US20230295344A1 - Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress - Google Patents
Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress Download PDFInfo
- Publication number
- US20230295344A1 US20230295344A1 US17/984,611 US202217984611A US2023295344A1 US 20230295344 A1 US20230295344 A1 US 20230295344A1 US 202217984611 A US202217984611 A US 202217984611A US 2023295344 A1 US2023295344 A1 US 2023295344A1
- Authority
- US
- United States
- Prior art keywords
- dpp3
- binder
- disease
- seq
- oxidative stress
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000011230 binding agent Substances 0.000 title claims abstract description 202
- 230000036542 oxidative stress Effects 0.000 title claims abstract description 174
- 230000027455 binding Effects 0.000 title claims abstract description 154
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 121
- 201000010099 disease Diseases 0.000 title claims abstract description 118
- 230000001154 acute effect Effects 0.000 title claims abstract description 95
- 238000011282 treatment Methods 0.000 title claims abstract description 61
- 230000002265 prevention Effects 0.000 title abstract description 40
- 101150036343 Dpp3 gene Proteins 0.000 title 1
- 101000931862 Homo sapiens Dipeptidyl peptidase 3 Proteins 0.000 claims abstract description 335
- 102100020750 Dipeptidyl peptidase 3 Human genes 0.000 claims abstract description 329
- 238000000034 method Methods 0.000 claims abstract description 54
- 150000001413 amino acids Chemical class 0.000 claims abstract description 42
- 206010028980 Neoplasm Diseases 0.000 claims description 47
- 210000004027 cell Anatomy 0.000 claims description 47
- 201000011510 cancer Diseases 0.000 claims description 38
- 230000004054 inflammatory process Effects 0.000 claims description 34
- 208000020832 chronic kidney disease Diseases 0.000 claims description 32
- 239000012634 fragment Substances 0.000 claims description 32
- 206010061218 Inflammation Diseases 0.000 claims description 31
- 208000018737 Parkinson disease Diseases 0.000 claims description 30
- 208000023105 Huntington disease Diseases 0.000 claims description 25
- 208000009304 Acute Kidney Injury Diseases 0.000 claims description 23
- 208000024827 Alzheimer disease Diseases 0.000 claims description 23
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 23
- 208000033626 Renal failure acute Diseases 0.000 claims description 23
- 201000011040 acute kidney failure Diseases 0.000 claims description 23
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 claims description 23
- 206010040070 Septic Shock Diseases 0.000 claims description 21
- 230000002757 inflammatory effect Effects 0.000 claims description 21
- 201000006417 multiple sclerosis Diseases 0.000 claims description 21
- 230000036303 septic shock Effects 0.000 claims description 21
- 206010019280 Heart failures Diseases 0.000 claims description 20
- 206010040047 Sepsis Diseases 0.000 claims description 20
- 230000004770 neurodegeneration Effects 0.000 claims description 20
- 208000015122 neurodegenerative disease Diseases 0.000 claims description 20
- 208000023275 Autoimmune disease Diseases 0.000 claims description 19
- 208000001145 Metabolic Syndrome Diseases 0.000 claims description 19
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 claims description 19
- 239000000427 antigen Substances 0.000 claims description 19
- 108091007433 antigens Proteins 0.000 claims description 19
- 102000036639 antigens Human genes 0.000 claims description 19
- 208000017169 kidney disease Diseases 0.000 claims description 19
- 208000019423 liver disease Diseases 0.000 claims description 19
- 208000035473 Communicable disease Diseases 0.000 claims description 17
- 206010020772 Hypertension Diseases 0.000 claims description 16
- 208000019693 Lung disease Diseases 0.000 claims description 16
- 241001465754 Metazoa Species 0.000 claims description 16
- 206010033128 Ovarian cancer Diseases 0.000 claims description 16
- 206010006187 Breast cancer Diseases 0.000 claims description 15
- 208000026310 Breast neoplasm Diseases 0.000 claims description 15
- 241000725303 Human immunodeficiency virus Species 0.000 claims description 15
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 15
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 15
- 208000010643 digestive system disease Diseases 0.000 claims description 15
- 206010017758 gastric cancer Diseases 0.000 claims description 15
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 15
- 201000011549 stomach cancer Diseases 0.000 claims description 15
- 230000003612 virological effect Effects 0.000 claims description 15
- 206010051379 Systemic Inflammatory Response Syndrome Diseases 0.000 claims description 14
- 206010012601 diabetes mellitus Diseases 0.000 claims description 14
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 claims description 13
- 208000008469 Peptic Ulcer Diseases 0.000 claims description 13
- 208000015181 infectious disease Diseases 0.000 claims description 13
- 230000008569 process Effects 0.000 claims description 13
- 206010016654 Fibrosis Diseases 0.000 claims description 12
- 208000007882 Gastritis Diseases 0.000 claims description 12
- 206010061535 Ovarian neoplasm Diseases 0.000 claims description 12
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 12
- 208000011906 peptic ulcer disease Diseases 0.000 claims description 12
- 206010005003 Bladder cancer Diseases 0.000 claims description 11
- 206010009944 Colon cancer Diseases 0.000 claims description 11
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 11
- 208000007342 Diabetic Nephropathies Diseases 0.000 claims description 11
- 206010019799 Hepatitis viral Diseases 0.000 claims description 11
- 206010060862 Prostate cancer Diseases 0.000 claims description 11
- 208000000236 Prostatic Neoplasms Diseases 0.000 claims description 11
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 claims description 11
- 208000006673 asthma Diseases 0.000 claims description 11
- 208000033679 diabetic kidney disease Diseases 0.000 claims description 11
- 201000000523 end stage renal failure Diseases 0.000 claims description 11
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 11
- 201000005112 urinary bladder cancer Diseases 0.000 claims description 11
- 201000001862 viral hepatitis Diseases 0.000 claims description 11
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 10
- 208000032928 Dyslipidaemia Diseases 0.000 claims description 10
- 208000017170 Lipid metabolism disease Diseases 0.000 claims description 10
- 208000000453 Skin Neoplasms Diseases 0.000 claims description 10
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 10
- 208000010125 myocardial infarction Diseases 0.000 claims description 10
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 10
- 206010022489 Insulin Resistance Diseases 0.000 claims description 9
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 9
- 208000008589 Obesity Diseases 0.000 claims description 9
- 241000700605 Viruses Species 0.000 claims description 9
- 230000007882 cirrhosis Effects 0.000 claims description 9
- 201000005202 lung cancer Diseases 0.000 claims description 9
- 208000020816 lung neoplasm Diseases 0.000 claims description 9
- 235000020824 obesity Nutrition 0.000 claims description 9
- 201000000849 skin cancer Diseases 0.000 claims description 9
- 208000011231 Crohn disease Diseases 0.000 claims description 8
- 230000002526 effect on cardiovascular system Effects 0.000 claims description 8
- 208000006454 hepatitis Diseases 0.000 claims description 8
- 231100000283 hepatitis Toxicity 0.000 claims description 8
- 241000701044 Human gammaherpesvirus 4 Species 0.000 claims description 7
- 208000032382 Ischaemic stroke Diseases 0.000 claims description 7
- 206010033645 Pancreatitis Diseases 0.000 claims description 7
- 241000725643 Respiratory syncytial virus Species 0.000 claims description 7
- 230000002490 cerebral effect Effects 0.000 claims description 7
- 201000001421 hyperglycemia Diseases 0.000 claims description 7
- 208000037797 influenza A Diseases 0.000 claims description 7
- 208000037906 ischaemic injury Diseases 0.000 claims description 7
- 208000028867 ischemia Diseases 0.000 claims description 7
- 201000007270 liver cancer Diseases 0.000 claims description 7
- 208000014018 liver neoplasm Diseases 0.000 claims description 7
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 4
- 210000003719 b-lymphocyte Anatomy 0.000 claims description 4
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims 2
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 323
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 97
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 97
- 239000003642 reactive oxygen metabolite Substances 0.000 description 77
- 230000000694 effects Effects 0.000 description 53
- 108090000623 proteins and genes Proteins 0.000 description 51
- 238000012552 review Methods 0.000 description 49
- 239000003814 drug Substances 0.000 description 48
- 108090000765 processed proteins & peptides Proteins 0.000 description 47
- 230000001965 increasing effect Effects 0.000 description 41
- 229940079593 drug Drugs 0.000 description 35
- 102000004169 proteins and genes Human genes 0.000 description 35
- 150000003254 radicals Chemical class 0.000 description 35
- 235000018102 proteins Nutrition 0.000 description 33
- 239000003963 antioxidant agent Substances 0.000 description 32
- 235000006708 antioxidants Nutrition 0.000 description 26
- 102000004196 processed proteins & peptides Human genes 0.000 description 24
- 238000004519 manufacturing process Methods 0.000 description 23
- -1 tynorphin Chemical class 0.000 description 22
- 230000001590 oxidative effect Effects 0.000 description 21
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 19
- 102000004190 Enzymes Human genes 0.000 description 19
- 108090000790 Enzymes Proteins 0.000 description 19
- 241001529936 Murinae Species 0.000 description 19
- 238000003556 assay Methods 0.000 description 19
- 229940088598 enzyme Drugs 0.000 description 19
- 230000002401 inhibitory effect Effects 0.000 description 19
- 239000007845 reactive nitrogen species Substances 0.000 description 19
- 101000588302 Homo sapiens Nuclear factor erythroid 2-related factor 2 Proteins 0.000 description 18
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical group OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 18
- 108060003951 Immunoglobulin Proteins 0.000 description 18
- 102100031701 Nuclear factor erythroid 2-related factor 2 Human genes 0.000 description 18
- 230000003078 antioxidant effect Effects 0.000 description 18
- 230000014509 gene expression Effects 0.000 description 18
- 102000018358 immunoglobulin Human genes 0.000 description 18
- 230000005764 inhibitory process Effects 0.000 description 18
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 16
- 230000003053 immunization Effects 0.000 description 16
- 238000002649 immunization Methods 0.000 description 16
- 230000001413 cellular effect Effects 0.000 description 15
- 230000006378 damage Effects 0.000 description 15
- 230000008506 pathogenesis Effects 0.000 description 15
- 239000008194 pharmaceutical composition Substances 0.000 description 15
- 108700032225 Antioxidant Response Elements Proteins 0.000 description 14
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 230000004913 activation Effects 0.000 description 14
- 239000000243 solution Substances 0.000 description 14
- 238000009825 accumulation Methods 0.000 description 13
- 230000001684 chronic effect Effects 0.000 description 13
- 238000011161 development Methods 0.000 description 13
- 230000018109 developmental process Effects 0.000 description 13
- 239000007800 oxidant agent Substances 0.000 description 13
- 238000011160 research Methods 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 12
- 206010019851 Hepatotoxicity Diseases 0.000 description 12
- 230000015572 biosynthetic process Effects 0.000 description 12
- 239000000872 buffer Substances 0.000 description 12
- 231100000504 carcinogenesis Toxicity 0.000 description 12
- 231100000304 hepatotoxicity Toxicity 0.000 description 12
- 230000007686 hepatotoxicity Effects 0.000 description 12
- 230000007246 mechanism Effects 0.000 description 12
- 231100000417 nephrotoxicity Toxicity 0.000 description 12
- 239000011780 sodium chloride Substances 0.000 description 12
- 210000001519 tissue Anatomy 0.000 description 12
- 208000005623 Carcinogenesis Diseases 0.000 description 11
- 239000008280 blood Substances 0.000 description 11
- 229940098773 bovine serum albumin Drugs 0.000 description 11
- 230000036952 cancer formation Effects 0.000 description 11
- 210000004185 liver Anatomy 0.000 description 11
- 230000004065 mitochondrial dysfunction Effects 0.000 description 11
- 210000002966 serum Anatomy 0.000 description 11
- 239000007790 solid phase Substances 0.000 description 11
- JVJFIQYAHPMBBX-UHFFFAOYSA-N 4-hydroxynonenal Chemical compound CCCCCC(O)C=CC=O JVJFIQYAHPMBBX-UHFFFAOYSA-N 0.000 description 10
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 10
- 210000004556 brain Anatomy 0.000 description 10
- 230000003834 intracellular effect Effects 0.000 description 10
- 230000004792 oxidative damage Effects 0.000 description 10
- 238000005406 washing Methods 0.000 description 10
- 208000015943 Coeliac disease Diseases 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- 108010002998 NADPH Oxidases Proteins 0.000 description 9
- 102000004722 NADPH Oxidases Human genes 0.000 description 9
- 230000032683 aging Effects 0.000 description 9
- 230000001404 mediated effect Effects 0.000 description 9
- 230000006318 protein oxidation Effects 0.000 description 9
- 230000001105 regulatory effect Effects 0.000 description 9
- 239000000758 substrate Substances 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 230000007423 decrease Effects 0.000 description 8
- LZDSILRDTDCIQT-UHFFFAOYSA-N dinitrogen trioxide Chemical compound [O-][N+](=O)N=O LZDSILRDTDCIQT-UHFFFAOYSA-N 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 208000002672 hepatitis B Diseases 0.000 description 8
- 102000054672 human DPP3 Human genes 0.000 description 8
- 229910052742 iron Inorganic materials 0.000 description 8
- 150000002632 lipids Chemical class 0.000 description 8
- 230000035772 mutation Effects 0.000 description 8
- 230000002018 overexpression Effects 0.000 description 8
- 239000001301 oxygen Substances 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 239000002953 phosphate buffered saline Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 230000001225 therapeutic effect Effects 0.000 description 8
- 241000711549 Hepacivirus C Species 0.000 description 7
- 102000004034 Kelch-Like ECH-Associated Protein 1 Human genes 0.000 description 7
- 108090000484 Kelch-Like ECH-Associated Protein 1 Proteins 0.000 description 7
- 241000699670 Mus sp. Species 0.000 description 7
- 102000035195 Peptidases Human genes 0.000 description 7
- 108091005804 Peptidases Proteins 0.000 description 7
- 230000006907 apoptotic process Effects 0.000 description 7
- 239000000090 biomarker Substances 0.000 description 7
- 238000004113 cell culture Methods 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 208000037976 chronic inflammation Diseases 0.000 description 7
- 230000006020 chronic inflammation Effects 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 7
- 230000003247 decreasing effect Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 230000007613 environmental effect Effects 0.000 description 7
- 230000002255 enzymatic effect Effects 0.000 description 7
- 239000003112 inhibitor Substances 0.000 description 7
- 230000003859 lipid peroxidation Effects 0.000 description 7
- 230000004060 metabolic process Effects 0.000 description 7
- 210000004165 myocardium Anatomy 0.000 description 7
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 description 7
- 238000012261 overproduction Methods 0.000 description 7
- 241000894007 species Species 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 231100000419 toxicity Toxicity 0.000 description 7
- 230000001988 toxicity Effects 0.000 description 7
- 102000016938 Catalase Human genes 0.000 description 6
- 108010053835 Catalase Proteins 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 6
- 108090000695 Cytokines Proteins 0.000 description 6
- 230000005778 DNA damage Effects 0.000 description 6
- 231100000277 DNA damage Toxicity 0.000 description 6
- 241000700721 Hepatitis B virus Species 0.000 description 6
- 208000005176 Hepatitis C Diseases 0.000 description 6
- 206010021143 Hypoxia Diseases 0.000 description 6
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 6
- 208000007571 Ovarian Epithelial Carcinoma Diseases 0.000 description 6
- 210000001744 T-lymphocyte Anatomy 0.000 description 6
- 239000002671 adjuvant Substances 0.000 description 6
- 210000000601 blood cell Anatomy 0.000 description 6
- 210000003169 central nervous system Anatomy 0.000 description 6
- 238000010168 coupling process Methods 0.000 description 6
- 238000005859 coupling reaction Methods 0.000 description 6
- 230000034994 death Effects 0.000 description 6
- 231100000517 death Toxicity 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 6
- 229940118019 malondialdehyde Drugs 0.000 description 6
- 239000003550 marker Substances 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 210000003470 mitochondria Anatomy 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 210000002569 neuron Anatomy 0.000 description 6
- 230000003647 oxidation Effects 0.000 description 6
- 238000007254 oxidation reaction Methods 0.000 description 6
- 210000002381 plasma Anatomy 0.000 description 6
- 238000000159 protein binding assay Methods 0.000 description 6
- 239000000700 radioactive tracer Substances 0.000 description 6
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 5
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 102000002933 Thioredoxin Human genes 0.000 description 5
- 230000002159 abnormal effect Effects 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 210000001175 cerebrospinal fluid Anatomy 0.000 description 5
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 5
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 5
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 5
- 230000007954 hypoxia Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 230000002438 mitochondrial effect Effects 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 230000002611 ovarian Effects 0.000 description 5
- 230000037361 pathway Effects 0.000 description 5
- CMFNMSMUKZHDEY-UHFFFAOYSA-M peroxynitrite Chemical compound [O-]ON=O CMFNMSMUKZHDEY-UHFFFAOYSA-M 0.000 description 5
- 230000000750 progressive effect Effects 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 230000011664 signaling Effects 0.000 description 5
- 230000009885 systemic effect Effects 0.000 description 5
- 108060008226 thioredoxin Proteins 0.000 description 5
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 5
- 238000001262 western blot Methods 0.000 description 5
- 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 4
- 208000030090 Acute Disease Diseases 0.000 description 4
- 208000003918 Acute Kidney Tubular Necrosis Diseases 0.000 description 4
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 4
- 201000001320 Atherosclerosis Diseases 0.000 description 4
- 206010008342 Cervix carcinoma Diseases 0.000 description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 4
- 206010029155 Nephropathy toxic Diseases 0.000 description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 108091023040 Transcription factor Proteins 0.000 description 4
- 102000040945 Transcription factor Human genes 0.000 description 4
- 206010048302 Tubulointerstitial nephritis Diseases 0.000 description 4
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 4
- 239000000730 antalgic agent Substances 0.000 description 4
- 239000012131 assay buffer Substances 0.000 description 4
- 230000033228 biological regulation Effects 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 201000010881 cervical cancer Diseases 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 230000008878 coupling Effects 0.000 description 4
- 239000012228 culture supernatant Substances 0.000 description 4
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 4
- 230000004064 dysfunction Effects 0.000 description 4
- 239000012636 effector Substances 0.000 description 4
- 239000002158 endotoxin Substances 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 4
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 210000004969 inflammatory cell Anatomy 0.000 description 4
- 230000000977 initiatory effect Effects 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 238000007912 intraperitoneal administration Methods 0.000 description 4
- 208000032839 leukemia Diseases 0.000 description 4
- 210000000265 leukocyte Anatomy 0.000 description 4
- 238000002761 liquid phase assay Methods 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- 230000036210 malignancy Effects 0.000 description 4
- 230000003211 malignant effect Effects 0.000 description 4
- 238000013507 mapping Methods 0.000 description 4
- 230000007694 nephrotoxicity Effects 0.000 description 4
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 4
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 4
- 230000007170 pathology Effects 0.000 description 4
- 229940097156 peroxyl Drugs 0.000 description 4
- 238000002823 phage display Methods 0.000 description 4
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 4
- ZJAOAACCNHFJAH-UHFFFAOYSA-N phosphonoformic acid Chemical compound OC(=O)P(O)(O)=O ZJAOAACCNHFJAH-UHFFFAOYSA-N 0.000 description 4
- 229920001223 polyethylene glycol Polymers 0.000 description 4
- 230000006950 reactive oxygen species formation Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 210000003523 substantia nigra Anatomy 0.000 description 4
- 239000013589 supplement Substances 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- 238000013518 transcription Methods 0.000 description 4
- 230000035897 transcription Effects 0.000 description 4
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 3
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 3
- 108091023037 Aptamer Proteins 0.000 description 3
- 201000009030 Carcinoma Diseases 0.000 description 3
- 208000008964 Chemical and Drug Induced Liver Injury Diseases 0.000 description 3
- 208000000419 Chronic Hepatitis B Diseases 0.000 description 3
- 208000006154 Chronic hepatitis C Diseases 0.000 description 3
- 108020004635 Complementary DNA Proteins 0.000 description 3
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 3
- 206010072268 Drug-induced liver injury Diseases 0.000 description 3
- 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 3
- 206010048554 Endothelial dysfunction Diseases 0.000 description 3
- 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 3
- 102000006587 Glutathione peroxidase Human genes 0.000 description 3
- 108700016172 Glutathione peroxidases Proteins 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 3
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 3
- 206010067125 Liver injury Diseases 0.000 description 3
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 3
- 239000004365 Protease Substances 0.000 description 3
- 206010038540 Renal tubular necrosis Diseases 0.000 description 3
- 102100033220 Xanthine oxidase Human genes 0.000 description 3
- 108010093894 Xanthine oxidase Proteins 0.000 description 3
- 229940035676 analgesics Drugs 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 239000000935 antidepressant agent Substances 0.000 description 3
- 229940005513 antidepressants Drugs 0.000 description 3
- 239000004599 antimicrobial Substances 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 210000004534 cecum Anatomy 0.000 description 3
- 230000030833 cell death Effects 0.000 description 3
- 230000007910 cell fusion Effects 0.000 description 3
- 239000013592 cell lysate Substances 0.000 description 3
- 230000005754 cellular signaling Effects 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 3
- 229960004316 cisplatin Drugs 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 230000002939 deleterious effect Effects 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 108090000283 dipeptidyl peptidase III Proteins 0.000 description 3
- 238000010494 dissociation reaction Methods 0.000 description 3
- 230000005593 dissociations Effects 0.000 description 3
- 208000028208 end stage renal disease Diseases 0.000 description 3
- 230000002357 endometrial effect Effects 0.000 description 3
- 230000008694 endothelial dysfunction Effects 0.000 description 3
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 229910001385 heavy metal Inorganic materials 0.000 description 3
- 210000004024 hepatic stellate cell Anatomy 0.000 description 3
- 208000010710 hepatitis C virus infection Diseases 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- 230000016784 immunoglobulin production Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 238000001990 intravenous administration Methods 0.000 description 3
- 210000001503 joint Anatomy 0.000 description 3
- 238000002372 labelling Methods 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- UPSFMJHZUCSEHU-JYGUBCOQSA-N n-[(2s,3r,4r,5s,6r)-2-[(2r,3s,4r,5r,6s)-5-acetamido-4-hydroxy-2-(hydroxymethyl)-6-(4-methyl-2-oxochromen-7-yl)oxyoxan-3-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-3-yl]acetamide Chemical compound CC(=O)N[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@H](O)[C@@H](NC(C)=O)[C@H](OC=2C=C3OC(=O)C=C(C)C3=CC=2)O[C@@H]1CO UPSFMJHZUCSEHU-JYGUBCOQSA-N 0.000 description 3
- 210000000440 neutrophil Anatomy 0.000 description 3
- 230000008520 organization Effects 0.000 description 3
- 230000010627 oxidative phosphorylation Effects 0.000 description 3
- 230000036407 pain Effects 0.000 description 3
- 230000002085 persistent effect Effects 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 230000000770 proinflammatory effect Effects 0.000 description 3
- 235000019833 protease Nutrition 0.000 description 3
- 235000019419 proteases Nutrition 0.000 description 3
- 238000006479 redox reaction Methods 0.000 description 3
- 230000010282 redox signaling Effects 0.000 description 3
- 230000035806 respiratory chain Effects 0.000 description 3
- 208000023504 respiratory system disease Diseases 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 239000000779 smoke Substances 0.000 description 3
- 238000002764 solid phase assay Methods 0.000 description 3
- 230000009870 specific binding Effects 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- 230000035882 stress Effects 0.000 description 3
- 238000006467 substitution reaction Methods 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 208000011580 syndromic disease Diseases 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- QBLFFOHVDMBOPS-DYEKYZERSA-N (2s)-2-[[(2s)-1-[(2r)-2-[2-(hydroxyamino)-2-oxoethyl]pentanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoic acid Chemical compound ONC(=O)C[C@@H](CCC)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O QBLFFOHVDMBOPS-DYEKYZERSA-N 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 2
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 2
- VAZJLPXFVQHDFB-UHFFFAOYSA-N 1-(diaminomethylidene)-2-hexylguanidine Polymers CCCCCCN=C(N)N=C(N)N VAZJLPXFVQHDFB-UHFFFAOYSA-N 0.000 description 2
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical class C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 description 2
- FBTSQILOGYXGMD-LURJTMIESA-N 3-nitro-L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C([N+]([O-])=O)=C1 FBTSQILOGYXGMD-LURJTMIESA-N 0.000 description 2
- MGWGWNFMUOTEHG-UHFFFAOYSA-N 4-(3,5-dimethylphenyl)-1,3-thiazol-2-amine Chemical compound CC1=CC(C)=CC(C=2N=C(N)SC=2)=C1 MGWGWNFMUOTEHG-UHFFFAOYSA-N 0.000 description 2
- XYJODUBPWNZLML-UHFFFAOYSA-N 5-ethyl-6-phenyl-6h-phenanthridine-3,8-diamine Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2N(CC)C1C1=CC=CC=C1 XYJODUBPWNZLML-UHFFFAOYSA-N 0.000 description 2
- HCAJQHYUCKICQH-VPENINKCSA-N 8-Oxo-7,8-dihydro-2'-deoxyguanosine Chemical compound C1=2NC(N)=NC(=O)C=2NC(=O)N1[C@H]1C[C@H](O)[C@@H](CO)O1 HCAJQHYUCKICQH-VPENINKCSA-N 0.000 description 2
- 239000005541 ACE inhibitor Substances 0.000 description 2
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 108010064733 Angiotensins Proteins 0.000 description 2
- 102000015427 Angiotensins Human genes 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 206010004146 Basal cell carcinoma Diseases 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 2
- 108010017384 Blood Proteins Proteins 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 206010048610 Cardiotoxicity Diseases 0.000 description 2
- 208000017667 Chronic Disease Diseases 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 229930105110 Cyclosporin A Natural products 0.000 description 2
- PMATZTZNYRCHOR-CGLBZJNRSA-N Cyclosporin A Chemical compound CC[C@@H]1NC(=O)[C@H]([C@H](O)[C@H](C)C\C=C\C)N(C)C(=O)[C@H](C(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](CC(C)C)N(C)C(=O)[C@@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)N(C)C(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)N(C)C(=O)CN(C)C1=O PMATZTZNYRCHOR-CGLBZJNRSA-N 0.000 description 2
- 108010036949 Cyclosporine Proteins 0.000 description 2
- 231100001074 DNA strand break Toxicity 0.000 description 2
- 208000002249 Diabetes Complications Diseases 0.000 description 2
- 108010016626 Dipeptides Proteins 0.000 description 2
- 108050001925 Dipeptidyl-peptidase 3 Proteins 0.000 description 2
- 206010013975 Dyspnoeas Diseases 0.000 description 2
- 206010014733 Endometrial cancer Diseases 0.000 description 2
- 206010014759 Endometrial neoplasm Diseases 0.000 description 2
- 201000009273 Endometriosis Diseases 0.000 description 2
- 108010092674 Enkephalins Proteins 0.000 description 2
- 102000018389 Exopeptidases Human genes 0.000 description 2
- 108010091443 Exopeptidases Proteins 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 208000002705 Glucose Intolerance Diseases 0.000 description 2
- 108010024636 Glutathione Proteins 0.000 description 2
- 108010023302 HDL Cholesterol Proteins 0.000 description 2
- 229940121710 HMGCoA reductase inhibitor Drugs 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101150043003 Htt gene Proteins 0.000 description 2
- HEFNNWSXXWATRW-UHFFFAOYSA-N Ibuprofen Chemical compound CC(C)CC1=CC=C(C(C)C(O)=O)C=C1 HEFNNWSXXWATRW-UHFFFAOYSA-N 0.000 description 2
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 2
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 2
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 2
- URLZCHNOLZSCCA-VABKMULXSA-N Leu-enkephalin Chemical class C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=CC=C1 URLZCHNOLZSCCA-VABKMULXSA-N 0.000 description 2
- 108010006035 Metalloproteases Proteins 0.000 description 2
- 102000005741 Metalloproteases Human genes 0.000 description 2
- 206010052641 Mitochondrial DNA mutation Diseases 0.000 description 2
- BYHJHXPTQMMKCA-QMMMGPOBSA-N N-formyl-L-kynurenine Chemical compound [O-]C(=O)[C@@H]([NH3+])CC(=O)C1=CC=CC=C1NC=O BYHJHXPTQMMKCA-QMMMGPOBSA-N 0.000 description 2
- MQUQNUAYKLCRME-INIZCTEOSA-N N-tosyl-L-phenylalanyl chloromethyl ketone Chemical compound C1=CC(C)=CC=C1S(=O)(=O)N[C@H](C(=O)CCl)CC1=CC=CC=C1 MQUQNUAYKLCRME-INIZCTEOSA-N 0.000 description 2
- XJLXINKUBYWONI-NNYOXOHSSA-N NADP zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-N 0.000 description 2
- 102000003945 NF-kappa B Human genes 0.000 description 2
- 108010057466 NF-kappa B Proteins 0.000 description 2
- CMWTZPSULFXXJA-UHFFFAOYSA-N Naproxen Natural products C1=C(C(C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-UHFFFAOYSA-N 0.000 description 2
- 108700019961 Neoplasm Genes Proteins 0.000 description 2
- 102000048850 Neoplasm Genes Human genes 0.000 description 2
- 208000012902 Nervous system disease Diseases 0.000 description 2
- 208000025966 Neurological disease Diseases 0.000 description 2
- 241000208125 Nicotiana Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 2
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 229920002413 Polyhexanide Polymers 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 206010061481 Renal injury Diseases 0.000 description 2
- 108010003723 Single-Domain Antibodies Proteins 0.000 description 2
- 108010021188 Superoxide Dismutase-1 Proteins 0.000 description 2
- 102100038836 Superoxide dismutase [Cu-Zn] Human genes 0.000 description 2
- 206010043118 Tardive Dyskinesia Diseases 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 108010018242 Transcription Factor AP-1 Proteins 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 2
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229960004150 aciclovir Drugs 0.000 description 2
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 description 2
- 239000012190 activator Substances 0.000 description 2
- 238000003915 air pollution Methods 0.000 description 2
- 230000033115 angiogenesis Effects 0.000 description 2
- 229940044094 angiotensin-converting-enzyme inhibitor Drugs 0.000 description 2
- 230000001430 anti-depressive effect Effects 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 230000000118 anti-neoplastic effect Effects 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 229940125715 antihistaminic agent Drugs 0.000 description 2
- 239000000739 antihistaminic agent Substances 0.000 description 2
- 239000003443 antiviral agent Substances 0.000 description 2
- 229940121357 antivirals Drugs 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- AUJRCFUBUPVWSZ-XTZHGVARSA-M auranofin Chemical compound CCP(CC)(CC)=[Au]S[C@@H]1O[C@H](COC(C)=O)[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@H]1OC(C)=O AUJRCFUBUPVWSZ-XTZHGVARSA-M 0.000 description 2
- 229960005207 auranofin Drugs 0.000 description 2
- 230000004900 autophagic degradation Effects 0.000 description 2
- 230000003376 axonal effect Effects 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 229920001222 biopolymer Polymers 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 210000005013 brain tissue Anatomy 0.000 description 2
- 230000000711 cancerogenic effect Effects 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 231100000315 carcinogenic Toxicity 0.000 description 2
- 231100000259 cardiotoxicity Toxicity 0.000 description 2
- 239000002327 cardiovascular agent Substances 0.000 description 2
- 229940125692 cardiovascular agent Drugs 0.000 description 2
- 230000007681 cardiovascular toxicity Effects 0.000 description 2
- 150000003943 catecholamines Chemical class 0.000 description 2
- 230000001364 causal effect Effects 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000007541 cellular toxicity Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000000038 chest Anatomy 0.000 description 2
- 235000019504 cigarettes Nutrition 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 239000000994 contrast dye Substances 0.000 description 2
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 230000006735 deficit Effects 0.000 description 2
- 210000004443 dendritic cell Anatomy 0.000 description 2
- 230000001627 detrimental effect Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- OQALFHMKVSJFRR-UHFFFAOYSA-N dityrosine Chemical compound OC(=O)C(N)CC1=CC=C(O)C(C=2C(=CC=C(CC(N)C(O)=O)C=2)O)=C1 OQALFHMKVSJFRR-UHFFFAOYSA-N 0.000 description 2
- 239000002934 diuretic Substances 0.000 description 2
- 229940030606 diuretics Drugs 0.000 description 2
- 229960003638 dopamine Drugs 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 230000008482 dysregulation Effects 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 239000000262 estrogen Substances 0.000 description 2
- 229940011871 estrogen Drugs 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 230000004761 fibrosis Effects 0.000 description 2
- 229930186097 fluostatin Natural products 0.000 description 2
- 229960005102 foscarnet Drugs 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 108020001507 fusion proteins Proteins 0.000 description 2
- 102000037865 fusion proteins Human genes 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 229960002518 gentamicin Drugs 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 208000038003 heart failure with preserved ejection fraction Diseases 0.000 description 2
- 208000038002 heart failure with reduced ejection fraction Diseases 0.000 description 2
- 230000000004 hemodynamic effect Effects 0.000 description 2
- 231100000753 hepatic injury Toxicity 0.000 description 2
- 230000004730 hepatocarcinogenesis Effects 0.000 description 2
- 230000003284 homeostatic effect Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 239000002471 hydroxymethylglutaryl coenzyme A reductase inhibitor Substances 0.000 description 2
- 229960001680 ibuprofen Drugs 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 239000003018 immunosuppressive agent Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 210000001865 kupffer cell Anatomy 0.000 description 2
- YGPSJZOEDVAXAB-UHFFFAOYSA-N kynurenine Chemical compound OC(=O)C(N)CC(=O)C1=CC=CC=C1N YGPSJZOEDVAXAB-UHFFFAOYSA-N 0.000 description 2
- 229920006008 lipopolysaccharide Polymers 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 210000005228 liver tissue Anatomy 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 201000001441 melanoma Diseases 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 210000000274 microglia Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 229940126619 mouse monoclonal antibody Drugs 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 229960002009 naproxen Drugs 0.000 description 2
- CMWTZPSULFXXJA-VIFPVBQESA-N naproxen Chemical compound C1=C([C@H](C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-VIFPVBQESA-N 0.000 description 2
- 230000017074 necrotic cell death Effects 0.000 description 2
- 230000003589 nefrotoxic effect Effects 0.000 description 2
- 230000000926 neurological effect Effects 0.000 description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 2
- 238000006396 nitration reaction Methods 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- CKNAQFVBEHDJQV-UHFFFAOYSA-N oltipraz Chemical compound S1SC(=S)C(C)=C1C1=CN=CC=N1 CKNAQFVBEHDJQV-UHFFFAOYSA-N 0.000 description 2
- 229950008687 oltipraz Drugs 0.000 description 2
- 229960005489 paracetamol Drugs 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000001991 pathophysiological effect Effects 0.000 description 2
- 230000037050 permeability transition Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 201000009104 prediabetes syndrome Diseases 0.000 description 2
- 230000003244 pro-oxidative effect Effects 0.000 description 2
- 150000003180 prostaglandins Chemical class 0.000 description 2
- 229940126409 proton pump inhibitor Drugs 0.000 description 2
- 239000000612 proton pump inhibitor Substances 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000007348 radical reaction Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000000392 somatic effect Effects 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- SUVMJBTUFCVSAD-UHFFFAOYSA-N sulforaphane Chemical compound CS(=O)CCCCN=C=S SUVMJBTUFCVSAD-UHFFFAOYSA-N 0.000 description 2
- IMCGHZIGRANKHV-AJNGGQMLSA-N tert-butyl (3s,5s)-2-oxo-5-[(2s,4s)-5-oxo-4-propan-2-yloxolan-2-yl]-3-propan-2-ylpyrrolidine-1-carboxylate Chemical compound O1C(=O)[C@H](C(C)C)C[C@H]1[C@H]1N(C(=O)OC(C)(C)C)C(=O)[C@H](C(C)C)C1 IMCGHZIGRANKHV-AJNGGQMLSA-N 0.000 description 2
- 229940094937 thioredoxin Drugs 0.000 description 2
- 229940113082 thymine Drugs 0.000 description 2
- 230000000451 tissue damage Effects 0.000 description 2
- 231100000827 tissue damage Toxicity 0.000 description 2
- 238000010967 transthoracic echocardiography Methods 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 230000002485 urinary effect Effects 0.000 description 2
- 210000002700 urine Anatomy 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 230000002861 ventricular Effects 0.000 description 2
- YFGBQHOOROIVKG-BHDDXSALSA-N (2R)-2-[[(2R)-2-[[2-[[2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]acetyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]-4-methylsulfanylbutanoic acid Chemical compound C([C@H](C(=O)N[C@H](CCSC)C(O)=O)NC(=O)CNC(=O)CNC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=CC=C1 YFGBQHOOROIVKG-BHDDXSALSA-N 0.000 description 1
