the end product.
- Pretreatment processes are known per se to the person skilled in the art.
- further treatment methods also include, for example, the final compositions and / or formulation of the recognition molecule obtained by the production method in suitable use and / or administration forms.
- the type of use or dosage form eg solution, lyophilisate or tablet, depends on the intended use. The person skilled in the art knows here which dosage form is suitable for which purpose.
- that produced by the method according to the invention can Detection molecule is present together with auxiliaries, carriers or other active ingredients.
- Auxiliary substances are preferably adjuvants, further active substances, preferably immunostimulatory molecules, such as interleukins.
- the recognition molecule produced with the method according to the invention can also be chemically modified in further processing steps.
- the recognition molecule is preferably linked to one or more other molecules in a suitable manner, ie by chemical or physical interaction.
- Other proteins in the sense of the invention are preferably other proteins or peptides which are covalently or non-covalently linked to the recognition molecule produced by the method according to the invention, for example in order to produce bispecific recognition molecules by using an inventive recognition molecule which specifically recognizes the core-1 antigen "linked to a second molecule " which, for example, specifically binds an immune effector cell (e.g.
- immune effectors are understood to mean those components of the invention which can directly or indirectly combat and / or destroy Core-1 positive tumor cells, for example immune effector cells such as macrophages, NK cells, Dendritic cells, or effector molecules, such as proteins or peptides of the complement system.
- Substances which have a therapeutic or diagnostic effect for example radioisotopes or toxins, are particularly suitable as further molecules in the process according to the invention. These substances are linked to the recognition molecules by methods known per se, for example radioisotopes are either incorporated directly (for example iodine) or bound via a covalently coupled chelator (for example yttrium, indium, bismuth). The steps of the further treatment process are known to the person skilled in the art.
- the cells used according to the invention for expressing the recognition molecules can be prokaryotic or eukaryotic cells, for example bacteria, yeast (preferably S. cerevisiae or P. pastoris), insect (D. melanogaster), plant, mammalian (preferably hamster, mouse) cells - or human cell lines) or organisms such as transgenic animals and plants.
- prokaryotic or eukaryotic cells for example bacteria, yeast (preferably S. cerevisiae or P. pastoris), insect (D. melanogaster), plant, mammalian (preferably hamster, mouse) cells - or human cell lines) or organisms such as transgenic animals and plants.
- the mammalian cell lines NS0, SP2 / 0, CHO-Kl, CHOdhfr-, COS-1, COS-7, HEK293, K562, Namalwa or Percy 6 used.
- the present invention relates to host cells which were produced by the method described above and with the aid of which recognition molecules according to the invention can be produced.
- the host cells can be part of a clone or represent it itself.
- the invention also relates to organisms which comprise host cells according to the invention. The techniques and methods to be used for producing these organisms are known to the person skilled in the art.
- the invention further relates to compositions for therapeutic, prophylactic or diagnostic purposes comprising at least one recognition molecule according to the invention in a suitable, in particular a pharmaceutically suitable form or composition.
- the pharmaceutical composition includes, in particular, additional substances and substances, for example medical and / or pharmaceutical-technical auxiliaries.
- compositions are both those pharmaceutical compositions which are used for therapeutic and prophylactic purposes and those pharmaceutical compositions which are used in vivo as a diagnostic agent.
- these are compositions for ex vivo diagnostics which can contain additional substances and substances. This embodiment is explained in more detail under the description for the diagnostics.
- Medical auxiliaries are those substances which are used for production as active ingredients of medicinal products.
- Pharmaceutical-technical auxiliary substances serve for the suitable formulation of the medicinal product or the * pharmaceutical composition and can even, if they are only required during the manufacturing process, be subsequently removed or may be part of the pharmaceutical composition as pharmaceutically acceptable carriers, examples of pharmaceutically acceptable carriers are below listed.
- the pharmaceutical formulation or formulation of the pharmaceutical composition is optionally carried out in combination with a pharmaceutically acceptable carrier and / or diluent.
- suitable pharmaceutically acceptable carriers include, for example, phosphate-buffered saline solutions, water, emulsions such as, for example, oil / water emulsions, various types of detergents, sterile solutions, etc.
- Medicaments or pharmaceutical compositions which comprise such carriers be formulated using known conventional methods. These medicaments or pharmaceutical compositions can be administered to an individual in a suitable dose, for example in a range from I ⁇ g to 10 g of recognition molecules per day and patient. Doses of 1 mg to 1 g are preferred.
- the administration can be carried out in various ways, for example intravenously, intraperitoneally, intrarectally, intragastrointestinally, intranodally, intramuscularly, locally, for example in the tumor, but also subcutaneously, intradermally or on the skin or via the mucous membranes.
- Nucleic acids can also be administered in the form of gene therapy, for example via viral vectors described above.
- the type of dosage and route of administration can be determined by the attending physician according to the clinical factors. It is known to the person skilled in the art that the type of dosage depends on various factors, such as, for example, the size, the body surface, the age, the sex or the general health of the patient, but also on the special agent which is administered, the duration and mode of administration and other drugs that may be administered in parallel.
- the pharmaceutical compositions or the medicament in particular comprises a pharmacological substance which is a or more or more recognition molecules according to the invention and / or nucleic acid molecules encoding them in a suitable solution or administration form.
- a pharmacological substance which is a or more or more recognition molecules according to the invention and / or nucleic acid molecules encoding them in a suitable solution or administration form.
- adjuvants for example QS-21, GPI-0100 or other saponins, water-oil emulsions such as, for example, Montanide adjuvants, polylysine, polyarginine compounds, DNA Compounds such as, for example, CpG, detox, bacterial vaccines such as, for example, thyroid vaccine or BCG vaccine, and / or another suitable substance for strengthening the action are administered; preferably immunostimulatory molecules, such as interleukins, for example IL-2, IL-12, IL-4 and / or growth actuators, for example GM-CSF.
- immunostimulatory molecules such
- the pharmaceutical composition or the medicament can of course also be a combination of 2 or more of the pharmaceutical compositions or medicaments according to the invention, as well as a combination with other medicaments, tumor vaccines or tumor treatments, such as, for example, antibody therapies, chemotherapy or radiotherapy, which together in a suitable manner in a timely manner or administered or applied separately.
- the pharmaceuticals or pharmaceutical compositions are prepared by methods known per se.
- the medicaments or pharmaceutical compositions can be used in particular for the treatment of core-1 positive Tumor diseases are used, such as breast cancer, cervical cancer, ovarian cancer, colon cancer, gastrointestinal cancer, pancreatic cancer, lung cancer, prostate cancer. These tumor diseases can also include core-1 and / or core-2 positive tumor diseases.
- the treatment is for example against primary tumors, minimal residual tumor diseases, relapses and / or metastases.
- the tumors can also be treated as adjunctive treatment.
- the drugs can also be used for the prophylaxis of core-1 positive tumor diseases. Prophylactic use is aimed, for example, at preventing the tumor and metastases.
- the tumor agents are administered in a suitable form according to known methods.
- a preferred variant is the injection or administration of the drugs intravenously, locally in body cavities, for example intraperitoneally, intrarectally, intragastrointestinally, locally, for example directly into the tumor, organs or lymphatic vessels (intranodally), but also subcutaneously, intradermally or on the skin, intramuscularly.
- Methods of administration can preferably also be combined, it being possible to administer them on different treatment days or on one treatment day.
- 2 or more of the pharmaceuticals or pharmaceutical compositions according to the invention can also be combined, or one or more pharmaceuticals according to the invention with one or more pharmaceuticals or tumor treatments, such as antibody therapies, chemotherapy or radiotherapy, which are administered or applied at the same time or separately.
- the present invention also relates to a method for producing a medicament or a pharmaceutical composition comprising the steps of producing Recognition molecules and further comprising the step of formulating the recognition molecules according to the invention in pharmaceutically acceptable form.
- the preferred recognition molecules according to the invention for this purpose are described in more detail above as embodiments for the treatment of tumor diseases and prophylaxis, and further below under in vivo diagnostics.
- the recognition molecules according to the invention and substances and compositions produced by the method according to the invention can accordingly preferably be used for the prophylaxis, diagnosis, follow-up and / or treatment of tumor diseases.
- the cancer or the tumor which is treated or prevented is selected from the group of cancers or tumors of the ear, nose and throat area, the lungs, the mediastinum, the gastrointestinal tract, the urogenital system, the gynecological System, breast, endocrine system, skin, bone and soft tissue sarcomas, mesotheliomas, melanomas, neoplasms of the central nervous system, cancer or childhood cancer, lymphomas, leukaemias, paraneoplastic syndromes, metastases with no known primary tumor (CUP syndrome), peritoneal Carcinoma mastosis, immunosuppression-related malignancies and / or tumor metastases.