- PYYCBPABFFOXLZ-IPYPFGDCSA-N (2s)-2-[[(2s)-1-[(2r)-2-[2-(hydroxyamino)-2-oxoethyl]-4-methylpentanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoic acid Chemical compound ONC(=O)C[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O PYYCBPABFFOXLZ-IPYPFGDCSA-N 0.000 description 1
- WLWBANYDSJWZBI-IIZANFQQSA-N (2s)-2-[[(2s)-1-[(2s)-2-[[(2s)-2-[[(2s)-2-azaniumyl-3-methylbutanoyl]amino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]pyrrolidine-2-carbonyl]amino]-3-(1h-indol-3-yl)propanoate Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CC=C(O)C=C1 WLWBANYDSJWZBI-IIZANFQQSA-N 0.000 description 1
- WMYLYYNMCFINGV-CKCBUVOCSA-N (2s)-2-amino-5-[[(2r)-1-(carboxymethylamino)-1-oxo-3-sulfanylpropan-2-yl]amino]-5-oxopentanoic acid Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O.OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O WMYLYYNMCFINGV-CKCBUVOCSA-N 0.000 description 1
- GIANIJCPTPUNBA-QMMMGPOBSA-N (2s)-3-(4-hydroxyphenyl)-2-nitramidopropanoic acid Chemical compound [O-][N+](=O)N[C@H](C(=O)O)CC1=CC=C(O)C=C1 GIANIJCPTPUNBA-QMMMGPOBSA-N 0.000 description 1
- CUKWUWBLQQDQAC-VEQWQPCFSA-N (3s)-3-amino-4-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s,3s)-1-[[(2s)-1-[(2s)-2-[[(1s)-1-carboxyethyl]carbamoyl]pyrrolidin-1-yl]-3-(1h-imidazol-5-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-methyl-1-ox Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 CUKWUWBLQQDQAC-VEQWQPCFSA-N 0.000 description 1
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- VHRUMKCAEVRUBK-GODQJPCRSA-N 15-deoxy-Delta(12,14)-prostaglandin J2 Chemical compound CCCCC\C=C\C=C1/[C@@H](C\C=C/CCCC(O)=O)C=CC1=O VHRUMKCAEVRUBK-GODQJPCRSA-N 0.000 description 1
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical class C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 1
- CGVPQHIRIGIDLE-UHFFFAOYSA-N 3-(2-aminooxyphenyl)propanoic acid Chemical compound NOC1=CC=CC=C1CCC(O)=O CGVPQHIRIGIDLE-UHFFFAOYSA-N 0.000 description 1
- UPXRTVAIJMUAQR-UHFFFAOYSA-N 4-(9h-fluoren-9-ylmethoxycarbonylamino)-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidine-2-carboxylic acid Chemical compound C1C(C(O)=O)N(C(=O)OC(C)(C)C)CC1NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 UPXRTVAIJMUAQR-UHFFFAOYSA-N 0.000 description 1
- SUVMJBTUFCVSAD-JTQLQIEISA-N 4-Methylsulfinylbutyl isothiocyanate Natural products C[S@](=O)CCCCN=C=S SUVMJBTUFCVSAD-JTQLQIEISA-N 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 208000004611 Abdominal Obesity Diseases 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 208000007788 Acute Liver Failure Diseases 0.000 description 1
- 206010000804 Acute hepatic failure Diseases 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 108010051842 Advanced Oxidation Protein Products Proteins 0.000 description 1
- 206010001497 Agitation Diseases 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 1
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 102400000347 Angiotensin 1-7 Human genes 0.000 description 1
- QMMRCKSBBNJCMR-KMZPNFOHSA-N Angiotensin III Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CCCN=C(N)N)C(C)C)C1=CC=C(O)C=C1 QMMRCKSBBNJCMR-KMZPNFOHSA-N 0.000 description 1
- QSBGWDDCOJYQGY-KOQODJNWSA-N Angiotensin IV Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)NC(=O)[C@@H](N)C(C)C)C1=CC=C(O)C=C1 QSBGWDDCOJYQGY-KOQODJNWSA-N 0.000 description 1
- 102400000345 Angiotensin-2 Human genes 0.000 description 1
- 101800000733 Angiotensin-2 Proteins 0.000 description 1
- 102400000348 Angiotensin-3 Human genes 0.000 description 1
- 101800000738 Angiotensin-3 Proteins 0.000 description 1
- 102400000349 Angiotensin-4 Human genes 0.000 description 1
- 101800000737 Angiotensin-4 Proteins 0.000 description 1
- 102000008102 Ankyrins Human genes 0.000 description 1
- 108010049777 Ankyrins Proteins 0.000 description 1
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 1
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- TXGZJQLMVSIZEI-UQMAOPSPSA-N Bardoxolone Chemical compound C1=C(C#N)C(=O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5[C@H]4C(=O)C=C3[C@]21C TXGZJQLMVSIZEI-UQMAOPSPSA-N 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 206010006100 Bradykinesia Diseases 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 206010007556 Cardiac failure acute Diseases 0.000 description 1
- 102000011727 Caspases Human genes 0.000 description 1
- 108010076667 Caspases Proteins 0.000 description 1
- 206010057248 Cell death Diseases 0.000 description 1
- 206010065941 Central obesity Diseases 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 241001573498 Compacta Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 206010071503 Crystal nephropathy Diseases 0.000 description 1
- 108010015742 Cytochrome P-450 Enzyme System Proteins 0.000 description 1
- 102000003849 Cytochrome P450 Human genes 0.000 description 1
- 102100030497 Cytochrome c Human genes 0.000 description 1
- 108010075031 Cytochromes c Proteins 0.000 description 1
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 1
- 230000009946 DNA mutation Effects 0.000 description 1
- 230000033616 DNA repair Effects 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 206010012655 Diabetic complications Diseases 0.000 description 1
- 102100029857 Dipeptidase 3 Human genes 0.000 description 1
- 108030006877 Dipeptidyl-dipeptidases Proteins 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 102100021218 Dual oxidase 1 Human genes 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 description 1
- 206010015108 Epstein-Barr virus infection Diseases 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 description 1
- 108010068370 Glutens Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 241001326189 Gyrodactylus prostae Species 0.000 description 1
- 208000023661 Haematological disease Diseases 0.000 description 1
- 206010019196 Head injury Diseases 0.000 description 1
- 241000590002 Helicobacter pylori Species 0.000 description 1
- 102000002737 Heme Oxygenase-1 Human genes 0.000 description 1
- 108010018924 Heme Oxygenase-1 Proteins 0.000 description 1
- 108010017480 Hemosiderin Proteins 0.000 description 1
- 206010019663 Hepatic failure Diseases 0.000 description 1
- 208000005331 Hepatitis D Diseases 0.000 description 1
- 208000033640 Hereditary breast cancer Diseases 0.000 description 1
- 101000968308 Homo sapiens Dual oxidase 1 Proteins 0.000 description 1
- 101000968305 Homo sapiens Dual oxidase 2 Proteins 0.000 description 1
- 101000935587 Homo sapiens Flavin reductase (NADPH) Proteins 0.000 description 1
- 101001046870 Homo sapiens Hypoxia-inducible factor 1-alpha Proteins 0.000 description 1
- 101001095266 Homo sapiens Prolyl endopeptidase Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 206010058490 Hyperoxia Diseases 0.000 description 1
- 208000006083 Hypokinesia Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 102100022875 Hypoxia-inducible factor 1-alpha Human genes 0.000 description 1
- PVHLMTREZMEJCG-GDTLVBQBSA-N Ile(5)-angiotensin II (1-7) Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H](CCCNC(N)=[NH2+])NC(=O)[C@@H]([NH3+])CC([O-])=O)C(C)C)C1=CC=C(O)C=C1 PVHLMTREZMEJCG-GDTLVBQBSA-N 0.000 description 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 description 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 description 1
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 description 1
- 102000012745 Immunoglobulin Subunits Human genes 0.000 description 1
- 108010079585 Immunoglobulin Subunits Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 206010056997 Impaired fasting glucose Diseases 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- ZEXLJFNSKAHNFH-SYKYGTKKSA-N L-Phenylalaninamide, L-tyrosyl-L-prolyl-L-tryptophyl- Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)C1=CC=C(O)C=C1 ZEXLJFNSKAHNFH-SYKYGTKKSA-N 0.000 description 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 description 1
- 102100038609 Lactoperoxidase Human genes 0.000 description 1
- 108010023244 Lactoperoxidase Proteins 0.000 description 1
- 102400000243 Leu-enkephalin Human genes 0.000 description 1
- 108010022337 Leucine Enkephalin Proteins 0.000 description 1
- GDBQQVLCIARPGH-UHFFFAOYSA-N Leupeptin Natural products CC(C)CC(NC(C)=O)C(=O)NC(CC(C)C)C(=O)NC(C=O)CCCN=C(N)N GDBQQVLCIARPGH-UHFFFAOYSA-N 0.000 description 1
- 102000019298 Lipocalin Human genes 0.000 description 1
- 108050006654 Lipocalin Proteins 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 102000003820 Lipoxygenases Human genes 0.000 description 1
- 108090000128 Lipoxygenases Proteins 0.000 description 1
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102400000988 Met-enkephalin Human genes 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 108010042237 Methionine Enkephalin Proteins 0.000 description 1
- 206010027525 Microalbuminuria Diseases 0.000 description 1
- 208000034486 Multi-organ failure Diseases 0.000 description 1
- 208000002740 Muscle Rigidity Diseases 0.000 description 1
- 208000021642 Muscular disease Diseases 0.000 description 1
- 102000003896 Myeloperoxidases Human genes 0.000 description 1
- 108090000235 Myeloperoxidases Proteins 0.000 description 1
- 206010069140 Myocardial depression Diseases 0.000 description 1
- 201000009623 Myopathy Diseases 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 1
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 description 1
- 206010028851 Necrosis Diseases 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 102000008299 Nitric Oxide Synthase Human genes 0.000 description 1
- 108010021487 Nitric Oxide Synthase Proteins 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 102000015636 Oligopeptides Human genes 0.000 description 1
- 108010038807 Oligopeptides Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 102000043276 Oncogene Human genes 0.000 description 1
- 206010033109 Ototoxicity Diseases 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 102000017323 Peptidase family M49 Human genes 0.000 description 1
- 108050005420 Peptidase family M49 Proteins 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 102000009097 Phosphorylases Human genes 0.000 description 1
- 108010073135 Phosphorylases Proteins 0.000 description 1
- 206010034972 Photosensitivity reaction Diseases 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 241000283080 Proboscidea <mammal> Species 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 102000056251 Prolyl Oligopeptidases Human genes 0.000 description 1
- QBLFFOHVDMBOPS-UHFFFAOYSA-N Propioxatin A Natural products ONC(=O)CC(CCC)C(=O)N1CCCC1C(=O)NC(C(C)C)C(O)=O QBLFFOHVDMBOPS-UHFFFAOYSA-N 0.000 description 1
- 102000004005 Prostaglandin-endoperoxide synthases Human genes 0.000 description 1
- 108090000459 Prostaglandin-endoperoxide synthases Proteins 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 102000052575 Proto-Oncogene Human genes 0.000 description 1
- 108700020978 Proto-Oncogene Proteins 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 206010039020 Rhabdomyolysis Diseases 0.000 description 1
- 102000000395 SH3 domains Human genes 0.000 description 1
- 108050008861 SH3 domains Proteins 0.000 description 1
- 108010017324 STAT3 Transcription Factor Proteins 0.000 description 1
- 206010040026 Sensory disturbance Diseases 0.000 description 1
- 102100024040 Signal transducer and activator of transcription 3 Human genes 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 101800001235 Spinorphin Proteins 0.000 description 1
- 208000007107 Stomach Ulcer Diseases 0.000 description 1
- 241000187180 Streptomyces sp. Species 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 102100024554 Tetranectin Human genes 0.000 description 1
- 208000034841 Thrombotic Microangiopathies Diseases 0.000 description 1
- 206010070863 Toxicity to various agents Diseases 0.000 description 1
- 102100023132 Transcription factor Jun Human genes 0.000 description 1
- 208000030886 Traumatic Brain injury Diseases 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102400000757 Ubiquitin Human genes 0.000 description 1
- 208000012931 Urologic disease Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- ZDVWPPFTKCGYBB-UHFFFAOYSA-N [4-[3-(2,5-dioxopyrrolidin-1-yl)oxy-3-oxopropyl]phenyl] 10-methylacridin-10-ium-9-carboxylate Chemical compound C12=CC=CC=C2[N+](C)=C2C=CC=CC2=C1C(=O)OC(C=C1)=CC=C1CCC(=O)ON1C(=O)CCC1=O ZDVWPPFTKCGYBB-UHFFFAOYSA-N 0.000 description 1
- CTCBPRXHVPZNHB-VQFZJOCSSA-N [[(2r,3s,4r,5r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate;(2r,3r,4s,5r)-2-(6-aminopurin-9-yl)-5-(hydroxymethyl)oxolane-3,4-diol Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O.C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O CTCBPRXHVPZNHB-VQFZJOCSSA-N 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- DZBUGLKDJFMEHC-UHFFFAOYSA-O acridine;hydron Chemical compound C1=CC=CC2=CC3=CC=CC=C3[NH+]=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-O 0.000 description 1
- 231100000836 acute liver failure Toxicity 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000004103 aerobic respiration Effects 0.000 description 1
- 239000002115 aflatoxin B1 Substances 0.000 description 1
- OQIQSTLJSLGHID-WNWIJWBNSA-N aflatoxin B1 Chemical compound C=1([C@@H]2C=CO[C@@H]2OC=1C=C(C1=2)OC)C=2OC(=O)C2=C1CCC2=O OQIQSTLJSLGHID-WNWIJWBNSA-N 0.000 description 1
- 229930020125 aflatoxin-B1 Natural products 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 210000001552 airway epithelial cell Anatomy 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- 206010001584 alcohol abuse Diseases 0.000 description 1
- 208000025746 alcohol use disease Diseases 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 108010052590 amastatin Proteins 0.000 description 1
- QFAADIRHLBXJJS-ZAZJUGBXSA-N amastatin Chemical compound CC(C)C[C@@H](N)[C@H](O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC(O)=O QFAADIRHLBXJJS-ZAZJUGBXSA-N 0.000 description 1
- 208000007769 amastia Diseases 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 239000003098 androgen Substances 0.000 description 1
- 108010021281 angiotensin I (1-7) Proteins 0.000 description 1
- 239000002416 angiotensin derivative Substances 0.000 description 1
- 229950006323 angiotensin ii Drugs 0.000 description 1
- 108010058865 angiotensinase Proteins 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 230000001760 anti-analgesic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000561 anti-psychotic effect Effects 0.000 description 1
- 230000000798 anti-retroviral effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000002221 antipyretic Substances 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 229910052785 arsenic Inorganic materials 0.000 description 1
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000002567 autonomic effect Effects 0.000 description 1
- 210000003050 axon Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000008952 bacterial invasion Effects 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 230000001588 bifunctional effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000002715 bioenergetic effect Effects 0.000 description 1
- 208000029028 brain injury Diseases 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 244000309464 bull Species 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000025084 cell cycle arrest Effects 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000006567 cellular energy metabolism Effects 0.000 description 1
- 230000004098 cellular respiration Effects 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 208000015114 central nervous system disease Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000002962 chemical mutagen Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- ZPEIMTDSQAKGNT-UHFFFAOYSA-N chlorpromazine Chemical compound C1=C(Cl)C=C2N(CCCN(C)C)C3=CC=CC=C3SC2=C1 ZPEIMTDSQAKGNT-UHFFFAOYSA-N 0.000 description 1
- 229960001076 chlorpromazine Drugs 0.000 description 1
- 230000007881 chronic fibrosis Effects 0.000 description 1
- 208000023652 chronic gastritis Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 208000024207 chronic leukemia Diseases 0.000 description 1
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 1
- 208000009060 clear cell adenocarcinoma Diseases 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 230000001149 cognitive effect Effects 0.000 description 1
- 231100000870 cognitive problem Toxicity 0.000 description 1
- 230000037319 collagen production Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000003340 combinatorial analysis Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 229940109239 creatinine Drugs 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 208000031513 cyst Diseases 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 230000001120 cytoprotective effect Effects 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000007682 dermal toxicity Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- HWOLQKJPMRZMEX-PJKMHFRUSA-N diazonio-[(2r,4s,5r)-4-hydroxy-5-(hydroxymethyl)-2-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]azanide Chemical compound O=C1NC(=O)C(C)=CN1[C@]1(N=[N+]=[N-])O[C@H](CO)[C@@H](O)C1 HWOLQKJPMRZMEX-PJKMHFRUSA-N 0.000 description 1
- 229960001259 diclofenac Drugs 0.000 description 1
- DCOPUUMXTXDBNB-UHFFFAOYSA-N diclofenac Chemical compound OC(=O)CC1=CC=CC=C1NC1=C(Cl)C=CC=C1Cl DCOPUUMXTXDBNB-UHFFFAOYSA-N 0.000 description 1
- 150000001993 dienes Chemical class 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 108010034479 digoxin antibodies Fab fragments Proteins 0.000 description 1
- 238000006471 dimerization reaction Methods 0.000 description 1
- 230000003292 diminished effect Effects 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
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 230000009266 disease activity Effects 0.000 description 1
- 210000005064 dopaminergic neuron Anatomy 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 208000000718 duodenal ulcer Diseases 0.000 description 1
- 235000006694 eating habits Nutrition 0.000 description 1
- 229960001484 edetic acid Drugs 0.000 description 1
- 230000027721 electron transport chain Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 208000023965 endometrium neoplasm Diseases 0.000 description 1
- 108010015205 endomorphin 1 Proteins 0.000 description 1
- 108010015198 endomorphin 2 Proteins 0.000 description 1
- XIJHWXXXIMEHKW-LJWNLINESA-N endomorphin-2 Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)C1=CC=C(O)C=C1 XIJHWXXXIMEHKW-LJWNLINESA-N 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 231100000317 environmental toxin Toxicity 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000004149 ethanol metabolism Effects 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 230000004090 etiopathogenesis Effects 0.000 description 1
- 230000001610 euglycemic effect Effects 0.000 description 1
- 230000003090 exacerbative effect Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 238000002637 fluid replacement therapy Methods 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000001156 gastric mucosa Anatomy 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 230000000762 glandular Effects 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 230000004190 glucose uptake Effects 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 108091005996 glycated proteins Proteins 0.000 description 1
- 208000027096 gram-negative bacterial infections Diseases 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 229940037467 helicobacter pylori Drugs 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 201000010284 hepatitis E Diseases 0.000 description 1
- 231100000334 hepatotoxic Toxicity 0.000 description 1
- 230000003082 hepatotoxic effect Effects 0.000 description 1
- 239000012245 hepatotoxicant Substances 0.000 description 1
- 231100001114 hepatotoxicant Toxicity 0.000 description 1
- 208000025581 hereditary breast carcinoma Diseases 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000013427 histology analysis Methods 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 210000004408 hybridoma Anatomy 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000000222 hyperoxic effect Effects 0.000 description 1
- 230000004046 hyporesponsiveness Effects 0.000 description 1
- 210000003767 ileocecal valve Anatomy 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 229940088592 immunologic factor Drugs 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 238000012744 immunostaining Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000000266 injurious effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 210000005061 intracellular organelle Anatomy 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000005865 ionizing radiation Effects 0.000 description 1
- 230000004171 ischemic cascade Effects 0.000 description 1
- 150000002535 isoprostanes Chemical class 0.000 description 1
- 229960004184 ketamine hydrochloride Drugs 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 208000037806 kidney injury Diseases 0.000 description 1
- 238000012933 kinetic analysis Methods 0.000 description 1
- 229940057428 lactoperoxidase Drugs 0.000 description 1
- URLZCHNOLZSCCA-UHFFFAOYSA-N leu-enkephalin Chemical compound C=1C=C(O)C=CC=1CC(N)C(=O)NCC(=O)NCC(=O)NC(C(=O)NC(CC(C)C)C(O)=O)CC1=CC=CC=C1 URLZCHNOLZSCCA-UHFFFAOYSA-N 0.000 description 1
- 150000002617 leukotrienes Chemical class 0.000 description 1
- GDBQQVLCIARPGH-ULQDDVLXSA-N leupeptin Chemical compound CC(C)C[C@H](NC(C)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C=O)CCCN=C(N)N GDBQQVLCIARPGH-ULQDDVLXSA-N 0.000 description 1
- 108010052968 leupeptin Proteins 0.000 description 1
- 210000004558 lewy body Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000029226 lipidation Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000012317 liver biopsy Methods 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 231100000835 liver failure Toxicity 0.000 description 1
- 208000007903 liver failure Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 208000012866 low blood pressure Diseases 0.000 description 1
- 230000004758 lung carcinogenesis Effects 0.000 description 1
- 201000005243 lung squamous cell carcinoma Diseases 0.000 description 1
- 230000000527 lymphocytic effect Effects 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000037323 metabolic rate Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000008811 mitochondrial respiratory chain Effects 0.000 description 1
- 230000008965 mitochondrial swelling Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000000302 molecular modelling Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 210000002161 motor neuron Anatomy 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 208000029744 multiple organ dysfunction syndrome Diseases 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 229950006238 nadide Drugs 0.000 description 1
- 230000006654 negative regulation of apoptotic process Effects 0.000 description 1
- 231100000381 nephrotoxic Toxicity 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000008599 nitrosative stress Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 108091008819 oncoproteins Proteins 0.000 description 1
- 102000027450 oncoproteins Human genes 0.000 description 1
- 230000008816 organ damage Effects 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 231100000262 ototoxicity Toxicity 0.000 description 1
- 230000008789 oxidative DNA damage Effects 0.000 description 1
- 230000004783 oxidative metabolism Effects 0.000 description 1
- 230000010421 oxidative-nitrosative stress Effects 0.000 description 1
- 108010071584 oxidized low density lipoprotein Proteins 0.000 description 1
- 230000000242 pagocytic effect Effects 0.000 description 1
- 229940073067 panoxyl Drugs 0.000 description 1
- FIKAKWIAUPDISJ-UHFFFAOYSA-L paraquat dichloride Chemical compound [Cl-].[Cl-].C1=C[N+](C)=CC=C1C1=CC=[N+](C)C=C1 FIKAKWIAUPDISJ-UHFFFAOYSA-L 0.000 description 1
- 239000013618 particulate matter Substances 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000007310 pathophysiology Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- BRWWKYJBIHJVNQ-UYXJWNHNSA-N phosphoric acid;(3s,4r)-3,4,5-trihydroxypentanal Chemical compound OP(O)(O)=O.OC[C@@H](O)[C@@H](O)CC=O BRWWKYJBIHJVNQ-UYXJWNHNSA-N 0.000 description 1
- 208000007578 phototoxic dermatitis Diseases 0.000 description 1
- 231100000018 phototoxicity Toxicity 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 108010040003 polyglutamine Proteins 0.000 description 1
- 229920000155 polyglutamine Polymers 0.000 description 1
- 229940093158 polyhexanide Drugs 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 208000037920 primary disease Diseases 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108010071870 propioxatin A Proteins 0.000 description 1
- 108010071872 propioxatin B Proteins 0.000 description 1
- 230000004224 protection Effects 0.000 description 1
- 230000022558 protein metabolic process Effects 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 239000012557 regeneration buffer Substances 0.000 description 1
- 230000028926 regulation of MAPK cascade Effects 0.000 description 1
- 230000029865 regulation of blood pressure Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000010410 reperfusion Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 229920002477 rna polymer Polymers 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 208000037921 secondary disease Diseases 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 210000000582 semen Anatomy 0.000 description 1
- 230000009758 senescence Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 208000013220 shortness of breath Diseases 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 231100000438 skin toxicity Toxicity 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000004071 soot Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 208000020431 spinal cord injury Diseases 0.000 description 1
- VEKPWANJVWWTMM-DYDSHOKNSA-N spinorphin Chemical class C([C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC=1[C]2C=CC=CC2=NC=1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C1=CC=C(O)C=C1 VEKPWANJVWWTMM-DYDSHOKNSA-N 0.000 description 1
- 201000010700 sporadic breast cancer Diseases 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000007863 steatosis Effects 0.000 description 1
- 231100000240 steatosis hepatitis Toxicity 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 239000003774 sulfhydryl reagent Substances 0.000 description 1
- 229960005559 sulforaphane Drugs 0.000 description 1
- 235000015487 sulforaphane Nutrition 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000000946 synaptic effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 108010013645 tetranectin Proteins 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 231100000588 tumorigenic Toxicity 0.000 description 1
- 230000000381 tumorigenic effect Effects 0.000 description 1
- 108010084595 tynorphin Proteins 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
- 238000010798 ubiquitination Methods 0.000 description 1
- 230000034512 ubiquitination Effects 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 208000014001 urinary system disease Diseases 0.000 description 1
- 230000006492 vascular dysfunction Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000002676 xenobiotic agent Substances 0.000 description 1
- 230000002034 xenobiotic effect Effects 0.000 description 1
- 229960001600 xylazine Drugs 0.000 description 1
- BPICBUSOMSTKRF-UHFFFAOYSA-N xylazine Chemical compound CC1=CC=CC(C)=C1NC1=NCCCS1 BPICBUSOMSTKRF-UHFFFAOYSA-N 0.000 description 1
- 239000011592 zinc chloride Substances 0.000 description 1
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
-
- 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
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/51—Complete heavy chain or Fd fragment, i.e. VH + CH1
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/515—Complete light chain, i.e. VL + CL
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/565—Complementarity determining region [CDR]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2318/00—Antibody mimetics or scaffolds
- C07K2318/20—Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
Definitions
- subject matter of the invention is a binder being directed to and binding to a dipeptidyl peptidase 3 (DPP3) protein or functional derivative thereof.
- DPP3 dipeptidyl peptidase 3
- the aforementioned binder is provided for the use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress.
- the aforementioned binder is provided for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress, and wherein said diseases are selected from a group comprising neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
- a binder is provided that is directed to and binding to an epitope according to SEQ ID NO.: 2, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 3, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 4, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1..
- binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- the above mentioned binder of the second major aspect of the invention that are directed to and binding to an epitope according to SEQ ID NO.: 2 is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- DPP3 dipeptidyl peptidase 3
- binder being directed to and binding to an epitope according to SEQ ID NO.: 2 according to the third aspect of the invention, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDRs) in the heavy chain comprises the sequences:
- binder being directed to and binding to an epitope according to SEQ ID NO.: 2 according to the third aspect of the invention, wherein said binder is a humanized monoclonal antibody or humanized monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- Additional subject matter of the invention is anyone of the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- DPP3 binder specifically a dipeptidyl peptidase 3 (hereinafter DPP3) binder, directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- DPP3 binder specifically a dipeptidyl peptidase 3 (hereinafter DPP3) binder, directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- a binder specifically a DPP3 binder, directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2, for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- aa amino acids
- subject matter of the present invention is a binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment, binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof or an anti-DPP3 non-Ig scaffold binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- aa amino acids
- a binder in accordance with the invention specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment, binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof or an anti-DPP3 non-Ig scaffold binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2, for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- aa amino acids
- Further subject matter of the present invention is a method of prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress, characterized in that a binder directed to and binding to DPP3, or a binder being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof is administered to said patient in pharmaceutically effective amounts.
- Subject matter of the present invention is further a pharmaceutical composition
- a pharmaceutical composition comprising a binder directed to and binding to DPP3, or a binder being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof for the use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Another subject of the present invention is a pharmaceutical composition
- a pharmaceutical composition comprising a binder of the invention, or a DPP3 binder in accordance with the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress as described above, and wherein said pharmaceutical composition comprises at least one additional pharmaceutically active drug which e.g.
- the said binder, or DPP3 binder, anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to DPP3 or the anti-DPP3 non-Ig scaffold binding to DPP3 may act as secondary medicament, which reduces or regulates the said induced oxidative stress.
- a further embodiment of the invention is a kit comprising a binder, or a DPP3 binder in accordance with the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress as described above, and wherein said pharmaceutical composition optionally comprises at least one additional pharmaceutically active drug which e.g.
- DPP3 binder, anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to DPP3 or the anti-DPP3 non-Ig scaffold binding to DPP3 may act as secondary medicament, which reduces or regulates the said induced oxidative stress.
- subject matter of the present invention is a binder in accordance with the invention, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 as described above for the use as a medicament, wherein said binder, or antibody or said antibody fragment or said non-Ig scaffold is a modulating binder, antibody or fragment or scaffold.
- said modulating anti-DPP3 antibody or a modulating anti-DPP3 fragment or a modulating non-Ig scaffold is used in the prevention or treatment of diseases or an acute condition in a patient, whereby said disease or acute condition is associated with oxidative stress.
- said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or a modulating non-Ig scaffold of the invention regulates the bioactivity of DPP3.
- DPP3 bioactivity may be defined as the DPP3 enzyme activity or the regulating activity of DPP3 in the oxidative stress pathway.
- said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or modulating non-Ig scaffold of the invention may enhance the bioactivity of DPP3.
- said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or modulating non-Ig scaffold of the invention may reduce the bioactivity of DPP3.
- a “modulating” anti-DPP3 antibody or a modulating anti-DPP3 antibody fragment or a modulating non-Ig scaffold as described above is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or a modulating anti-DPP3 non-Ig scaffold blocks the bioactivity of DPP3 at least 10 %, preferably at least 50 %, more preferably > 50 %, most preferably 100%.
- a modulating binder, or modulating anti-DPP3 antibody or a modulating anti-DPP3 antibody fragment or a modulating anti-DPP3 non-Ig scaffold according to the present invention is used for the prevention or treatment of diseases or an acute condition of a patient, wherein said disease or acute condition is associated with oxidative stress.
- kits or an assay comprising the above described binder, or anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress.
- DPP3 Dipeptidyl Peptidase 3
- Dipeptidyl peptidase 3 also known as Dipeptidyl aminopeptidase III, Dipeptidyl arylamidase III, Dipeptidyl peptidase III, Enkephalinase B or red cell angiotensinase; short name: DPP3, or DPPIII — is a metallopeptidase that removes dipeptides from physiologically active peptides, such as enkephalins and angiotensins.
- DPP3 will be used throughout the text as abbreviated form of the above described dipeptidyl peptidase 3.
- DPP3 was first identified and its activity measured in extracts of purified bovine anterior pituitary by Ellis & Nuenke, 1967.
- the enzyme which is listed as EC 3.4.14.4, has a molecular mass of about 83 kDa and is highly conserved in procaryotes and eucaryotes (Prajapati & Chauhan, 2011).
- DPP3 is a mainly cytosolic peptidase, which is ubiquitously expressed. Despite lacking a signal sequence, a few studies reported membranous activity (Lee & Snyder, 1982).
- DPP3 is a zinc-depending exo-peptidase belonging to the peptidase family M49. It has broad substrate specificity for oligopeptides from three or four to ten amino acids of various compositions and is also capable of cleaving after proline. DPP3 is known to hydrolyze dipeptides from the N-terminus of its substrates, including angiotensin II, III and IV; angiotensin 1-7 (Cruz-Diaz et al., 2016); Leu- and Met-enkephalin; endomorphin 1 and 2.
- the metallopeptidase DPP3 has its activity optimum at pH 8.0 — 9.0 and can be activated by addition of divalent metal ions, such as Co 2+ and Mg 2+ .
- Structural analysis of DPP3 revealed the catalytic motifs HELLGH (SEQ ID NO: 14) (hDPP3 450 — 455) and EECRAE (SEQ ID NO: 15) (hDPP3 507 — 512), as well as the following amino acids, that are important for substrate binding and hydrolysis: Glu 316, Tyr 318, Asp 366, Asn 391, Asn 394, His 568, Arg 572, Arg 577, Lys 666 and Arg 669 (Prajapati & Chauhan, 2011; Kumar et al., 2016; numbering refers to the sequence of human DPP3, see SEQ ID No.: 1). Considering all known amino acids or sequence regions that are involved in substrate binding and hydrolysis, the active site of human DPP3 can be defined as the region
- DPP3 has been also shown to be a promising biomarker in several publications. It has been shown that DPP3 activity is elevated in homogenates of ovarian and endometrial tumors. DPP3 activity even increases with the severity/malignancy of said tumors ( ⁇ imaga et al., 1998 and 2003). Immune histology and western blot analysis of glioblastoma cell lines also revealed elevated DPP3 levels (Singh et al., 2014).
- DPP3 was also proposed to be a potential arterio-risk marker (US 2011008805) and as a marker for rheumatoid arthritis (US 2006177886).
- the patent application WO 2005/106486 describes DPP3-expression and activity as diagnostic marker and DPP3 as therapeutic target in all kinds of diseases, due to ubiquitous expression of DPP3 in or at surface of cell.
- EP 1498480 mentions the potential diagnostic and therapeutic use of hydrolytic enzymes, including DPP3.
- WO 2005/106486 describes in a general manner a method of screening for therapeutic agents which may be useful in the treatment of diseases, comprising cardiovascular diseases, infections, respiratory diseases, cancer, endocrinological diseases, metabolic diseases, gastroenterological diseases, inflammation, haematological diseases, muscle skeleton diseases, neurological and urological diseases.
- a test compound is contacted with a DPP3 polynucleotide and the binding between said test compound and said DPP3 polynucleotide is detected.
- the document describes in a general manner compounds, which may bind to and / or activate or inhibit the activity of DPP3.
- the invention describes pharmaceutical compositions, which comprise such compounds.
- Hast et al. 2013 describe a comparison of the spectrum of KEAP1 interacting proteins with the genomic profile of 178 squamous cell lung carcinomas characterized by The Cancer Genome Atlas and reveal amplification and mRNA over-expression of the DPP3 gene in tumors with high Nrf2activity but lacking Nrf2 stabilizing mutations. They further describe that tumor-derived mutations in KEAP1 are hypomorphic with respect to Nrf2 inhibition and that DPP3 over-expression in the presence of these mutants further promotes Nrf2 activation.
- DPP3 intracellular DPP3 was identified as an activator of the antioxidant response element (ARE) in an unbiased screen of a cDNA library consisting of approximately 15,000 full-length human expression cDNAs (Liu et al. 2007).
- ARE antioxidant response element
- DPP3 disrupt the KEAP1-Nrf2 complex by competing with Nrf2 about the KEAP1 binding site (Hast et al. 2013). This disruption prevents NRF2 degradation and subsequently leads to translocalization of Nrf2 into the nucleus and ARE activation.
- Overexpression of DPP3 in neuroblastoma cells (Liu et al. 2007), in HEK293T cells (Hast et al. 2013) or in MCF7 breast cancer cells (Lu et al. 2017) activates Nrf2-mediated transcription. Active and inactive variants of DPP3 were overexpressed in MCF7 cells and showed the same regulatory effect on oxidative stress (Lu et al. 2017). Hast et al. (2013) also showed a loss-of-function effect: silencing of DPP3 using specific siRNA Nrf2-mediated transcription was decreased down to levels of Nrf2-silencing.
- DPP3 is known as an intracellular protein
- DPP3 activity was detected in some bodily fluids as well: retroplacental serum (Shimamori et al. 1986), seminal plasma (Vanha-Perttula et al. 1988) and CSF (Aoyagi et al. 1993).