- CUP syndrome primary tumor
- peritoneal Carcinoma mastosis immunosuppression-related malignancies and / or tumor metastases.
- the tumors can be the following types of cancer: adenocarcinoma of the breast, prostate and of the large intestine; all forms of lung cancer that originate from the bronchi; Bone marrow cancer, melanoma, hepatoma, neuroblastoma; the papilloma; the apudome, the choristome, the branchchioma; the malignant carcinoid syndrome; the carcinoid heart disease; the carcinoma (e.g.
- Walker carcinoma basal cell carcinoma, basosquamous carcinoma, Brown Pearce carcinoma, ductal carcinoma, Ehrlich tumor, in situ carcinoma, cancer 2 carcinoma, Merkel cell carcinoma, mucus cancer, non-small cell cancer Bronchial carcinoma, oat cell carcinoma, papillary carcinoma, scirrhous carcinoma, bronchioloalveolar carcinoma, bronchial carcinoma,
- Leukemia e.g. in connection with B-cell leukemia, mixed-cell leukemia, zero-cell leukemia, T-cell leukemia, chronic T-cell leukemia, HTLV-II-associated leukemia, acute lymphocytic leukemia, chronic-lymphocytic leukemia , Mast line leukemia and myeloid leukemia
- malignant histiocytosis Hodgkin's disease, non-Hodgkin's lymphoma, solitary plasma cell tumor; Reticuloendotheliosis, chondroblastoma; Chondroma, chondrosarcoma; fibroma; fibrosarcoma; Giant cell tumors; histiocytoma; lipoma; liposarcoma; Leukosarkom; mesothelioma; myxoma; myxosarcoma; osteo
- the central nervous system the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system, the central nervous system and the central nervous system, the central nervous system, the central nervous system and the central nervous system, the central nervous system, the central nervous system and the central nervous system, the central nervous system, the central nervous system and the central nervous system, the central nervous system, the central nervous system and the central nervous system, the central nervous system, the central nervous system and the central nervous system,
- the cancer or the tumor which is treated or prevented is selected from the group of cancerous diseases or tumorous diseases which comprise cells which comprise the Core-1 in the definition according to the invention, selected from the group: tumors of the ear, nose and throat area including tumors of the inner nose, paranasal sinuses, nasopharynx, lips, oral cavity, oropharynx, larynx, hypopharynx, ear, salivary glands and paragangliomas, tumors of the lung including non-small cell Bronchial carcinomas, small-cell bronchial carcinomas, tumors of the mediastinum, tumors of the gastrointestinal tract including tumors of the esophagus, stomach, pancreas, liver, gallbladder and biliary tract, small intestine, colon and rectal cancer and kidney cancer, including urogenital cancer, including urogenital cancer, and urogenital cancer, including urogenital cancer, including urogenital cancer, including urogenital cancer, and
- the cancer or the tumor which is treated or prevented is selected from the group comprising cancer or tumor diseases of breast cancer, gastrointestinal cancer, including colon cancer, gastric cancer, pancreatic cancer, colon cancer, Small bowel cancer, ovarian cancer, cervical cancer, lung cancer, prostate cancer, renal cell carcinoma and / or liver metastases.
- effector structures are understood to mean those chemical or biochemical compounds, molecules or atoms which directly or indirectly kill or damage, including, for example, growth retardation or growth inhibition of tumor cells.
- effector molecules include, for example, radioisotopes, toxins, cytostatics and other effector molecules such as cytokines and Chemokines or other structures which themselves are effectors or to which effector molecules are coupled, for example liposomes loaded with toxins or cytostatics, which carry recognition molecules according to the invention.
- the liposomes in particular also those effector structures are meant which, in addition to the recognition molecule for the tumor specificity, also include those Carrying molecules that are responsible for the absorption of the effector structures or parts thereof in the cells, such as antibodies against receptors that have a prescription or-mediated endocytosis.
- the recognition molecules preferably comprise a transmembrane domain which allows them to be inserted into the liposome membrane or, in another preferred embodiment, the recognition molecules are chemically coupled to the liposome surface. The techniques used for this are known to the person skilled in the art, including the production. the liposomes.
- the recognition molecules are also connected to the other effector structures by methods known per se.
- the couplings can as already stated above, for example, directly by covalent or non-covalent loading, by chemical coupling, an additional chemical or biological molecule may be necessary, for example a chelator or a linker, or in the form of fusion proteins or peptides by fusion.
- the recognition molecules are used in the treatment of tumor diseases with tumors carrying Core-1, and / or for a subset of recognition molecules according to the invention, which are described above about their specificity for Core-1 and Core-2, Core-2 and / or Core -1 bearing tumor cells or for prophylaxis, which prevents, for example, the formation of primary tumors or metastases.
- the preferred goal is the treatment of minimal residual disease and metastases.
- Another preferred application is the inhibition of liver metastasis of core-1 and / or core-2 positive tumor cells.
- the recognition molecules according to the invention are administered once or repeatedly in a suitable formulation at suitable time intervals and doses.
- the core-1 antigen also means core-1 and / or core-2 and core-1 positive cells or tumor cells and / or tissues also core-1 and / or core-2 positive cells or tumor cells and / or tissues.
- the radioactive recognition molecules according to the invention described above are combined with an application of unmarked core-1-specific recognition molecules according to the invention.
- This serves to improve the background and thus a more specific binding to the tumor by saturating potential Core-1-bearing molecules in the blood become.
- IgM-derived recognition molecules are preferably used, for example the clgM described in the examples or the humanized form thereof, since they bind to core-1 antigen in the blood in particular and thus reduce the background and the serum load with radioactivity and increase the relative tumor targeting, while penetration into tissues and tumors is limited due to the size of the molecules.
- the methods and technologies used for this are known to the person skilled in the art, and the person skilled in the art can also prepare a suitable dose, formulations, application route and time of administration of the unmarked recognition molecules.
- the invention further relates to methods using the recognition molecules according to the invention, which make it possible to identify and / or obtain core-1-bearing molecules from a large pool of different molecules, which molecules can be used advantageously for use in tumor treatment, tumor prophylaxis and tumor diagnosis.
- core-1-bearing molecules are understood to mean molecules which carry core-1 and / or core-2 structures and are specifically bound by the recognition molecules according to the invention.
- core-1-bearing molecules are glycoproteins, glycopeptides and / or glycolipids and also cells or other carrier substances, such as viruses, bacteria, parts of cells, such as exosomes or cell lysates, or liposomes, which contain one or more core-1 structures.
- the molecules carrying Core-1 can be obtained from cells or cell lines, from culture supernatants, from tumor tissue, Tumor cells or body fluids such as blood, blood serum, lymph, urine, spinal fluid or sperm are enriched or isolated.
- molecules bearing core-1 are identified and / or isolated and obtained by binding to the above-described core-1-specific recognition molecules.
- the above-described core-1-bearing molecules can be obtained from body fluids or from supernatants from cell cultures by affinity chromatography. Further purification and / or concentration steps can be combined with one or more affinity chromatography steps according to methods known per se.
- Tumor-associated core-1-bearing molecules can also be obtained from tumor cells, tumor tissues or tumor cell lines by connecting a suitable step according to methods known per se, which makes it possible to make the line-associated core-1-bearing molecules accessible for affinity purification, for example by solubilization with suitable detergents or by cleavage by proteolysis or by cell lysis.
- core-1-carrying molecules or cells are obtained from tissues.
- the tissue is disrupted according to methods known per se in order to make the molecules or cells carrying the Core 1 accessible, for example by proteolytic or mechanical methods. These methods are known to the person skilled in the art.
- core-1 positive cells or cell lines are also isolated or enriched using the core-1-specific recognition molecules "and separated from those cells which carry no or small amounts of core-1 structures.
- isolation or enrichment of the cells means all measures for the separation of cells that have formed a complex with the recognition molecules according to the invention by carrying core-1 structures.
- Enrichment takes place by binding recognition molecules according to the invention to the core-1 structure on the cell surface and then selecting the cells labeled in this way by binding to carrier materials which specifically interact with the recognition molecule, for example anti-mouse IgM antibodies coupled to magnetic beads (MACS sorting)
- carrier materials which specifically interact with the recognition molecule
- the core-1-specific recognition molecules can themselves be covalently coupled to a support.
- FACS sorter which sorts cells that carry the recognition molecules that have been fluorescence-labeled. Both methods are known to the person skilled in the art.
- These core-1 positive cells enriched in this way can be used for the production of vaccines, for example for loading dendritic cells or directly as tumor cell lysate in a vaccine composition.