- CSF cerebral spastic spasmodic spasmodic spasmodic spasmodic spasmodic s, and CSF (Aoyagi et al. 1993).
- AD Alzheimer’s disease
- DPP3 is known for being expressed as membranous, intracellular or circulating DPP3.
- DPP3 has been not only proposed as potential biomarker but also as potential therapeutic target due to its ability to cleave several bioactive peptides. Influenca A virus changes host DPP3 levels for own replication (cell culture studies, Meliopoulos et al. 2012). Enkephalin and/or angiotensin degrading enzymes in general, including DPP3, have a therapeutic potential as targets for treatment of pain, cardiovascular diseases (CVD) and cancer and the corresponding inhibitors as potential treatments of pain, mental illnesses and CVD (Khaket et al. 2012, Patel et al. 1993, Igic et al. 2007).
- CVD cardiovascular diseases
- DPP3 can be inhibited unspecifically by different general protease inhibitors (e.g. PMSF, TPCK), sulfhydryl reagents (e.g. pHMB, DTNB) and metal chelators (EDTA, o-phenantroline) (Abrami ⁇ et al. 2000, EP 2949332).
- general protease inhibitors e.g. PMSF, TPCK
- sulfhydryl reagents e.g. pHMB, DTNB
- EDTA metal chelators
- DPP3 activity can be further inhibited specifically by different kinds of compounds: an endogenous DPP3-inhibitor is the peptide spinorphin.
- a synthetic derivatives of spinorphin e.g. tynorphin, have been produced and shown to inhibit DPP3 activity to varying extents (Yamamoto et al. 2000).
- Other published peptide inhibitors of DPP3 are propioxatin A and B (US 4804676) and propioxatin A analogues (Inaoka et al. 1988).
- DPP3 can also be inhibited by small molecules such as fluostatins and benzimidazol derivatives.
- Fluostatins A and B are antibiotics produced in Streptomyces sp. TA-3391 that are non-toxic and strongly inhibit DPP3 activity.
- 20 different derivatives of benzimidazol have been synthesized and published (Agi ⁇ et al., 2007; Rastija et al., 2015), of which the two compounds 1′ and 4′ show the strongest inhibitory effect (Agi ⁇ et al., 2007).
- Several dipeptidyl hydroxamic acids have been shown to inhibit DPP3 activity as well (Cvite ⁇ i ⁇ et al., 2016).
- Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species (hereinafter ROS) / reactive nitrogen species (hereinafter RNS) and antioxidants in favour of excessive generation of free radicals.
- ROS reactive oxygen species
- RNS reactive nitrogen species
- This process leads to the oxidation of biomolecules with consequent loss of its biological functions and/or homeostatic imbalances, whose manifestation is the potential oxidative damage to cells and tissues.
- Accumulation of ROS / RNS can result in a number of deleterious effects such as lipid peroxidation, protein oxidation and DNA damage (including base damage and strand breaks).
- some reactive oxidative species act as cellular messengers in redox signalling.
- oxidative stress can cause disruptions in normal mechanisms of cellular signalling.
- ROS and RNS are the terms collectively describing free radicals and other non-radical reactive derivatives, which are also called oxidants. Radicals are less stable than non-radical species, although their reactivity is generally stronger. A molecule with one or more unpaired electron in its outer shell is called a free radical. Free radicals are formed from molecules via the breakage of a chemical bond such that each fragment keeps one electron, by cleavage of a radical to give another radical and, also via redox reactions. Free radicals related to oxidative stress include hydroxyl (OH•), superoxide (O 2 • - ), nitric oxide (NO•), nitrogen dioxide (NO 2 •), peroxyl (ROO•) and lipid peroxyl (LOO•).
- hydrogen peroxide H 2 O 2
- ozone O 3
- singlet oxygen 1 O 2
- hypochlorous acid HOCl
- nitrous acid HNO 2
- peroxynitrite ONOO -
- dinitrogen trioxide N 2 O 3
- lipid peroxide LOOH
- ROS and RNS can occur in the cells by two ways: enzymatic and non-enzymatic reactions.
- Enzymatic reactions generating free radicals include those involved in the respiratory chain, the phagocytosis, the prostaglandin synthesis and the cytochrome P450 system.
- Free radicals can be produced from non-enzymatic reactions of oxygen with organic compounds as well as those initiated by ionizing radiations.
- the non-enzymatic process can also occur during oxidative phosphorylation (i.e. aerobic respiration) in the mitochondria.
- oxidative stress is linked to a number of diseases, including but not limited to neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer and inflammation, and thus associated therewith.
- diseases including but not limited to neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer and inflammation, and thus associated therewith.
- DPP3 intracellular DPP3 is known to be closely linked to oxidative stress regulation.
- DPP3 was identified as an activator of the antioxidant response element (ARE) in an unbiased screen of a cDNA library consisting of approximately 15,000 full-length human expression cDNAs (Liu et al. 2007; see also above).
- ARE antioxidant response element
- This antioxidant effect of DPP3 is due to the interference of DPP3 with the KEAP1-Nrf2 signalling pathway.
- Nrf2 is a transcription factor that controls the basal and induced expression of an array of antioxidant response element-dependent genes to regulate the physiological and pathophysiological outcomes of oxidant exposure.
- Nrf2 is bound to Kelch like-ECH-associated protein 1 (KEAP1) via its ETGE (SEQ ID NO: 4) and its DLG motif.
- KEAP1 Kelch like-ECH-associated protein 1
- ETGE SEQ ID NO: 4
- DLG motif DLG motif
- Nrf2 Within this protein cluster Nrf2 is kept in the cytoplasm, quickly ubiquinated and degraded by proteasome. Under oxidative stress, Nrf2 is not degraded, but instead translocates to the nucleus where it binds to a DNA promoter and induces expression of an array of antioxidant response element (ARE)-dependent genes.
- ARE antioxidant response element
- oxidative stress can be also reduced or regulated by a binder directed to and binding to a DPP3 protein or functional derivative thereof.
- oxidative stress can be also reduced or regulated by a binder directed to and binding to an epitope of SEQ ID NO.: 2, wherein the epitope is comprised in the DPP3 protein.
- DPP3 dipeptidyl peptidase 3
- SEQ ID NO. 2 SEQ ID NO. 2
- said epitope is comprised in a DPP3 protein or a functional derivative thereof
- said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- oxidative stress may be reduced or regulated by an anti-DPP3 antibody or an anti DPP3-antibody fragment directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2 or an anti-DPP3 non-Ig scaffold directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- the present invention provides the herein disclosed binder, and anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in methods of the preventive treatment or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- binder DPP3 binder, and specifically the anti-DPP3 antibody, or anti DPP3-antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3, the inventors have found binder to DPP3 which rapidly reduce or regulate oxidative stress in cells of a mammal when determined by the methods of respective biomarker measurements as further set out below.
- Another subject of the present invention is a pharmaceutical composition
- a pharmaceutical composition comprising the binder of the invention, DPP3 binder of the invention, and specifically comprising the anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 of the invention for use in methods of the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- compositions are also provided for use in the prevention or treatment of symptoms, or syndromes, or pathological and acute conditions and disease associated problems, which are mediated by oxidative stress.
- oxidative stress is linked to a number of diseases or disorders, which in accordance of the invention include:
- Oxidative stress is suspected to be important in neurological and neurodegenerative disorders including Amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Depression, Multiple sclerosis, tardive dyskinesia (TD), epilepsy and acute diseases of the central nervous system, such as spinal cord injuries and/or brain traumatic.
- the human brain is vulnerable to oxidative stress due to many facts such as (i) metabolism of catecholamines; (ii) decrease in antioxidants; (iii) presence of transition metals; (iv) occurrence of brain trauma/injury; and also (v) the brain is an organ that proportionally requires more oxygen and (vi) expresses low levels of antioxidant enzymes, which contribute to formation of ROS.
- lipid membrane As a consequence of redox unbalance in brain, one of the most affected structures is the lipid membrane (Rao and Balachandran 2002. Nutritional Neuroscience 5: 291-309). A common feature of these diseases is oxidative damage of neurons, which might be responsible for the dysfunction or death of neuronal cells that contributes to disease pathogenesis.
- AD Alzheimer’s disease
- a ⁇ ⁇ -amyloid
- a ⁇ proteins can directly initiate free radical formation via the activation of NADPH oxidase.
- inflammation is responsible for increased expression of cytokines, ROS levels, and cellular toxicity, thereby exacerbating AD progression.
- HD Huntington’s disease
- CAG cytosine, adenine, guanine
- the expansion of CAG repeats within the exon1 of the HTT gene gives rise to a mutation that leads to the elongation of polyglutamine tract, resulting in an HTT protein product that is susceptible to aggregation.
- the mHTT aggregates are accumulated throughout the brain of the affected individuals, which can interrupt protein quality control and transcription process. Those alterations are potentially responsible for the aberrant motor and cognitive problems in HD. Though oxidative damage is not much reported in the early stages of HD, it is proposed as one of the major mechanisms in HD as it progresses.
- Elevated oxidative stress plays a critical role in the late stage of HD pathogenesis. Impairment in the electron transport chain and mitochondrial dysfunction are the major mechanisms involved in the ROS mediated etiopathogenesis of HD. Dysfunction in the oxidative phosphorylation components has been documented in the brain tissues of HD patients. HD patients showed an increased level of oxidative stress markers accompanied by a decrease in antioxidant status compared to healthy subjects. For review see Liu et al. 2017. Oxidative Medicine and Cellular Longevity 2525967; Manoharan et al. 2016. Oxid Med Cell Longev 8590578.
- Parkinson’s disease the most common neurodegenerative disease of the elderly, is characterized by progressive loss of muscle control. PD is predominant at the 6 th decade of life and men are 1.5 to 2 times more likely to contract the disease than women. Head trauma, illness, or exposure to environmental toxins is identified as a risk factor. This neurodegenerative disorder is characterized by tremor, rigidity, bradykinesia, and impairment in balance. PD also causes cognitive, psychiatric, autonomic, and sensory disturbances. The pathology of PD is characterized by the gradual and selective loss of dopaminergic neurons in the substantia nigra pars compacta. Imbalance in dopamine metabolism due to oxidative stress has been recognised as a contributor to this disease.
- the major pathological findings include the presence of Lewy bodies in the substantia nigra and loss of nerve cells in the portions of its ventral tier.
- Several studies have reported impaired respiratory chain and somatic mitochondrial DNA mutations in the brain of patients with PD, which suggests the extensive role of oxidative metabolism in PD.
- Enhanced dopamine metabolism in the brain of patients with PD could account for the accumulation of toxic radicals such as hydroxyl in the brain.
- Iron accumulation in the neurons in the redox active form plays a crucial role in pathogenesis of this disease.
- Amyotrophic lateral sclerosis is characterized by progressive loss of motor neurons in the anterior horn of the spinal cord. It is classified as either familial or sporadic depending on whether there is a clearly defined, inherited genetic element. Sporadic ALS (sALS) typically emerges between 50 and 60 years old. The onset of sALS is unknown, and thus the identification of causal genes and environmental factors remains elusive. In familial ALS, about 20% of the cases resulted from mutations in SOD1. The functions of SOD1 are diverse and include scavenging excessive superoxide radical, modulating cellular respiration, energy metabolism, and posttranslational modification. SOD dysfunction leads to a loss of antioxidant capability.
- MS Multiple sclerosis
- CNS central nervous system
- ROS reactive oxygen species
- Oxidative stress damages the mitochondria, which disrupts the transport of adenosine triphosphate along the axon, and consequently leads to neurodegeneration.
- Oxidative stress is associated with the dysregulation of axonal bioenergetics, cytokine-induced synaptic hyperexcitability, abnormal iron accumulation, and the oxidant/antioxidant balance. Markers of oxidative stress assessed in the serum, erythrocytes CSF, saliva, and urine may have diagnostic properties whereas antioxidants may have clinical application in the future. For review see Adamczvk and Adamczyk-Sowa 2016. Oxidative Medicine and Cellular Longevity 1973834.
- Oxidative stress is related to metabolic syndrome and its individual component pathologies, e.g. obesity, insulin resistance, dyslipidemia, impaired glucose tolerance and high blood pressure.
- the metabolic syndrome was defined by the World Health Organization criteria (Alberti and Zimmet 1998. Diabet Med . 15:539-553; World Health Organization. 1999. Definition, diagnosis and classification of diabetes mellitus and its complications: report of a WHO Consultation. Part 1: diagnosis and classification of diabetes mellitus.
- oxidative stress has emerged as playing a central role in metabolic syndrome and its component pathologies and may be a unifying factor in the progression of this disease. Moreover, oxidative stress has been identified as a major mechanism of micro- and macrovascular complications in the metabolic syndrome. For review see Hutcheson and Rocic 2012. Exp Diabetes Res . 2012:271028.
- oxidative stress plays an important role in the pathogenesis and development of cardiovascular diseases, including hypertension, dyslipidemia, atherosclerosis, myocardial infarction, angina pectoris, and heart failure (Elahi et al. 2009. Oxidative Medicine and Cellular Longevity 2(5): 259-269).
- One of the key concepts of free radical mediated pathogenesis of cardiovascular disease is endothelial dysfunction, whereby the regulation of vascular wall microenvironment is disrupted.
- ROS activity in the vessel wall for example, is thought to contribute to the formation of oxidized LDL, a major contributor to the pathogenesis of atherosclerosis.
- Oxidative stress also plays a role in the ischemic cascade due to oxygen reperfusion injury following hypoxia.
- This cascade includes both stroke (Chen et al. 2011. Antioxidants and Redox Signaling 14(8): 1505-1517) and myocardial infarction (MI) ( Hori and Nishida et al. 2009. Cardiovascular Research 81: 457-464).
- MI myocardial infarction
- brain ischemia/reperfusion multiple detrimental processes take place, including overproduction of oxidants, inactivation of detoxification systems, and consumption of antioxidants. These changes cause the disruption of the normal antioxidative defense ability of brain tissue (Chen et al. 2011. Antioxidants and Redox Signaling 14(8): 1505-1517).
- Oxidative Medicine and Cellular Longevity 2(5): 259-269 For further review see Elahi et al. 2009. Oxidative Medicine and Cellular Longevity 2(5): 259-269.
- Oxidative stress is thought to have an important role in the pathogenesis of autoimmune diseases. Many studies have shown that T and B lymphocytes contribute to the pathogenesis of autoimmune diseases by the production of autoantibodies and ROS under environmental and genetic influence. Oxidative stress has been implicated in autoimmune disorders (rheumatoid arthritis, systemic lupus erythematosus, psoriasis, and celiac disease) where it plays an important role in the disease process. Oxidative stress is increased in systemic lupus erythematosus (SLE), and it contributes to immune system dysregulation, abnormal activation and processing of cell-death signals, autoantibody production and fatal comorbidities.
- SLE systemic lupus erythematosus
- RA Rheumatoid arthritis
- RA is an autoimmune disease characterized by chronic inflammation of the joints and tissue around the joints with infiltration of macrophages and activated T cells. The pathogenesis of this disease is due to the generation of ROS and RNS at the site of inflammation. RA is one of the conditions that induce oxidative stress.
- oxidative stress is a pathogenic hallmark in RA.
- Free radicals are indirectly implicated in joint damage because they also play an important role as secondary messengers in inflammatory and immunological cellular response in RA. T-cell exposure to increased oxidative stress becomes refractory to several stimuli including those for growth and death and may perpetuate the abnormal immune response.
- free radicals can degrade directly the joint cartilage, attacking its proteoglycan and inhibiting its synthesis (for review see Qui ⁇ onez-Flores et al. 2016. Biomed Res Int . 2016:6097417).
- inflammatory lung diseases such as asthma and chronic obstructive pulmonary disease (COPD) are characterized by systemic and local chronic inflammation and oxidative stress.
- An important source for increased airway oxidative stress is the recruitment of inflammatory cells into the airway after exposure to trigger factors. These activated cells can generate anion superoxide through reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathway.
- NADPH nicotinamide adenine dinucleotide phosphate
- Mitochondrial dysfunction in airway epithelial cells which occurs in response to mechanical and environmental stimuli, can also contribute to the formation of anion superoxide and airway oxidative stress.
- Subjects with asthma have greater systemic and airway increased oxidative stress, which is associated with worse asthma severity.
- IBD Inflammatory bowel disease
- GI gastrointestinal
- Crohn’s disease CD
- ulcerative colitis UC
- CD Crohn’s disease
- UC ulcerative colitis
- immune cells such as leukocytes, monocytes, and neutrophils augment ROS production during respiratory, prostaglandin, and leukotriene metabolism, resulting in further tissue damages.
- Celiac disease is an immune-mediated chronic inflammatory disorder of the upper small intestine induced by gluten and related prolamines in genetically susceptible individuals. As in other autoimmune conditions, environmental, genetic, and immunological factors may be involved in the pathogenesis of CD. In addition to this, oxidative stress is also implicated in the pathogenesis of CD. For example, activation of xanthine oxidase is one of the mechanisms of ROS overproduction in small intestinal mucosa of celiac patients. For review see Patlevic et al. 2016. Integr Med Res 5: 250-258.
- Gastritis is defined as inflammation of the stomach mucosal lining and occurs in several conditions including H. pylori infection, NSAID use, alcohol consumption, and stress.
- Peptic ulcer disease occurs in the proximal GI tract and is often associated with chronic gastritis. Gastric and duodenal ulcers represent the most common and chronic PUDs. Gastritis and peptic ulcer are caused by multiple factors, both endogenous and exogenous, and free radicals are closely linked to both conditions.
- ROS ROS
- Reduced antioxidant enzyme SOD levels and antioxidant vitamin intake contribute to the accumulation of ROS associated with gastroduodenal inflammatory diseases. Ethanol-induced gastric inflammation is associated with increased superoxide generation.
- Phagocytic leukocytes are the main source of ROS in chronic inflammation such as one observes in H. pylori induced gastritis and IBD. Significant numbers of neutrophils and/or macrophages infiltrate the gastric mucosa during inflammation, generating large amounts of ROS. For review see Bhattacharyya et al. 2014. Physiol Rev 94: 329-354.
- Oxidative stress also plays a critical role in liver diseases like viral Hepatitis (Type A, B and C) and liver cirrhosis. It has been clearly established that hepatitis C is associated with strong oxidative stress. This was revealed in liver tissues and in blood serum/plasma samples of Cchronic hepatitis C patients using a variety of techniques, including direct measurement of ROS, quantification of DNA, lipid and protein oxidation products, as well as by assessing the total oxidant/antioxidant status or the levels of individual antioxidants.
- liver biopsies of chronic hepatitis C virus carriers revealed significant elevation of the levels of oxygen radicals and stress markers malondialdehyde (MDA) and 4-hydroxynonenal- (HNE)-and other protein adducts.
- MDA malondialdehyde
- HNE 4-hydroxynonenal-
- serum/plasma of such patients is characterized by increased levels of a wide array of oxidative stress markers such as MDA, lipid peroxides, protein carbonyl content or thioredoxin (for review see Ivanov et al. 2017. Oncotarget , 2017, Vol. 8, (No. 3), pp: 3895-3932).
- Patients with chronic hepatitis B exhibit signs of pronounced oxidative stress. Levels of oxygen radicals in liver specimens from these patients exceed the levels in healthy people. Patients with hepatitis B exhibit signs of oxidative stress not only in the liver but also in plasma/sera. Chronic hepatitis B is accompanied by an increase in total oxidant status and a concomitant reduction of total antioxidant status. Plasma/serum of these patients was also characterized by the elevated levels of ROS, including H 2 O 2 , and oxidation products of lipids and proteins. Oxidative stress is not just a hallmark of chronic HBV infection and advanced liver disease; it is also observed in acute and occult hepatitis B, as well as in asymptomatic HBV infections.
- Occult hepatitis B infection is characterized by increased levels of ROS in lymphocytes and consequent DNA damage.
- the most dramatic changes have been described in hepatitis B patients with liver cirrhosis and with acute chronic hepatitis B liver failure (for review see Ivanov et al. 2017. Oncotarget , 2017, Vol. 8, (No. 3), pp: 3895-3932).
- Cirrhosis is a complication of many forms of chronic liver diseases and is a late stage of fibrosis, in which regenerative nodular formation surrounded by fibrous bands of the liver.
- oxidative stress induced mainly by an overproduction of reactive oxygen species, which is a critical determinant of endothelial dysfunction and is due to disturbed balance between oxidant and antioxidant enzymes.
- Increased superoxide formation in the presence of equimolar concentrations of NO will lead to the formation of the potent ROS and reactive nitrogen species.
- Hepatotoxicity implies chemical-driven liver damage. Drug-induced liver injury is a cause of acute and chronic liver disease. Drug-induced liver injury is responsible for 5% of all hospital admissions and 50% of all acute liver failures. The liver is the most frequently targeted organ in terms of drug toxicity.
- the production of radical species, specifically ROS and RNS, has been proposed as an early event of drugs hepatotoxicity and as an indicator of hepatotoxic potential. It has been discovered that a lot of drugs could induce oxidative stress including increase of cellular oxidants and lipid peroxidation, depletion of antioxidants in the liver, such as anti-inflammation drugs, anti-analgesic drugs, anti-cancer drugs and antidepressants (Li et al. 2015. Int. J. Mol. Sci .
- CKD chronic kidney disease
- ESRD end stage renal disease
- TRX1 urinary thioredoxin 1
- AKI acute kidney injury
- AKI drugs-induced kidney injury
- AKI causes a severe condition associated with high probabilities of developing progressive chronic kidney disease or end-stage renal disease, thus leading to high mortality rates.
- Most drugs found to cause nephrotoxicity exert toxic effects by one or more common pathogenic mechanisms. These include altered intraglomerular hemodynamics, tubular cell toxicity, inflammation, crystal nephropathy, rhabdomyolysis, and thrombotic microangiopathy.
- AKI includes acute tubular necrosis (ATN) and acute interstitial nephritis (AIN).
- a mechanism underlying ATN is oxidative stress.
- Proximal tubular toxicity develops due to direct nephrotoxic effects such as mitochondrial dysfunction, lysosomal hydrolase inhibition, phospholipid damage, and increased intracellular calcium concentration, leading to formation of reactive oxygen species (ROS) with injurious oxidative stress (Hosohata 2016. Int. J. Mol. Sci. 17: 1826).
- ROS reactive oxygen species
- Medications that are potentially harmful to the kidneys are, e.g. antimicrobials like antibiotics (for example streptomycin, gentamicin) or antivirals (for example acyclovir, foscarnet) or antifungal (for example amphotecerin B), analgesics, non-steroidal anti-inflammatory drugs (NSAID) (for example ibuprofen, naproxen), diuretics, proton pump inhibitors, chemotherapeutics (for example cisplatin), contrast dyes, cardiovascular agents like ACE-inhibitors or statins, anti-depressants, immune suppressants (for example cyclosporine A) and antihistamines (for reference see Naughton 2008. Am Fam Physician . 2008;78(6):743-750, Table 1; Hosohata 2016. Int. J. Mol. Sci . 17: 1826).
- antimicrobials like antibiotics (for example streptomycin, gentamicin) or antivirals (for example acyclovir,
- ROS generated by the respiratory chain in the mitochondria and by the Nox enzymes in the cytoplasm are particularly important.
- Nox proteins are now considered to be oncogenic proteins, and mitochondrial dysfunction is associated with tumorigenesis.
- Oxidative stress is involved in all stages of carcinogenesis and there is a dose-dependent association between level of the persistent or chronic oxidative stress and the tumor stage.
- the carcinogenesis process the normal cells are transformed into abnormal cells owing to a number of structural changes and mutations in genes expression.
- carcinogenesis is described by three main stages: initiation, promotion and progression. All these stages have been postulated to be linked with contribution of ROS and RNS.
- ROS have an important role in the pathophysiological states involved in neovascularization.
- ROS-generating enzymes such as NADPH oxidases (e.g., Nox: Nox1-5), activate redox signaling pathways that ultimately lead to angiogenesis.
- NADPH oxidases e.g., Nox: Nox1-5
- Prostate cancer is the most frequently diagnosed non-cutaneous malignancy in males. This is a multi-focal, filed-type disease, which forms solid tumors of glandular origin. Prostate cancer is mainly a disease of aging, with most cases occurring in men over the age of 55. Over the last decade association between prostate cancer risk and oxidative stress has been recognized, and epidemiological, experimental and clinical studies have unequivocally proven a role for oxidative stress in the development and progression of this disease, commonly associated with a shift in the antioxidant-prooxidant balance towards increased oxidative stress.
- the excessive production of ROS in breast cancer cells include: a strong expression of thymine phosphorylase leading to degradation of thymidine to thymine and 2-deoxy-D-ribose phosphate; oxidation of 17-estradiol panoxyl radicals to lactoperoxidase participating in metabolism of estrogens and inflammation.
- antioxidant changes are related to breast cancer risk, for example the levels of SOD and GPX were found to be higher in the blood of breast cancer cases compared to that of healthy women as a reply on the increased production of superoxide and hydrogen peroxide.
- the tumor suppressor gene breast cancer gene 1 (BRCA1) is mutated in 40-50% of hereditary breast cancers and absent or expressed at low levels in 30-40% of sporadic breast cancers.
- BRCA1 is a caretaker gene that is responsible for repairing DNA, and it is able to upregulate several genes involved in the antioxidant response by controlling the activity of the transcription factors Nrf2 and Nf ⁇ B.
- Nrf2 and Nf ⁇ B Apart from the inhibitory action of BRCA1 on ROS generation, BRCA1 also reduces the levels of protein nitration due to RNS accumulation in cells, and it enhances DNA repair processes that ultimately help to cope with oxidative stress.
- respirable particles or fibrous dusts to penetrate the respiratory system and reach the lung alveoli in order to generate ROS and other oxidants or free radicals is suggested to be the main factor involved in their pathogenic potential.
- Synergistic mechanisms of inhalable particulate matter (penetrating deep into the lung’s alveoli) and other components of air pollution (ozone, nitric oxide, soot, heavy metals, PAHs) and tobacco smoke have been studied.
- the porous surfaces of airborne particles provide a fertile ground for catalyzing the increased generation of ROS or other damaging oxidants, which are potential initiators of pulmonary carcinogenesis.
- Colorectal cancer is one of the most common cancers worldwide, with the highest incidence rates in western countries. Colon cancer originates from the epithelial cells that line the bowel. These cells divide rapidly and have a high metabolic rate, which has been found as a potential factor that may be responsible for increased oxidation of DNA. It was found that the human colorectal tumors (adenomasand carcinomas) have increased levels of different markers of oxidative stress, such as increased levels nitric oxide (NO), 8-oxodG in DNA, lipid peroxides, glutathione peroxidase (GPx), catalase (CAT), and decreased methylation of cytosine in DNA.
- NO nitric oxide
- 8-oxodG lipid peroxides
- GPx glutathione peroxidase
- CAT catalase
- bladder cancer is the fourth most frequently occurring malignant tumors. Recent studies indicate the involvement of oxidative and nitrosative stress in the formation and development of this disease. Red-ox disorders are characteristic for both, the initiation and progression of bladder cancer. There are observed changes in the activity of transcription factors, such as nuclear factor NF-kB; transcription factors: AP-1, Nrf2 and STAT3 and hypoxia-inducible factor HIF-1 ⁇ . In addition, studies indicate a role for oxidative stress in the regulation of MAPK cascade and its involvement in carcinogenesis consisting bladder. Nitric oxide also plays an important role in tumor biology. Numerous studies show that the bladder cancer is characterized by an intensified production of NO.
- Ovarian cancer is the fifth leading cause of cancer death; the leading cause of death from gynecologic malignancies, and the second most commonly diagnosed gynecologic malignancy.
- the overwhelming majority of ovarian cancers are derived from ovarian surface epithelium. Metastasis is achieved through detachment of single cells or clusters of cells from the primary tumor followed by implantation on peritoneal mesothelial lining.
- Ovarian, endometrial, and cervical cancer consist a great problem in oncology due to their diagnosis in advanced stage. Research finding have shown that oxidative stress plays a causal role in the carcinogenesis of two subtypes of ovarian cancer: clear cell carcinoma and endometriosis carcinoma.
- EOC epithelial ovarian cancer
- Endometrial cancer has been reported to be associated with endometriosis disease, and the high levels of free iron hemosiderin or heme in endometrial cysts are considered as a main factor responsible for the oxidative stress development and chronic inflammation.
- Cervical cancer is the second most common cancer in women worldwide being a subject of intensive research.
- oxidative stress in cervical, indicating that antioxidants can alter the redox balance in cervical cancer cells, inhibit transcription factors AP-1 and NF- ⁇ B or induce cell apoptosis.
- For review see Saed et al. 2017. Gynecologic Oncology 145: 595-602; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- Oxidative damage induced by OS has been also implicated in leukemia and the decreased levels of antioxidants and oxidatively modified DNA and lipids caused by high ROS production were found in serum of chronic lymphocytic leukemia patients. Moreover, it was found that the chronic leukemia cells were able to adapt to intracellular OS through upregulation the stress-responsive hemeoxygenase-1 confirming involvement of ROS in the pathogenesis of leukemia cancer. Also, GSH depletion in lymphocytes of the chronic lymphocytic has been demonstrated in leukemia B patients. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919.
- Gastric cancer is one of the most frequent diseases in human population. It is the fourth frequent cancer and the second most common cause of deaths from cancer in the world.
- the main risk factor for gastric cancer is chronic inflammation caused by bacterial growth. For example, infection by Helicobacter pylori, which increases the production of reactive oxygen and nitrogen species in human stomach, is thought to be important in the development of gastric cancer. It has been shown that protein oxidation products were significantly higher in GC patients. Moreover, it was found that the antioxidant potentials of SOD and catalase were lower in gastric cancer tissues compared to the control healthy tissues. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17 : 904-919; Ma et al. 2013. Oxidative Medicine and Cellular Longevity 543760.
- liver carcinogenesis There are many factors involved in liver carcinogenesis, including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection, alcohol abuse, and nonalcoholic fatty liver disease (NAFLD), aflatoxin B1, obesity, diabetes, dietary habits, and iron accumulation.
- HBV hepatitis B virus
- HCV hepatitis C virus
- NAFLD nonalcoholic fatty liver disease
- oxidative stress can be triggered by any dangerous or inflammatory signal and affects multiple cells in the liver.
- Liver injury can be either an acute or a chronic inflammatory process.
- liver sinusoidal endothelial cells (LSECs), hepatic stellate cells (HSCs), dendritic cells (DCs), and Kupffer cells (KCs) are activated.
- HBV- and HCV-related chronic inflammation and fibrosis of the liver are usually induced by oxidative stress, which contributes to the pathogenesis of hepatocarcinogenesis.
- HBV infection results in activation of macrophages to produce a variety of proinflammatory cytokines, such as IL-1 ⁇ , IL-6, CXCL-8, and TNF- ⁇ .
- cytokines such as IL-1 ⁇ , IL-6, CXCL-8, and TNF- ⁇ .
- HCV-induced oxidative stress contributes to the development of hepatocellular carcinoma (HCC).
- oxidative stress markers in chronic hepatitis C patients correlate positively with the probability of development of HCC and can serve as prognostic markers for HCC recurrence in chronic hepatitis C patients who underwent liver transplantation.
- Carcinogenesis is orchestrated by multiple ROS-mediated processes. For review see Wang et al. 2016. Oxidative Medicine and Cellular Longevity 7891574.
- Skin is a major environmental interface for the body, which accidentally or occupationally gets exposed to a number of chemical mutagens and carcinogens.
- Skin cancer represents a major and growing public health problem. It accounts for more than 40 % of all new cancer diagnosed. 80 % of skin cancers result from basal cell carcinomas (BCC); another 16 % are squamous cell carcinomas (SCC), and 4 % are melanomas.
- BCC basal cell carcinomas
- SCC squamous cell carcinomas
- melanomas melanomas.
- An important process in skin cancer is generation of hydrogen peroxide by melanocytes and decrease in catalase activity.
- mutations in several genes linked with melanoma result from oxidative stress. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17 : 904-919; Narendhirakannan. 2013. Ind J Clin Biochem 28(2):110-115.
- Reactive species generated during infection may have serious consequences for the disease once they are released to any degree.
- the oxidative stress can initiate adverse effects in different organs. Development of oxidative stress can be accelerated in the course of hypoxia. Hypoxia is a known complication of infectious diseases. Hypoxia is not peculiar to one disease. Influenza, viral hepatitis and tuberculosis are all examples of infectious diseases in which hypoxia takes place.
- Direct generation of reactive oxygen species can be initiated by metals.
- iron, cooper, and cadmium can catalyze the development of oxidative stress by Fenton reaction in which hydrogen peroxide is converted into hydroxyl radical and hydroxide anion. Heavy metals are involved in pathological processes linked to infections like other pathologies.
- livers damaged by viral hepatitis are vulnerable to heavy metals due to faulty elimination processes.
- Clinical studies on patients with viral hepatitis A, B, C, D, and E demonstrated that accumulation of copper and iron caused oxidative stress and oxidative damage to patients’ liver tissue.
- AIDS is accompanied by imbalance in oxidative homeostasis. Elevated markers of oxidative damage of targets in the body and accumulation of reactive oxygen species are common in HIV-infected patients. Blood antioxidants are reduced over the long term in the infected individuals. For review see Pohanka 2013. Folia Microbiol 58:503-513.
- ROS reactive oxygen species
- NO nitric oxide
- peroxynitrite peroxynitrite
- ROS and RNS During sepsis, excess production of ROS and RNS threatens the integrity of various biomolecules including proteins, lipids as well as lipoproteins, protein oxidation and DNA resulting in tissue damage, by lipid peroxidation of cell membranes, protein oxidation and DNA strand breaks. These mechanisms contribute to multi organ failure during sepsis resulting in myocardial depression, hepatocellular dysfunction, endothelial dysfunction, and vascular catecholamine hypo-responsiveness. As a major source of ROS production, mitochondria are especially prone to ROS-mediated damage. Such damage can induce the mitochondrial permeability transition caused by opening of nonspecific high conductance permeability transition pores in the mitochondrial inner membrane.
- ROS themselves also provide a signal leading to the induction of autophagy, apoptosis, and necrosis.
- Excessive ROS production and adenosine triphosphate depletion from uncoupling of oxidative phosphorylation promote necrotic cell death.
- Release of cytochrome-c after mitochondrial swelling activates caspases and initiates apoptotic cell death.
- the herein disclosed DPP3 binder specifically the herein provided anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold which are binding to an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof are provided for the use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress, said disease is selected from the group comprising the above described neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
- said disease is selected from the group comprising neurodegenerative diseases (e.g. Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)), metabolic syndrome (including insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes), cardiovascular disorders (e.g. atherosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury/ stroke and myocardial infarction), autoimmune diseases (e.g. rheumatoid arthritis and systemic lupus erythematosus), inflammatory lung diseases (e.g.
- neurodegenerative diseases e.g. Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)
- metabolic syndrome including insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes
- cardiovascular disorders e.g. at
- COPD chronic kidney disease
- CKD chronic kidney disease
- ESRD end-stage renal disease
- liver diseases e.g. hepatotoxicity, viral hepatitis, cirrhosis
- digestive diseases including inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease; gastritis, pancreatitis and peptic ulcer
- viral infectious diseases e.g. blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus)
- cancer e.g. prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer
- inflammation sepsis, septic shock and SIRS.
- the herein disclosed DPP3 binder specifically the herein provided anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold which are binding to an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof are provided for the use in the prevention or treatment of an acute condition, wherein said acute condition may be selected from a group comprising hepatotoxicity and kidney toxicity.
- oxidative stress and subsequent toxicities can also be induced by chronic alcohol consumption, chronic exposure to cigarette smoke and as a side-effect of different drug treatments (reviewed in Deavall et al. 2012, see table 2 below).
- Acetaminophen a widely used analgesic and antipyretic drug — is a prototypical hepatotoxicant for drug-induced liver injury that is connected to KEAP-Nrf2 signaling (Ma, 2013).
- Other therapeutics inducing oxidative stress include oltipraz and auranofin (Ma, 2013).