- the previous enrichment of Core-1 positive cells is said to lead to a higher tumor specificity of the vaccination.
- a diagnostic agent comprising the steps of the method according to the invention for producing the core-1-specific recognition molecules according to the invention and furthermore comprising the step of formulating the recognition molecules in a form that can be used diagnostically
- the term “diagnostic agent” defines substances and preparations made of substances which are intended to identify diseases, conditions, body damage or pathological complaints by application to or in the human body or parts thereof.
- Parts of the human body are preferably to be understood as body fluids, such as blood, blood serum, lymph, urine, spinal fluid or sperm, or tissue biopsies or samples.
- the formulation of the diagnostic agent preferably also includes the modification of the recognition molecules produced with substances which detect the core-1 antigen, in certain embodiments which depend on the fine specificity of the recognition molecule according to the invention. allow the antigen Core-2.
- Suitable substances are known in the prior art. Based on the choice of the substance, the person skilled in the art is able to use suitable measures for formulating the diagnostic agent.
- substances according to the invention can also be coupled to the recognition molecules according to methods known per se, which facilitate detection of the core 1 antigens and / or their carrier molecules and / or cells, for example by biotinylation, fluorescence labeling, radioactive labeling or enzyme coupling of the recognition molecules ,
- tumor diagnosis and prognosis recognition molecules which recognize core-1 antigens and / or their carrier molecules in human serum.
- the determination is preferably carried out qualitatively, quantitatively and / or in temporally relative quantities by methods known per se.
- the same methods are also used according to the invention for monitoring the course of tumor diseases and for monitoring treatment courses, including monitoring immune responses and for monitoring and dosing tumor treatments.
- the methods used in the methods are known per se, for example ELISA, Westerblot, FACS (fluorescence-activated cell sorting), MACS (magnet-mediated cell sorting), ADCC (antibody-mediated cell cytotoxicity), CDC (complement-mediated cytotoxicity), immunocytochemistry and immunohistochemistry.
- core-1-specific recognition molecules are used in methods known per se in order to detect the antigen core-1 in serum or in tissue preparations.
- the antigen Core-1 is detected on carrier molecules, core-1 present in immune complexes on carrier molecules and / or core-1 bound to cells and the presence of the core-1 antigen and / or the core-1 carrying molecules is qualitative, quantitative and / or determined in relative quantities by methods known per se.
- the same methods are also used for monitoring the course of tumor diseases and for checking the course of treatment.
- the methods used in the methods are known per se, for example ELISA, Western blot, FACS (fluorescence-activated cell sorting), MACS (magnet-mediated cell sorting), ADCC (antibody-mediated cell cytotoxicity), CDC (complement-mediated cytotoxicity), immunocytochemistry and immunohistochemistry.
- a preferred embodiment is a rapid tissue test in which the tissue samples are stained with fluorescence-labeled recognition molecules according to the invention in an immunohistological method.
- the recognition molecule according to the invention preferably an antibody of the IgM isotype, is combined with a further antibody which specifically recognizes the antigen MUC1, preferably isotype IgGl.
- the antibodies and recognition molecules are directly labeled with different fluorescent dyes, for example Cy3 and Cy5 or Cy3 and FITC.
- the antibodies and / or recognition molecules are amplified by labeled secondary antibodies or the biotin streptavidin. It is advantageous to use different isotypes and / or species sequences in the constant part of the antibodies.
- the technologies and methods used here, for example labeling and immunohistology, and the choice of the suitable formats of the recognition molecules are known to the person skilled in the art.
- the diagnostic procedure described is not limited to gastrointestinal tumors, but can be used for all tumor diseases carrying the antigen Core-1.
- a serological test is carried out using a sandwich ELISA as the method. This consists of a capture antibody, which binds carrier molecules of the core 1 antigen from the serum to a solid phase, and a detection antibody, which, according to the invention, also includes other recognition molecules according to the invention which recognize the core 1 antigen. This makes it possible to differentiate which carrier molecule carries the Core-1. In a preferred form, conclusions can be drawn as to the origin of the primary tumor.
- Various antibodies can serve as catcher antibodies. Recognize glycoproteins that carry O-glycosylations.
- a preferred embodiment uses antibodies against the epithelial mucin MUCl as a capture antibody, which is often a carrier of the Core-1 in the tumor case.
- all antigens in the blood that carry the Core-1 antigen are determined. This is possible because the Core-1 antigen usually occurs in several copies per carrier molecule.
- a core-1-specific recognition molecule according to the invention is used as the capture antibody and a labeled core-1-specific recognition molecule according to the invention as detection antibody, the recognition molecules not having to be antibodies.
- an IgM is used as a recognition molecule at least as a catcher or detection antibody.
- the detection antibody is labeled with biotin and the system is detected via streptavidin in combination with a suitable detection method.
- a suitable detection method is, for example, POD labels or fluorescent labels of streptavidin.
- the determination of the core-1 antigen is combined with the determination of other serological tumor markers, for example PSA, CEA or AFP.
- a preferred embodiment is the determination of the MUCl and the Core-1 antigen.
- the MUCl is immobilized from the serum on a solid phase with the aid of an MUCl-specific antibody and is detected as a detection antibody with a second anti-MUCl-specific antibody, preferably those which recognize the DTR region in a glycosylated form and the core-1 antigen was detected on the MUCl immobilized with the aid of an anti-MUCl capture antibody using a recognition molecule according to the invention.
- This diagnostic test combines early detection with a prognostic statement about the course of the disease and / or the likelihood of liver metastasis.
- the technologies used here for example labeling and serology, including the detection methods, are known to the person skilled in the art.
- the diagnostic methods described are not restricted to gastrointestinal tumors, but can be used for all tumors carrying the antigen Core-1.
- the serological tests described serve to diagnose, monitor the course of the tumor disease and to forecast Core-1 antigen-positive tumors.
- the core 1-specific recognition molecules according to the invention are used for in vivo diagnostics.
- the recognition molecules are marked with suitable methods known per se and thus made accessible for known imaging methods in humans, for example radioimmuno diagnostics, PET scanning methods or immunofluorescence endoscopy, for example by coupling and / or Loading with corresponding molecules, for example radioactive isotopes, for example indium, or fluorescent dyes, for example Cy3, Cy2, Cy5 or FITC.
- multibodies according to the invention are covalently coupled with a suitable chelator (for example DOTA or DTPA) and loaded with indium-111 and used for in vivo diagnosis.
- a suitable chelator for example DOTA or DTPA
- these are administered intravenously in a dose suitable for the individual and the location of the core-1 antigen and a potential tumor are measured according to methods known per se.
- the methods and technologies used for this, including the imaging methods, are known to the person skilled in the art, and the person skilled in the art can also prepare a suitable dose and formulations.
- immunoglobulins preferably IgM and IgG, as described above and described in more detail in the examples, are radioactively labeled, for example with indium-111, and blood vessels supplying or disposing locally to the tumor or to the tumor. In one embodiment, this serves to determine the size of the tumor and in a further embodiment to determine affected lymph nodes.
- the methods and technologies used for this are known to the person skilled in the art, and the person skilled in the art can also prepare a suitable dose and formulations.
- the radioactive-labeled recognition molecules according to the invention are also administered via other application routes.
- Preferred routes are intraperitoneal, intranodal or intrarectal or intragastrointestinal. Intraperitoneally is particularly advantageous for the determination of tumors that pass through the Perito ⁇ eum are accessible and / or metastasize in this, for example ovarian carcinomas and certain gastrointestinal carcinomas. Intrarectal or intragastrointestinal administration is advantageous for certain gastrointestinal tumors and their location and size determination. In certain cases, intranodal can be used to directly infiltrate individual lymph nodes.
- radioactive recognition molecules according to the invention described above are specific for in vivo diagnostics with an application of unmarked Core-1 according to the invention
- IgM-derived recognition molecules are preferably used, since these are primarily on core-1
- recognition molecules according to the invention are labeled with a fluorescent dye and administered in vivo.
- Preferred application routes here are intrarectal, intragastrointestinal, intraperitoneal, intravenous and into supplying or discharging blood vessels.
- a particularly preferred embodiment serves for the localization of gastrointestinal carcinomas, which is carried out by fluorescence endoscopy after application of the fluorescence-labeled recognition molecules.
- a recognition molecule according to the invention is combined with at least one antibody against a further tumor antigen, preferably anti-MUCl antibody.
- fluorescent dyes are preferably used, which allow a differentiation of the recognition molecules and antibodies, with which a prognostic statement is combined with early detection and a larger number of cases.
- Preferred fluorescent dyes are those with low background fluorescence, which are known to the person skilled in the art.
- the methods and technologies used for this purpose, including the imaging methods, for example fluorescence endoscopy, are known to the person skilled in the art, and the person skilled in the art can also prepare a suitable dose, formulations, route of application and time of administration of the unlabelled detection molecules.
- the invention has several advantages:
- the "core-1-specific recognition molecules according to the invention specifically recognize types of carcinoma, as a result of which they can advantageously be used for diagnosis and / or therapy in many tumor patients with various indications.
- the recognition molecules advantageously advantageously do not bind to normal tissues This is a particular advantage over the known tumor markers and an outstanding property of the recognition molecules according to the invention.
- the recognition molecules recognize the Core-1 antigen carrier independently.
- a particular advantage of the recognition molecules according to the invention is the high specificity for the tumor tissue is due in particular to the high specificity for definite carbohydrate antigens, since the non-specific recognition of other carbohydrate structures would increase the risk of non-specific recognition of Increase non-tumor tissue.
- the recognition molecules according to the invention have a high affinity. This in particular gives the possibility of constructing less valued fragments, such as IgG and multibodies. The possibility of these different formats is advantageous for the development of therapeutic agents.
- the core-1 and / or core-2 structure on the cell surface increases the probability of the formation of metastases, for example liver metastases; by blocking the Core-1 and / or Gore-2 structure with recognition molecules, the formation of metastases is reduced or inhibited.
- Multibodies with the sequences SEQ ID NO. 96 to 106 were obtained by shortening or deleting the linker between the V H and the V L of the single chain antibody with the sequence SEQ ID NO. 95 formed (Fig.la).
- the V H and the V L were amplified with specific primers in such a way that 22 nucleotides form a complementary region at the 3 "end of the V H and at the 5" end of the V L (Fig. Lb, PCR I and PCR II) , and then linked the two PCR fragments together after purification in a SOE-PCR (Fig. lb, PCR III).
- the PCR fragment was cloned into a prokaryotic expression vector via Ncol / Notl.
- This vector contains the lacZ promoter, a ribosome binding site (RBS), the M13 origin, the pelB signal sequence for secretion into the periplasm, an ampicillin resistance gene and a cloning cassette to be used to couple the C-terminal end of the scFv with a hexa-histidine tag for efficient purification and a c-myc tag (Fig. 2).
- the antibody fragments from Example 1 were expressed and purified in Escherichia coli.
- the corresponding plasmid was transformed into electrocompetent E. coli by electroporation and cultured overnight in 2xTY medium (10 g yeast extract, 16 g tryptone, 5 g NaCl per L) with 100 ⁇ g / mL ampicillin.
- This culture was diluted 1: 100 with 2xTY medium, to which 100 ⁇ g / ml ampicillin and 0.5% glucose was added, and incubated at 37 ° C. until an OD 60 o nm of approximately 0.6 was reached. Then 1 mM IPTG was added to the culture for induction and this was incubated at 25 ° C. for a further 5 h.
- the bacteria were harvested by centrifugation at 4000xg for 20 min, the cell pellet was resuspended in TES buffer (30 mM Tris-HCl, pH 8.0, 20% sucrose, 1 mM EDTA) and incubated on ice for 20 min. Then 5 mM MgSO 4 were added and the suspension was incubated on ice for a further 20 min. The periplasmic fraction was then obtained by centrifugation at 4000 ⁇ g for 60 min and dialyzed overnight at 4 ° C. against binding buffer (50 M phosphate buffer, pH 8.0, 300 mM NaCl, 10 mM imidazole).
- binding buffer 50 M phosphate buffer, pH 8.0, 300 mM NaCl, 10 mM imidazole.
- the antibody fragments contained in the periplasmic fraction were purified using the C-terminal His tag by metal ion affinity chromatography (HiTrap Chelating HP, Amersham Pharmacia Biotech). For this purpose, the dialyzed fraction was placed on the column previously equilibrated with binding buffer and the non-binding proteins were washed off the column with washing buffer (50 mM phosphate buffer, pH 8.0, 300 mM NaCl, 30 mM imidazole). Then the antibody fragments eluted with elution buffer (50 mM phosphate buffer, pH 8.0, 300 mM NaCl, 300 mM imidazole). This purification protocol was used for all Core-1-specific antibody fragments with a hexahistidine tag, for example the humanized single chain antibodies from Example 6.
- Asialoglycophorm was used as the antigen for the ELISA test
- the Ncol / Xhol DNA fragment from the scFv vector which codes for the V H was cloned into the Ncol / Sall-cut BS leader vector.
- the BS leader vector contains a cloning cassette for the introduction of the T cell receptor signal peptide sequence at the 5 "end and a splice donor
- FIG. 4 The V L sequence of the corresponding antibody was amplified with specific primers for the introduction of the Ncol site at the 5 "end and the Nhel site at the 3" end in the PCR using the scFv sequence as a template and after Ncol / Nhel digestion cloned into the same digested BS leader vector. Then the HindIII / BamHI fragment from the BS leader vector was cloned into the corresponding eukaryotic expression vector.
- vectors contain the EF-l ⁇ promoter and the HCMV enhancer, the SV40-origin, the BGH polyadenylation signal, the puromycin resistance gene in the vector for the heavy chain and the neomycin resistance gene or the dehydrofolate reductase gene in the vector for the light chain and the genomic sequences of the human constant ⁇ l region or ⁇ region for the heavy chain or the human constant K region for the light chain (primer for amplification from genomic human DNA and vector map see fig. 4).
- CHOdhfr cells ATCC No. CRL-9096
- electroporation 10 6 cells / ml, 500 V, 50 ⁇ s
- co-transfected in selection medium CHO-S-SFM II medium (Life Technologies)
- HT supplement biochrom
- the stably transfected CHO cells secreting the chimeric IgG or IgM were cultivated in spinner bottles in CHO-S-SFM II medium, supplemented by HT supplement, until a cell density of approximately 1 ⁇ 10 6 cells / ml was reached. After the cells had been separated from the cell culture supernatant by centrifugation (400 ⁇ g, 15 min), the chimeric antibody was purified using a Protein A column (HiTrap rProtein A FF, Amersham Pharmacia Biotech) for the chimeric IgG or an anti-human Fc5 ⁇ antibody affinity column.
- a Protein A column HiTrap rProtein A FF, Amersham Pharmacia Biotech
- the purified antibody fraction eluted by pH jump was buffered and concentrated in PBS using Centriprep centrifuge tubes (cut off 50 kDa, Millipore). 6. Alignment of the sequence of the core-1-specific antibody sequences with human germline sequences
- core-1 binding antibody sequences For the adaptation of the core-1 binding antibody sequences to human sequences, homologous sequences were searched in the database of human germline sequences, and humanized core-1 binding sequences were developed using the human consensus sequences and the knowledge of the canonical structure of human antibodies.
- the human germ line sequence VH1-46 served as template for the variable heavy chain, and the sequence A18 for the variable light chain.
- the humanized V H or V L sequences SEQ ID NO. 56 to 79 and 85 to 94 were produced using the gene assembly PCR (single overlap extension PCR).
- the PCR reaction was carried out according to the following scheme: first denaturation 94 ° C for 2 min, then 30 cycles denaturation at 94 ° C for 45 sec, annealing at 55 ° C for 45 sec and elongation at 73 ° C for 1.5 min and on End an elongation step at 73 ° C for 7 min.
- V H and V L chains thus produced were cut with the enzymes Ncol and Xhol or Notl and Xhol and cloned into a cloning vector (pLitmus 28 or pBluescript KS) for sequencing.
- the correct V H and V L chains were then amplified again in order to insert a Bbsl interface at the 3 "end of the V H and at the 5" end of the V L , in order to cover the V H and the V L with only one alanine Linker link.
- the complete scFv (the ligation products) were amplified using the flanking primers and cloned into a bacterial expression vector. 7.
- a dilution of 5 ⁇ g / ml in PBS was prepared from the respective stock solutions (1 mg in 1 ml Bi-dest. H 2 0), which are stored in portions at -20 ° C. 50 ⁇ l / well thereof were pipetted into a microtiter plate (NUNCLON-TC Microwell 96 F) and the test plate was incubated for 1 h at 37 ° C. and overnight at 4 ° C. The next day the test plate was washed 3 times with PBS / 0.2% Tween.
- a color reaction with TMB (3, 3 ", 5, 5" - tetramethylbenzidine) was carried out. After 15 minutes, the reaction was stopped by adding 2.5N H 2 S0. The measurement was carried out with a microtiter plate photometer with a 50nm filter in dual mode against a reference filter 630nm.
- Figure 5 compares two recognition molecules with varying loop sequences in IgM format.