- oxidative stress may be determined and quantified by suitable biomarker assays known in the art. Respective examples for these markers are given below, but these shall be not construed as limiting possibilities to measure oxidative stress in accordance with the invention:
- Free radicals can damage biological molecules including nucleic acids, proteins, and lipids.
- the products of these reactions can become markers of oxidative stress.
- Serum is the most common material for the evaluation of the components of oxidative stress. It allows the estimation of most enzymes, substrates, and products of redox reactions. These enzymes include xanthine oxidase, NOS, lipoxygenase, cyclooxygenase, myeloperoxidase, prolyl-oligopeptidase, nicotinamide adenine dinucleotide phosphate-oxidase 1 (NOX1), and NADPH-dependent oxidase.
- isoprostanes IsoP-prostaglandin like substances
- 8-iso-prostaglandin F2 ⁇ -8-iso-PGF2 ⁇
- MDA malondialdehyde
- Oxidative stress involves the oxidation of proteins and glycoxidation.
- the glycophore content the total level of advanced protein oxidation (AOPP), protein carbonyls, dityrosine level, N′-formylkynurenine, and a decreased level of serum protein thiol groups.
- Other specific markers of protein oxidation are tyrosine (a marker for hydroxyl radical) and 3-nitrotyrosine (a marker for RNS).
- 3-nitrotyrosine is a specific marker of peroxynitrite-induced cellular damage.
- Other indicators in the serum include kynurenine, N′-formylkynurenine, thioredoxin, and 8-hydroxy-2′-deoxyguanosine.
- Inhibition of DPP3 activity in a liquid phase assay by a binder may be determined as followed: Blood samples (e.g. serum, heparin-plasma, Li-plasma, citrate-plasma, whole blood) of patients before and after anti-DPP3 antibody treatment is incubated with specific DPP3 substrates in a liquid phase assay.
- the specific liquid phase DPP3 activity assay to determine the inhibitory ability of inhibitory DPP3 antibodies in blood samples comprises the following steps:
- a solid phase assay is an assay where the respective binding events take place at the solid phase.
- Inhibition of DPP3 activity in a solid phase assay by a binder may be determined as followed according: Blood samples (e.g. serum, plasma, whole blood) of patients before and after anti-DPP3 antibody treatment are contacted with an immobilized capture-binder for enzyme capture activity assay (ECA) on a solid phase.
- ECA enzyme capture activity assay
- as capture-binder for the ECA is chosen the one with the least inhibitory ability.
- the capture-binder should inhibit DPP3 activity less than 50 %, preferably less than 40 %, preferably less than 30 %.
- the specific liquid phase DPP3 activity assay to determine the inhibitory ability of possible capture-binders is described in detail in Example 1 below.
- the ECA to determine the inhibitory ability of inhibitory DPP3 antibodies in blood samples comprises the following steps:
- the method for determining active DPP3 may be conducted as liquid phase assay and as solid phase assay. Inhibition of DPP3 activity may be determined in a liquid assay nevertheless according to the above-described procedure.
- a capture or binding assay may be performed to detect and/or quantitate the protease activity of DPP3.
- an antibody reactive with DPP3 protein, but which does not interfere with peptidase activity may be immobilized upon a solid phase.
- the test sample is passed over the immobile antibody, and DPP3, if present, binds to the antibody and is itself immobilized for detection.
- a substrate may then be added, and the reaction product may be detected to indicate the presence or amount of DPP3 in the test sample.
- solid phase may used to include any material or vessel in which or on which the assay may be performed and includes, but is not limited to, porous materials, nonporous materials, test tubes, wells, slides, etc.
- the binding affinity of the herein disclosed DPP3 binder to DPP3 may be measured by various suitable assays known in the art. Respective examples are given below, but these shall be not construed as limiting possibilities to measure binding affinity of the herein disclosed DPP3 binder to DPP3:
- the binding affinity of the DPP3 binder to the epitope according to SEQ ID NO.: 2 in accordance with the invention may be determined in accordance with Example 1 and as further set out below:
- a binding assay may be performed to detect and/or quantitate antibody binding to the immunization peptide (i.e. SEQ ID NO.: 2).
- this immunization peptide may be immobilized upon a solid phase.
- the test sample e.g. antibody solution
- the term “solid phase” may be used to include any material or vessel in which or on which the assay may be performed and includes, but is not limited to, porous materials, nonporous materials, test tubes, wells, slides, etc.
- the herein disclosed binder of the invention, and DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or anti-DPP3 non-Ig scaffolds are capable to bind circulating DPP3, and thus are directed against circulating DPP3.
- the herein disclosed binder of the invention, and DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or antiDPP3 non-Ig scaffolds are capable to bind intracellular DPP3, and thus are directed against intracellular DPP3.
- the herein disclosed binder of the invention DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or anti-DPP3 non-Ig scaffolds are capable to bind membranous DPP3, and thus are directed against membranous DPP3.
- DPP3 binder specifically is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress
- said binder, DPP3 binder specifically is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in a circulating DPP3 protein or functional derivative thereof.
- binder of the invention DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress, and whereby said binder, DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in an intracellular DPP3 protein or functional derivative thereof.
- DPP3 binder specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress
- said DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in a membranous DPP3 protein or functional derivative thereof.
- Subject matter of the present invention is further a method for regulating and/or preventing or treatment of oxidative stress in a patient having a chronic or acute disease or acute condition, characterized in that to said patient a binder of the invention, or a DPP3 binder of the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold is administered in pharmaceutically effective amounts.
- said patient is a patient in need of regulating and/or preventing or in need of treatment of oxidative stress.
- Another subject of the present invention is a pharmaceutical composition
- a pharmaceutical composition comprising the herein disclosed binder of the invention, or DPP3 binder, specifically comprising an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions of a patient, wherein said disease or acute condition is associated with oxidative stress.
- said pharmaceutical composition is a solution, preferably a ready-to-use solution.
- said pharmaceutical composition is a solution, preferably a ready-to-use solution comprising PBS at a pH of 7.4.
- said pharmaceutical composition is in a dried state that is to be reconstituted before use.
- said pharmaceutical composition is in a freeze-dried state that is to be reconstituted before use.
- said pharmaceutical composition that is to be used in the prevention and/or treatment of a disease or an acute condition of a patient, wherein said disease or acute condition is associated with oxidative stress is administered orally, epicutaneously, subcutaneously, intradermally, sublingually, intramuscularly, intraarterially, intracerebrally, intracerebroventricularly, intravenously, or via the central nervous system (CNS) or via intraperitoneal administration.
- CNS central nervous system
- kits or an assay comprising the herein disclosed binder of the invention, or DPP3 binder, specifically comprising an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress.
- an “anti-DPP3 antibody” is an antibody that binds specifically to DPP3
- an “anti-DPP3 antibody fragment” is a fragment of said anti-DPP3 antibody, wherein said fragment binds specifically to DPP3.
- An “anti-DPP3 non-Ig scaffold” is a non-Ig scaffold that binds specifically to DPP3.
- DPP3 specifically binding to DPP3 may also allow binding to other antigens as well. This means, this specificity would not exclude that the binder may cross-react with other proteins or polypeptides or peptides that contain the epitope according to SEQ ID NO.: 2 against which the binder has been raised. This specifically includes functional variants of DPP3, which also comprise an epitope according to SEQ ID NO.: 2. This also pertains to the specificity of the anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold in accordance with the invention.
- an “antibody” is a protein including one or more polypeptides substantially encoded by immunoglobulin genes that specifically binds an antigen.
- the recognized immunoglobulin genes include the kappa, lambda, alpha (IgA), gamma (IgG 1 , IgG 2 , IgG 3 , IgG 4 ), delta (IgD), epsilon (IgE) and mu (IgM) constant region genes, as well as the myriad immunoglobulin variable region genes.
- Full-length immunoglobulin light chains are generally about 25 kDa or 214 amino acids in length.
- Full-length immunoglobulin heavy chains are generally about 50 kDa or 446 amino acids in length.
- Light chains are encoded by a variable region gene at the NH 2 -terminus (about 110 amino acids in length) and a kappa or lambda constant region gene at the COOH-terminus.
- Heavy chains are similarly encoded by a variable region gene (about 116 amino acids in length) and one of the other constant region genes.
- the basic structural unit of an antibody is generally a tetramer that consists of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions bind to an antigen, and the constant regions mediate effector functions.
- Immunoglobulins also exist in a variety of other forms including, for example, Fv, Fab, and F(ab′) 2 , as well as bifunctional hybrid antibodies and single chains (e.g., Lanzavecchia et al., Eur. J. Immunol . 17:105,1987; Huston et al., Proc. Natl. Acad. Sci.
- An immunoglobulin light or heavy chain variable region includes a framework region interrupted by three hypervariable regions, also called complementarity determining regions (CDR’s) (see, Sequences of Proteins of Immunological Interest, E. Kabat et al., U.S. Department of Health and Human Services, 1983). As noted above, the CDRs are primarily responsible for binding to an epitope of an antigen.
- An immune complex is an antibody, such as a monoclonal antibody, chimeric antibody, humanized antibody or human antibody, or functional antibody fragment, specifically bound to the antigen.
- “Chimeric antibodies” are antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from immunoglobulin variable and constant region genes belonging to different species.
- the variable segments of the genes from a mouse monoclonal antibody can be joined to human constant segments, such as kappa and gamma 1 or gamma 3.
- a therapeutic chimeric antibody is thus a hybrid protein composed of the variable or antigen-binding domain from a mouse antibody and the constant or effector domain from a human antibody, although other mammalian species can be used, or the variable region can be produced by molecular techniques. Methods of making chimeric antibodies are well known in the art, e.g., see U.S. Pat. No. 5,807,715.
- a “humanized” immunoglobulin is an immunoglobulin including a human framework region and one or more CDRs from a non-human (such as a mouse, rat, or synthetic) immunoglobulin.
- the non-human immunoglobulin providing the CDRs is termed a “donor” and the human immunoglobulin providing the framework is termed an “acceptor.”
- all the CDRs are from the donor immunoglobulin in a humanized immunoglobulin. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, such as about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDRs, are substantially identical to corresponding parts of natural human immunoglobulin sequences.
- a “humanized antibody” in accordance with the invention is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin.
- a humanized antibody binds to the same antigen as the donor antibody that provides the CDRs.
- the acceptor framework of a humanized immunoglobulin or antibody may have a limited number of substitutions by amino acids taken from the donor framework.
- Humanized or other monoclonal antibodies can have additional conservative amino acid substitutions which have substantially no effect on antigen binding or other immunoglobulin functions. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, glu; asn, gln; ser, thr; lys, arg; and phe, tyr.
- Humanized immunoglobulins can be constructed by means of genetic engineering (e.g., see U.S. Pat. No. 5,585,089).
- a human antibody is an antibody wherein the light and heavy chain genes are of human origin.
- Human antibodies can be generated using methods known in the art. Human antibodies can be produced by immortalizing a human B cell secreting the antibody of interest. Immortalization can be accomplished, for example, by EBV infection or by fusing a human B cell with a myeloma or hybridoma cell to produce a trioma cell. Human antibodies can also be produced by phage display methods (see, e.g., Dower et al., PCT Publication No.
- Human antibodies can also be prepared by using transgenic animals carrying a human immunoglobulin gene (for example, see Lonberg et al., PCT Publication No. WO 93/12227; and Kucherlapati, PCT Publication No. WO 91/10741).
- the anti-DPP3 antibody or anti-DPP3 antibody fragment in accordance with the invention may have the formats known in the art. Examples are human antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, CDR-grafted antibodies or antibody fragments thereof, but not limited to.
- the anti-DPP3 antibody is a monoclonal antibody or a fragment thereof.
- the anti-DPP3 antibody or the antiDPP3 antibody fragment is a human or humanized antibody or derived therefrom.
- one or more (murine) CDR’s are grafted into a human antibody or antibody fragment.
- antibodies according to the present invention are recombinantly produced antibodies as e.g. IgG, a typical full-length immunoglobulin, or antibody fragments containing at least the F-variable domain of heavy and/or light chain as e.g. chemically coupled antibodies (fragment antigen binding) including but not limited to Fab-fragments including Fab minibodies, single chain Fab antibody, monovalent Fab antibody with epitope tags, e.g. Fab-V5Sx2; bivalent Fab (mini-antibody) dimerized with the CH3 domain; bivalent Fab or multivalent Fab, e.g. formed via multimerization with the aid of a heterologous domain, e.g.
- chemically coupled antibodies fragment antigen binding
- fragment antigen binding including but not limited to Fab-fragments including Fab minibodies, single chain Fab antibody, monovalent Fab antibody with epitope tags, e.g. Fab-V5Sx2; bivalent Fab (mini-antibody) dimerized
- dHLX domains e.g. Fab-dHLX-FSx2; F(ab′)2-fragments, scFv-fragments, multimerized multivalent and/or multispecific scFv-fragments, bivalent and/or bispecific diabodies, BITE ® (bispecific T-cell engager), trifunctional antibodies, polyvalent antibodies, e.g. from a different class than G; single-domain antibodies, e.g. nanobodies derived from camelid or fish immunoglobulines and numerous others.
- biopolymer scaffolds are well known in the art to complex a target molecule and have been used for the generation of highly target specific biopolymers. Examples are aptamers, spiegelmers, anticalins and conotoxins.
- Non-Ig scaffolds with the context of the invention may be protein scaffolds and may be used as antibody mimics as they are capable to bind to ligands or antigens.
- Non-Ig scaffolds may be selected from the group comprising tetranectin-based non-Ig scaffolds (e.g. described in US 2010/0028995), fibronectin scaffolds (e.g. described in EP 1266 025; lipocalin-based scaffolds (e.g. described in WO 2011/154420); ubiquitin scaffolds (e.g. described in WO 2011/073214), transferring scaffolds (e.g. described in US 2004/0023334), protein A scaffolds (e.g.
- Non-Ig scaffolds may be peptide or oligonucleotide aptamers.
- Aptamers are usually created by selecting them from a large random sequence pool and are either short strands of oligonucleotides (DNA, RNA or XNA; Xu et al. 2010, Deng et al. 2014) or short variable peptide domains attached to a protein scaffold (Li et al. 2011).
- the anti-DPP3 antibody format is selected from the group comprising Fv fragment, scFv fragment, Fab fragment, scFab fragment, F(ab) 2 fragment and scFv-Fc Fusion protein.
- the antibody format is selected from the group comprising scFab fragment, Fab fragment, scFv fragment and bioavailability optimized conjugates thereof, such as PEGylated fragments.
- antibody generally comprises monoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called “single-chain-antibodies” (Bird et al. 1988), chimeric, humanized, in particular CDR-grafted antibodies, and di- or tetrabodies (Holliger et al. 1993). Also comprised are immunoglobulin-like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample.
- specific binding refers to antibodies raised against the molecule of interest or a fragment thereof.
- An antibody is considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least preferably 50-fold higher, more preferably 100-fold higher, most preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity.
- said anti-DPP3 antibody or anti-DPP3 antibody fragment binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or functional derivative thereof is a monoclonal antibody or a monoclonal antibody fragment thereof.
- the anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or functional derivative thereof is a human or humanized antibody or derived therefrom or humanized antibody fragment or derived therefrom.
- one or more (murine) CDR’s are grafted into a human antibody or antibody fragment.
- said DPP3 binder of the invention specifically said anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold is a modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold.
- a modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold of the invention may act inhibitory and may block the bioactivity of DPP3 to nearly 100%, preferably to at least more than 90%, more preferably to at least 80, or 70, or 60, or 50, or 40, or 30, or 20, or 10 % when determined by means of the above described method for detecting and measuring the inhibition of DPP3; i.e. measuring the DPP3 binder influence on DPP-3 bioactivity.
- a modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold of the invention may act upregulating and thus may enhance the bioactivity of DPP3 to at least 50 %, preferably to at least more than 60 %, more preferably to at least more than 70 %, more preferably to at least more than 80 %, even more preferably to at least more than 90 %, even more so preferably to at least 95 % when determined by means of the above described method for detecting and measuring the inhibition of DPP3; i.e. measuring the DPP3 binder influence on DPP-3 bioactivity.
- Anti-DPP3 antibodies according to the present invention may be synthesised as follows:
- DPP3 peptides for immunization were synthesized, see table 3 below, (JPT Technologies, Berlin, Germany) with an additional N-terminal cystein (if no cystein is present within the selected DPP3-sequence) residue for conjugation of the peptides to Bovine Serum Albumin (BSA).
- BSA Bovine Serum Albumin
- the peptides were covalently linked to BSA by using Sulfolink-coupling gel (Perbio-science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio. Recombinant GST-hDPP3 was produced by USBio.
- mice were intraperitoneally (i.p.) injected with 84 ⁇ g GST-hDPP3 or 100 ⁇ g DPP3-peptide-BSA-conjugates at day 0 (emulsified in TiterMax Gold Adjuvant), 84 ⁇ g or 100 ⁇ g at day 14 (emulsified in complete Freund’s adjuvant) and 42 ⁇ g or 50 ⁇ g at day 21 and 28 (in incomplete Freund’s adjuvant).
- the animal received an intravenous (i.v.) injection of 42 ⁇ g GST-hDPP3 or 50 ⁇ g DPP3-peptide-BSA-conjugates dissolved in saline. Three days later the mice were sacrificed and the immune cell fusion was performed.
- Splenocytes from the immunized mice and cells of the myeloma cell line SP2/0 were fused with 1 ml 50% polyethylene glycol for 30 s at 37° C. After washing, the cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement]. After one week, the HAT medium was replaced with HT Medium for three passages followed by returning to the normal cell culture medium.
- the cell culture supernatants were primarily screened for recombinant DPP3 binding IgG antibodies two weeks after fusion. Therefore, recombinant GST-tagged DPP3 (USBiologicals, Salem, USA) was immobilized in 96-well plates (100 ng/ well) and incubated with 50 ⁇ l cell culture supernatant per well for 2 hours at room temperature. After washing of the plate, 50 ⁇ l / well POD-rabbit anti mouse IgG was added and incubated for 1h at RT.
- a chromogen solution (3,7 mM o-phenylendiamin in citrate/hydrogen phosphate buffer, 0.012 % H 2 O 2 ) were added to each well, incubated for 15 minutes at RT and the chromogenic reaction stopped by the addition of 50 ⁇ l 4 N sulfuric acid. Absorption was detected at 490 mm.
- the positive tested microcultures were transferred into 24-well plates for propagation. After retesting the selected cultures were cloned and recloned using the limiting-dilution technique and the isotypes were determined.
- Antibodies raised against GST-tagged human DPP3 or DPP3-peptides were produced via standard antibody production methods (Marx et al. 1997) and purified via Protein A. The antibody purities were ⁇ 90 % based on SDS gel electrophoresis analysis.
- Humanization of murine antibodies may be conducted according to the following procedure:
- the antibody sequence is analyzed for the structural interaction of framework regions (FR) with the complementary determining regions (CDR) and the antigen. Based on structural modelling an appropriate FR of human origin is selected and the murine CDR sequences are transplanted into the human FR. Variations in the amino acid sequence of the CDRs or FRs may be introduced to regain structural interactions, which were abolished by the species switch for the FR sequences. This recovery of structural interactions may be achieved by random approach using phage display libraries or via directed approach guided by molecular modeling (Almagro JC, Fransson J., 2008. Humanization of antibodies. Front Biosci. ⁇ 2008 Jan 1;13:1619-33).
- the provided subject matter is a human CDR-grafted antiDPP3 antibody or anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said human CDR-grafted anti-DPP3 antibody or antiDPP3 antibody fragment thereof comprises an antibody heavy chain variable region (H chain) comprising
- a human CDR-grafted antiDPP3 antibody or anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein the said human CDR-grafted anti-DPP3 antibody or anti-DPP3 antibody fragment thereof comprises an antibody heavy chain variable region (H chain) comprising:
- subject matter of the present invention is a human monoclonal anti-DPP3 antibody or monoclonal anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein the heavy chain comprises at least one CDR of:
- variable region can be connected to any subclass of constant regions (IgG, IgM, IgE. IgA), or only scaffolds, Fab fragments, Fv, Fab and F(ab)2.
- IgG constant regions
- IgM constant regions
- IgE. IgA only scaffolds
- Fab fragments fragments
- Fv fragments
- Fab fragments
- F(ab)2 the murine antibody variant with an IgG2a backbone was used.
- chimerization and humanization a human IgG1 ⁇ backbone was used.
- CDRs Complementarity Determining Regions
- AK1967 The CDRs for the heavy chain and the light chain of the murine anti-DPP3 antibody of the present invention (AK1967) are shown in SEQ ID NO. 7, SEQ ID NO. 8 and SEQ ID NO. 9 for the heavy chain and SEQ ID NO. 10, sequence KVS and SEQ ID NO. 11 for the light chain, respectively.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to SEQ ID NO.: 1, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and anti-DPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably at least 6 aa of said SEQ ID NO.:1.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and anti-DPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably at least 6 aa of SEQ ID NO.: 2.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said epitope according to SEQ ID NO.: 3 is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and antiDPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably to 6 aa of SEQ ID NO.: 3.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said epitope according to SEQ ID NO.: 4 is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and antiDPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably to four aa of SEQ ID NO.: 4.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, may act as inhibitor or effector of the bioactivity of DPP3.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof are useful in the prevention or treatment of a disease or acute condition in a patient, wherein said disease or acute condition is associated with oxidative stress in accordance with the invention.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, exhibit an affinity towards DPP3 in such that the affinity constant is at least 10 -7 M -1 , preferably at least 10 -8 M -1 , more preferably the affinity constant is at least 10 -9 M -1 , most preferred the affinity constant is at least 10 -10 M -1 when determined by means of the methods for measuring the binding affinity of the DPP3 binder of the invention to the epitope according to sequence SEQ ID NO.: 2 as described above.
- binder e.g. an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold, and this measure would not lead out-of-the-scope of the invention.
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibody or anti-DPP3 antibody fragment thereof or anti-DPP3 non-Ig scaffold may be used in combination with at least one additional drug that induces oxidative stress as side effect.
- Such drugs are administered as primary medicament for use in the prevention or treatment of a primary disease and may be selected from a group comprising antimicrobials like antibiotics (for example streptomycin, gentamicin) or antivirals (for example acyclovir, foscarnet) or antifungal (for example amphotecerin B), analgesics, non-steroidal anti-inflammatory drugs (NSAID) (for example ibuprofen, naproxen), diuretics, proton pump inhibitors, chemotherapeutics (for example cisplatin), contrast dyes, cardiovascular agents like ACEinhibitors or statins, anti-depressants, immune suppressants (for example cyclosporine A) and antihistamines.
- antibiotics for example streptomycin, gentamicin
- antivirals for example acyclovir, foscarnet
- antifungal for example amphotecerin B
- analgesics non-steroidal anti-inflammatory drugs (NSAID) (for example ibuprofen
- the herein provided DPP3 binder specifically the herein provided anti-DPP3 antibody or anti-DPP3 antibody fragment thereof or a DPP3 non-Ig scaffold binding to DPP3 may be used as secondary medicament either in combination or as stand-alone drug in the prevention or treatment of the induced oxidative stress and resultant toxicities as secondary diseases.
- the herein provided DPP3 binder are pharmaceutically acceptable, selective and/or specific for an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- the herein provided DPP3 binder is an inhibitory binder that is pharmaceutically acceptable, selective and/or specific for an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- selective and specific inhibitors of DPP3 do not bind to other proteins/peptides/enzymes or are bound by other proteins/peptides/enzymes, and do not inhibit any other enzyme/protease/peptidase other than DPP3. Therefore, the preferred inhibitors of DPP3 bioactivity with the context of the invention are specific anti-DPP3 antibodies, antibody fragments or non-Ig scaffolds binding to DPP3.
- Monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold with the context of the invention means that said antibody or antibody fragment or non-Ig scaffold binds specifically to one specific region encompassing at least 3 amino acids, preferably at least 4 aa within the target DPP3.
- monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or antiDPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or antiDPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 3, which is comprised in a DPP3 protein or a functional derivative thereof.
- monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or anti-DPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 4, which is comprised in a DPP3 protein or a functional derivative thereof.
- Monospecific antibodies may also be produced by other means than producing them from a common germ cell.
- binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the embodiments 1 to 3, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold, and wherein said epitope is comprised in SEQ ID NO.: 1, which corresponds to the amino acid sequence of DPP3.
- binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- DPP3 dipeptidyl peptidase 3
- binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDR ⁇ s) in the heavy chain comprises the sequences:
- binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a human monoclonal antibody or human monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- the binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the embodiments 8 to 10, wherein said disease is selected from a group comprising sepsis, septic shock, and SIRS.
- the binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to embodiment 8, wherein said acute condition is selected from a group comprising renal toxicity and hepatotoxicity.
- the binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the embodiments 8 and 12, wherein said acute condition is hepatotoxicity which is drug-induced or alcohol-induced hepatotoxicity.
- the binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to embodiment 8, wherein said acute condition is renal toxicity which is drug-induced renal toxicity.
- the binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the embodiments 8 to 11, wherein said disease is associated with oxidative stress in the myocard.
- composition comprising a binder according to any of the embodiments 1 to 7 for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress.
- a kit comprising a binder according to any of the embodiments 1 to 16.
- binder according to any of the embodiments 19 to 24, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold.
- binder according to any of the embodiments 19 to 25, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDR ⁇ s) in the heavy chain comprises the sequences:
- binder according any of the embodiments 19 to 26, wherein said binder is a humanized monoclonal antibody or humanized monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- binder according to any of the embodiments 19 to 27, wherein said binder is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- DPP3 dipeptidyl peptidase 3
- an “DPP3 binder” is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three aa of SEQ ID NO.: 2 or a respective subsequence thereof according to the SEQ ID NO’S.: 3 or 4.
- a DPP3 binder is preferably an anti-DPP3 antibody, or an anti-DPP3 antibody fragment, or an anti-DPP3 non-Ig scaffold directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three aa of SEQ ID NO.: 2 or a respective subsequence thereof according to the SEQ ID NO’S.: 3 or 4.
- a “functional derivative” of a DPP3 protein denotes a peptide, polypeptide or protein that differs from the sequence of SEQ ID NO.: 1 by means of deletion of aa, addition of aa or changes of specific aa, but remains the bioactivity and function of a native DPP3 protein.
- the bioactivity and function may be influenced to a certain extent, but the enzymatic protease reaction catalysed by DDP 3 is still maintained when assay by a suitable bioactivity assay as described above or commonly known by the skilled person.
- a dipeptidyl peptidase 3 (DPP3) antibody or an antiDPP3 antibody fragment or anti-DPP3 non-Ig scaffold is synonymous to dipeptidyl peptidase 3 (DPP3) antibody or a dipeptidyl peptidase 3 antibody fragment or DPP3 non-Ig scaffold and means anti- dipeptidyl peptidase 3 (DPP3) antibody or an anti- dipeptidyl peptidase 3 antibody fragment or anti-DPP3 non-Ig scaffold binding to DPP3, respectively.
- antibody generally comprises monoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called “single-chain-antibodies” (Bird et al. 1988), chimeric, humanized, in particular CDR-grafted antibodies, and di- or tetrabodies (Holliger et al. 1993). Also comprised are immunoglobulin-like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample.
- the term “specific binding” refers to antibodies raised against the molecule of interest or a fragment thereof.
- An antibody is considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least preferably 50-fold higher, more preferably 100-fold higher, most preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity.
- Diseases associated with oxidative stress include, but are not limited to, neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, and inflammation, sepsis, septic shock, SIRS.
- neurodegenerative diseases comprise Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS).
- AD Alzheimer’s disease
- PD Parkinson’s disease
- HD Huntington’s disease
- ALS amyotrophic lateral sclerosis
- MS multiple sclerosis
- metabolic syndrome comprises insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes.
- cardiovascular disorders comprise aterosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury/ stroke and myocardial infarction.
- autoimmune diseases comprise rheumatoid arthritis and systemic lupus erythematosus.
- inflammatory lung diseases comprise COPD and asthma.
- kidney diseases comprise renal toxicity (drug-induced kidney disease), acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy and end-stage renal disease (ESRD).
- AKI acute kidney injury
- CKD chronic kidney disease
- ESRD end-stage renal disease
- liver diseases comprise hepatotoxicity, viral hepatitis, cirrhosis.
- digestive diseases comprise inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease; gastritis, pancreatitis and peptic ulcer.
- viral infectious diseases comprise blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus and respiratory syncytial virus.
- cancer comprises prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer and liver cancer.
- Acute condition associated with oxidative stress with the context of the present invention denote symptoms that appear and change or worsen rapidly due to the occurrence of oxidative stress.
- An acute condition associated with oxidative stress is sudden in onset.
- An acute condition associated with oxidative stress may lead to a chronic syndrome, if untreated.
- chronic condition or a “chronic syndrome”, respectively, with the context of the present invention denote a condition or symptom that develops and worsens over an extended period of time, and may be persistent, even if treated.
- Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species (ROS)/ reactive nitrogen species (RNS) and antioxidants in favour of excessive generation of free radicals. This process leads to the oxidation of biomolecules with consequent loss of its biological functions and/or homeostatic imbalances, whose manifestation is the potential oxidative damage to cells and tissues. Accumulation of ROS/RNS can result in a number of deleterious effects such as lipid peroxidation, protein oxidation and DNA damage (including base damage and strand breaks). Further, some reactive oxidative species act as cellular messengers in redox signalling. Thus, oxidative stress can cause disruptions in normal mechanisms of cellular signalling.
- ROS reactive oxygen species
- RNS reactive nitrogen species
- a “free radical in the context of the present invention is a molecule with one or more unpaired electron in its outer shell. Free radicals are formed from molecules via the breakage of a chemical bond such that each fragment keeps one electron, by cleavage of a radical to give another radical and, also via redox reactions. Free radicals related to oxidative stress include hydroxyl (OH•), superoxide (O 2 • - ), nitric oxide (NO•), nitrogen dioxide (NO 2 •), peroxyl (ROO•) and lipid peroxyl (LOO•).
- hydrogen peroxide H 2 O 2
- ozone O 3
- singlet oxygen 1O2
- hypochlorous acid HOCl
- nitrous acid HNO 2
- peroxynitrite ONOO -
- dinitrogen trioxide N 2 O 3
- lipid peroxide LOOH
- Primary medicament means a medicament that acts against the primary cause of said disease or condition.
- “Secondary medication” is a medication that improves the condition of the patient in a supportive way; e.g. reduces or regulates oxidative stress which is induced by the administration of a primary medicament.
- bioactivity is defined as the effect that a substance takes on a living organism or tissue or organ or functional unit in vivo or in vitro (e.g. in an assay) after its interaction.
- DPP3 bioactivity may be defined as the DPP3 enzyme activity or the regulating activity of DPP3 in the oxidative stress pathway.
- ABBREVIATIONS Abbreviation Meaning aa amino acid(s) AD Alzheimer’s disease AHF acute heart failure AIN Acute interstitial nephritis AKI acute kidney injury ALD Alcoholic liver disease ALS amyotrophic lateral sclerosis ARE antioxidant response element ATN Acute tubular necrosis ATP adenosine triphosphate AZT Azidothymidin BRCA1 Breast cancer gene 1 BP blood pressure BSA bovine serum albumin cDNA complementary DNA CAT catalase CD Celiac disease CDR complementarity determining region CKD chronic kidney disease CLP cecal ligation and puncture CNS central nervous system COPD Chronic obstructive pulmonary disease CSF Cerebrospinal fluid CVD cardiovascular diseases DHE dihydroethidium DN diabetic nephropathy DNA Deoxyribonucleic acid DPP3, DPPIII dipeptidyl dipeptidase 3 (5,5′-dithiobis-(2-nitrobenzoic acid), Ellman’s
- DPP3 peptides for immunization were synthesized, see table 3, (JPT Technologies, Berlin, Germany) with an additional N-terminal cystein (if no cystein is present within the selected DPP3-sequence) residue for conjugation of the peptides to Bovine Serum Albumin (BSA).
- BSA Bovine Serum Albumin
- the peptides were covalently linked to BSA by using Sulfolink-coupling gel (Perbio-science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio. Recombinant GST-hDPP3 was produced by USBio (United States Biological, Salem, MA, USA).
- mice were intraperitoneally (i.p.) injected with 84 ⁇ g GST-hDPP3 or 100 ⁇ g DPP3-peptide-BSA-conjugates at day 0 (emulsified in TiterMax Gold Adjuvant), 84 ⁇ g or 100 ⁇ g at day 14 (emulsified in complete Freund’s adjuvant) and 42 ⁇ g or 50 ⁇ g at day 21 and 28 (in incomplete Freund’s adjuvant).
- the animal received an intravenous (i.v.) injection of 42 ⁇ g GST-hDPP3 or 50 ⁇ g DPP3-peptide-BSA-conjugates dissolved in saline. Three days later the mice were sacrificed and the immune cell fusion was performed.
- Splenocytes from the immunized mice and cells of the myeloma cell line SP2/0 were fused with 1 ml 50% polyethylene glycol for 30 s at 37° C. After washing, the cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement]. After one week, the HAT medium was replaced with HT Medium for three passages followed by returning to the normal cell culture medium.
- the cell culture supernatants were primarily screened for recombinant DPP3 binding IgG antibodies two weeks after fusion. Therefore, recombinant GST-tagged hDPP3 (USBiologicals, Salem, USA) was immobilized in 96-well plates (100 ng/ well) and incubated with 50 ⁇ l cell culture supernatant per well for 2 hours at room temperature. After washing of the plate, 50 ⁇ l / well POD-rabbit anti mouse IgG was added and incubated for 1 h at RT.
- a chromogen solution (3,7 mM o-phenylendiamin in citrate/ hydrogen phosphate buffer, 0.012% H 2 O 2 ) were added to each well, incubated for 15 minutes at RT and the chromogenic reaction stopped by the addition of 50 ⁇ l 4 N sulfuric acid. Absorption was detected at 490 mm.
- the positive tested microcultures were transferred into 24-well plates for propagation. After retesting the selected cultures were cloned and recloned using the limiting-dilution technique and the isotypes were determined.
- Antibodies raised against GST-tagged human DPP3 or DPP3-peptides were produced via standard antibody production methods (Marx et al. 1997) and purified via Protein A. The antibody purities were ⁇ 90% based on SDS gel electrophoresis analysis.
- Recombinant GST-tagged hDPP3 (SEQ ID No. 1) or a DPP3 peptide (immunization peptide, SEQ ID No. 2) was immobilized onto a high binding microtiter plate surface (96-Well polystyrene microplates, Greiner Bio-One international AG, Austria, 1 ⁇ g/well in coupling buffer [50 mM Tris, 100 mM NaCl, pH7,8], 1h at RT). After blocking with 5% bovine serum albumin, the microplates were vacuum dried.
- the purified labeled antibody was diluted in assay buffer (50 mmol/l potassium phosphate, 100 mmol/l NaCl, 10 mmol/l Na 2 -EDTA, 5 g/l bovine serum albumin, 1 g/l murine IgG, 1 g/l bovine IgG, 50 ⁇ mol/l amastatin, 100 ⁇ mol/l leupeptin, pH 7.4).
- the final concentration was approx. 5-7*10 6 relative light units (RLU) of labelled compound (approx. 20 ng labeled antibody) per 200 ⁇ 1.
- acridinium ester chemiluminescence was measured by using a Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG).
- the plates were filled with 200 ⁇ l of labeled and diluted detection antibody (tracer) and incubated for 2-4 h at 2-8° C. Unbound tracer was removed by washing 4 times with 350 ⁇ l washing solution (20 mM PBS, pH 7.4, 0.1 % Triton X-100). Well-bound chemiluminescence was measured by using the Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG).