- the antibody constructs mIgM-Karo2 (SEQ ID NO. 107 and SEQ ID NO. 109) and mIgM-Karo4 (SEQ ID NO. 108 and SEQ ID NO. 110) bind highly specifically to the antigen Core-1, preferably to the alpha-anomer Galßl-3GalNAc ⁇ and weaker to the beta anomer Galßl-3GalNAcß.
- the recognition molecules according to the invention can also bind only the alpha anomer Galßl-3GalNAc ⁇ or both anomers Galßl-3GalNAc ⁇ and Galßl-3GalNAcß in the same way.
- mIgM-Karo4 binds the core 2 structure Galßl-3 (GlcNAcßl-6) GalNAc ⁇ . All other carbohydrate structures tested, including structurally strongly related structures, are not recognized by the binding proteins claimed here.
- AGP shows a strong signal with both variants, whereby the also core-1-bearing glycoprotein asialofetuin reacts much more strongly with the Karo2 variant, which is very likely related to the different core-1 density in both proteins.
- Figure 6 shows the specificity pattern of the exemplary humanized recognition molecules Karoll (SEQ ID NO. 56 and SEQ ID NO. 90), Karo21 (SEQ ID NO. 59 and SEQ ID NO.
- the exemplary recognition molecule mlgM-Karo4 according to the invention only reacts with very few structures in normal tissue. However, these are in areas that are not accessible to an antibody (Table 3).
- the claimed recognition molecules react positively with a variety of carcinomas.
- the data in Table 4 show that the core-1 specific recognition molecules have a large size Identify the percentage of tumor patients in an indication that differs from indication to indication.
- FIG. 8 shows an example of an immunohistochemical staining of a xenograft preparation with the core-1-specific antibody cIgG-Karo4.
- anti-mouse or anti-human IgG or IgM for whole antibodies
- anti-Myc-Tag or anti-His-Tag antibodies for single chain
- Figure 9 shows an example of fluorescent labeling of KG-1 cells, an acute myeloid leukemia cell line, with different antibody constructs, a murine IgM and two scFv antibodies with different linker lengths (SEQ ID NO. 95 with 18 amino acids and SEQ ID NO. 104 with an amino acid as a linker). All three constructs show a specific staining of the tumor cell line, the monovalent antibody fragment SEQ ID NO. 95 shows the weakest signal.
- the antibody cIgG-Karo4 or the multibody with the sequence SEQ ID NO. 104 a chelator covalently bound, which enables the binding of a radiometal.
- the conjugation took place by reaction of the isothiocyanate group of the chelator with a free ⁇ -amino group of the amino acid lysine on the antibody. A covalent N-C bond is created between the chelator and the antibody.
- the purified antibody or the • purified antibody fragment must first be buffered in pH 8.7 coupling buffer.
- ultrafiltration was carried out in a filtration sleeve (Centriprep YM50 (Amicon)). This was done by repeated dilution with a 10-fold volume and filtration through a membrane with a defined pore size by centrifugation. This replaced PBS with the alkaline coupling buffer (0.05 M sodium carbonate, 0.15 M sodium chloride, pH 8.7).
- the chelation was carried out with the bifunctional chelators p-SCN-Bz-DTPA or p-SCN-Bz-DOTA.
- protein (1-10 mg / ml) in coupling buffer and a solution of the chelator of 1 mg / ml in 2% DMSO / water were mixed in such a way that a molar excess of the chelator was ensured.
- the mixture was then incubated at 37 ° C. for 1 h.
- the unbound chelator was then separated by ultrafiltration in the same vessel (Centriprep YM50 (Amicon)) and, as described above, buffered to pH 4.2 in the loading buffer required for radioactive labeling (0.15 M sodium acetate, 0.15 M sodium chloride, pH 4, 2).
- the protein concentration during and after this step was reset to 1-10 mg / ml by means of a UV measurement at 280 nm. Conditions for the chelation reaction were found which allowed the antibody to be radiolabeled without significantly reducing its bioactivity.
- the chelated antibody was loaded with a radio metal, thereby generating the radio antibody.
- the isotopes 11: L indium and 90 yttrium were used for loading. Both have chemically and physicochemically comparable properties and are bound by the chelator as trivalent ions ( 11: L In 3+ , 90 ⁇ 3+ ).
- Antibody is a ⁇ -emitter and is used in the clinic for individual dose determination for the patient, while 90 yttrium is a ß-emitter that is used therapeutically.
- the half-lives are 1: L1 in 67 hours and 90 Y 64 hours.
- L1 indium chloride from NEN (Perkin Elmer, Belgium) was used for loading.
- the radio metal is supplied in a hydrochloric acid solution.
- This 11: L InCl3 solution was briefly brought to an HCl concentration of IM.
- the mixture was then diluted with 0.05M HCl to a specific activity of 80-320mCi / ml and an aliquot thereof was used for incorporation into the chelated antibody, the added volume of HCl acidic 1: L1 InCl 3 solution being equal to the volume of the antibody solution presented should be pH 4.2 in the coupling buffer to ensure pH stability. Incubation was 1 hour at 37 ° C with occasional gentle mixing.
- the filter insert was then reinserted into the filtration sleeve and, as described above, buffered in phosphate buffer pH 7.2 with a physiological content of sodium chloride. A separation of high molecular weight took place radiolabeled antibody and unbound 1: InCl3. The incorporation of 11: L into the chelated antibody was quantified by thin layer chromatography. The incorporation rate of the radio metal was 70-99% of the radioactivity used.
- Core-1 positive, secretory MUCl can be detected in the sandwich ELISA.
- a MUCl-specific antibody serves as a capture antibody of MUCl and a Core-1-specific antibody for the detection of the Core-1 antigen.
- a third enzyme or fluorescence-coupled antibody must be used to detect the second antibody.
- the supernatants of two tumor cell lines were analyzed as an example (K562 and T47D). The results are shown in Table 6. 10 5 cells per ml of cell culture medium were sown, cultivated for 4 days without changing the medium, then an aliquot was removed and the cell culture supernatant was separated from the cell pellet by centrifugation. 50 ⁇ l of these supernatants were used undiluted in the ELISA.
- the anti-MUCl-anti-Core-1 sandwich ELISA was carried out by coating the microtiter plate with the capture antibody (1 ⁇ g / ml) in PBS at 4 ° C. overnight.
- the neuraminidase treatment was carried out in the wells provided.
- the neuraminidase solution (DADE Behring, Germany) in immidazole buffer (0.68 g immidazole, 0.19 g CaCl 2 and 0.4 g NaCl in 100 ml H 2 O, pH 6.8) was 1: 5 diluted and 50 ul / well incubated at 37 ° C for 30 min.
- the imidazole buffer was incubated in an appropriate well without neuraminidase solution.
- the wells were then washed three times and the mlgM-Karo4 antibody for the detection of the core-1 antigen was added in a 1: 500 dilution in 2% BSA in washing buffer and incubated for a further hour at room temperature.
- a peroxidase-coupled anti-mouse IgM ( ⁇ ) antibody (Dianova) was diluted 1: 5000 in 2% BSA in washing buffer and incubated for 1 h at room temperature. Finally, the plates were washed twice in wash buffer and once in PBS.
- the dyeing reaction was carried out in 25 mM citric acid, phosphate buffer pH 5.0 with 0.04% H 2 0 2 and 0.4 mg / ml o-phenylenediamine (Sigma) in the dark at room temperature.
- the color reaction was stopped by adding 2.5 N sulfuric acid (final concentration 0.07 N) and measured in an ELISA reader at 492 nm with a reference filter of 620 nm.
- the core-1 positive tumor cell line NM-D4 [DSMZ deposit NO. DSM ACC2605] (see Table 5) were used to test the binding ability of the radiolabeled recognition molecules to Core-1 positive tumor cells. A certain number of cells were placed in a 1.5 ml tube in duplicate determinations and incubated with increasing amounts of antibody. After washing, the count rate was used to determine how much antibody had bound.
- Table 7 summarizes the association constants and the number of cell binding sites of different Core-1-specific multibodies on NM-D4 cells.
- Table 7 Cell binding test and Scatchard analysis with 1: L1 unmarked recognition molecules on NM-D4 cells.
- ZR-75-1 cells were injected subcutaneously into nude mice (Ncr: nu / nu, female). After about 3-4 weeks the tumor is palpable under the skin.
- Table 8 shows the specific high Enrichment of the multibodies in the tumor (in ⁇ ID / g tumor based on the injected dose and the tumor weight) compared to the serum and the organs.
- the therapy studies were carried out using the same established ZR-75-1 tumor model as described for the biodistribution studies (see example 12).
- the chelated recognition molecules (see Example 9) were loaded with 90 yttrium (a .beta.-emitter to destroy the tumor cells) (pH 4.5, 37.degree. C., 30 min; compare incorporation of 1: L1 indium) and in thin-layer chromatography Stability controlled.
- the tumor-bearing mice (about three weeks after subcutaneous injection of the ZR-75-1 cells) were given 200 ⁇ l iv in the tail vein.
- the solution for injection contained the 90 Y-labeled multibody (up to a maximum of 100 ⁇ Ci per dose) in Ca / Mg-PBS with 0.2 to 4% fetal calf serum to protect against radiolysis.
- Control groups received the same injection without a radiolabeled recognition molecule.
- Body weight and tumor size were measured and compared twice a week. The relative tumor growth was determined taking into account the respective tumor size at the beginning of the treatment.
- a second injection was injected three weeks after the first treatment. By suitable. Treatment significantly reduced tumor growth compared to the control group.
- Fig. La Sequences of the linkers in the different multibody single chain antibody fragments.
- Fig. Lb Cloning scheme for the production of single chain antibody fragments with different linker lengths.
- Fig. 2 Vector for cloning and bacterial expression of single chain antibody fragments.
- Fig. 3 Analysis of multibodies in scFv format with different linker lengths in the ELISA.
- Multibodies with the amino acid sequences SEQ ID NO. 95, 96, 97, 98, 99, 100, 101, 103, 104 and 105 were expressed in E. coli as described above and the periplasm fractions were obtained. Asialoglykophorm, was used as the antigen for the ELISA test Core-1-bearing glycoprotein used. A gradual shortening of the linker leads to an increase in binding to the asialoglycophorm. The variants with the SEQ ID NO show the best binding properties. 104 and 105. These multivalent dia / triabody format constructs are preferred embodiments of the invention.
- Fig. 4 Vector system for cloning and eukaryotic expression of chimeric antibodies in IgGl or IgM format.
- Fig. 5 and 6 Specificity analysis in the ELISA.
- PAA [6]; Galßl-3GalNAcßl-OC 3 H 6 NH-PAA [7]; Gal ⁇ l-3GalNAcßl-
- Galßl-3 (Neu5Ac ⁇ 2-6) GalNAc ⁇ l-OC 3 H 6 NH-PAA [12]; GlcNAcßl-
- Figure 6 shows the specificity pattern of three humanized
- Fig. 7 Specific binding of various preferred formats and combinations of recognition molecules according to the invention in the ELISA, for example to the Antigens AGP, GP and / or Core-1-PAA (Galßl-3GalNAcocl-OC 3 H 6 NH-PAA).
- Fig. 8 Immunohistochemical staining of xenograft preparations.
- Human colon carcinoma tissue was transplanted to nude mice and passed through after reaching a certain size.
- the tumor tissue was embedded and cut and used for immunohistochemical staining.
- the tissue was labeled with cIgG-Karo4 as the primary antibody and coupled with an anti-human Fc ⁇ antibody - POD as the secondary antibody.
- the brown color indicates the Core 1 positive structures.
- Fig. 9 Fluorescent labeling of cells of the tumor cell line KG-1 with different core-1 specific recognition molecules.
- Fig. 10 Scatchard diagram for the analysis of cell binding of radioactively labeled Core-1-specific recognition molecules.
- the binding data of the multibody SEQ ID NO are exemplary here.
- 104 with a linker length of one amino acid (Prl and Pr2 correspond to two different preparations).
- F free binding as the difference between the total amount and the bound amount of antibody [M]. The corresponding one is above
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Cell Biology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Physics & Mathematics (AREA)
- Oncology (AREA)
- Optics & Photonics (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Compounds Of Unknown Constitution (AREA)
- Polyesters Or Polycarbonates (AREA)
Abstract
Description
Claims
Priority Applications (8)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/536,834 US8088357B2 (en) | 2002-11-29 | 2003-12-01 | Tumor-specific recognition molecules |
AU2003293270A AU2003293270A1 (en) | 2002-11-29 | 2003-12-01 | Tumor-specific recognition molecules |
SI200331372T SI1572747T1 (sl) | 2002-11-29 | 2003-12-01 | Molekule za prepoznavanje specifičnih tumorjev |
DE50310081T DE50310081D1 (de) | 2002-11-29 | 2003-12-01 | Tumorspezifische erkennungsmoleküle |
DK03788853T DK1572747T3 (da) | 2002-11-29 | 2003-12-01 | Tumorspecifikke genkendelsesmolekyler |
EP03788853A EP1572747B1 (de) | 2002-11-29 | 2003-12-01 | Tumorspezifische erkennungsmoleküle |
US13/302,698 US8617846B2 (en) | 2002-11-29 | 2011-11-22 | Tumor-specific recognition molecules |
US14/102,214 US9345794B2 (en) | 2002-11-29 | 2013-12-10 | Tumor-specific recognition molecules |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE10256900A DE10256900A1 (de) | 2002-11-29 | 2002-11-29 | Tumorspezifische Erkennungsmoleküle |
DE10256900.2 | 2002-11-29 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/536,834 A-371-Of-International US8088357B2 (en) | 2002-11-29 | 2003-12-01 | Tumor-specific recognition molecules |
US13/302,698 Continuation US8617846B2 (en) | 2002-11-29 | 2011-11-22 | Tumor-specific recognition molecules |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2004050707A2 true WO2004050707A2 (de) | 2004-06-17 |
WO2004050707A3 WO2004050707A3 (de) | 2004-09-23 |
Family
ID=32336015
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/DE2003/003994 WO2004050707A2 (de) | 2002-11-29 | 2003-12-01 | Tumorspezifische erkennungsmoleküle |
Country Status (11)
Country | Link |
---|---|
US (3) | US8088357B2 (de) |
EP (3) | EP1572747B1 (de) |
AT (2) | ATE555132T1 (de) |
AU (1) | AU2003293270A1 (de) |
CY (3) | CY1108389T1 (de) |
DE (2) | DE10256900A1 (de) |
DK (3) | DK1572747T3 (de) |
ES (3) | ES2523761T3 (de) |
PT (3) | PT1572747E (de) |
SI (3) | SI1572747T1 (de) |
WO (1) | WO2004050707A2 (de) |
Cited By (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1937815A1 (de) * | 2005-09-13 | 2008-07-02 | National Research Council of Canada | Verfahren und zusammensetzungen zur modulation der tumorzellaktivität |
US7772375B2 (en) * | 2005-12-12 | 2010-08-10 | Ac Immune S.A. | Monoclonal antibodies that recognize epitopes of amyloid-beta |
EP2347769A1 (de) * | 2010-01-20 | 2011-07-27 | Glycotope GmbH | Krebsstammzellenmarker und Verwendungen damit |
EP2428223A2 (de) | 2006-09-10 | 2012-03-14 | Glycotope GmbH | Verwendung von menschlichen Zellen von Myeloischer Leukämie zur Expression von Antikörpern |
WO2013026887A1 (en) | 2011-08-22 | 2013-02-28 | Glycotope Gmbh | Microorganisms carrying a tumor antigen |
US8592165B2 (en) | 2006-11-10 | 2013-11-26 | Glycotope Gmbh | Carbohydrate specific cellular immunity inducing microorganisms and fractions thereof |
US8796439B2 (en) | 2006-07-14 | 2014-08-05 | Ac Immune S.A. | Nucleic acid molecules encoding a humanized antibody |
US8802826B2 (en) | 2009-11-24 | 2014-08-12 | Alethia Biotherapeutics Inc. | Anti-clusterin antibodies and antigen binding fragments and their use to reduce tumor volume |
US9062101B2 (en) | 2010-08-14 | 2015-06-23 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
US9175094B2 (en) | 2007-06-12 | 2015-11-03 | Ac Immune S.A. | Monoclonal antibody |