- fluorogenic substrate Arg-Arg- ⁇ NA (20 ⁇ l, 2 mM) was added to the solution and the generation of free ⁇ NA over time was monitored using the Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH & Co. KG) at 37° C. Fluorescence of ⁇ NA is detected by exciting at 340 nm and measuring emission at 410 nm. Slopes (in RFU/ min) of increasing fluorescence of the different samples are calculated. The slope of GST-hDPP3 with buffer control is appointed as 100 % activity. The inhibitory ability of a possible capture-binder is defined as the decrease of GST-hDPP3 activity by incubation with said capture-binder in percent.
- the following table represents a selection of obtained antibodies and their binding rate in Relative Light Units (RLU) as well as their relative inhibitory ability (%; table 3).
- RLU Relative Light Unit
- % relative inhibitory ability
- Antibodies raised against SEQ ID NO. 2 were characterized in more detail (epitope mapping, binding affinities, specificity, inhibitory potential). Here the results for clone 1967 of SEQ ID NO. 2 (“AK1967”) are shown as an example.
- peptides&elephants GmbH For epitope mapping of AK1967 a number of N- or C-terminally biotinylated peptides were synthesized (peptides&elephants GmbH, Hennigsdorf, Germany). These peptides include the sequence of the full immunization peptide (SEQ ID No. 2) or fragments thereof, with stepwise removal of one amino acid from either C- or N-terminus (see table 5 for a complete list of peptides).
- High binding 96 well plates were coated with 2 ⁇ g Avidin per well (Greiner Bio-One international AG, Austria) in coupling buffer (500 mM Tris-HCl, pH 7.8, 100 mM NaCl). Afterwards plate were washed and filled with specific solutions of biotinylated peptides (10 ng/ well; buffer – 1xPBS with 0.5% BSA)
- AntiDPP3 antibody AK1967 was labelled with a chemiluminescence label according to Example 1.
- the experiment was performed using Octet Red96 (ForteBio). AK1967 was captured on kinetic grade anti-humanFc (AHC) biosensors. The loaded biosensors were then dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, 3.7 nM). Association was observed for 120 seconds followed by 180 seconds of dissociation. The buffers used for the experiment are depicted in table 4. Kinetic analysis was performed using a 1:1 binding model and global fitting.
- Buffer Composition Assay Buffer PBS with 0.1% BSA, 0.02% Tween-21 Regeneration Buffer 10 mM Glycine buffer (pH 1.7) Neutralization Buffer PBS with 0.1% BSA, 0.02% Tween-21
- Blood cells from human EDTA-blood were washed (3x in PBS), diluted in PBS and lysed by repeated freeze-thaw-cycles.
- the blood cell lysate had a total protein concentration of 250 ⁇ g/ml, and a DPP3 concentration of 10 ⁇ g/ml.
- Dilutions of blood cell lysate (1:40, 1:80, 1:160 and 1:320) and of purified recombinant human His-DPP3 (31.25-500 ng/ml) were subjected to SDS-PAGE and Western Blot.
- the blots were incubated in 1.) blocking buffer (1xPBS-T with 5% skim milk powder), 2.) primary antibody solution (AK1967 1:2.000 in blocking buffer) and 3.) HRP labelled secondary antibody (goat anti mouse IgG, 1:1.000 in blocking buffer). Bound secondary antibody was detected using the Amersham ECL Western Blotting Detection Reagent and the Amersham Imager 600 UV (both from GE Healthcare).
- a DPP3 activity assay with known procedure (Jones et al., 1982) was performed. Recombinant GST-tagged hDPP3 was diluted in assay buffer (25 ng/ ml GST-DPP3 in 50 mM Tris-HCl, pH7,5) and increasing concentrations of AK1967 were added. Fluorogenic substrate Arg-Arg- ⁇ NA was added to the solution and the generation of free ⁇ NA over time was monitored using the Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH & Co. KG) at 37° C. Fluorescence of ⁇ NA is detected by exciting at 340 nm and measuring emission at 410 nm.
- AK1967 binds with an affinity of 2.2*10 -9 M to recombinant GST-hDPP3 (for more details see table 6 and for kinetic curves see FIG. 1 A ).
- the only protein detected with AK1967 as primary antibody in lysate of blood cells was DPP3 at 80 kDa ( FIG. 1 B ).
- the total protein concentration of the lysate was 250 ⁇ g/ml whereas the estimated DPP3 concentration is about 10 ⁇ g/ml. Even though there is 25 times more unspecific protein in the lysate, AK1967 binds and detects specifically DPP3 and no other unspecific binding takes place.
- AK1967 inhibits 15 ng/ ml DPP3 in a specific DPP3 activity assay with an IC50 of about 15 ng/ml ( FIG. 1 C ).
- a septic shock model was used to induce heart failure in rats and then to characterize AK1967′s influence on oxidative stress in myocardium.
- mice Male Wistar rats (2-3 months, 300 to 400 g, group size refer to table 7) from the Centre d′élevage Janvier (France) were allocated randomly to one of three groups. All the animals were anesthetized using ketamine hydrochloride (90 mg/ kg) and xylazine (9 mg/ kg) intraperitoneally (i.p.). For induction of polymicrobial sepsis, cecal ligation and puncture (CLP) was performed using Rittirsch’s protocol with minor modification. A ventral midline incision (1.5 cm) was made to allow exteriorization of the cecum. The cecum is then ligated just below the ileocecal valve and punctured once with an 18-gauge needle.
- CLP cecal ligation and puncture
- the abdominal cavity is then closed in two layers, followed by fluid resuscitation (3 ml/ 100 g body of weight of saline injected subcutaneously) and returning the animal to its cage. Sham animals were subjected to surgery, without getting their cecum punctured.
- Dihydroethidium (DHE; Sigma-Aldrich) staining was used to evaluate the in situ levels of superoxide anion in the myocardium. Cardiac cryostat sections (7 ⁇ m) of the ventricles were incubated with DHE (37 ⁇ M) for 30 min in a dark humidified chamber. Acquisition of fluorescent images of ethidium bromide with Leica fluorescence microscope was performed under identical setting whatever the block tissue. The stained area was measured with IPLab software and expressed as a percentage of area of interest (% of ROI).
- ROS reactive oxygen species
- Human dipeptidyl peptidase III acts as a post-proline-cleaving enzyme on endomorphins. Biological Chemistry, 388(3), pp.343-348.
- Oxidative stress An essential factor in pathogenesis of gastrointestinal mucosal diseases. Physio.l Rev. 94, pp. 329-354.
- Nrf2 p21Cip1/WAF1 upregulates the Nrf2-mediated antioxidant response. Mol Cell., 34(6), pp. 663-673.
- Tumor cytosol dipeptidyl peptidase III activity is increased with histological aggressiveness of ovarian primary carcinomas. Gynecologic Oncology, 91(1), pp.194-200.
- Oxidative stress and cancer An overview. Ageing Research Reviews, 12, pp. 376-390.
- SEQ ID No. 12 humanized AK1967 — heavy chain sequence (IgG1 ⁇ : backbone)
- FIG. 1 A illustrates Association- and dissociation curve of the AK1967-DPP3 binding analysis using Octet.
- AK1967 loaded biosensors were dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, 3.7 nM) and association and dissociation monitored.
- FIG. 1 B is a Western Blot of dilutions of blood cell lysate and detection of DPP3 with AK1967 as primary antibody.
- FIG. 1 C shows an Inhibition curve of native DPP3 from blood cells with inhibitory antibody AK1967. Inhibition of DPP3 by a specific antibody is concentration dependent, with an IC 50 at ⁇ 15 ng/ml when analyzed against 15 ng/ml DPP3.
- FIG. 2 A is an Experimental design of heart failure study of rats in septic shock.
- FIG. 2 B illustrates Fluorescence images of DHE labelled myocardium of sham, CLP and CLP AK1967 animals.
- FIG. 2 C illustrates Quantification of DHE stained areas and expression as percentage of area of interest (% of ROI).
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
The present invention provides binder directed to and binding to a DPP3 protein or functional derivative thereof and its use in a method of prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress. With this context, specifically the present invention provides a binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
Description
- The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on Nov. 28, 2022, is named BOEHMERP-0285-C01_SL.xml and is 36,712 bytes in size.
- In a first major aspect of the invention, subject matter of the invention is a binder being directed to and binding to a dipeptidyl peptidase 3 (DPP3) protein or functional derivative thereof.
- In another embodiment of the first major aspect of the invention, the aforementioned binder is provided for the use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress.
- In another embodiment of the first major aspect of the invention, the aforementioned binder is provided for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress, and wherein said diseases are selected from a group comprising neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
- In a second major aspect of the present invention, a binder is provided that is directed to and binding to an epitope according to SEQ ID NO.: 2, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- The below text refers to the above second major aspect of the invention:
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 3, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 4, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1..
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- In a third major aspect of the invention, the above mentioned binder of the second major aspect of the invention that are directed to and binding to an epitope according to SEQ ID NO.: 2 is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2 according to the third aspect of the invention, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDRs) in the heavy chain comprises the sequences:
- SEQ ID NO.: 7, SEQ ID NO.: 8 and/ or SEQ ID NO.: 9
- and the complementarity determining regions in the light chain comprises the sequences:
- SEQ ID NO.: 10, KVS and/ or SEQ ID NO.: 11.
- Additional subject matter of the invention is the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2 according to the third aspect of the invention, wherein said binder is a humanized monoclonal antibody or humanized monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- SEQ ID NO.: 12
- and wherein the light chain comprises the sequence:
- SEQ ID NO.: 13.
- Additional subject matter of the invention is anyone of the aforementioned binder being directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress, and wherein the epitope is comprised in DPP3 as depicted in SEQ ID NO.: 1.
- Specific subject matter of the present invention is a binder, specifically a dipeptidyl peptidase 3 (hereinafter DPP3) binder, directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- Further subject matter of the present invention is a binder, specifically a DPP3 binder, directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2, for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Also, subject matter of the present invention is a binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment, binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof or an anti-DPP3 non-Ig scaffold binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- With the above context, also subject matter of the present invention is a binder in accordance with the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment, binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof or an anti-DPP3 non-Ig scaffold binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2, for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Further subject matter of the present invention is a method of prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress, characterized in that a binder directed to and binding to DPP3, or a binder being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof is administered to said patient in pharmaceutically effective amounts.
- Subject matter of the present invention is further a pharmaceutical composition comprising a binder directed to and binding to DPP3, or a binder being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold being directed to and binding to SEQ ID.: 2 as epitope that is comprised in DPP3 protein or a functional derivative thereof for the use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Another subject of the present invention is a pharmaceutical composition comprising a binder of the invention, or a DPP3 binder in accordance with the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress as described above, and wherein said pharmaceutical composition comprises at least one additional pharmaceutically active drug which e.g. may be used as primary medicament in methods of treatment of a disease or acute condition and wherein said treatment induces oxidative stress as side effect, and thus the said binder, or DPP3 binder, anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to DPP3 or the anti-DPP3 non-Ig scaffold binding to DPP3 may act as secondary medicament, which reduces or regulates the said induced oxidative stress.
- A further embodiment of the invention is a kit comprising a binder, or a DPP3 binder in accordance with the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress as described above, and wherein said pharmaceutical composition optionally comprises at least one additional pharmaceutically active drug which e.g. may be used as primary medicament in methods of treatment of a disease or acute condition and wherein said treatment induces oxidative stress as side effect, and thus the said DPP3 binder, anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to DPP3 or the anti-DPP3 non-Ig scaffold binding to DPP3 may act as secondary medicament, which reduces or regulates the said induced oxidative stress.
- Moreover, subject matter of the present invention is a binder in accordance with the invention, or an anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 as described above for the use as a medicament, wherein said binder, or antibody or said antibody fragment or said non-Ig scaffold is a modulating binder, antibody or fragment or scaffold.
- In a preferred embodiment said modulating anti-DPP3 antibody or a modulating anti-DPP3 fragment or a modulating non-Ig scaffold is used in the prevention or treatment of diseases or an acute condition in a patient, whereby said disease or acute condition is associated with oxidative stress.
- Further, in accordance with the invention, said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or a modulating non-Ig scaffold of the invention regulates the bioactivity of DPP3.
- With the context of the invention, DPP3 bioactivity may be defined as the DPP3 enzyme activity or the regulating activity of DPP3 in the oxidative stress pathway.
- In accordance with the invention, said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or modulating non-Ig scaffold of the invention may enhance the bioactivity of DPP3.
- In another embodiment of the invention, said modulating anti-DPP3 antibody or an anti-DPP3 antibody fragment or modulating non-Ig scaffold of the invention may reduce the bioactivity of DPP3.
- In another specific embodiment with the context of the present invention, a “modulating” anti-DPP3 antibody or a modulating anti-DPP3 antibody fragment or a modulating non-Ig scaffold as described above is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or a modulating anti-DPP3 non-Ig scaffold blocks the bioactivity of DPP3 at least 10 %, preferably at least 50 %, more preferably > 50 %, most preferably 100%.
- In a specific embodiment a modulating binder, or modulating anti-DPP3 antibody or a modulating anti-DPP3 antibody fragment or a modulating anti-DPP3 non-Ig scaffold according to the present invention is used for the prevention or treatment of diseases or an acute condition of a patient, wherein said disease or acute condition is associated with oxidative stress.
- Another embodiment of the present invention is a kit or an assay comprising the above described binder, or anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Dipeptidyl peptidase 3 — also known as Dipeptidyl aminopeptidase III, Dipeptidyl arylamidase III, Dipeptidyl peptidase III, Enkephalinase B or red cell angiotensinase; short name: DPP3, or DPPIII — is a metallopeptidase that removes dipeptides from physiologically active peptides, such as enkephalins and angiotensins. Hereinafter, the expression “DPP3” will be used throughout the text as abbreviated form of the above described dipeptidyl peptidase 3.
- DPP3 was first identified and its activity measured in extracts of purified bovine anterior pituitary by Ellis & Nuenke, 1967. The enzyme, which is listed as EC 3.4.14.4, has a molecular mass of about 83 kDa and is highly conserved in procaryotes and eucaryotes (Prajapati & Chauhan, 2011).
- The amino acid sequence of the human variant of DPP3 is depicted in SEQ ID No.: 1. DPP3 is a mainly cytosolic peptidase, which is ubiquitously expressed. Despite lacking a signal sequence, a few studies reported membranous activity (Lee & Snyder, 1982).
- DPP3 is a zinc-depending exo-peptidase belonging to the peptidase family M49. It has broad substrate specificity for oligopeptides from three or four to ten amino acids of various compositions and is also capable of cleaving after proline. DPP3 is known to hydrolyze dipeptides from the N-terminus of its substrates, including angiotensin II, III and IV; angiotensin 1-7 (Cruz-Diaz et al., 2016); Leu- and Met-enkephalin;
endomorphin - Recent findings for DPP3 indicate its role for being a part of the protein metabolism but also playing a role in blood pressure regulation, pain modulation, inflammatory processes and oxidative stress regulation (Prajapati & Chauhan, 2011).
- DPP3 has been also shown to be a promising biomarker in several publications. It has been shown that DPP3 activity is elevated in homogenates of ovarian and endometrial tumors. DPP3 activity even increases with the severity/malignancy of said tumors (Šimaga et al., 1998 and 2003). Immune histology and western blot analysis of glioblastoma cell lines also revealed elevated DPP3 levels (Singh et al., 2014).
- DPP3 was also proposed to be a potential arterio-risk marker (US 2011008805) and as a marker for rheumatoid arthritis (US 2006177886). The patent application WO 2005/106486 describes DPP3-expression and activity as diagnostic marker and DPP3 as therapeutic target in all kinds of diseases, due to ubiquitous expression of DPP3 in or at surface of cell. EP 1498480 mentions the potential diagnostic and therapeutic use of hydrolytic enzymes, including DPP3.
- The relevant prior art can be further summarized as follows:
- WO 2005/106486 describes in a general manner a method of screening for therapeutic agents which may be useful in the treatment of diseases, comprising cardiovascular diseases, infections, respiratory diseases, cancer, endocrinological diseases, metabolic diseases, gastroenterological diseases, inflammation, haematological diseases, muscle skeleton diseases, neurological and urological diseases. In said method of screening, a test compound is contacted with a DPP3 polynucleotide and the binding between said test compound and said DPP3 polynucleotide is detected. Further, the document describes in a general manner compounds, which may bind to and / or activate or inhibit the activity of DPP3. Further, the invention describes pharmaceutical compositions, which comprise such compounds.
- Liu et al. 2007 describe a relation between activation of the antioxidant response element (ARE) and overexpression of DPP3 and
Sequestome 1 in IMR-32 cells. Overexpression of DPP3 andSequestome 1 stimulated the Nrf2 translocation and led to increased levels of NAD(P)H:quinone oxireductase 1, a protein which is transcriptionally regulated by the ARE. - Hast et al. 2013 describe a comparison of the spectrum of KEAP1 interacting proteins with the genomic profile of 178 squamous cell lung carcinomas characterized by The Cancer Genome Atlas and reveal amplification and mRNA over-expression of the DPP3 gene in tumors with high Nrf2activity but lacking Nrf2 stabilizing mutations. They further describe that tumor-derived mutations in KEAP1 are hypomorphic with respect to Nrf2 inhibition and that DPP3 over-expression in the presence of these mutants further promotes Nrf2 activation.
- Thus, according to the prior art, overexpression of intracellular DPP3 is known to be closely linked to oxidative stress regulation. DPP3 was identified as an activator of the antioxidant response element (ARE) in an unbiased screen of a cDNA library consisting of approximately 15,000 full-length human expression cDNAs (Liu et al. 2007).
- DPP3 disrupt the KEAP1-Nrf2 complex by competing with Nrf2 about the KEAP1 binding site (Hast et al. 2013). This disruption prevents NRF2 degradation and subsequently leads to translocalization of Nrf2 into the nucleus and ARE activation. Overexpression of DPP3 in neuroblastoma cells (Liu et al. 2007), in HEK293T cells (Hast et al. 2013) or in MCF7 breast cancer cells (Lu et al. 2017) activates Nrf2-mediated transcription. Active and inactive variants of DPP3 were overexpressed in MCF7 cells and showed the same regulatory effect on oxidative stress (Lu et al. 2017). Hast et al. (2013) also showed a loss-of-function effect: silencing of DPP3 using specific siRNA Nrf2-mediated transcription was decreased down to levels of Nrf2-silencing.
- Although DPP3 is known as an intracellular protein, DPP3 activity was detected in some bodily fluids as well: retroplacental serum (Shimamori et al. 1986), seminal plasma (Vanha-Perttula et al. 1988) and CSF (Aoyagi et al. 1993). In CSF there were elevated DPP3 activity levels measured in patients suffering from Alzheimer’s disease (AD, Aoyagi et al., 1993).
- DPP3 is known for being expressed as membranous, intracellular or circulating DPP3.
- DPP3 has been not only proposed as potential biomarker but also as potential therapeutic target due to its ability to cleave several bioactive peptides. Influenca A virus changes host DPP3 levels for own replication (cell culture studies, Meliopoulos et al. 2012). Enkephalin and/or angiotensin degrading enzymes in general, including DPP3, have a therapeutic potential as targets for treatment of pain, cardiovascular diseases (CVD) and cancer and the corresponding inhibitors as potential treatments of pain, mental illnesses and CVD (Khaket et al. 2012, Patel et al. 1993, Igic et al. 2007).
- The activity of DPP3 can be inhibited unspecifically by different general protease inhibitors (e.g. PMSF, TPCK), sulfhydryl reagents (e.g. pHMB, DTNB) and metal chelators (EDTA, o-phenantroline) (Abramić et al. 2000, EP 2949332).
- DPP3 activity can be further inhibited specifically by different kinds of compounds: an endogenous DPP3-inhibitor is the peptide spinorphin. Several synthetic derivatives of spinorphin, e.g. tynorphin, have been produced and shown to inhibit DPP3 activity to varying extents (Yamamoto et al. 2000). Other published peptide inhibitors of DPP3 are propioxatin A and B (US 4804676) and propioxatin A analogues (Inaoka et al. 1988).
- DPP3 can also be inhibited by small molecules such as fluostatins and benzimidazol derivatives. Fluostatins A and B are antibiotics produced in Streptomyces sp. TA-3391 that are non-toxic and strongly inhibit DPP3 activity. So far, 20 different derivatives of benzimidazol have been synthesized and published (Agić et al., 2007; Rastija et al., 2015), of which the two
compounds 1′ and 4′ show the strongest inhibitory effect (Agić et al., 2007). Several dipeptidyl hydroxamic acids have been shown to inhibit DPP3 activity as well (Cvitešić et al., 2016). - Oxidative stress reflects an imbalance between the systemic manifestation of reactive oxygen species (hereinafter ROS) / reactive nitrogen species (hereinafter RNS) and antioxidants in favour of excessive generation of free radicals. This process leads to the oxidation of biomolecules with consequent loss of its biological functions and/or homeostatic imbalances, whose manifestation is the potential oxidative damage to cells and tissues. Accumulation of ROS / RNS can result in a number of deleterious effects such as lipid peroxidation, protein oxidation and DNA damage (including base damage and strand breaks). Further, some reactive oxidative species act as cellular messengers in redox signalling. Thus, oxidative stress can cause disruptions in normal mechanisms of cellular signalling.
- “ROS” and “RNS” are the terms collectively describing free radicals and other non-radical reactive derivatives, which are also called oxidants. Radicals are less stable than non-radical species, although their reactivity is generally stronger. A molecule with one or more unpaired electron in its outer shell is called a free radical. Free radicals are formed from molecules via the breakage of a chemical bond such that each fragment keeps one electron, by cleavage of a radical to give another radical and, also via redox reactions. Free radicals related to oxidative stress include hydroxyl (OH•), superoxide (O2•-), nitric oxide (NO•), nitrogen dioxide (NO2•), peroxyl (ROO•) and lipid peroxyl (LOO•). Also, hydrogen peroxide (H2O2), ozone (O3), singlet oxygen (1O2), hypochlorous acid (HOCl), nitrous acid (HNO2), peroxynitrite (ONOO-), dinitrogen trioxide (N2O3), lipid peroxide (LOOH), are not free radicals and generally called oxidants, but can easily lead to free radical reactions in living organisms.
- Formation of ROS and RNS can occur in the cells by two ways: enzymatic and non-enzymatic reactions. Enzymatic reactions generating free radicals include those involved in the respiratory chain, the phagocytosis, the prostaglandin synthesis and the cytochrome P450 system. Free radicals can be produced from non-enzymatic reactions of oxygen with organic compounds as well as those initiated by ionizing radiations. The non-enzymatic process can also occur during oxidative phosphorylation (i.e. aerobic respiration) in the mitochondria. For a review see Pham-Huy et al. 2008. Int J Biomed Sci 4 (2): 89-96.
- In view of the stated above and with the context of the present invention, oxidative stress is linked to a number of diseases, including but not limited to neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer and inflammation, and thus associated therewith.
- According to the prior art, intracellular DPP3 is known to be closely linked to oxidative stress regulation. DPP3 was identified as an activator of the antioxidant response element (ARE) in an unbiased screen of a cDNA library consisting of approximately 15,000 full-length human expression cDNAs (Liu et al. 2007; see also above). ARE regulates the expression of a number of cytoprotective antioxidant enzymes and scavengers, contributing to endogenous defence against oxidative stress. This antioxidant effect of DPP3 is due to the interference of DPP3 with the KEAP1-Nrf2 signalling pathway. Nrf2 is a transcription factor that controls the basal and induced expression of an array of antioxidant response element-dependent genes to regulate the physiological and pathophysiological outcomes of oxidant exposure. Under normal or unstressed conditions, Nrf2 is bound to Kelch like-ECH-associated protein 1 (KEAP1) via its ETGE (SEQ ID NO: 4) and its DLG motif. Within this protein cluster Nrf2 is kept in the cytoplasm, quickly ubiquinated and degraded by proteasome. Under oxidative stress, Nrf2 is not degraded, but instead translocates to the nucleus where it binds to a DNA promoter and induces expression of an array of antioxidant response element (ARE)-dependent genes. A variety of chemicals, including phytochemicals and derivatives (CDDO, sulforaphane), therapeutics (oltipraz, auranofin), environmental agents (paraquat, arsenic), and endogenous chemicals [NO, 15d-PGJ2, nitro-fatty acids, and 4-hydroxynonenal (4-HNE)], induce ARE genes through Nrf2 (Ma 2013).
- In view of the stated above, the inventors surprisingly and unexpectedly found that oxidative stress can be also reduced or regulated by a binder directed to and binding to a DPP3 protein or functional derivative thereof.
- The inventors also found that oxidative stress can be also reduced or regulated by a binder directed to and binding to an epitope of SEQ ID NO.: 2, wherein the epitope is comprised in the DPP3 protein.
- Moreover, the present inventors found a dipeptidyl peptidase 3 (hereinafter DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- In the above context, the inventors specifically found that oxidative stress may be reduced or regulated by an anti-DPP3 antibody or an anti DPP3-antibody fragment directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2 or an anti-DPP3 non-Ig scaffold directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids (aa), preferably at least 4 aa of SEQ ID NO.: 2.
- Thus, the present invention provides the herein disclosed binder, and anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 for use in methods of the preventive treatment or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Moreover, with the herein provided binder, DPP3 binder, and specifically the anti-DPP3 antibody, or anti DPP3-antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3, the inventors have found binder to DPP3 which rapidly reduce or regulate oxidative stress in cells of a mammal when determined by the methods of respective biomarker measurements as further set out below.
- Another subject of the present invention is a pharmaceutical composition comprising the binder of the invention, DPP3 binder of the invention, and specifically comprising the anti-DPP3 antibody or an anti-DPP3 antibody fragment binding to DPP3 or an anti-DPP3 non-Ig scaffold binding to DPP3 of the invention for use in methods of the prevention or treatment of diseases or acute conditions of a patient, whereby said disease or acute condition is associated with oxidative stress.
- Hence, the herein disclosed pharmaceutical compositions are also provided for use in the prevention or treatment of symptoms, or syndromes, or pathological and acute conditions and disease associated problems, which are mediated by oxidative stress.
- As mentioned above, the occurrence of oxidative stress is linked to a number of diseases or disorders, which in accordance of the invention include:
- neurodegenerative diseases, wherein said neurodegenerative diseases may be selected from a group comprising Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)),
- metabolic syndrome, wherein said metabolic syndrome may be selected from a group comprising insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes,
- cardiovascular disorders, wherein said cardiovascular disorders may be selected from a group comprising atherosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury, stroke and myocardial infarction,
- autoimmune diseases, wherein said autoimmune diseases may be selected from a group comprising rheumatoid arthritis, systemic lupus erythematosus,
- inflammatory lung diseases, wherein said inflammatory lung diseases may be selected from a group comprising COPD, asthma,
- kidney diseases wherein said kidney diseases may be selected from a group comprising renal toxicity (drug-induced kidney disease), acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD),
- liver diseases wherein said liver diseases may be selected from a group comprising hepatotoxicity, viral hepatitis, cirrhosis,
- digestive diseases wherein said digestive diseases may be selected from a group comprising inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease, gastritis, pancreatitis and peptic ulcer,
- viral infectious diseases wherein said viral infectious diseases may be selected from a group comprising blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus,
- cancer, wherein said cancer may be selected from a group comprising prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer,
- and inflammation, and
- sepsis, septic shock, and SIRS.
- In view of the stated above and with the context of the present invention, a detailed list of diseases and their association with oxidative stress is depicted in Table 1 below:
-
TABLE 1 The association of oxidative stress with diseases in accordance with the invention Disease group Diseases Reference Neurodegenerative disorders Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) Liu et al. 2017; Manoharan et al. 2016 Multiple sclerosis (MS) Adamczyk and Adamczyk-Sowa 2016 Metabolic Syndrome Insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension Hutcheson and Rocic 2012 Diabetes Pitocco et al. 2013; Ullah et al. 2016 Cardiovascular disorders aterosclerosis, hypertension, heart failure, cardiovascular ischemia Elahi et al. 2009 heart failure Tsutsui et al. 2011 cerebral ischemic injury/ stroke Chen et al. 2011 myocardial infarction Hori and Nishida 2009 Autoimmune disease rheumatoid arthritis Quiñonez-Flores et al. 2016 systemic lupus erythematosus Perl 2013 Inflammatory lung disease COPD, asthma Holguin 2013 Kidney disease renal toxicity (drug-induced kidney disease) Hosohata 2016; Naughton 2008 acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD) Sureshbabu et al. 2015 Liver diseases Hepatotoxicity (drug-induced, alcohol-induced) Li et al. 2015 Viral Hepatitis Ivanov et al. 2017 Cirrhosis Vairappan 2015 Digestive diseases Inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease Tian et al. 2017; Bhattacharyya et al. 2014 Gastritis, Pancreatitis, Peptic ulcer Bhattacharyya et al. 2014 Viral infectious diseases blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus(HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus Pohanka 2013; Schwarz 1996 Cancer general cancer Sosa et al. 2013; Kruk and Aboul-Enein 2017 prostate cancer Khandrika et al. 2009 breast cancer Nourazarian et al. 2014 lung cancer Valavanidis et al. 2013 colorectal cancer Perse 2013 bladder cancer Sawicka et al. 2015 ovarian, endometrial, cervical cancer Saed et al. 2017 skin cancer Narendhirakannan 2013 gastric cancer Ma et al. 2013 liver cancer Wang et al. 2013 leukemia Kruk and Aboul-Enein 2017 Inflammation /sepsis, septic shock, SIRS sepsis Kaymak et al. 2011 - In more detail:
- Oxidative stress is suspected to be important in neurological and neurodegenerative disorders including Amyotrophic lateral sclerosis (ALS), Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, Depression, Multiple sclerosis, tardive dyskinesia (TD), epilepsy and acute diseases of the central nervous system, such as spinal cord injuries and/or brain traumatic. The human brain is vulnerable to oxidative stress due to many facts such as (i) metabolism of catecholamines; (ii) decrease in antioxidants; (iii) presence of transition metals; (iv) occurrence of brain trauma/injury; and also (v) the brain is an organ that proportionally requires more oxygen and (vi) expresses low levels of antioxidant enzymes, which contribute to formation of ROS. As a consequence of redox unbalance in brain, one of the most affected structures is the lipid membrane (Rao and Balachandran 2002. Nutritional Neuroscience 5: 291-309). A common feature of these diseases is oxidative damage of neurons, which might be responsible for the dysfunction or death of neuronal cells that contributes to disease pathogenesis.
- Alzheimer’s disease (AD), the most prevalent neurodegenerative disorder, is characterized by the progressive deterioration of behaviour, cognition and functionality, which significantly impairs daily living activities. Numerous experimental and clinical studies have demonstrated that oxidative damage plays a key role in the loss of neurons and the progression to dementia in Alzheimer’s disease. The production of β-amyloid (Aβ), a toxic peptide often found in Alzheimer’s patients’ brain, is due to oxidative stress and plays an important role in the neurodegenerative processes. In addition, Aβ proteins can directly initiate free radical formation via the activation of NADPH oxidase. Moreover, inflammation is responsible for increased expression of cytokines, ROS levels, and cellular toxicity, thereby exacerbating AD progression. For review see Liu et al. 2017. Oxidative Medicine and Cellular Longevity 2525967; Manoharan et al. 2016. Oxid Med Cell Longev 8590578.
- Huntington’s disease (HD) is a progressive neurodegenerative disease linked with unstable expansion of cytosine, adenine, guanine (CAG) repeats in the HTT gene. The expansion of CAG repeats within the exon1 of the HTT gene gives rise to a mutation that leads to the elongation of polyglutamine tract, resulting in an HTT protein product that is susceptible to aggregation. The mHTT aggregates are accumulated throughout the brain of the affected individuals, which can interrupt protein quality control and transcription process. Those alterations are potentially responsible for the aberrant motor and cognitive problems in HD. Though oxidative damage is not much reported in the early stages of HD, it is proposed as one of the major mechanisms in HD as it progresses. Elevated oxidative stress plays a critical role in the late stage of HD pathogenesis. Impairment in the electron transport chain and mitochondrial dysfunction are the major mechanisms involved in the ROS mediated etiopathogenesis of HD. Dysfunction in the oxidative phosphorylation components has been documented in the brain tissues of HD patients. HD patients showed an increased level of oxidative stress markers accompanied by a decrease in antioxidant status compared to healthy subjects. For review see Liu et al. 2017. Oxidative Medicine and Cellular Longevity 2525967; Manoharan et al. 2016. Oxid Med Cell Longev 8590578.
- Parkinson’s disease (PD), the most common neurodegenerative disease of the elderly, is characterized by progressive loss of muscle control. PD is predominant at the 6th decade of life and men are 1.5 to 2 times more likely to contract the disease than women. Head trauma, illness, or exposure to environmental toxins is identified as a risk factor. This neurodegenerative disorder is characterized by tremor, rigidity, bradykinesia, and impairment in balance. PD also causes cognitive, psychiatric, autonomic, and sensory disturbances. The pathology of PD is characterized by the gradual and selective loss of dopaminergic neurons in the substantia nigra pars compacta. Imbalance in dopamine metabolism due to oxidative stress has been recognised as a contributor to this disease. The major pathological findings include the presence of Lewy bodies in the substantia nigra and loss of nerve cells in the portions of its ventral tier. Several studies have reported impaired respiratory chain and somatic mitochondrial DNA mutations in the brain of patients with PD, which suggests the extensive role of oxidative metabolism in PD. Enhanced dopamine metabolism in the brain of patients with PD could account for the accumulation of toxic radicals such as hydroxyl in the brain. Iron accumulation in the neurons in the redox active form plays a crucial role in pathogenesis of this disease. Accumulation of iron has been reported in the substantia nigra in patients diagnosed with PD, which suggests the critical role of iron-induced lipid peroxidation in pathogenesis of PD A twofold increase in protein oxidation has been shown in the substantia nigra of PD patients compared to healthy subjects. Accumulation of hydroxyl radical due to lowered glutathione content in the brain has been reported in PD patients. Lowered activities of antioxidant enzymes and non-enzymatic antioxidants could be responsible for the progression of PD. For review see Liu et al. 2017. Oxidative Medicine and Cellular Longevity 2525967; Manoharan et al. 2016. Oxid Med Cell Longev 8590578.
- Amyotrophic lateral sclerosis (ALS) is characterized by progressive loss of motor neurons in the anterior horn of the spinal cord. It is classified as either familial or sporadic depending on whether there is a clearly defined, inherited genetic element. Sporadic ALS (sALS) typically emerges between 50 and 60 years old. The onset of sALS is unknown, and thus the identification of causal genes and environmental factors remains elusive. In familial ALS, about 20% of the cases resulted from mutations in SOD1. The functions of SOD1 are diverse and include scavenging excessive superoxide radical, modulating cellular respiration, energy metabolism, and posttranslational modification. SOD dysfunction leads to a loss of antioxidant capability. Moreover, increased levels of ROS and ROS-associated damage have been widely reported in ALS. Increased markers of ROS damage have been found in biofluids of patients with sporadic ALS as well as in post-mortem tissue. For review see Liu et al. 2017. Oxidative Medicine and Cellular Longevity 2525967.
- Multiple sclerosis (MS) is a multifactorial disease of the central nervous system (CNS) in which both inflammatory and neurodegenerative processes occur simultaneously. In the course of the disease inflammation is decreased whereas the degeneration of the CNS progresses. The inflammatory component in MS is important not only due to axonal and neuronal loss but also due to the fact that it starts the degenerative cascade in the early stage of MS. The induction of the activation of microglia and mitochondrial dysfunction plays a particular role in inflammatory processes. Microglia activated by T-lymphocytes release proteolytic enzymes, cytokines, oxidative products, and free radicals. It is also important that mitochondrial dysfunction results in an increased production of reactive oxygen species (ROS), which is detrimental to neurons and glia. On the other hand, oxidative stress damages the mitochondria, which disrupts the transport of adenosine triphosphate along the axon, and consequently leads to neurodegeneration. Oxidative stress is associated with the dysregulation of axonal bioenergetics, cytokine-induced synaptic hyperexcitability, abnormal iron accumulation, and the oxidant/antioxidant balance. Markers of oxidative stress assessed in the serum, erythrocytes CSF, saliva, and urine may have diagnostic properties whereas antioxidants may have clinical application in the future. For review see Adamczvk and Adamczyk-Sowa 2016. Oxidative Medicine and Cellular Longevity 1973834.