US9822171B2 (en) | 2010-04-15 | 2017-11-21 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
US9822170B2 (en) | 2012-02-22 | 2017-11-21 | Alethia Biotherapeutics Inc. | Co-use of a clusterin inhibitor with an EGFR inhibitor to treat cancer |
US9951125B2 (en) | 2006-11-30 | 2018-04-24 | Abbvie Inc. | Aβ conformer selective anti-Aβ globulomer monoclonal antibodies |
WO2018178046A1 (en) | 2017-03-29 | 2018-10-04 | Glycotope Gmbh | Humanized anti-cd40 antibodies |
US10208109B2 (en) | 2005-11-30 | 2019-02-19 | Abbvie Inc. | Monoclonal antibodies against amyloid beta protein and uses thereof |
US10464976B2 (en) | 2003-01-31 | 2019-11-05 | AbbVie Deutschland GmbH & Co. KG | Amyloid β(1-42) oligomers, derivatives thereof and antibodies thereto, methods of preparation thereof and use thereof |
CN110684740A (zh) * | 2019-08-09 | 2020-01-14 | 无锡傲锐东源生物科技有限公司 | 一种抗人泛素羧基末端水解酶-1(uch-l1)的单克隆抗体及其应用 |
US11872289B2 (en) | 2018-05-18 | 2024-01-16 | Daiichi Sankyo Co., Ltd. | Anti-MUC1 antibody-drug conjugate |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7658924B2 (en) * | 2001-10-11 | 2010-02-09 | Amgen Inc. | Angiopoietin-2 specific binding agents |
CN101432302A (zh) | 2005-11-30 | 2009-05-13 | 艾博特公司 | 抗-Aβ球聚体抗体,其抗原结合部分,相应的杂交瘤、核酸、载体、宿主细胞,生产所述抗体的方法,包含所述抗体的组合物,所述抗体的应用以及使用所述抗体的方法 |
EP2124952A2 (de) | 2007-02-27 | 2009-12-02 | Abbott GmbH & Co. KG | Verfahren zur behandlung von amyloidosen |
US8048420B2 (en) | 2007-06-12 | 2011-11-01 | Ac Immune S.A. | Monoclonal antibody |
CA2701793C (en) | 2007-10-05 | 2017-04-25 | Genentech, Inc. | Use of anti-amyloid beta antibody in ocular diseases |
JO2913B1 (en) | 2008-02-20 | 2015-09-15 | امجين إنك, | Antibodies directed towards angiopoietin-1 and angiopoietin-2 proteins and their uses |
SG187173A1 (en) | 2010-07-30 | 2013-02-28 | Ac Immune Sa | Safe and functional humanized anti beta-amyloid antibody |
US8987421B2 (en) | 2013-02-15 | 2015-03-24 | Immunomedics, Inc. | Chimeric and humanized anti-histone antibodies |
EP3157950A4 (de) * | 2014-06-20 | 2018-01-10 | Stephen D. Gillies | Influenza-impfstoffe und verfahren zur verwendung davon |
WO2015200260A1 (en) | 2014-06-24 | 2015-12-30 | Immunomedics, Inc. | Anti-histone therapy for vascular necrosis in severe glomerulonephritis |
US10351621B2 (en) | 2014-06-24 | 2019-07-16 | Immunomedics, Inc. | Anti-histone therapy in acute kidney injury |
WO2019191563A1 (en) * | 2018-03-29 | 2019-10-03 | Remd Biotherapeutics, Inc. | Treatment of autoimmune and inflammatory disorders using antibodies that bind interleukin-17a (il-17a) |
EP3887386A4 (de) * | 2018-11-26 | 2022-11-30 | North Carolina State University | Peptidliganden zur einfangen von wirtszellproteinen |
WO2021191865A1 (en) * | 2020-03-26 | 2021-09-30 | Dusa Pharmaceuticals, Inc. | Management of dermal neurofibromatosis lesions |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE4329004A1 (de) * | 1993-08-28 | 1995-03-09 | Max Delbrueck Centrum | Monoklonale Antikörper gegen das Thomsen-Friedenreich-Antigen, ihre Herstellung und ihre Verwendung zum Tumornachweis |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5683674A (en) * | 1987-01-07 | 1997-11-04 | Imperial Cancer Research Technology Ltd. | Antibody against human mucin core protein and method of preparing and using same |
DE69329643T2 (de) * | 1992-04-13 | 2001-03-01 | Dana-Farber Cancer Institute, Inc. | Gegen karzinom-assoziierte antigene gerichtete antikörper |
US5804187A (en) * | 1992-11-16 | 1998-09-08 | Cancer Research Fund Of Contra Costa | Modified antibodies with human milk fat globule specificity |
US5739277A (en) * | 1995-04-14 | 1998-04-14 | Genentech Inc. | Altered polypeptides with increased half-life |
US5902582A (en) * | 1995-09-05 | 1999-05-11 | Chiron Corporation | Use of TFPI inhibitor for treatment of cancer |
JP2003519096A (ja) | 1999-08-18 | 2003-06-17 | アルタレックス コーポレーション | Muc−1抗原に対する治療用抗体およびその使用方法 |
US7147850B2 (en) * | 1999-08-18 | 2006-12-12 | Altarex Medical Corp. | Therapeutic binding agents against MUC-1 antigen and methods for their use |
GB0029360D0 (en) | 2000-12-01 | 2001-01-17 | Univ Nottingham | Humanised antibodies and uses thereof |
DE10303664A1 (de) | 2003-01-23 | 2004-08-12 | Nemod Immuntherapie Ag | Erkennungsmoleküle zur Behandlung und Detektion von Tumoren |
-
2002
- 2002-11-29 DE DE10256900A patent/DE10256900A1/de not_active Withdrawn
-
2003
- 2003-12-01 ES ES10012760.4T patent/ES2523761T3/es not_active Expired - Lifetime
- 2003-12-01 SI SI200331372T patent/SI1572747T1/sl unknown
- 2003-12-01 EP EP03788853A patent/EP1572747B1/de not_active Expired - Lifetime
- 2003-12-01 DE DE50310081T patent/DE50310081D1/de not_active Expired - Lifetime
- 2003-12-01 AT AT08158547T patent/ATE555132T1/de active
- 2003-12-01 ES ES03788853T patent/ES2312842T3/es not_active Expired - Lifetime
- 2003-12-01 AT AT03788853T patent/ATE399797T1/de active
- 2003-12-01 PT PT03788853T patent/PT1572747E/pt unknown
- 2003-12-01 PT PT08158547T patent/PT1975183E/pt unknown
- 2003-12-01 PT PT100127604T patent/PT2386576E/pt unknown
- 2003-12-01 DK DK03788853T patent/DK1572747T3/da active
- 2003-12-01 WO PCT/DE2003/003994 patent/WO2004050707A2/de active IP Right Grant
- 2003-12-01 DK DK08158547.3T patent/DK1975183T3/da active
- 2003-12-01 DK DK10012760.4T patent/DK2386576T3/da active
- 2003-12-01 US US10/536,834 patent/US8088357B2/en not_active Expired - Fee Related
- 2003-12-01 SI SI200332172T patent/SI1975183T1/sl unknown
- 2003-12-01 AU AU2003293270A patent/AU2003293270A1/en not_active Abandoned
- 2003-12-01 SI SI200332394T patent/SI2386576T1/sl unknown
- 2003-12-01 EP EP08158547A patent/EP1975183B1/de not_active Expired - Lifetime
- 2003-12-01 ES ES08158547T patent/ES2387072T3/es not_active Expired - Lifetime
- 2003-12-01 EP EP10012760.4A patent/EP2386576B1/de not_active Expired - Lifetime
-
2008
- 2008-10-02 CY CY20081101089T patent/CY1108389T1/el unknown
-
2011
- 2011-11-22 US US13/302,698 patent/US8617846B2/en not_active Expired - Lifetime
-
2012
- 2012-06-27 CY CY20121100569T patent/CY1112940T1/el unknown
-
2013
- 2013-12-10 US US14/102,214 patent/US9345794B2/en not_active Expired - Fee Related
-
2014
- 2014-11-10 CY CY20141100933T patent/CY1115859T1/el unknown
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE4329004A1 (de) * | 1993-08-28 | 1995-03-09 | Max Delbrueck Centrum | Monoklonale Antikörper gegen das Thomsen-Friedenreich-Antigen, ihre Herstellung und ihre Verwendung zum Tumornachweis |
Non-Patent Citations (4)
Title |
---|
JESCHKE U ET AL: "Expression of the Thomsen-Friedenreich antigen and of its putative carrier protein mucin 1 in the human placenta and in trophoblast cells in vitro" HISTOCHEMISTRY AND CELL BIOLOGY, Bd. 117, Nr. 3, März 2002 (2002-03), Seiten 219-226, XP002290192 ISSN: 0948-6143 * |
KARSTEN UWE ET AL: "A New Monoclonal Antibody (A78-G/A7) to the Thomsen-Friedenreich Pan-Tumor Antigen" HYBRIDOMA, Bd. 14, Nr. 1, 1995, Seiten 37-44, XP009034408 ISSN: 0272-457X * |
PRICE M R ET AL: "SUMMARY REPORT ON THE ISOBM TD-4 WORKSHOP: ANALYSIS OF 56 MONOCLONAL ANTIBODIES AGAINST THE MUC1 MUCIN" TUMOR BIOLOGY, KARGER, BASEL, CH, Bd. 19, Nr. SUPPL 1, 1998, Seiten 1-20, XP002071245 ISSN: 1010-4283 * |