- Oxidative stress is related to metabolic syndrome and its individual component pathologies, e.g. obesity, insulin resistance, dyslipidemia, impaired glucose tolerance and high blood pressure. The metabolic syndrome was defined by the World Health Organization criteria (Alberti and Zimmet 1998. Diabet Med. 15:539-553; World Health Organization. 1999. Definition, diagnosis and classification of diabetes mellitus and its complications: report of a WHO Consultation. Part 1: diagnosis and classification of diabetes mellitus. Geneva, Switzerland: World Health Organization) that require the presence of insulin resistance identified by one of the following: (1) type II diabetes; (2) impaired fasting glucose; (3) impaired glucose tolerance or (4) for those with normal fasting glucose levels (< 110 mg/dL), glucose uptake below the lowest quartile for background population under investigation under hyperinsulemic, euglycemic conditions, AND two of the following: (1) blood pressure: ≥ 140/90 mmHg; (2) dyslipidemia: triglycerides (TG): ≥ 1.695 mmol/L and high-density lipoprotein cholesterol (HDL-C) ≤ 0.9 mmol/L (male), ≤ 1.0 mmol/L (female); (3) central obesity: waist:hip ratio > 0.90 (male); > 0.85 (female), or body mass index > 30 kg/m2; (4) microalbuminuria: urinary albumin excretion ratio ≥ 20 µg/min or albumin:creatinine ratio ≥ 30 mg/g.
- Increased oxidative stress has emerged as playing a central role in metabolic syndrome and its component pathologies and may be a unifying factor in the progression of this disease. Moreover, oxidative stress has been identified as a major mechanism of micro- and macrovascular complications in the metabolic syndrome. For review see Hutcheson and Rocic 2012. Exp Diabetes Res. 2012:271028.
- There is a bulk of evidence demonstrating that mitochondrial ROS (predominantly superoxide anion) overproduction is involved in diabetes and diabetic complications. It was suggested that glucose can directly stimulate ROS overproduction, and it was also shown that high glucose (HG) activates various enzymatic cascades in mitochondria, including activation of NADPH oxidase, uncoupling of NO synthases and stimulation of xanthine oxidase. Glycated proteins can also be the promoters of ROS formation, thus suggesting that different sources may be responsible for ROS overproduction and oxidative stress in diabetes. For review see Pitocco et al. 2013. Int. J. Mol. Sci. 2013, 14, 21525-21550.
- Moreover, oxidative stress plays an important role in the pathogenesis and development of cardiovascular diseases, including hypertension, dyslipidemia, atherosclerosis, myocardial infarction, angina pectoris, and heart failure (Elahi et al. 2009. Oxidative Medicine and Cellular Longevity 2(5): 259-269). One of the key concepts of free radical mediated pathogenesis of cardiovascular disease is endothelial dysfunction, whereby the regulation of vascular wall microenvironment is disrupted. ROS activity in the vessel wall, for example, is thought to contribute to the formation of oxidized LDL, a major contributor to the pathogenesis of atherosclerosis. Oxidative stress also plays a role in the ischemic cascade due to oxygen reperfusion injury following hypoxia. This cascade includes both stroke (Chen et al. 2011. Antioxidants and Redox Signaling 14(8): 1505-1517) and myocardial infarction (MI) (Hori and Nishida et al. 2009. Cardiovascular Research 81: 457-464). During brain ischemia/reperfusion, multiple detrimental processes take place, including overproduction of oxidants, inactivation of detoxification systems, and consumption of antioxidants. These changes cause the disruption of the normal antioxidative defense ability of brain tissue (Chen et al. 2011. Antioxidants and Redox Signaling 14(8): 1505-1517). For further review see Elahi et al. 2009. Oxidative Medicine and Cellular Longevity 2(5): 259-269.
- A number of experimental and clinical studies have demonstrated the increased generation of ROS in heart failure (HF) and showed that oxidative stress is involved in the pathophysiology of HF in the heart as well as in the skeletal muscle. The high metabolic activity of the mitochondria-rich myocardium makes these findings seem intuitively obvious. Oxidative stress clearly activates processes in isolated heart cells such as changes in gene expression and cell death that are now accepted components of myocardial remodeling and heart failure. Moreover, many studies have been performed in animal models that demonstrate therapeutic effects of antioxidants on progression of heart failure. For review see Tsutsui et al. 2011. Am J Physiol Heart Circ Physiol 301: H2181-H2190.
- Excessive oxidative stress is thought to have an important role in the pathogenesis of autoimmune diseases. Many studies have shown that T and B lymphocytes contribute to the pathogenesis of autoimmune diseases by the production of autoantibodies and ROS under environmental and genetic influence. Oxidative stress has been implicated in autoimmune disorders (rheumatoid arthritis, systemic lupus erythematosus, psoriasis, and celiac disease) where it plays an important role in the disease process. Oxidative stress is increased in systemic lupus erythematosus (SLE), and it contributes to immune system dysregulation, abnormal activation and processing of cell-death signals, autoantibody production and fatal comorbidities. Mitochondrial dysfunction in T cells promotes the release of highly diffusible inflammatory lipid hydroperoxides, which spread oxidative stress to other intracellular organelles and through the bloodstream. Oxidative modification of self antigens triggers autoimmunity, and the degree of such modification of serum proteins shows striking correlation with disease activity and organ damage in SLE (Perl 2013. Nat Rev Rheumatol. 9(11): 674-686). Rheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation of the joints and tissue around the joints with infiltration of macrophages and activated T cells. The pathogenesis of this disease is due to the generation of ROS and RNS at the site of inflammation. RA is one of the conditions that induce oxidative stress. A fivefold increase in mitochondrial ROS production in whole blood and monocytes of RA patients-compared with healthy subjects suggests that oxidative stress is a pathogenic hallmark in RA. Free radicals are indirectly implicated in joint damage because they also play an important role as secondary messengers in inflammatory and immunological cellular response in RA. T-cell exposure to increased oxidative stress becomes refractory to several stimuli including those for growth and death and may perpetuate the abnormal immune response. On the other hand, free radicals can degrade directly the joint cartilage, attacking its proteoglycan and inhibiting its synthesis (for review see Quiñonez-Flores et al. 2016. Biomed Res Int. 2016:6097417).
- There is now substantial evidence that inflammatory lung diseases such as asthma and chronic obstructive pulmonary disease (COPD) are characterized by systemic and local chronic inflammation and oxidative stress. An important source for increased airway oxidative stress is the recruitment of inflammatory cells into the airway after exposure to trigger factors. These activated cells can generate anion superoxide through reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase pathway. Mitochondrial dysfunction in airway epithelial cells, which occurs in response to mechanical and environmental stimuli, can also contribute to the formation of anion superoxide and airway oxidative stress. Subjects with asthma have greater systemic and airway increased oxidative stress, which is associated with worse asthma severity. As with asthma, subjects with COPD have increased airway oxidative stress and nitrosative stress. Patients with COPD have a greater degree of immunostaining for nitrotyrosine in the airway epithelium and inflammatory cells in sputumA significant imbalance in the airway thiol metabolism has also been described for patients with COPD, which may be associated with downstream redox transcription changes and proinflammatory events. For review see Holguin 2013. Ann
Am Thorac Soc 10 Supplement: S150-S157. - Inflammatory bowel disease (IBD) is an incurable chronic inflammatory intestinal disorder of the gastrointestinal (GI) tract that dramatically impacts quality of life. Crohn’s disease (CD) and ulcerative colitis (UC) are the principal types of IBD. CD may occur in any region of the GI tract involving the ileum and colon in a discontinuous pattern by transmural inflammation, while UC affects only the colon and rectum continuously and is restricted to the mucosa. Accumulating data from both experimental models and clinical studies indicate that oxidative stress signalling is involved in and contributes to the development of IBD through multiple levels of function. Oxidative stress leads to damages of the mucosal layer in the GI tract and bacterial invasion, which in turn stimulates the immune response and initiates IBD. During inflammation, immune cells, such as leukocytes, monocytes, and neutrophils augment ROS production during respiratory, prostaglandin, and leukotriene metabolism, resulting in further tissue damages. For review see Tian et al. 2017. Oxid Med Cell Longev 4535194.
- Celiac disease (CD) is an immune-mediated chronic inflammatory disorder of the upper small intestine induced by gluten and related prolamines in genetically susceptible individuals. As in other autoimmune conditions, environmental, genetic, and immunological factors may be involved in the pathogenesis of CD. In addition to this, oxidative stress is also implicated in the pathogenesis of CD. For example, activation of xanthine oxidase is one of the mechanisms of ROS overproduction in small intestinal mucosa of celiac patients. For review see Patlevic et al. 2016. Integr Med Res 5: 250-258.
- Gastritis is defined as inflammation of the stomach mucosal lining and occurs in several conditions including H. pylori infection, NSAID use, alcohol consumption, and stress. Peptic ulcer disease (PUD) occurs in the proximal GI tract and is often associated with chronic gastritis. Gastric and duodenal ulcers represent the most common and chronic PUDs. Gastritis and peptic ulcer are caused by multiple factors, both endogenous and exogenous, and free radicals are closely linked to both conditions. There are several factors contributing to the accumulation of ROS in the stomach. Reduced antioxidant enzyme SOD levels and antioxidant vitamin intake contribute to the accumulation of ROS associated with gastroduodenal inflammatory diseases. Ethanol-induced gastric inflammation is associated with increased superoxide generation. Phagocytic leukocytes are the main source of ROS in chronic inflammation such as one observes in H. pylori induced gastritis and IBD. Significant numbers of neutrophils and/or macrophages infiltrate the gastric mucosa during inflammation, generating large amounts of ROS. For review see Bhattacharyya et al. 2014. Physiol Rev 94: 329-354.
- Oxidative stress also plays a critical role in liver diseases like viral Hepatitis (Type A, B and C) and liver cirrhosis. It has been clearly established that hepatitis C is associated with strong oxidative stress. This was revealed in liver tissues and in blood serum/plasma samples of Cchronic hepatitis C patients using a variety of techniques, including direct measurement of ROS, quantification of DNA, lipid and protein oxidation products, as well as by assessing the total oxidant/antioxidant status or the levels of individual antioxidants. Screening of the liver biopsies of chronic hepatitis C virus carriers revealed significant elevation of the levels of oxygen radicals and stress markers malondialdehyde (MDA) and 4-hydroxynonenal- (HNE)-and other protein adducts. In addition, serum/plasma of such patients is characterized by increased levels of a wide array of oxidative stress markers such as MDA, lipid peroxides, protein carbonyl content or thioredoxin (for review see Ivanov et al. 2017. Oncotarget, 2017, Vol. 8, (No. 3), pp: 3895-3932).
- Patients with chronic hepatitis B exhibit signs of pronounced oxidative stress. Levels of oxygen radicals in liver specimens from these patients exceed the levels in healthy people. Patients with hepatitis B exhibit signs of oxidative stress not only in the liver but also in plasma/sera. Chronic hepatitis B is accompanied by an increase in total oxidant status and a concomitant reduction of total antioxidant status. Plasma/serum of these patients was also characterized by the elevated levels of ROS, including H2O2, and oxidation products of lipids and proteins. Oxidative stress is not just a hallmark of chronic HBV infection and advanced liver disease; it is also observed in acute and occult hepatitis B, as well as in asymptomatic HBV infections. Occult hepatitis B infection is characterized by increased levels of ROS in lymphocytes and consequent DNA damage. However, the most dramatic changes have been described in hepatitis B patients with liver cirrhosis and with acute chronic hepatitis B liver failure (for review see Ivanov et al. 2017. Oncotarget, 2017, Vol. 8, (No. 3), pp: 3895-3932).
- Cirrhosis is a complication of many forms of chronic liver diseases and is a late stage of fibrosis, in which regenerative nodular formation surrounded by fibrous bands of the liver. In cirrhosis, oxidative stress induced mainly by an overproduction of reactive oxygen species, which is a critical determinant of endothelial dysfunction and is due to disturbed balance between oxidant and antioxidant enzymes. Increased superoxide formation in the presence of equimolar concentrations of NO will lead to the formation of the potent ROS and reactive nitrogen species. For review see Vairappan 2015. World J Hepatol 27; 7(3): 443-459.
- Hepatotoxicity implies chemical-driven liver damage. Drug-induced liver injury is a cause of acute and chronic liver disease. Drug-induced liver injury is responsible for 5% of all hospital admissions and 50% of all acute liver failures. The liver is the most frequently targeted organ in terms of drug toxicity. The production of radical species, specifically ROS and RNS, has been proposed as an early event of drugs hepatotoxicity and as an indicator of hepatotoxic potential. It has been discovered that a lot of drugs could induce oxidative stress including increase of cellular oxidants and lipid peroxidation, depletion of antioxidants in the liver, such as anti-inflammation drugs, anti-analgesic drugs, anti-cancer drugs and antidepressants (Li et al. 2015. Int. J. Mol. Sci. 16: 26087-26124). More than 900 drugs have been implicated in causing liver injury, which are hereby incorporated by reference (https://livertox.nlm.nih.gov/; Björnsson 2016. Int. J. Mol. Sci. 17: 224).
- In the pathogenesis of alcoholic liver disease (ALD), the direct consequence of ethanol metabolism also seems to be related to ROS production, mitochondrial injury and steatosis, which are the common features of acute and chronic alcohol exposure (Li et al. 2015. Int. J. Mol. Sci. 16: 26087-26124).
- Both acute kidney injury (AKI) and chronic kidney disease (CKD) that lead to diminished kidney function are interdependent risk factors for increased mortality. If untreated over time, end stage renal disease (ESRD) is an inevitable outcome. Acute and chronic kidney diseases occur partly due to imbalance between the molecular mechanisms that govern oxidative stress, inflammation, autophagy and cell death. Numerous studies suggest oxidative stress and its systemic effects play a pivotal role in the development of AKI. A recent study demonstrated an increased urinary thioredoxin 1 (TRX1) expression as an oxidative stress biomarker with respect to renal injury. Diabetic nephropathy (DN) is a devastating complication of diabetes and a major cause of CKD. In kidney, mitochondrial respiratory chain and NADPH oxidases (NOX) are the major common sources of ROS and NOX have been demonstrated to produce oxidative stress by enhancing vascular dysfunction and fibrosis in CKD. For review see Sureshbabu et al. 2015. Redox Biology 4: 208-214.
- Drug-induced kidney injury (nephrotoxicity) is a serious problem in clinical practice and accounts for 19%-26% of cases with acute kidney injury (AKI) among hospitalized patients. Moreover, AKI causes a severe condition associated with high probabilities of developing progressive chronic kidney disease or end-stage renal disease, thus leading to high mortality rates. Most drugs found to cause nephrotoxicity exert toxic effects by one or more common pathogenic mechanisms. These include altered intraglomerular hemodynamics, tubular cell toxicity, inflammation, crystal nephropathy, rhabdomyolysis, and thrombotic microangiopathy. AKI includes acute tubular necrosis (ATN) and acute interstitial nephritis (AIN). A mechanism underlying ATN is oxidative stress. Proximal tubular toxicity develops due to direct nephrotoxic effects such as mitochondrial dysfunction, lysosomal hydrolase inhibition, phospholipid damage, and increased intracellular calcium concentration, leading to formation of reactive oxygen species (ROS) with injurious oxidative stress (Hosohata 2016. Int. J. Mol. Sci. 17: 1826).
- Medications that are potentially harmful to the kidneys (nephrotoxic) are, e.g. antimicrobials like antibiotics (for example streptomycin, gentamicin) or antivirals (for example acyclovir, foscarnet) or antifungal (for example amphotecerin B), analgesics, non-steroidal anti-inflammatory drugs (NSAID) (for example ibuprofen, naproxen), diuretics, proton pump inhibitors, chemotherapeutics (for example cisplatin), contrast dyes, cardiovascular agents like ACE-inhibitors or statins, anti-depressants, immune suppressants (for example cyclosporine A) and antihistamines (for reference see Naughton 2008. Am Fam Physician. 2008;78(6):743-750, Table 1; Hosohata 2016. Int. J. Mol. Sci. 17: 1826).
- In carcinogenesis, high reactive hydroxyl radicals cause oxidative DNA damage and peroxynitrite, which causes both oxidative damage and nitration of DNA bases. The majority of mutations induced by ROS appear to involve guanine modification, causing guanine (G) → thymin (T) transversions. If it relates to critical genes such as oncogenes or tumor suppressor genes, initiation or progression of cancer can result. The high metabolism of cancer cells is generally associated with an increase in ROS; however, such levels are less deleterious in cancer cells than they would be in normal cells. For example, although the ROS level increases by a modest degree, tumorigenic cells can induce a new redox balance, resulting in cellular adaptation and proliferation. The ROS generated by the respiratory chain in the mitochondria and by the Nox enzymes in the cytoplasm are particularly important. In fact, Nox proteins are now considered to be oncogenic proteins, and mitochondrial dysfunction is associated with tumorigenesis. Oxidative stress is involved in all stages of carcinogenesis and there is a dose-dependent association between level of the persistent or chronic oxidative stress and the tumor stage. During the carcinogenesis process the normal cells are transformed into abnormal cells owing to a number of structural changes and mutations in genes expression. In the subject literature carcinogenesis is described by three main stages: initiation, promotion and progression. All these stages have been postulated to be linked with contribution of ROS and RNS. ROS have an important role in the pathophysiological states involved in neovascularization. For example ROS-generating enzymes, such as NADPH oxidases (e.g., Nox: Nox1-5), activate redox signaling pathways that ultimately lead to angiogenesis. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- Prostate cancer is the most frequently diagnosed non-cutaneous malignancy in males. This is a multi-focal, filed-type disease, which forms solid tumors of glandular origin. Prostate cancer is mainly a disease of aging, with most cases occurring in men over the age of 55. Over the last decade association between prostate cancer risk and oxidative stress has been recognized, and epidemiological, experimental and clinical studies have unequivocally proven a role for oxidative stress in the development and progression of this disease, commonly associated with a shift in the antioxidant-prooxidant balance towards increased oxidative stress. Environmental factors like diet, inflammation, and changes in cellular functions pertaining to NAD(P)H Oxidase, androgen signalling, mitochondrial DNA mutations, aging, and redox imbalance are possible mechanisms that contribute to increase ROS generation. This increased ROS may further stimulate cell proliferation, cause somatic DNA mutations and promote genetic instability, cell cycle arrest, senescence, and in cancer cells can cause increased angiogenesis, and motility. Especially increased Nox expression driven ROS generation in prostate cancer could lead to the generation of a malignant phenotype by modulating various signalling cascades. For review see Khandrika et al. 2009. Cancer Lett. 282(2): 125-136; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- The excessive production of ROS in breast cancer cells include: a strong expression of thymine phosphorylase leading to degradation of thymidine to thymine and 2-deoxy-D-ribose phosphate; oxidation of 17-estradiol panoxyl radicals to lactoperoxidase participating in metabolism of estrogens and inflammation. In addition to the increase in free radicals, antioxidant changes are related to breast cancer risk, for example the levels of SOD and GPX were found to be higher in the blood of breast cancer cases compared to that of healthy women as a reply on the increased production of superoxide and hydrogen peroxide. In breast cancer, the tumor suppressor gene breast cancer gene 1 (BRCA1) is mutated in 40-50% of hereditary breast cancers and absent or expressed at low levels in 30-40% of sporadic breast cancers. BRCA1 is a caretaker gene that is responsible for repairing DNA, and it is able to upregulate several genes involved in the antioxidant response by controlling the activity of the transcription factors Nrf2 and NfκB. Apart from the inhibitory action of BRCA1 on ROS generation, BRCA1 also reduces the levels of protein nitration due to RNS accumulation in cells, and it enhances DNA repair processes that ultimately help to cope with oxidative stress. For review see Nourazarian et al. 2014 Asian Pac J Cancer Prev, 15 (12): 4745-4751; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- The human lungs are exposed continuously to air pollution oxidants in addition to endogenously generated ROS and RNS, which are involved in physiological biochemical mechanisms and normal cellular signalling pathways. It is a commonly considered hypothesis that tobacco smoking is a key risk factor in lung cancer development. Both, clinical and experimental research has consistently shown the important role of OS in lung cancer. Evidence is available that supports the importance of oxidative stress and its correlation with increased incidence of malignant respiratory diseases due to inflammation, activation of transcriptional factors and DNA damage. During inflammation, enhanced ROS production induces DNA damage, inhibition of apoptosis, and activation of protooncogenes by initiating signal transduction pathways. Inflammatory cells are particularly effective in generating ROS and other reactive species, thus increasing oxidative damage and promoting mechanisms of carcinogenesis. The ability of respirable particles or fibrous dusts to penetrate the respiratory system and reach the lung alveoli in order to generate ROS and other oxidants or free radicals is suggested to be the main factor involved in their pathogenic potential. Synergistic mechanisms of inhalable particulate matter (penetrating deep into the lung’s alveoli) and other components of air pollution (ozone, nitric oxide, soot, heavy metals, PAHs) and tobacco smoke have been studied. The porous surfaces of airborne particles provide a fertile ground for catalyzing the increased generation of ROS or other damaging oxidants, which are potential initiators of pulmonary carcinogenesis. For review see Valavanidis et al. 2013. Int. J. Environ. Res. Public Health 10: 3886-3907; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- Colorectal cancer (CRC) is one of the most common cancers worldwide, with the highest incidence rates in western countries. Colon cancer originates from the epithelial cells that line the bowel. These cells divide rapidly and have a high metabolic rate, which has been found as a potential factor that may be responsible for increased oxidation of DNA. It was found that the human colorectal tumors (adenomasand carcinomas) have increased levels of different markers of oxidative stress, such as increased levels nitric oxide (NO), 8-oxodG in DNA, lipid peroxides, glutathione peroxidase (GPx), catalase (CAT), and decreased methylation of cytosine in DNA. Besides lipid modifications also increased leukocyte activation in carcinogenic tissue was found, which indicates possible contribution of inflammatory cells to a further oxidative stress. Moreover, it was shown that levels of anti-oxidant vitamins A, C, E in blood plasma of colorectal cancer patients were statistically lower compared to healthy individuals. For review see Perse 2013. BioMed Research International 725710; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- In the industrialized countries, bladder cancer is the fourth most frequently occurring malignant tumors. Recent studies indicate the involvement of oxidative and nitrosative stress in the formation and development of this disease. Red-ox disorders are characteristic for both, the initiation and progression of bladder cancer. There are observed changes in the activity of transcription factors, such as nuclear factor NF-kB; transcription factors: AP-1, Nrf2 and STAT3 and hypoxia-inducible factor HIF-1α. In addition, studies indicate a role for oxidative stress in the regulation of MAPK cascade and its involvement in carcinogenesis consisting bladder. Nitric oxide also plays an important role in tumor biology. Numerous studies show that the bladder cancer is characterized by an intensified production of NO. In contrast to the ROS, which overproduction result from exposure to carcinogenic xenobiotic, nitrogen oxide in high level is produced during inflammation. Sustained iNOS activity therefore plays an important role in carcinogenesis associated with the inflammatory response, characteristic also for bladder cancer. For review see Sawicka et al. 2015. Postepy Hig Med Dosw 69: 744-752.
- Ovarian cancer is the fifth leading cause of cancer death; the leading cause of death from gynecologic malignancies, and the second most commonly diagnosed gynecologic malignancy. The overwhelming majority of ovarian cancers are derived from ovarian surface epithelium. Metastasis is achieved through detachment of single cells or clusters of cells from the primary tumor followed by implantation on peritoneal mesothelial lining. Ovarian, endometrial, and cervical cancer consist a great problem in oncology due to their diagnosis in advanced stage. Research finding have shown that oxidative stress plays a causal role in the carcinogenesis of two subtypes of ovarian cancer: clear cell carcinoma and endometriosis carcinoma. Evidence suggests that ovarian cancer patients have decreased levels of circulating antioxidants and higher levels of oxidative stress. It has been reported that epithelial ovarian cancer (EOC) tissues and cells manifest a pro-oxidant state characterized by an increased expression of key pro-oxidant enzymes and decreased expression of antioxidant enzymes. Specifically, EOC cells and tissues manifested an increased expression of iNOS, MPO, NAD(P)H oxidase, as well as an increase in NO levels. Moreover, EOC cells manifested lower apoptosis.
- Endometrial cancer has been reported to be associated with endometriosis disease, and the high levels of free iron hemosiderin or heme in endometrial cysts are considered as a main factor responsible for the oxidative stress development and chronic inflammation.
- Cervical cancer is the second most common cancer in women worldwide being a subject of intensive research. Several experimental studies suggested the participation of oxidative stress in cervical, indicating that antioxidants can alter the redox balance in cervical cancer cells, inhibit transcription factors AP-1 and NF-κB or induce cell apoptosis. For review see Saed et al. 2017. Gynecologic Oncology 145: 595-602; Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Sosa et al. 2013. Ageing Research Reviews 12: 376- 390.
- Oxidative damage induced by OS has been also implicated in leukemia and the decreased levels of antioxidants and oxidatively modified DNA and lipids caused by high ROS production were found in serum of chronic lymphocytic leukemia patients. Moreover, it was found that the chronic leukemia cells were able to adapt to intracellular OS through upregulation the stress-responsive hemeoxygenase-1 confirming involvement of ROS in the pathogenesis of leukemia cancer. Also, GSH depletion in lymphocytes of the chronic lymphocytic has been demonstrated in leukemia B patients. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919.
- Gastric cancer (GC) is one of the most frequent diseases in human population. It is the fourth frequent cancer and the second most common cause of deaths from cancer in the world. The main risk factor for gastric cancer is chronic inflammation caused by bacterial growth. For example, infection by Helicobacter pylori, which increases the production of reactive oxygen and nitrogen species in human stomach, is thought to be important in the development of gastric cancer. It has been shown that protein oxidation products were significantly higher in GC patients. Moreover, it was found that the antioxidant potentials of SOD and catalase were lower in gastric cancer tissues compared to the control healthy tissues. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Ma et al. 2013. Oxidative Medicine and Cellular Longevity 543760.
- There are many factors involved in liver carcinogenesis, including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection, alcohol abuse, and nonalcoholic fatty liver disease (NAFLD), aflatoxin B1, obesity, diabetes, dietary habits, and iron accumulation. In general, oxidative stress can be triggered by any dangerous or inflammatory signal and affects multiple cells in the liver. Liver injury can be either an acute or a chronic inflammatory process. In the environment of local inflammation, many types of liver cells, such as liver sinusoidal endothelial cells (LSECs), hepatic stellate cells (HSCs), dendritic cells (DCs), and Kupffer cells (KCs), are activated. These cells produce many kinds of immune mediators, cytokines, and chemokines that may also lead to the production of oxidative stress. In recent years, studies on the relationship between oxidative stress and hepatic stellate cells have been increasing. These cells have been proven to play a central part in the process of liver fibrosis and can induce collagen production after activation in the body by free radicals, which are produced by ROS and superoxide anions, and further induce damage to liver cells.
- It is known that over 80% of cases of HCC are associated with chronic HBV or HCV infection. HBV- and HCV-related chronic inflammation and fibrosis of the liver are usually induced by oxidative stress, which contributes to the pathogenesis of hepatocarcinogenesis. HBV infection results in activation of macrophages to produce a variety of proinflammatory cytokines, such as IL-1β, IL-6, CXCL-8, and TNF-α. Such persistent abnormal production of cytokines and the resulting production of ROS have an influence on hepatocarcinogenesis. HCV-induced oxidative stress contributes to the development of hepatocellular carcinoma (HCC). Moreover, levels of oxidative stress markers in chronic hepatitis C patients correlate positively with the probability of development of HCC and can serve as prognostic markers for HCC recurrence in chronic hepatitis C patients who underwent liver transplantation. Carcinogenesis is orchestrated by multiple ROS-mediated processes. For review see Wang et al. 2016. Oxidative Medicine and Cellular Longevity 7891574.
- Skin is a major environmental interface for the body, which accidentally or occupationally gets exposed to a number of chemical mutagens and carcinogens. Skin cancer represents a major and growing public health problem. It accounts for more than 40 % of all new cancer diagnosed. 80 % of skin cancers result from basal cell carcinomas (BCC); another 16 % are squamous cell carcinomas (SCC), and 4 % are melanomas. A large number of evidence indicates that UV can induce DNA damage and as a consequence DNA strand breaks and DNA cross links are detected. An important process in skin cancer is generation of hydrogen peroxide by melanocytes and decrease in catalase activity. Moreover, there are findings showing that mutations in several genes linked with melanoma result from oxidative stress. For review see Kruk and Aboul-Einein 2017. Mini-Reviews in Medicinal Chemistry 17: 904-919; Narendhirakannan. 2013. Ind J Clin Biochem 28(2):110-115.
- Reactive species generated during infection may have serious consequences for the disease once they are released to any degree. The oxidative stress can initiate adverse effects in different organs. Development of oxidative stress can be accelerated in the course of hypoxia. Hypoxia is a known complication of infectious diseases. Hypoxia is not peculiar to one disease. Influenza, viral hepatitis and tuberculosis are all examples of infectious diseases in which hypoxia takes place. Direct generation of reactive oxygen species can be initiated by metals. In an example, iron, cooper, and cadmium can catalyze the development of oxidative stress by Fenton reaction in which hydrogen peroxide is converted into hydroxyl radical and hydroxide anion. Heavy metals are involved in pathological processes linked to infections like other pathologies. As mentioned, livers damaged by viral hepatitis are vulnerable to heavy metals due to faulty elimination processes. Clinical studies on patients with viral hepatitis A, B, C, D, and E demonstrated that accumulation of copper and iron caused oxidative stress and oxidative damage to patients’ liver tissue. Like hepatitis, AIDS is accompanied by imbalance in oxidative homeostasis. Elevated markers of oxidative damage of targets in the body and accumulation of reactive oxygen species are common in HIV-infected patients. Blood antioxidants are reduced over the long term in the infected individuals. For review see Pohanka 2013. Folia Microbiol 58:503-513.
- Sepsis and septic shock remain as leading cause of death in adult intensive care units. It is widely accepted that sepsis and septic shock are caused predominantly by gram-negative bacteria and their endotoxins. Endotoxin or Lipopolysaccharide (LPS) have important roles as host responses and trigger the inflammatory processes, caused by gram-negative bacterial infection. Production of oxygen radicals by neutrophils and macrophages such as reactive oxygen species (ROS), NO (nitric oxide) and peroxynitrite promote gene expression of proinflammatory mediators. ROS and RNS are antimicrobial agents produced by these leukocytes that can directly destroy microbial pathogens. During sepsis, excess production of ROS and RNS threatens the integrity of various biomolecules including proteins, lipids as well as lipoproteins, protein oxidation and DNA resulting in tissue damage, by lipid peroxidation of cell membranes, protein oxidation and DNA strand breaks. These mechanisms contribute to multi organ failure during sepsis resulting in myocardial depression, hepatocellular dysfunction, endothelial dysfunction, and vascular catecholamine hypo-responsiveness. As a major source of ROS production, mitochondria are especially prone to ROS-mediated damage. Such damage can induce the mitochondrial permeability transition caused by opening of nonspecific high conductance permeability transition pores in the mitochondrial inner membrane. ROS themselves also provide a signal leading to the induction of autophagy, apoptosis, and necrosis. Excessive ROS production and adenosine triphosphate depletion from uncoupling of oxidative phosphorylation promote necrotic cell death. Release of cytochrome-c after mitochondrial swelling activates caspases and initiates apoptotic cell death. For review see Kaymak et al. 2011. FABAD J. Pharm. Sci. 36: 41-47.
- Thus, in another specific embodiment of the invention the herein disclosed DPP3 binder, specifically the herein provided anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold which are binding to an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof are provided for the use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress, said disease is selected from the group comprising the above described neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
- In another specific embodiment said disease is selected from the group comprising neurodegenerative diseases (e.g. Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)), metabolic syndrome (including insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes), cardiovascular disorders (e.g. atherosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury/ stroke and myocardial infarction), autoimmune diseases (e.g. rheumatoid arthritis and systemic lupus erythematosus), inflammatory lung diseases (e.g. COPD, asthma), kidney diseases (including renal toxicity (drug-induced kidney disease), acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD)), liver diseases (e.g. hepatotoxicity, viral hepatitis, cirrhosis), digestive diseases (including inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease; gastritis, pancreatitis and peptic ulcer), viral infectious diseases (e.g. blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus)), cancer (e.g. prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer) and inflammation, sepsis, septic shock and SIRS.
- In another specific embodiment of the invention the herein disclosed DPP3 binder, specifically the herein provided anti-DDP3 antibody, and/or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold which are binding to an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof are provided for the use in the prevention or treatment of an acute condition, wherein said acute condition may be selected from a group comprising hepatotoxicity and kidney toxicity.
- In the context of the present invention, oxidative stress and subsequent toxicities can also be induced by chronic alcohol consumption, chronic exposure to cigarette smoke and as a side-effect of different drug treatments (reviewed in Deavall et al. 2012, see table 2 below).
- Acetaminophen — a widely used analgesic and antipyretic drug — is a prototypical hepatotoxicant for drug-induced liver injury that is connected to KEAP-Nrf2 signaling (Ma, 2013). Other therapeutics inducing oxidative stress include oltipraz and auranofin (Ma, 2013).
-
TABLE 2 Examples of toxicities associated with drug-induced oxidative stress (from Deavall et al. 2012) Therapeutic class Drug Example toxicities Evidence for oxidative stress Antineoplastic (anthracycline) Doxorubicin Cardiac toxicity Reduction of doxorubicin to free radical increases ROS in cardiomyocytes. Lipid peroxidation, mitochondrial dysfunction, apoptosis Antiretroviral AZT Skeletal myopathy, cardiac toxicity Increased ROS and NOS (peroxide and peroxynitrate). Overexpression of superoxidase dismutase/catalase protects against toxicity, apoptosis Anti-inflammatory Diclofenac Nephrotoxicity, hepatotoxicity Oxidative stress generated by a cation radical or redox cycling of intermediates derived from hydroxylation. Multifactorial perturbations in mitochondrial dysfunction Analgesia Paracetamol Hepatotoxicity Formation of reactive metabolite, depletion of glutathione, activation of proapoptotic proteins. Mitochondrial dysfunction, inflammation Antineoplastic (platinum) Cisplatin Nephrotoxicity, ototoxicity Increases in superoxide anion, hydrogen peroxide, and hydroxyl radical. Depletion of antioxidants GSH-peroxidase and GSH-reductase. Mitochondrial dysfunction, apoptosis Antipsychotic Chlorpromazine Dermal toxicity (due to phototoxicity) Generation of singlet oxygen and superoxide in response to UVA/B irradiation - In accordance with the present invention, the person skilled in the art is well aware that the presence and degree of oxidative stress may be determined and quantified by suitable biomarker assays known in the art. Respective examples for these markers are given below, but these shall be not construed as limiting possibilities to measure oxidative stress in accordance with the invention:
- Free radicals can damage biological molecules including nucleic acids, proteins, and lipids. The products of these reactions can become markers of oxidative stress. Serum is the most common material for the evaluation of the components of oxidative stress. It allows the estimation of most enzymes, substrates, and products of redox reactions. These enzymes include xanthine oxidase, NOS, lipoxygenase, cyclooxygenase, myeloperoxidase, prolyl-oligopeptidase, nicotinamide adenine dinucleotide phosphate-oxidase 1 (NOX1), and NADPH-dependent oxidase. The following are markers of oxidative lipid damage: isoprostanes (IsoP-prostaglandin like substances), for example, 8-iso-prostaglandin (F2α-8-iso-PGF2α) which constitutes the product of lipid peroxidation of arachidonic acid, malondialdehyde (MDA), the formation of fluorescent peroxidized lipid-protein covalent adducts, and the increase in conjugated diene. Oxidative stress involves the oxidation of proteins and glycoxidation. The following are the results of this reaction: the glycophore content, the total level of advanced protein oxidation (AOPP), protein carbonyls, dityrosine level, N′-formylkynurenine, and a decreased level of serum protein thiol groups. Other specific markers of protein oxidation are tyrosine (a marker for hydroxyl radical) and 3-nitrotyrosine (a marker for RNS). Furthermore, 3-nitrotyrosine is a specific marker of peroxynitrite-induced cellular damage. Other indicators in the serum include kynurenine, N′-formylkynurenine, thioredoxin, and 8-hydroxy-2′-deoxyguanosine.
- Respective measurements of biomarkers for oxidative stress in humans are summarized in Ilaria Marrocco, Fabio Altieri, and Ilaria Peluso; Review Article: Measurement and Clinical Significance of Biomarkers of Oxidative Stress in Humans; Oxidative Medicine and Cellular Longevity Volume 2017 (2017), Article ID 6501046, 32 pages.
- Moreover, in accordance with the present invention, the person skilled in the art is well aware that the bioactivity of DPP3, which is influenced by the herein disclosed DPP3 binder, can be measured for e.g. inhibition via suitable assays known in the art. Respective examples are given below, but these shall be not construed as limiting possibilities to determine DPP3 bioactivity:
- Inhibition of DPP3 activity in a liquid phase assay by a binder may be determined as followed: Blood samples (e.g. serum, heparin-plasma, Li-plasma, citrate-plasma, whole blood) of patients before and after anti-DPP3 antibody treatment is incubated with specific DPP3 substrates in a liquid phase assay. The specific liquid phase DPP3 activity assay to determine the inhibitory ability of inhibitory DPP3 antibodies in blood samples comprises the following steps:
- Addition of 20 µl blood sample in 200
µl 50 mM Tris-HCl, pH 7,5 in a black non-binding microtiter plate (96-well). Hereby, the person skilled in the art is aware that buffering conditions, concentrations and pH etc. can be varied. - Addition of the fluorogenic substrate Arg-Arg-βNA (20 µl, 2 mM).
- Incubation at 37° C. and monitoring the generation of free βNA in a Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH) over 1 hour. Fluorescence of βNA is detected by exciting at 340 nm and measuring emission at 410 nm.
- Slopes (in RFU/ min) of increasing fluorescence of the different samples are calculated.
- Analysis of DPP3 activity values before and after anti-DPP3 antibody treatment.
- In contrast thereto a solid phase assay is an assay where the respective binding events take place at the solid phase. Inhibition of DPP3 activity in a solid phase assay by a binder may be determined as followed according: Blood samples (e.g. serum, plasma, whole blood) of patients before and after anti-DPP3 antibody treatment are contacted with an immobilized capture-binder for enzyme capture activity assay (ECA) on a solid phase. Preferably, as capture-binder for the ECA is chosen the one with the least inhibitory ability. The capture-binder should inhibit DPP3 activity less than 50 %, preferably less than 40 %, preferably less than 30 %. The specific liquid phase DPP3 activity assay to determine the inhibitory ability of possible capture-binders is described in detail in Example 1 below.
- The ECA to determine the inhibitory ability of inhibitory DPP3 antibodies in blood samples comprises the following steps:
- Contacting said sample with a capture-binder that binds to full-length DPP3 but preferably inhibits DPP3 activity in a liquid phase assay less than 50%, preferably less than 40%, more preferably 30%
- Separating DPP3 bound to said capture binder from bodily fluid sample,
- Adding substrate of DPP3 to said separated DPP3,
- Quantifying DPP3 activity by measuring the conversion of the substrate of DPP3,
- Evaluation of measured signals and analysis of DPP3 activity values before and after anti-DPP3 antibody treatment.
- The method for determining active DPP3 may be conducted as liquid phase assay and as solid phase assay. Inhibition of DPP3 activity may be determined in a liquid assay nevertheless according to the above-described procedure.
- In yet another embodiment, a capture or binding assay may be performed to detect and/or quantitate the protease activity of DPP3. For example, an antibody reactive with DPP3 protein, but which does not interfere with peptidase activity, may be immobilized upon a solid phase. The test sample is passed over the immobile antibody, and DPP3, if present, binds to the antibody and is itself immobilized for detection. A substrate may then be added, and the reaction product may be detected to indicate the presence or amount of DPP3 in the test sample. For the purposes of the present description, the term “solid phase” may used to include any material or vessel in which or on which the assay may be performed and includes, but is not limited to, porous materials, nonporous materials, test tubes, wells, slides, etc.
- Moreover, in accordance with the present invention, the person skilled in the art is well aware that the binding affinity of the herein disclosed DPP3 binder to DPP3 may be measured by various suitable assays known in the art. Respective examples are given below, but these shall be not construed as limiting possibilities to measure binding affinity of the herein disclosed DPP3 binder to DPP3:
- The binding affinity of the DPP3 binder to the epitope according to SEQ ID NO.: 2 in accordance with the invention may be determined in accordance with Example 1 and as further set out below:
- A binding assay may be performed to detect and/or quantitate antibody binding to the immunization peptide (i.e. SEQ ID NO.: 2). For example, this immunization peptide may be immobilized upon a solid phase. The test sample (e.g. antibody solution) is passed over the immobile immunization peptide, and bound antibody can be detected. For the purposes of the present description, the term “solid phase” may be used to include any material or vessel in which or on which the assay may be performed and includes, but is not limited to, porous materials, nonporous materials, test tubes, wells, slides, etc.
- Exemplary detection methods:
- Label antibody before contacting with solid phase and detect respective label (fluorescence, chemiluminescence, enzymatic etc.)
- Use labeled secondary antibody against specific Fc part of sample-antibody. Incubate solid phase bound antibody with secondary antibody (e.g. anti human IgG, anti murine IgG) and detect respective label (fluorescence, chemiluminescence, enzymatic etc.)
- Use a labeled antibody as competitor for solid phase binding (e.g. labeled AK1967).
- Quantifiy binding affinity by decrease of signal.
- In another embodiment of the invention, the herein disclosed binder of the invention, and DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or anti-DPP3 non-Ig scaffolds are capable to bind circulating DPP3, and thus are directed against circulating DPP3.
- In yet another embodiment of the invention, the herein disclosed binder of the invention, and DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or antiDPP3 non-Ig scaffolds are capable to bind intracellular DPP3, and thus are directed against intracellular DPP3.
- In yet another embodiment of the invention, the herein disclosed binder of the invention, DPP3 binder, specifically the anti-DPP3 antibodies, anti-DPP3 antibody fragments, or anti-DPP3 non-Ig scaffolds are capable to bind membranous DPP3, and thus are directed against membranous DPP3.
- Also subject matter of the present invention, are the herein disclosed binder of the invention, DPP3 binder, specifically is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress, and whereby said binder, DPP3 binder, specifically is an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in a circulating DPP3 protein or functional derivative thereof.
- Also subject matter of the present invention, are the herein disclosed binder of the invention, DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress, and whereby said binder, DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in an intracellular DPP3 protein or functional derivative thereof.
- Also subject matter of the present invention, are the herein disclosed binder of the invention, DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an antiDPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said diseases or acute conditions are associated with oxidative stress, and whereby said DPP3 binder, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold are directed to and binding to an epitope of SEQ ID NO.: 2, wherein said epitope is comprised in a membranous DPP3 protein or functional derivative thereof.
- Subject matter of the present invention is further a method for regulating and/or preventing or treatment of oxidative stress in a patient having a chronic or acute disease or acute condition, characterized in that to said patient a binder of the invention, or a DPP3 binder of the invention, specifically an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold is administered in pharmaceutically effective amounts. According to the invention said patient is a patient in need of regulating and/or preventing or in need of treatment of oxidative stress.
- Another subject of the present invention is a pharmaceutical composition comprising the herein disclosed binder of the invention, or DPP3 binder, specifically comprising an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of diseases or acute conditions of a patient, wherein said disease or acute condition is associated with oxidative stress.
- In another embodiment of the present invention said pharmaceutical composition is a solution, preferably a ready-to-use solution.
- In another embodiment of the present invention said pharmaceutical composition is a solution, preferably a ready-to-use solution comprising PBS at a pH of 7.4.
- In another embodiment of the present invention said pharmaceutical composition is in a dried state that is to be reconstituted before use.
- In another embodiment of the present invention said pharmaceutical composition is in a freeze-dried state that is to be reconstituted before use.
- In another embodiment of the present invention said pharmaceutical composition that is to be used in the prevention and/or treatment of a disease or an acute condition of a patient, wherein said disease or acute condition is associated with oxidative stress is administered orally, epicutaneously, subcutaneously, intradermally, sublingually, intramuscularly, intraarterially, intracerebrally, intracerebroventricularly, intravenously, or via the central nervous system (CNS) or via intraperitoneal administration.
- Another embodiment of the present invention is a kit or an assay comprising the herein disclosed binder of the invention, or DPP3 binder, specifically comprising an anti-DPP3 antibody or an anti-DPP3 antibody fragment or an anti-DPP3 non-Ig scaffold for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress.
- In accordance with the invention the “anti-DPP3 antibody” is an antibody that binds specifically to DPP3, an “anti-DPP3 antibody fragment” is a fragment of said anti-DPP3 antibody, wherein said fragment binds specifically to DPP3. An “anti-DPP3 non-Ig scaffold” is a non-Ig scaffold that binds specifically to DPP3.
- With the context of the invention, “specifically binding to DPP3” may also allow binding to other antigens as well. This means, this specificity would not exclude that the binder may cross-react with other proteins or polypeptides or peptides that contain the epitope according to SEQ ID NO.: 2 against which the binder has been raised. This specifically includes functional variants of DPP3, which also comprise an epitope according to SEQ ID NO.: 2. This also pertains to the specificity of the anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold in accordance with the invention.
- An “antibody” according to the present invention is a protein including one or more polypeptides substantially encoded by immunoglobulin genes that specifically binds an antigen. The recognized immunoglobulin genes include the kappa, lambda, alpha (IgA), gamma (IgG1, IgG2, IgG3, IgG4), delta (IgD), epsilon (IgE) and mu (IgM) constant region genes, as well as the myriad immunoglobulin variable region genes. Full-length immunoglobulin light chains are generally about 25 kDa or 214 amino acids in length.
- Full-length immunoglobulin heavy chains are generally about 50 kDa or 446 amino acids in length. Light chains are encoded by a variable region gene at the NH2-terminus (about 110 amino acids in length) and a kappa or lambda constant region gene at the COOH-terminus. Heavy chains are similarly encoded by a variable region gene (about 116 amino acids in length) and one of the other constant region genes.
- The basic structural unit of an antibody is generally a tetramer that consists of two identical pairs of immunoglobulin chains, each pair having one light and one heavy chain. In each pair, the light and heavy chain variable regions bind to an antigen, and the constant regions mediate effector functions. Immunoglobulins also exist in a variety of other forms including, for example, Fv, Fab, and F(ab′)2, as well as bifunctional hybrid antibodies and single chains (e.g., Lanzavecchia et al., Eur. J. Immunol. 17:105,1987; Huston et al., Proc. Natl. Acad. Sci. U.S.A., 85:5879-5883, 1988; Bird et al., Science 242:423-426, 1988; Hood et al., Immunology, Benjamin, N.Y., 2nd ed., 1984; Hunkapiller and Hood, Nature 323:15-16,1986).
- An immunoglobulin light or heavy chain variable region includes a framework region interrupted by three hypervariable regions, also called complementarity determining regions (CDR’s) (see, Sequences of Proteins of Immunological Interest, E. Kabat et al., U.S. Department of Health and Human Services, 1983). As noted above, the CDRs are primarily responsible for binding to an epitope of an antigen. An immune complex is an antibody, such as a monoclonal antibody, chimeric antibody, humanized antibody or human antibody, or functional antibody fragment, specifically bound to the antigen.
- “Chimeric antibodies” are antibodies whose light and heavy chain genes have been constructed, typically by genetic engineering, from immunoglobulin variable and constant region genes belonging to different species. For example, the variable segments of the genes from a mouse monoclonal antibody can be joined to human constant segments, such as kappa and
gamma 1 or gamma 3. In one example, a therapeutic chimeric antibody is thus a hybrid protein composed of the variable or antigen-binding domain from a mouse antibody and the constant or effector domain from a human antibody, although other mammalian species can be used, or the variable region can be produced by molecular techniques. Methods of making chimeric antibodies are well known in the art, e.g., see U.S. Pat. No. 5,807,715. A “humanized” immunoglobulin is an immunoglobulin including a human framework region and one or more CDRs from a non-human (such as a mouse, rat, or synthetic) immunoglobulin. The non-human immunoglobulin providing the CDRs is termed a “donor” and the human immunoglobulin providing the framework is termed an “acceptor.” - In one embodiment of the invention, all the CDRs are from the donor immunoglobulin in a humanized immunoglobulin. Constant regions need not be present, but if they are, they must be substantially identical to human immunoglobulin constant regions, i.e., at least about 85-90%, such as about 95% or more identical. Hence, all parts of a humanized immunoglobulin, except possibly the CDRs, are substantially identical to corresponding parts of natural human immunoglobulin sequences.
- A “humanized antibody” in accordance with the invention is an antibody comprising a humanized light chain and a humanized heavy chain immunoglobulin. A humanized antibody binds to the same antigen as the donor antibody that provides the CDRs. The acceptor framework of a humanized immunoglobulin or antibody may have a limited number of substitutions by amino acids taken from the donor framework. Humanized or other monoclonal antibodies can have additional conservative amino acid substitutions which have substantially no effect on antigen binding or other immunoglobulin functions. Exemplary conservative substitutions are those such as gly, ala; val, ile, leu; asp, glu; asn, gln; ser, thr; lys, arg; and phe, tyr. Humanized immunoglobulins can be constructed by means of genetic engineering (e.g., see U.S. Pat. No. 5,585,089). A human antibody is an antibody wherein the light and heavy chain genes are of human origin. Human antibodies can be generated using methods known in the art. Human antibodies can be produced by immortalizing a human B cell secreting the antibody of interest. Immortalization can be accomplished, for example, by EBV infection or by fusing a human B cell with a myeloma or hybridoma cell to produce a trioma cell. Human antibodies can also be produced by phage display methods (see, e.g., Dower et al., PCT Publication No. WO 91/17271; McCafferty et al., PCT Publication No. WO 92/001047; and Winter, PCT Publication No. WO 92/20791), or selected from a human combinatorial monoclonal antibody library (see the Morphosys website). Human antibodies can also be prepared by using transgenic animals carrying a human immunoglobulin gene (for example, see Lonberg et al., PCT Publication No. WO 93/12227; and Kucherlapati, PCT Publication No. WO 91/10741).
- Thus, the anti-DPP3 antibody or anti-DPP3 antibody fragment in accordance with the invention may have the formats known in the art. Examples are human antibodies, monoclonal antibodies, humanized antibodies, chimeric antibodies, CDR-grafted antibodies or antibody fragments thereof, but not limited to.
- In a specific embodiment of the invention the anti-DPP3 antibody is a monoclonal antibody or a fragment thereof. In one embodiment of the invention the anti-DPP3 antibody or the antiDPP3 antibody fragment is a human or humanized antibody or derived therefrom. In one specific embodiment one or more (murine) CDR’s are grafted into a human antibody or antibody fragment.
- In a preferred embodiment antibodies according to the present invention are recombinantly produced antibodies as e.g. IgG, a typical full-length immunoglobulin, or antibody fragments containing at least the F-variable domain of heavy and/or light chain as e.g. chemically coupled antibodies (fragment antigen binding) including but not limited to Fab-fragments including Fab minibodies, single chain Fab antibody, monovalent Fab antibody with epitope tags, e.g. Fab-V5Sx2; bivalent Fab (mini-antibody) dimerized with the CH3 domain; bivalent Fab or multivalent Fab, e.g. formed via multimerization with the aid of a heterologous domain, e.g. via dimerization of dHLX domains, e.g. Fab-dHLX-FSx2; F(ab′)2-fragments, scFv-fragments, multimerized multivalent and/or multispecific scFv-fragments, bivalent and/or bispecific diabodies, BITE® (bispecific T-cell engager), trifunctional antibodies, polyvalent antibodies, e.g. from a different class than G; single-domain antibodies, e.g. nanobodies derived from camelid or fish immunoglobulines and numerous others.
- In addition to anti-DPP3 antibodies or anti-DPP3 antibody fragments, other biopolymer scaffolds, so called non-Ig scaffolds, are well known in the art to complex a target molecule and have been used for the generation of highly target specific biopolymers. Examples are aptamers, spiegelmers, anticalins and conotoxins.
- Non-Ig scaffolds with the context of the invention may be protein scaffolds and may be used as antibody mimics as they are capable to bind to ligands or antigens. Non-Ig scaffolds may be selected from the group comprising tetranectin-based non-Ig scaffolds (e.g. described in US 2010/0028995), fibronectin scaffolds (e.g. described in EP 1266 025; lipocalin-based scaffolds (e.g. described in WO 2011/154420); ubiquitin scaffolds (e.g. described in WO 2011/073214), transferring scaffolds (e.g. described in US 2004/0023334), protein A scaffolds (e.g. described in EP 2231860), ankyrin repeat based scaffolds (e.g. described in WO 2010/060748), microprotein (preferably microproteins forming a cystine knot) scaffolds (e.g. described in EP 2314308), Fyn SH3 domain based scaffolds (e.g. described in WO 2011/023685), EGFR-A-domain based scaffolds (e.g. described in WO 2005/040229) and Kunitz domain based scaffolds (e.g. described in EP 1941867). Non-Ig scaffolds may be peptide or oligonucleotide aptamers. Aptamers are usually created by selecting them from a large random sequence pool and are either short strands of oligonucleotides (DNA, RNA or XNA; Xu et al. 2010, Deng et al. 2014) or short variable peptide domains attached to a protein scaffold (Li et al. 2011).
- In an alternative embodiment the anti-DPP3 antibody format is selected from the group comprising Fv fragment, scFv fragment, Fab fragment, scFab fragment, F(ab)2 fragment and scFv-Fc Fusion protein. In another preferred embodiment the antibody format is selected from the group comprising scFab fragment, Fab fragment, scFv fragment and bioavailability optimized conjugates thereof, such as PEGylated fragments.
- With the context of the invention, the term “antibody” generally comprises monoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called “single-chain-antibodies” (Bird et al. 1988), chimeric, humanized, in particular CDR-grafted antibodies, and di- or tetrabodies (Holliger et al. 1993). Also comprised are immunoglobulin-like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample. In this context the term “specific binding” refers to antibodies raised against the molecule of interest or a fragment thereof. An antibody is considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least preferably 50-fold higher, more preferably 100-fold higher, most preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity.
- In a specific embodiment of the invention said anti-DPP3 antibody or anti-DPP3 antibody fragment binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or functional derivative thereof is a monoclonal antibody or a monoclonal antibody fragment thereof. In one embodiment of the invention the anti-DPP3 antibody or the anti-DPP3 antibody fragment binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or functional derivative thereof is a human or humanized antibody or derived therefrom or humanized antibody fragment or derived therefrom.
- In one specific embodiment one or more (murine) CDR’s are grafted into a human antibody or antibody fragment.
- In a specific embodiment said DPP3 binder of the invention, specifically said anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold is a modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold.
- A modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold of the invention may act inhibitory and may block the bioactivity of DPP3 to nearly 100%, preferably to at least more than 90%, more preferably to at least 80, or 70, or 60, or 50, or 40, or 30, or 20, or 10 % when determined by means of the above described method for detecting and measuring the inhibition of DPP3; i.e. measuring the DPP3 binder influence on DPP-3 bioactivity.
- In another specific embodiment, a modulating DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold of the invention may act upregulating and thus may enhance the bioactivity of DPP3 to at least 50 %, preferably to at least more than 60 %, more preferably to at least more than 70 %, more preferably to at least more than 80 %, even more preferably to at least more than 90 %, even more so preferably to at least 95 % when determined by means of the above described method for detecting and measuring the inhibition of DPP3; i.e. measuring the DPP3 binder influence on DPP-3 bioactivity.
- Anti-DPP3 antibodies according to the present invention may be synthesised as follows:
- DPP3 peptides for immunization were synthesized, see table 3 below, (JPT Technologies, Berlin, Germany) with an additional N-terminal cystein (if no cystein is present within the selected DPP3-sequence) residue for conjugation of the peptides to Bovine Serum Albumin (BSA). The peptides were covalently linked to BSA by using Sulfolink-coupling gel (Perbio-science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio. Recombinant GST-hDPP3 was produced by USBio.
- Balb/c mice were intraperitoneally (i.p.) injected with 84 µg GST-hDPP3 or 100 µg DPP3-peptide-BSA-conjugates at day 0 (emulsified in TiterMax Gold Adjuvant), 84 µg or 100 µg at day 14 (emulsified in complete Freund’s adjuvant) and 42 µg or 50 µg at
day 21 and 28 (in incomplete Freund’s adjuvant). At day 49 the animal received an intravenous (i.v.) injection of 42 µg GST-hDPP3 or 50 µg DPP3-peptide-BSA-conjugates dissolved in saline. Three days later the mice were sacrificed and the immune cell fusion was performed. - Splenocytes from the immunized mice and cells of the myeloma cell line SP2/0 were fused with 1
ml 50% polyethylene glycol for 30 s at 37° C. After washing, the cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement]. After one week, the HAT medium was replaced with HT Medium for three passages followed by returning to the normal cell culture medium. - The cell culture supernatants were primarily screened for recombinant DPP3 binding IgG antibodies two weeks after fusion. Therefore, recombinant GST-tagged DPP3 (USBiologicals, Salem, USA) was immobilized in 96-well plates (100 ng/ well) and incubated with 50 µl cell culture supernatant per well for 2 hours at room temperature. After washing of the plate, 50 µl / well POD-rabbit anti mouse IgG was added and incubated for 1h at RT.
- After a next washing step, 50 µ1 of a chromogen solution (3,7 mM o-phenylendiamin in citrate/hydrogen phosphate buffer, 0.012 % H2O2) were added to each well, incubated for 15 minutes at RT and the chromogenic reaction stopped by the addition of 50 µl 4 N sulfuric acid. Absorption was detected at 490 mm.
- The positive tested microcultures were transferred into 24-well plates for propagation. After retesting the selected cultures were cloned and recloned using the limiting-dilution technique and the isotypes were determined.
- Antibodies raised against GST-tagged human DPP3 or DPP3-peptides were produced via standard antibody production methods (Marx et al. 1997) and purified via Protein A. The antibody purities were ≥ 90 % based on SDS gel electrophoresis analysis.
- Humanization of murine antibodies may be conducted according to the following procedure:
- For humanization of an antibody of murine origin the antibody sequence is analyzed for the structural interaction of framework regions (FR) with the complementary determining regions (CDR) and the antigen. Based on structural modelling an appropriate FR of human origin is selected and the murine CDR sequences are transplanted into the human FR. Variations in the amino acid sequence of the CDRs or FRs may be introduced to regain structural interactions, which were abolished by the species switch for the FR sequences. This recovery of structural interactions may be achieved by random approach using phage display libraries or via directed approach guided by molecular modeling (Almagro JC, Fransson J., 2008. Humanization of antibodies. Front Biosci. \2008
Jan 1;13:1619-33). - In another aspect of the invention, the provided subject matter is a human CDR-grafted antiDPP3 antibody or anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said human CDR-grafted anti-DPP3 antibody or antiDPP3 antibody fragment thereof comprises an antibody heavy chain variable region (H chain) comprising
- SEQ ID NO.: 5
- and/or further comprises an antibody light chain variable region (L chain) comprising:
- SEQ ID NO.: 6.
- Further subject matter of the present invention in another aspect is a human CDR-grafted antiDPP3 antibody or anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein the said human CDR-grafted anti-DPP3 antibody or anti-DPP3 antibody fragment thereof comprises an antibody heavy chain variable region (H chain) comprising:
- SEQ ID NO.: 12
- and/or further comprises an antibody light chain variable region (L chain) comprising:
- SEQ ID NO.: 13.
- In one specific embodiment of the invention subject matter of the present invention is a human monoclonal anti-DPP3 antibody or monoclonal anti-DPP3 antibody fragment thereof that is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein the heavy chain comprises at least one CDR of:
- SEQ ID NO.: 7, SEQ ID NO.: 8 or SEQ ID NO.: 9
- and wherein the light chain comprises at least one CDR of:
- SEQ ID NO.: 8, KVS or SEQ ID NO.: 11.
- With the above context, the variable region can be connected to any subclass of constant regions (IgG, IgM, IgE. IgA), or only scaffolds, Fab fragments, Fv, Fab and F(ab)2. In example 3 below, the murine antibody variant with an IgG2a backbone was used. For chimerization and humanization a human IgG1 κ backbone was used.
- For epitope binding only the Complementarity Determining Regions (CDRs) are of importance. The CDRs for the heavy chain and the light chain of the murine anti-DPP3 antibody of the present invention (AK1967) are shown in SEQ ID NO. 7, SEQ ID NO. 8 and SEQ ID NO. 9 for the heavy chain and SEQ ID NO. 10, sequence KVS and SEQ ID NO. 11 for the light chain, respectively.
- In accordance with the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to SEQ ID NO.: 1, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and anti-DPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably at least 6 aa of said SEQ ID NO.:1.
- In accordance with the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and anti-DPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably at least 6 aa of SEQ ID NO.: 2.
- In another aspect of the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said epitope according to SEQ ID NO.: 3 is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and antiDPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably at least 4 aa, more preferably at least 5 aa, even more preferably to 6 aa of SEQ ID NO.: 3.
- In another aspect of the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds are directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said epitope according to SEQ ID NO.: 4 is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder, anti-DPP3 antibody, anti-DPP3 antibody fragment and antiDPP3 non Ig-scaffold recognizes and binds to at least three aa, preferably to four aa of SEQ ID NO.: 4.
- In a specific embodiment of the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, may act as inhibitor or effector of the bioactivity of DPP3.
- Thus, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof are useful in the prevention or treatment of a disease or acute condition in a patient, wherein said disease or acute condition is associated with oxidative stress in accordance with the invention.
- In a specific embodiment of the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibodies, anti-DPP3 antibody fragments and anti-DPP3 non Ig-scaffolds which are directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, exhibit an affinity towards DPP3 in such that the affinity constant is at least 10-7 M-1, preferably at least 10-8 M-1, more preferably the affinity constant is at least 10-9 M-1, most preferred the affinity constant is at least 10-10 M-1 when determined by means of the methods for measuring the binding affinity of the DPP3 binder of the invention to the epitope according to sequence SEQ ID NO.: 2 as described above.
- Thereby, a person skilled in the art knows that it may be considered to compensate lower affinity by applying a higher dose of binder; e.g. an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold, and this measure would not lead out-of-the-scope of the invention.
- In another embodiment of the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibody or anti-DPP3 antibody fragment thereof or anti-DPP3 non-Ig scaffold may be used in combination with at least one additional drug that induces oxidative stress as side effect.
- Such drugs are administered as primary medicament for use in the prevention or treatment of a primary disease and may be selected from a group comprising antimicrobials like antibiotics (for example streptomycin, gentamicin) or antivirals (for example acyclovir, foscarnet) or antifungal (for example amphotecerin B), analgesics, non-steroidal anti-inflammatory drugs (NSAID) (for example ibuprofen, naproxen), diuretics, proton pump inhibitors, chemotherapeutics (for example cisplatin), contrast dyes, cardiovascular agents like ACEinhibitors or statins, anti-depressants, immune suppressants (for example cyclosporine A) and antihistamines. Thereby, and in accordance with the invention, the herein provided DPP3 binder, specifically the herein provided anti-DPP3 antibody or anti-DPP3 antibody fragment thereof or a DPP3 non-Ig scaffold binding to DPP3 may be used as secondary medicament either in combination or as stand-alone drug in the prevention or treatment of the induced oxidative stress and resultant toxicities as secondary diseases.
- In a preferred embodiment of the invention the herein provided DPP3 binder are pharmaceutically acceptable, selective and/or specific for an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- In a more preferred embodiment of the invention the herein provided DPP3 binder is an inhibitory binder that is pharmaceutically acceptable, selective and/or specific for an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- In one aspect of the invention, selective and specific inhibitors of DPP3 do not bind to other proteins/peptides/enzymes or are bound by other proteins/peptides/enzymes, and do not inhibit any other enzyme/protease/peptidase other than DPP3. Therefore, the preferred inhibitors of DPP3 bioactivity with the context of the invention are specific anti-DPP3 antibodies, antibody fragments or non-Ig scaffolds binding to DPP3.
- Monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold with the context of the invention means that said antibody or antibody fragment or non-Ig scaffold binds specifically to one specific region encompassing at least 3 amino acids, preferably at least 4 aa within the target DPP3.
- With the context of the invention, monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or antiDPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 2, which is comprised in a DPP3 protein or a functional derivative thereof.
- In another specific embodiment, monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or antiDPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 3, which is comprised in a DPP3 protein or a functional derivative thereof.
- In another embodiment, monospecific anti-DPP3 antibody or monospecific anti-DPP3 antibody fragment or monospecific anti-DPP3 non-Ig scaffold are anti-DPP3 antibodies or anti-DPP3 antibody fragments or anti-DPP3 non-Ig scaffolds all have affinity for the same antigen as a target which is in accordance with the invention an epitope according to SEQ ID NO.: 4, which is comprised in a DPP3 protein or a functional derivative thereof.
- Monospecific antibodies may also be produced by other means than producing them from a common germ cell.
- With the above context, further preferred embodiments within the scope of the present invention are consecutively numbered below:
- 1. A binder directed to and binding to an epitope according to SEQ ID NO.: 2, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2, and wherein said epitope is comprised in SEQ ID NO.: 1, which corresponds to the amino acid sequence of DPP3.
- 2. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of
embodiment 1, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 3, and wherein said epitope is comprised in SEQ ID NO.: 1, which corresponds to the amino acid sequence of DPP3. - 3. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of
embodiment 1 orembodiment 2, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 4, and wherein said epitope is comprised in SEQ ID NO.: 1, which corresponds to the amino acid sequence of DPP3. - 4. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the
embodiments 1 to 3, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold, and wherein said epitope is comprised in SEQ ID NO.: 1, which corresponds to the amino acid sequence of DPP3. - 5. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- 6. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDR´s) in the heavy chain comprises the sequences:
- SEQ ID NO.: 7, SEQ ID NO.: 8 and/ or SEQ ID NO.: 9
- and the complementarity determining regions in the light chain comprises the sequences:
- SEQ ID NO.: 10, KVS and/ or SEQ ID NO.: 11.
- 7. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments, wherein said binder is a human monoclonal antibody or human monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- SEQ ID NO.: 12
- and wherein the light chain comprises the sequence:
- SEQ ID NO.: 13.
- 8. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 of any of the preceding embodiments for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress.
- 9. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to
embodiment 8, wherein said diseases are selected from a group comprising neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS. - 10. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to
embodiment 8 or 9, and wherein said: - o neurodegenerative disease may be selected from a group comprising Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)),
- o metabolic syndrome may be selected from a group comprising insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes,
- o cardiovascular disorder may be selected from a group comprising aterosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury, stroke and myocardial infarction,
- o autoimmune disease may be selected from a group comprising rheumatoid arthritis, systemic lupus erythematosus,
- o inflammatory lung disease may be selected from a group comprising COPD, asthma,
- o kidney disease may be selected from a group comprising acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD),
- o liver disease may be selected from a group comprising viral hepatitis, and cirrhosis,
- o digestive disease may be selected from a group comprising inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease, gastritis, pancreatitis and peptic ulcer,
- o viral infectious disease may be selected from a group comprising blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus,
- o cancer may be selected from a group comprising prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer,
- o inflammation,
- o sepsis, septic shock, SIRS.
- 11. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the
embodiments 8 to 10, wherein said disease is selected from a group comprising sepsis, septic shock, and SIRS. - 12. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to
embodiment 8, wherein said acute condition is selected from a group comprising renal toxicity and hepatotoxicity. - 13. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the
embodiments 8 to 12, wherein the binder is an anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold binding to an epitope according to SEQ ID NO.: 2, and wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said anti-DPP3 antibody or anti-DPP3 antibody fragment or anti-DPP3 non-Ig scaffold exhibits a binding affinity to DPP3 of at least 10-7 M. - 14. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the
embodiments 8 and 12, wherein said acute condition is hepatotoxicity which is drug-induced or alcohol-induced hepatotoxicity. - 15. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to
embodiment 8, wherein said acute condition is renal toxicity which is drug-induced renal toxicity. - 16. The binder directed to and binding to an epitope according to SEQ ID NO.: 2 for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the
embodiments 8 to 11, wherein said disease is associated with oxidative stress in the myocard. - 17. Pharmaceutical composition comprising a binder according to any of the
embodiments 1 to 7 for use in the prevention or treatment of a disease or acute condition of a patient, whereby said disease or acute condition is associated with oxidative stress. - 18. A kit comprising a binder according to any of the
embodiments 1 to 16. - 19. A binder directed to and binding to a DPP3 protein or functional derivative thereof for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress.
- 20. The binder for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to embodiment 19, wherein said diseases are selected from a group comprising neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
- 21. The binder for use in the prevention or treatment of diseases or acute conditions in a patient, wherein said disease or acute condition is associated with oxidative stress according to any of the
embodiments 19 or 20, and wherein said: - o neurodegenerative disease may be selected from a group comprising Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)),
- o metabolic syndrome may be selected from a group comprising insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes,
- o cardiovascular disorder may be selected from a group comprising aterosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury, stroke and myocardial infarction,
- o autoimmune disease may be selected from a group comprising rheumatoid arthritis, systemic lupus erythematosus,
- o inflammatory lung disease may be selected from a group comprising COPD, asthma,
- o kidney disease may be selected from a group comprising acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD),
- o liver disease may be selected from a group comprising viral hepatitis, and cirrhosis,
- o digestive disease may be selected from a group comprising inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease, gastritis, pancreatitis and peptic ulcer,
- o viral infectious disease may be selected from a group comprising blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus,
- o cancer may be selected from a group comprising prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer,
- o inflammation,
- o sepsis, septic shock, SIRS.
- 22. A binder according to any of the embodiments 19 to 21, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 2, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- 23. The binder according any of the embodiments 19 to 22, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 3, and wherein said binder recognizes and binds to at least three amino acids of SEQ ID NO.: 3.
- 24. The binder according to any of the embodiments 19 to 23, wherein said binder is directed to and binding to an epitope according to SEQ ID NO.: 4, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 4.
- 25. The binder according to any of the embodiments 19 to 24, wherein said binder is selected from a group comprising an antibody or antibody fragment or non-Ig scaffold.
- 26. The binder according to any of the embodiments 19 to 25, wherein said binder is a monoclonal antibody or monoclonal antibody fragment, and wherein the complementarity determining regions (CDR´s) in the heavy chain comprises the sequences:
- SEQ ID NO.: 7, SEQ ID NO.: 8 and/ or SEQ ID NO.: 9
- and the complementarity determining regions in the light chain comprises the sequences:
- SEQ ID NO.: 10, KVS and/ or SEQ ID NO.: 11.
- 27. The binder according any of the embodiments 19 to 26, wherein said binder is a humanized monoclonal antibody or humanized monoclonal antibody fragment, wherein the heavy chain comprises the sequence:
- SEQ ID NO.: 12
- and wherein the light chain comprises the sequence:
- SEQ ID NO.: 13.
- 28. The binder according to any of the embodiments 19 to 27, wherein said binder is a dipeptidyl peptidase 3 (DPP3) binder directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three amino acids of SEQ ID NO.: 2.
- In accordance with the invention, an “DPP3 binder” is directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three aa of SEQ ID NO.: 2 or a respective subsequence thereof according to the SEQ ID NO’S.: 3 or 4.