SCHNEIDER F ET AL: "Overexpression of sialyltransferase CMP-sialic acid:Galbeta1,3GalNAc-R alpha6-Sialyltransferase is related to poor patient survival in human colorectal carcinomas" CANCER RESEARCH, AMERICAN ASSOCIATION FOR CANCER RESEARCH, BALTIMORE, MD, US, Bd. 61, Nr. 11, 1. Juni 2001 (2001-06-01), Seiten 4605-4611, XP002232470 ISSN: 0008-5472 * |
Cited By (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10464976B2 (en) | 2003-01-31 | 2019-11-05 | AbbVie Deutschland GmbH & Co. KG | Amyloid β(1-42) oligomers, derivatives thereof and antibodies thereto, methods of preparation thereof and use thereof |
EP1937815A1 (de) * | 2005-09-13 | 2008-07-02 | National Research Council of Canada | Verfahren und zusammensetzungen zur modulation der tumorzellaktivität |
US8426562B2 (en) | 2005-09-13 | 2013-04-23 | National Research Council Of Canada | Methods and compositions for modulating tumor cell activity |
US8044179B2 (en) | 2005-09-13 | 2011-10-25 | National Research Council Of Canada | Methods and compositions for modulating tumor cell activity |
EP1937815A4 (de) * | 2005-09-13 | 2010-03-17 | Ca Nat Research Council | Verfahren und zusammensetzungen zur modulation der tumorzellaktivität |
US10208109B2 (en) | 2005-11-30 | 2019-02-19 | Abbvie Inc. | Monoclonal antibodies against amyloid beta protein and uses thereof |
US10323084B2 (en) | 2005-11-30 | 2019-06-18 | Abbvie Inc. | Monoclonal antibodies against amyloid beta protein and uses thereof |
US7772375B2 (en) * | 2005-12-12 | 2010-08-10 | Ac Immune S.A. | Monoclonal antibodies that recognize epitopes of amyloid-beta |
US8796439B2 (en) | 2006-07-14 | 2014-08-05 | Ac Immune S.A. | Nucleic acid molecules encoding a humanized antibody |
EP2428223A2 (de) | 2006-09-10 | 2012-03-14 | Glycotope GmbH | Verwendung von menschlichen Zellen von Myeloischer Leukämie zur Expression von Antikörpern |
EP2428224A2 (de) | 2006-09-10 | 2012-03-14 | Glycotope GmbH | Verwendung von menschlichen Zellen von Myeloischer Leukämie zur Expression von Antikörpern |
US10280230B2 (en) | 2006-09-10 | 2019-05-07 | Glycotope Gmbh | Use of human cells of myeloid leukemia origin for expression of antibodies |
EP2428225A2 (de) | 2006-09-10 | 2012-03-14 | Glycotope GmbH | Verwendung von menschlichen Zellen von Myeloischer Leukämie zur Expression von Antikörpern |
US9051356B2 (en) | 2006-09-10 | 2015-06-09 | Glycotope Gmbh | Use of human cells of myeloid leukaemia origin for expression of antibodies |
EP3539981A1 (de) | 2006-09-10 | 2019-09-18 | Glycotope GmbH | Verwendung von menschlichen zellen myeloischen leukämieursprungs zur expression von antikörpern |
US9494587B2 (en) | 2006-11-10 | 2016-11-15 | Glycotope Gmbh | Microorganisms or fractions thereof capable of activating cellular immunity against carbohydrates |
US8592165B2 (en) | 2006-11-10 | 2013-11-26 | Glycotope Gmbh | Carbohydrate specific cellular immunity inducing microorganisms and fractions thereof |
US9951125B2 (en) | 2006-11-30 | 2018-04-24 | Abbvie Inc. | Aβ conformer selective anti-Aβ globulomer monoclonal antibodies |
US9585956B2 (en) | 2007-06-12 | 2017-03-07 | Ac Immune S.A. | Polynucleotides encoding anti-amyloid beta monoclonal antibodies |
US9175094B2 (en) | 2007-06-12 | 2015-11-03 | Ac Immune S.A. | Monoclonal antibody |
US9512211B2 (en) | 2009-11-24 | 2016-12-06 | Alethia Biotherapeutics Inc. | Anti-clusterin antibodies and antigen binding fragments and their use to reduce tumor volume |
US8802826B2 (en) | 2009-11-24 | 2014-08-12 | Alethia Biotherapeutics Inc. | Anti-clusterin antibodies and antigen binding fragments and their use to reduce tumor volume |
EP2347769A1 (de) * | 2010-01-20 | 2011-07-27 | Glycotope GmbH | Krebsstammzellenmarker und Verwendungen damit |
US9822171B2 (en) | 2010-04-15 | 2017-11-21 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
US9062101B2 (en) | 2010-08-14 | 2015-06-23 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
US10047121B2 (en) | 2010-08-14 | 2018-08-14 | AbbVie Deutschland GmbH & Co. KG | Amyloid-beta binding proteins |
WO2013026887A1 (en) | 2011-08-22 | 2013-02-28 | Glycotope Gmbh | Microorganisms carrying a tumor antigen |
US9700610B2 (en) | 2011-08-22 | 2017-07-11 | Glycotope Gmbh | Microorganisms carrying a tumor antigen |
US9822170B2 (en) | 2012-02-22 | 2017-11-21 | Alethia Biotherapeutics Inc. | Co-use of a clusterin inhibitor with an EGFR inhibitor to treat cancer |
WO2018178046A1 (en) | 2017-03-29 | 2018-10-04 | Glycotope Gmbh | Humanized anti-cd40 antibodies |
US11872289B2 (en) | 2018-05-18 | 2024-01-16 | Daiichi Sankyo Co., Ltd. | Anti-MUC1 antibody-drug conjugate |
CN110684740A (zh) * | 2019-08-09 | 2020-01-14 | 无锡傲锐东源生物科技有限公司 | 一种抗人泛素羧基末端水解酶-1(uch-l1)的单克隆抗体及其应用 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1975183B1 (de) | Tumorspezifische Erkennungsmoleküle | |
EP1585770B1 (de) | Erkennungsmoleküle zur behandlung und detektion von tumoren | |
DE69734109T2 (de) | Humanisierung von anti-carcinoembryonalen Antigen anti-idiotypischen Antikörper und dessen Verwendung als Tumorvakzin und zur Markierung | |
DE69333807T2 (de) | Marker für krebs und biosynthetisches bindeprotein dafür | |
RU2673908C2 (ru) | Мини-антитела j591 и цис-диатела для направленной доставки простата-специфичного мембранного антигена (psma) человека и способы их применения | |
DE69222956T2 (de) | Kleinste erkennungseinheit eines pem-mucin-"tandem repeat"-spezifischen monoklonalen antikoerpers | |
EP1478667B1 (de) | Monoklonaler anti-mensch-tenascin-Antikörper | |
EP0699756B1 (de) | BR96 Mutantantikörper die mit menschlichen Karzinomen reagieren | |
DE69927268T2 (de) | Spezifische antikörper enthaltende bindungsproteine die das nekrotische tumorzentrum binden und deren verwendung | |
DE69331735T2 (de) | Gegen das A33 Antigen gerichtete Humanisierte Antikörper | |
DE69020182T2 (de) | Vernetzte Antikörper und Verfahren zu ihrer Herstellung. | |
DE69332930T2 (de) | Peptide mit breiter neoplastischer spezifizität | |
EP0585570B1 (de) | Granulozytenbindende Antikörperkonstrukte, ihre Herstellung und Verwendung | |
DE69334036T2 (de) | Hybridom und humanisierter momoklonaler Anti-KC-4-Antikörper, dafür kodierende DNA und RNA, Kit und diagnostische Verfahren | |
EP1090927A1 (de) | Polypeptide (scFv) zur Detektion und Elimination CA19-9 antigen positiver Zellen | |
EP2193147B1 (de) | Nukleotid- und proteinsequenzen eines gegen ein humanen sauren und basischen ferritinen eigenen epitop gerichteten antikörpers, monoklonale antikörper oder antikörperähnliche moleküle, die diese sequenzen enthalten, und deren verwendung | |
WO2001080904A2 (de) | Mittel zur diagnose und therapie von karzinomen |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): BW GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2003788853 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 2003788853 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2006251668 Country of ref document: US Ref document number: 10536834 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: JP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: JP |
|
WWP | Wipo information: published in national office |
Ref document number: 10536834 Country of ref document: US |
|
WWG | Wipo information: grant in national office |
Ref document number: 2003788853 Country of ref document: EP |