- In accordance with the invention, a DPP3 binder is preferably an anti-DPP3 antibody, or an anti-DPP3 antibody fragment, or an anti-DPP3 non-Ig scaffold directed to and binding to an epitope according to SEQ ID NO.: 2, wherein said epitope is comprised in a DPP3 protein or a functional derivative thereof, and wherein said DPP3 binder recognizes and binds to at least three aa of SEQ ID NO.: 2 or a respective subsequence thereof according to the SEQ ID NO’S.: 3 or 4.
- With the context of the invention, a “functional derivative” of a DPP3 protein denotes a peptide, polypeptide or protein that differs from the sequence of SEQ ID NO.: 1 by means of deletion of aa, addition of aa or changes of specific aa, but remains the bioactivity and function of a native DPP3 protein. Thereby, due to the modifications of the SEQ ID NO.: 1 the bioactivity and function may be influenced to a certain extent, but the enzymatic protease reaction catalysed by DDP 3 is still maintained when assay by a suitable bioactivity assay as described above or commonly known by the skilled person.
- A person skilled in the art understands that a dipeptidyl peptidase 3 (DPP3) antibody or an antiDPP3 antibody fragment or anti-DPP3 non-Ig scaffold is synonymous to dipeptidyl peptidase 3 (DPP3) antibody or a dipeptidyl peptidase 3 antibody fragment or DPP3 non-Ig scaffold and means anti- dipeptidyl peptidase 3 (DPP3) antibody or an anti- dipeptidyl peptidase 3 antibody fragment or anti-DPP3 non-Ig scaffold binding to DPP3, respectively.
- Throughout the text, the term “antibody” generally comprises monoclonal and polyclonal antibodies and binding fragments thereof, in particular Fc-fragments as well as so called “single-chain-antibodies” (Bird et al. 1988), chimeric, humanized, in particular CDR-grafted antibodies, and di- or tetrabodies (Holliger et al. 1993). Also comprised are immunoglobulin-like proteins that are selected through techniques including, for example, phage display to specifically bind to the molecule of interest contained in a sample.
- In this context the term “specific binding” refers to antibodies raised against the molecule of interest or a fragment thereof. An antibody is considered to be specific, if its affinity towards the molecule of interest or the aforementioned fragment thereof is at least preferably 50-fold higher, more preferably 100-fold higher, most preferably at least 1000-fold higher than towards other molecules comprised in a sample containing the molecule of interest. It is well known in the art how to make antibodies and to select antibodies with a given specificity.
- “Diseases associated with oxidative stress” with the context of the present invention include, but are not limited to, neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, and inflammation, sepsis, septic shock, SIRS.
- In the context of the present invention, neurodegenerative diseases comprise Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS).
- In the context of the present invention, metabolic syndrome comprises insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes.
- In the context of the present invention, cardiovascular disorders comprise aterosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury/ stroke and myocardial infarction.
- In the context of the present invention, autoimmune diseases comprise rheumatoid arthritis and systemic lupus erythematosus.
- In the context of the present invention inflammatory lung diseases comprise COPD and asthma.
- In the context of the present invention, kidney diseases comprise renal toxicity (drug-induced kidney disease), acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy and end-stage renal disease (ESRD).
- In the context of the present invention, liver diseases comprise hepatotoxicity, viral hepatitis, cirrhosis.
- In the context of the present invention, digestive diseases comprise inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease; gastritis, pancreatitis and peptic ulcer. In this context, viral infectious diseases comprise blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus and respiratory syncytial virus.
- In the context of the present invention, cancer comprises prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer and liver cancer.
- “Acute condition associated with oxidative stress” with the context of the present invention denote symptoms that appear and change or worsen rapidly due to the occurrence of oxidative stress. An acute condition associated with oxidative stress is sudden in onset. An acute condition associated with oxidative stress may lead to a chronic syndrome, if untreated.
- By contrast, a “chronic condition” or a “chronic syndrome”, respectively, with the context of the present invention denote a condition or symptom that develops and worsens over an extended period of time, and may be persistent, even if treated.
- “Oxidative stress” reflects an imbalance between the systemic manifestation of reactive oxygen species (ROS)/ reactive nitrogen species (RNS) and antioxidants in favour of excessive generation of free radicals. This process leads to the oxidation of biomolecules with consequent loss of its biological functions and/or homeostatic imbalances, whose manifestation is the potential oxidative damage to cells and tissues. Accumulation of ROS/RNS can result in a number of deleterious effects such as lipid peroxidation, protein oxidation and DNA damage (including base damage and strand breaks). Further, some reactive oxidative species act as cellular messengers in redox signalling. Thus, oxidative stress can cause disruptions in normal mechanisms of cellular signalling.
- A “free radical in the context of the present invention is a molecule with one or more unpaired electron in its outer shell. Free radicals are formed from molecules via the breakage of a chemical bond such that each fragment keeps one electron, by cleavage of a radical to give another radical and, also via redox reactions. Free radicals related to oxidative stress include hydroxyl (OH•), superoxide (O2•-), nitric oxide (NO•), nitrogen dioxide (NO2•), peroxyl (ROO•) and lipid peroxyl (LOO•). Also, hydrogen peroxide (H2O2), ozone (O3), singlet oxygen (1O2), hypochlorous acid (HOCl), nitrous acid (HNO2), peroxynitrite (ONOO-), dinitrogen trioxide (N2O3), lipid peroxide (LOOH), are not free radicals and generally called oxidants, but can easily lead to free radical reactions in living organisms.
- “Primary medicament” means a medicament that acts against the primary cause of said disease or condition.
- “Secondary medication” is a medication that improves the condition of the patient in a supportive way; e.g. reduces or regulates oxidative stress which is induced by the administration of a primary medicament.
- With the context of the invention, generally the “bioactivity” is defined as the effect that a substance takes on a living organism or tissue or organ or functional unit in vivo or in vitro (e.g. in an assay) after its interaction.
- In this regard and specifically with the context of the invention, DPP3 bioactivity may be defined as the DPP3 enzyme activity or the regulating activity of DPP3 in the oxidative stress pathway.
-
ABBREVIATIONS Abbreviation Meaning aa amino acid(s) AD Alzheimer’s disease AHF acute heart failure AIN Acute interstitial nephritis AKI acute kidney injury ALD Alcoholic liver disease ALS amyotrophic lateral sclerosis ARE antioxidant response element ATN Acute tubular necrosis ATP adenosine triphosphate AZT Azidothymidin BRCA1 Breast cancer gene 1BP blood pressure BSA bovine serum albumin cDNA complementary DNA CAT catalase CD Celiac disease CDR complementarity determining region CKD chronic kidney disease CLP cecal ligation and puncture CNS central nervous system COPD Chronic obstructive pulmonary disease CSF Cerebrospinal fluid CVD cardiovascular diseases DHE dihydroethidium DN diabetic nephropathy DNA Deoxyribonucleic acid DPP3, DPPIII dipeptidyl dipeptidase 3 (5,5′-dithiobis-(2-nitrobenzoic acid), Ellman’s DTNB reagent EBV Eppstein Barr virus EC enzyme category EDTA Ethylene diamine tetraacetic acid EF ejection fraction EOC epithelial ovarian cancer ESRD end-stage renal disease Fab Fragment antigen binding Fc crystallisable fragment FR framework region GC Gastric cancer GSH glutathione GPx Glutathione peroxidase HBV Hepatitis B virus HCV Hepatitis C virus HD Huntington’s disease hDPP3 human DPP3 HF heart failure HFmrEF heart failure with mid-range ejection fraction HFpEF heart failure with preserved ejection fraction HFrEF heart failure with reduced ejection fraction HIV human immunodeficiency virus HNE 4-hydroxynonenal IBD Inflammatory bowel disease i.p intraperitoneally i.v. intravenous Ig immunoglobulin KEAP1 Kelch like-ECH-associated protein 1LV left ventricular LVEF left ventricular ejection fraction MDA Malon dialdehyde MS multiple sclerosis NAFLD Non-alcoholic fatty liver disease NHS N-Hydroxysuccinimid non-Ig non-immunoglobulin NOS nitric oxide species Nrf2 nuclear factor erythroid 2-related factor 2NSAID Non-steroidal anti-inflammatory drugs o- ortho OS Oxidative stress PBS phosphate buffered saline PD parkinson’s disease PEG Polyethylene glycole PEG polyethylene glycole pHMB polyhexanide, polyhexamethylene biguanide PMSF phenlymethylsulfonyl fluoride PUD Peptic ulcer disease RLU relative light units RNA Ribonucleic acid RNS reactive nitrogen species ROI reactive oxygen intermediates ROS reactive oxygen species RT room temperature scFv single chain variable fragment SDS sodium dodecyl sulfate SOB shortness of breath TPCK tosyl phenylalanin chloromethyl ketone TRX1 thioreduxin 1 TTE transthoracic echocardiography UV ultraviolet XNA xeno nucleic acid - Generation of antibodies and determination DPP3 binding ability: Several murine antibodies were produced and screened by their ability of binding human DPP3 in a specific binding assay (see table 3).
- DPP3 peptides for immunization were synthesized, see table 3, (JPT Technologies, Berlin, Germany) with an additional N-terminal cystein (if no cystein is present within the selected DPP3-sequence) residue for conjugation of the peptides to Bovine Serum Albumin (BSA). The peptides were covalently linked to BSA by using Sulfolink-coupling gel (Perbio-science, Bonn, Germany). The coupling procedure was performed according to the manual of Perbio. Recombinant GST-hDPP3 was produced by USBio (United States Biological, Salem, MA, USA).
- Balb/c mice were intraperitoneally (i.p.) injected with 84 µg GST-hDPP3 or 100 µg DPP3-peptide-BSA-conjugates at day 0 (emulsified in TiterMax Gold Adjuvant), 84 µg or 100 µg at day 14 (emulsified in complete Freund’s adjuvant) and 42 µg or 50 µg at
day 21 and 28 (in incomplete Freund’s adjuvant). At day 49 the animal received an intravenous (i.v.) injection of 42 µg GST-hDPP3 or 50 µg DPP3-peptide-BSA-conjugates dissolved in saline. Three days later the mice were sacrificed and the immune cell fusion was performed. - Splenocytes from the immunized mice and cells of the myeloma cell line SP2/0 were fused with 1
ml 50% polyethylene glycol for 30 s at 37° C. After washing, the cells were seeded in 96-well cell culture plates. Hybrid clones were selected by growing in HAT medium [RPMI 1640 culture medium supplemented with 20% fetal calf serum and HAT-Supplement]. After one week, the HAT medium was replaced with HT Medium for three passages followed by returning to the normal cell culture medium. - The cell culture supernatants were primarily screened for recombinant DPP3 binding IgG antibodies two weeks after fusion. Therefore, recombinant GST-tagged hDPP3 (USBiologicals, Salem, USA) was immobilized in 96-well plates (100 ng/ well) and incubated with 50 µl cell culture supernatant per well for 2 hours at room temperature. After washing of the plate, 50 µl / well POD-rabbit anti mouse IgG was added and incubated for 1 h at RT. After a next washing step, 50 µ1 of a chromogen solution (3,7 mM o-phenylendiamin in citrate/ hydrogen phosphate buffer, 0.012% H2O2) were added to each well, incubated for 15 minutes at RT and the chromogenic reaction stopped by the addition of 50 µl 4 N sulfuric acid. Absorption was detected at 490 mm.
- The positive tested microcultures were transferred into 24-well plates for propagation. After retesting the selected cultures were cloned and recloned using the limiting-dilution technique and the isotypes were determined.
- Antibodies raised against GST-tagged human DPP3 or DPP3-peptides were produced via standard antibody production methods (Marx et al. 1997) and purified via Protein A. The antibody purities were ≥ 90% based on SDS gel electrophoresis analysis.
- To analyze the capability of DPP3/ immunization peptide binding by the different antibodies and antibody clones a binding assay was performed:
- Recombinant GST-tagged hDPP3 (SEQ ID No. 1) or a DPP3 peptide (immunization peptide, SEQ ID No. 2) was immobilized onto a high binding microtiter plate surface (96-Well polystyrene microplates, Greiner Bio-One international AG, Austria, 1 µg/well in coupling buffer [50 mM Tris, 100 mM NaCl, pH7,8], 1h at RT). After blocking with 5% bovine serum albumin, the microplates were vacuum dried.
- 100 µg (100 µl) of the different antiDPP3 antibodies (detection antibody, 1 mg/ ml in PBS, pH 7.4) were mixed with 10 µl acridinium NHS-ester (1 mg/ml in acetonitrile, InVent GmbH, Germany;
EP 0 353 971) and incubated for 30 min at room temperature. Labelled antiDPP3 antibody was purified by gel-filtration HPLC on Shodex Protein 5 µm KW-803 (Showa Denko, Japan). The purified labeled antibody was diluted in assay buffer (50 mmol/l potassium phosphate, 100 mmol/l NaCl, 10 mmol/l Na2-EDTA, 5 g/l bovine serum albumin, 1 g/l murine IgG, 1 g/l bovine IgG, 50 µmol/l amastatin, 100 µmol/l leupeptin, pH 7.4). The final concentration was approx. 5-7*106 relative light units (RLU) of labelled compound (approx. 20 ng labeled antibody) per 200 µ1. acridinium ester chemiluminescence was measured by using a Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG). - The plates were filled with 200 µl of labeled and diluted detection antibody (tracer) and incubated for 2-4 h at 2-8° C. Unbound tracer was removed by washing 4 times with 350 µl washing solution (20 mM PBS, pH 7.4, 0.1 % Triton X-100). Well-bound chemiluminescence was measured by using the Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG).
- To analyze the capability of DPP3 inhibition by the different antibodies and antibody clones a DPP3 activity assay with known procedure (Jones et al., 1982) was performed. Recombinant GST-tagged hDPP3 was diluted in assay buffer (25 ng/ ml GST-DPP3 in 50 mM Tris-HCl, pH7,5 and 100 µM ZnCl2) and 200 µl of this solution incubated with 10 µg of the respective antibody at room temperature. After 1 hour of pre-incubation, fluorogenic substrate Arg-Arg-βNA (20 µl, 2 mM) was added to the solution and the generation of free βNA over time was monitored using the Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH & Co. KG) at 37° C. Fluorescence of βNA is detected by exciting at 340 nm and measuring emission at 410 nm. Slopes (in RFU/ min) of increasing fluorescence of the different samples are calculated. The slope of GST-hDPP3 with buffer control is appointed as 100 % activity. The inhibitory ability of a possible capture-binder is defined as the decrease of GST-hDPP3 activity by incubation with said capture-binder in percent.
- The following table represents a selection of obtained antibodies and their binding rate in Relative Light Units (RLU) as well as their relative inhibitory ability (%; table 3). The monoclonal antibodies raised against the below depicted DPP3 regions, were selected by their ability to bind recombinant DPP3 and/ or immunization peptide, as well as by their inhibitory potential.
- All antibodies raised against the GST-tagged, full length form of recombinant hDPP3 show a strong binding to immobilized GST-tagged hDPP3. Also antibodies raised against the
SEQ ID 2 peptide bind to GST-hDPP3. TheSEQ ID 2 antibodies also strongly bind to the immunization peptide. Those antibodies were characterized in more detail (see example 2). The monoclonal antibody AK1967, with the ability of inhibiting DPP3 activity by 70 %, was chosen as possible therapeutic antibody and was also used as template for chimerization and humanization. -
TABLE 3 list of antibodies raised against full-length or sequences of hDPP3 and their ability to bind hDPP3 (SEQ ID No. 1) or immunization peptide (SEQ ID No. 2) in RLU, as well as the maximum inhibition of recombinant GST-hDPP3. Sequence number Antigen/ Immunogen hDPP3 region Clone hDPP3 binding [RLU] immunization peptide binding [RLU] Max. inhibition of hDPP3 SEQ ID: 1 GST tagged recombinant FLhDPP3 1-737 2552 3.053.621 0 65% 2553 3.777.985 0 35% 2554 1.733.815 0 30% 2555 3.805.363 0 25% SEQ ID: 2 CETVINPETGEQIQSWYRSGE 474-493 1963 141.822 2.163.038 60% 1964 100.802 2.041.928 60% 1965 99.493 1.986.794 70% 1966 118.097 1.990.702 65% 1967 113.736 1.909.954 70% 1968 105.696 2.017.731 65% 1969 82.558 2.224.025 70% - Antibodies raised against SEQ ID NO. 2 were characterized in more detail (epitope mapping, binding affinities, specificity, inhibitory potential). Here the results for clone 1967 of SEQ ID NO. 2 (“AK1967”) are shown as an example.
- For epitope mapping of AK1967 a number of N- or C-terminally biotinylated peptides were synthesized (peptides&elephants GmbH, Hennigsdorf, Germany). These peptides include the sequence of the full immunization peptide (SEQ ID No. 2) or fragments thereof, with stepwise removal of one amino acid from either C- or N-terminus (see table 5 for a complete list of peptides).
- High binding 96 well plates were coated with 2 µg Avidin per well (Greiner Bio-One international AG, Austria) in coupling buffer (500 mM Tris-HCl, pH 7.8, 100 mM NaCl). Afterwards plate were washed and filled with specific solutions of biotinylated peptides (10 ng/ well; buffer – 1xPBS with 0.5% BSA)
- AntiDPP3 antibody AK1967 was labelled with a chemiluminescence label according to Example 1.
- The plates were filled with 200 µl of labeled and diluted detection antibody (tracer) and incubated for 4 h at room temperature. Unbound tracer was removed by washing 4 times with 350 µl washing solution (20 mM PBS, pH 7.4, 0.1 % Triton X-100). Well-bound chemiluminescence was measured by using the Centro LB 960 luminometer (Berthold Technologies GmbH & Co. KG). Binding of AK1967 to the respective peptides is determined by evaluation of the relative light units (RLU). Any peptide that shows a significantly higher RLU signal than the unspecific binding of AK1967 is defined as AK1967 binder. The combinatorial analysis of binding and non-binding peptides reveals the specific DPP3 epitope of AK1967.
- The experiment was performed using Octet Red96 (ForteBio). AK1967 was captured on kinetic grade anti-humanFc (AHC) biosensors. The loaded biosensors were then dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, 3.7 nM). Association was observed for 120 seconds followed by 180 seconds of dissociation. The buffers used for the experiment are depicted in table 4. Kinetic analysis was performed using a 1:1 binding model and global fitting.
-
TABLE 4 Buffers used for Octet measurements Buffer Composition Assay Buffer PBS with 0.1% BSA, 0.02% Tween-21 Regeneration Buffer 10 mM Glycine buffer (pH 1.7) Neutralization Buffer PBS with 0.1% BSA, 0.02% Tween-21 - Blood cells from human EDTA-blood were washed (3x in PBS), diluted in PBS and lysed by repeated freeze-thaw-cycles. The blood cell lysate had a total protein concentration of 250 µg/ml, and a DPP3 concentration of 10 µg/ml. Dilutions of blood cell lysate (1:40, 1:80, 1:160 and 1:320) and of purified recombinant human His-DPP3 (31.25-500 ng/ml) were subjected to SDS-PAGE and Western Blot. The blots were incubated in 1.) blocking buffer (1xPBS-T with 5% skim milk powder), 2.) primary antibody solution (AK1967 1:2.000 in blocking buffer) and 3.) HRP labelled secondary antibody (goat anti mouse IgG, 1:1.000 in blocking buffer). Bound secondary antibody was detected using the Amersham ECL Western Blotting Detection Reagent and the Amersham Imager 600 UV (both from GE Healthcare).
- To analyze the capability of DPP3 inhibition by AK1967 a DPP3 activity assay with known procedure (Jones et al., 1982) was performed. Recombinant GST-tagged hDPP3 was diluted in assay buffer (25 ng/ ml GST-DPP3 in 50 mM Tris-HCl, pH7,5) and increasing concentrations of AK1967 were added. Fluorogenic substrate Arg-Arg-βNA was added to the solution and the generation of free βNA over time was monitored using the Twinkle LB 970 microplate fluorometer (Berthold Technologies GmbH & Co. KG) at 37° C. Fluorescence of βNA is detected by exciting at 340 nm and measuring emission at 410 nm. Slopes (in RFU/ min) of increasing fluorescence of the different samples are calculated. The slope of GST-hDPP3 with buffer control is appointed as 100 % activity. The inhibitory ability AK1967 is defined as the decrease of GST-hDPP3 activity by incubation with said antibody in percent. The resulting lowered DPP3 activities are shown in an inhibition curve in
FIG. 1C . - The analysis of peptides that AK1967 binds to and does not bind to revealed the DPP3 sequence INPETG (SEQ ID No. 3) as necessary epitope for AK1967 binding (see table 5).
-
TABLE 5 Peptides used for Epitope mapping of AK1967 peptide ID peptide sequence AK1967 binding #1 bio a f n f d q e t v i n p e t g e q i q s w y r s g yes #2 bio a f n f d q e t v i n p e t g e q i q yes #3 bio a f n f d q e t v i n p e t g e q i yes #4 bio a f n f d q e t v i n p e t g e q yes #5 bio a f n f d q e t v i n p e t g e yes #6 bio a f n f d q e t v i n p e t g yes #7 bio a f n f d q e t v i n p e no #8 bio a f n f d q e t v i n p e no #9 bio a f n f d q e t v i n p no #10 bio a f n f d q e t v i n no #11 e t g e q i q s w y k bio no #12 p e t g e q i q s w y k bio no #13 n p e t g e q i q s w y k bio no #14 i n p e t g e q i q s w y k bio yes #15 v i n p e t g e q i q s w y k bio yes #16 t v i n p e t g e q i q s w y k bio yes #17 e t v i n p e t g e q i q s w y k bio yes - AK1967 binds with an affinity of 2.2*10-9 M to recombinant GST-hDPP3 (for more details see table 6 and for kinetic curves see
FIG. 1A ). -
TABLE 6 Kinetic Constants of AK1967 affinity measurements KD (M) kon(⅟Ms) kdis(⅟s) Full X^2 Full R^2 2.2E-09 1.6E+05 3.5E-04 0.0413 0.9987 - The only protein detected with AK1967 as primary antibody in lysate of blood cells was DPP3 at 80 kDa (
FIG. 1B ). The total protein concentration of the lysate was 250 µg/ml whereas the estimated DPP3 concentration is about 10 µg/ml. Even though there is 25 times more unspecific protein in the lysate, AK1967 binds and detects specifically DPP3 and no other unspecific binding takes place. - AK1967 inhibits 15 ng/ ml DPP3 in a specific DPP3 activity assay with an IC50 of about 15 ng/ml (
FIG. 1C ). - A septic shock model was used to induce heart failure in rats and then to characterize AK1967′s influence on oxidative stress in myocardium.
- The study flow is depicted in
FIG. 2A below. After CLP or sham surgery the animals were allowed to rest for 20 hours with free access to water and food. Afterwards they were anesthetized, tracheotomy done and arterial and venous line laid. At 24 hours after CLP surgery either AK1967 or vehicle (saline) were administered with 2 mg/kg. As a safety measure hemodynamics were monitored invasively and continuously from t = 0 till 3 h. - Male Wistar rats (2-3 months, 300 to 400 g, group size refer to table 7) from the Centre d′élevage Janvier (France) were allocated randomly to one of three groups. All the animals were anesthetized using ketamine hydrochloride (90 mg/ kg) and xylazine (9 mg/ kg) intraperitoneally (i.p.). For induction of polymicrobial sepsis, cecal ligation and puncture (CLP) was performed using Rittirsch’s protocol with minor modification. A ventral midline incision (1.5 cm) was made to allow exteriorization of the cecum. The cecum is then ligated just below the ileocecal valve and punctured once with an 18-gauge needle. The abdominal cavity is then closed in two layers, followed by fluid resuscitation (3 ml/ 100 g body of weight of saline injected subcutaneously) and returning the animal to its cage. Sham animals were subjected to surgery, without getting their cecum punctured.
- At t=0 (baseline) all CLP animals are in septic shock and developed a decrease in heart function (low blood pressure, low shortening fraction). At this time point AK1967 (2 mg/kg) or vehicle (saline) were injected (i.v.) and saline infusion was started. There were 1 control group and 2 CLP groups which are summarized in the table below (table 7). At the end of the experiment, the animals were euthanized, and organs (e.g. heart) harvested for subsequent analysis.
-
TABLE 7 list of experimental groups Group Group size CLP treatment 1 - sham 4 no saline 2 - CLP-saline 5 yes saline 3 - CLP-AK1967 5 yes AK1967 - Dihydroethidium (DHE; Sigma-Aldrich) staining was used to evaluate the in situ levels of superoxide anion in the myocardium. Cardiac cryostat sections (7 µm) of the ventricles were incubated with DHE (37 µM) for 30 min in a dark humidified chamber. Acquisition of fluorescent images of ethidium bromide with Leica fluorescence microscope was performed under identical setting whatever the block tissue. The stained area was measured with IPLab software and expressed as a percentage of area of interest (% of ROI).
- Rats with septic shock induced heart failure after CLP surgery develop high amounts of reactive oxygen species (ROS) in their myocardium, whereas sham operated animals show almost no oxidative stress (
FIGS. 2B and C ). Treatment of the sick (CLP) animals with AK1967 reduces the oxidative stress levels in the myocardium to levels of healthy (sham-operated) animals. This strong ROS decrease is achieved within only 3 hours of treatment (FIG. 2B andFIG. 2C ). - Abramić, M. et al., 2000. Human and rat dipeptidyl peptidase III: Biochemical and mass spectrometric arguments for similarities and differences. Biological Chemistry, 381(December), pp.1233-1243.
- Abramić, M., Zubanović, M. & Vitale, L., 1988. Dipeptidyl peptidase III from human erythrocytes. Biol Chem Hoppe Seyler, pp.29-38.
- Adamczyk B. et al. 2016. New Insights into the Role of Oxidative Stress Mechanisms in the Pathophysiology and Treatment of Multiple Sclerosis. Oxidative Medicine and Cellular Longevity, Vol. 2016, pp. 1-18, DOI: 10.1155/2016/1973834″
- Agić, D. et al., 2007. Novel amidino-substituted benzimidazoles: Synthesis of compounds and inhibition of dipeptidyl peptidase III. Bioorganic Chemistry, 35(2), pp.153-169.
- Almagro JC, Fransson J., 2008. Humanization of antibodies. Front Biosci. 2008
Jan 1;13:1619-33. - Aoyagi, T. et al., 1993. Enzymatic Changes in Cerebrospinal Fluid of Patients with Alzheimer-Type Dementia. J Clin Biochem Nutr, 14, pp.133-139.
- Balogun, R.A. et al., 2010. Clinical applications of therapeutic apheresis. Journal of clinical apheresis, 25(5), pp.250-64.
- Baršun, M. et al., 2007. Human dipeptidyl peptidase III acts as a post-proline-cleaving enzyme on endomorphins. Biological Chemistry, 388(3), pp.343-348.
- Bhattacharyya A. et al. 2014. Oxidative stress: An essential factor in pathogenesis of gastrointestinal mucosal diseases. Physio.l Rev. 94, pp. 329-354.
- Bird et al., 1988. Single-chain antigen-binding proteins. Science 242:423-426.
- Chen H. et al. 2011. Oxidative Stress in Ischemic Brain Damage: Mechanisms of Cell Death and Potential Molecular Targets for Neuroprotection. Antioxidants and redox signaling, 14, pp. 1505-1517.
- Chen, W. et al., 2009. Direct interaction between Nrf2 and p21Cip1/WAF1 upregulates the Nrf2-mediated antioxidant response. Mol Cell., 34(6), pp. 663-673.
- Chiba, T. et al., 2003. Inhibition of recombinant dipeptidyl peptidase III by synthetic hemorphin-like peptides. Peptides, 24(5), pp.773-778.
- Couston, R.G. et al., Adsorption behavior of a human monoclonal antibody at hydrophilic and hydrophobic surfaces. mAbs, 5(1), pp.126-39.
- Deavall, D. G. et al., 2012. Drug-induced oxidative stress and toxicity. J Toxicol, pp. 645460.
- Deng, B. et al., 2014. Aptamer binding assays for proteins: The thrombin example-A review. Analytica Chimica Acta, 837, pp.1-15.
- Dhanda, S., Singh, J. & Singh, H., 2008. Hydrolysis of various bioactive peptides by goat brain dipeptidylpeptidase-III homologue. Cell biochemistry and function, 26(3), pp.339-45.
- Elahi M. M. et al. 2009. Oxidative stress as a mediator of cardiovascular disease. Oxidative Medicine and Cellular Longevity, 2:5, pp. 259-269.
- Ellis, S. & Nuenke, J.M., 1967. Dipeptidyl Arylamidase III of the Pituitary: Purification and Characterization. Journal of Biological Chemistry, 242(20), pp.4623-4629.
- Gamrekelashvili, J. et al., 2013. Peptidases released by necrotic cells control CD8 + T cell cross-priming. journal of clinical investigation, 123(11), pp.4755-4768.
- Hartley et al, 1982. Radiology 143: 29-36
- Hast B. E. et al., 2013. Proteomic analysis of ubiquitin ligase KEAP1 reveals associated proteins that inhibit NRF2 ubiquitination. Cancer Res., 73(7), pp. 2199-2210.
- Hast B. E. et al., 2014. Cancer-Derived Mutations in KEAP1 Impair NRF2 Degradation but not Ubiquitination. Molecular and Cellular Pathobiology. Cancer research, 74 (3), pp. 808-817.
- Holguin F. 2013. Oxidative Stress in Airway Diseases. Ann. Am. Thorac. Soc.,
Vol 10, Supplement, pp. S150-S157. - Holliger P. et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90:6444-8
- Hood et al., Immunology, Benjamin, N.Y., 2nd ed., 1984
- Hori M. et al. 2009, Oxidative stress and left ventricular remodelling after myocardial infarction. Cardivascular research, 81, pp. 457-464.
- Hosohata K. 2016. Role of Oxidative Stress in Drug-Induced Kidney Injury. International Journal of Molecular Sciences, 17, pp. 1826-1836.
- Hultschig C et al., Curr Opin Chem Biol. 2006 February; 10(1):4-10. PMID: 16376134)
- Hunkapiller & Hood, 1986. The growing immunoglobulin gene superfamily. Nature 323:15-16.
- Hust, M., Meyer, T., Voedisch, B., Rülker, T., Thie, H., El-Ghezal, A., Kirsch, M.I., Schütte, M., Helmsing, S., Meier, D., Schirrmann, T., Dübel, S., 2011. A human scFv antibody generation pipeline for proteome research. Journal of Biotechnology 152, 159-170
- Huston et al., 1988. Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A., 85:5879-5883.
- Hutcheson R. et al. The Metabolic Syndrome, Oxidative Stress, Environment, and Cardiovascular Disease: The Great Exploration. Experimental Diabetes Research, Vol. 2012, Article ID 271028.
- Igic, R. & Behnia, R., 2007. Pharmacological, immunological, and gene targeting of the renin-angiotensin system for treatment of cardiovascular disease. Current pharmaceutical design, 13(12), pp.1199-214.
- Inaoka, Y. & Naruto, S., 1988. Propioxatins A and B, New Enkephalinase B Inhibitors, IV. Characterization of the Active Site of the Enzyme Using Synthetic Propioxatin Analogues. J. Biochem, 104(5), pp.706-711.
- Ivanov A. V. et al. 2017. Oxidative Stress in Infection and Consequent Disease. Oxidative Medicine and Cellular Longevity, Vol. 2017, Article ID 3496043.
- Ivanov A. V. et al. 2017. Oxidative stress, a trigger of hepatitis C and B virus-induced liver carcinogenesis. Oncotarget, 8, pp. 3895-3932.
- Jones, T.H. & Kapralou, A, 1982. A rapid assay for dipeptidyl aminopeptidase III in human erythrocytes. Analytical biochemistry, 119(2), pp.418-23.
- Kabat et al., 1983. Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services.
- Kaymak C. et al. 2011. Reactive Oxygen Species (Ros) Generation in Sepsis. FABAD J. Pharm. Sci., 36, pp. 41-47.
- Khaket, T.P. et al., 2012. Enkephalin degrading enzymes: metalloproteases with high potential for drug development. Current pharmaceutical design, 18(2), pp.220-30.
- Khandrika L. et al. 2009. Role of Oxidative Stress in Prostate Cancer. Cancer Lett., 282(2), pp. 125-136.
- Kim, D. & Herr, A.E., 2013. Protein immobilization techniques for microfluidic assays. Biomicrofluidics, 7(4), p.41501.Kirk-Othmer, Encyclopedia of chemical technology, 4th ed., executive editor, J. I. Kroschwitz; editor, M. Howe-Grant, John Wiley & Sons, 1993, vol. 15, p. 518-562
- Kruk J. et al. 2017. Reactive Oxygen and Nitrogen Species in Carcinogenesis: Implications of Oxidative Stress on the Progression and Development of Several Cancer Types. Mini-Reviews in Medicinal Chemistry, 17, pp. 904-919.
- Kumar, P. et al., 2016. Substrate complexes of human dipeptidyl peptidase III reveal the mechanism of enzyme inhibition. Scientific Reports, 6(March), p.23787.
- Lanzavecchia, A. & Scheidegger, D., 1987. The use of hybrid hybridomas to target human cytotoxic T lymphocytes, Eur. J. Immunol. 17:105.
- Lee, C.M. & Snyder, S.H., 1982. Dipeptidyl-aminopeptidase III of rat brain. Selective affinity for enkephalin and angiotensin. The Journal of biological chemistry, 257(20), pp.12043-50.
- Li S. et al. 2015. The Role of Oxidative Stress and Antioxidants in Liver Diseases. International Journal of Molecular Sciences, 16, pp. 26087-26124.
- Li, J. et al., 2011. Peptide aptamers with biological and therapeutic applications. Current medicinal chemistry, 18(27), pp.4215-22.
- Liu Z. et al. 2017. Oxidative Stress in Neurodegenerative Diseases: From Molecular Mechanisms to Clinical Applications. Oxidative Medicine and Cellular Longevity, Vol. 2017, Article ID 2525967.
- Liu, Y. et al., 2007. A genomic screen for activators of the antioxidant response element. Proceedings of the National Academy of Sciences of the United States of America, 104(12), pp.5205-10.
- Lu K. et al., 2017. NRF2 induction supporting breast cancer cell survival is enabled by oxidative stress-induced DPP3-KEAP2 interaction. OnlineFirst; DOI: 10.1158/0008-5472.CAN-16-2204.
- Ma Q. et al., Role of Nrf2 in Oxidative Stress and Toxicity. Annu Rev Pharmacol Toxicol., 53, pp. 401-426.
- Ma Y. et al. 2013, Relation between Gastric Cancer and Protein Oxidation, DNA Damage, and Lipid Peroxidation. Oxidative Medicine and Cellular Longevity, Vol. 2013, Article ID 543760.
- Marx et al., 1997. Monoclonal Antibody Production,
ATLA 25, 121. - Mazzocco, C. et al., 2006. Identification and characterization of two dipeptidyl-peptidase III isoforms in Drosophila melanogaster. FEBS Journal, 273(5), pp.1056-1064.
- Meliopoulos, V.A. et al., 2012. MicroRNA regulation of human protease genes essential for influenza virus replication. PloS one, 7(5), p.e37169.
- Müller, J. et al., 2012. Monitoring of plasma levels of activated protein C using a clinically applicable oligonucleotide-based enzyme capture assay. Journal of Thrombosis and Haemostasis, 10(3), pp.390-398.
- Müller, J. et al., 2016. Aptamer-Based Enzyme Capture Assay for Measurement of Plasma Thrombin Levels. Methods in molecular biology (Clifton, N.J.), 1380, pp.179-89.
- Narendhirakannan R. T. et al., 2012. Oxidative Stress and Skin Cancer: An Overview. Ind. J. Clin. Biochem., 28, pp. 110-115.
- Nourazarian A. R. et al., 2014. Roles of Oxidative Stress in the Development and Progression of Breast Cancer. Asian Pac. J. Cancer Prev., 15 (12), pp. 4745-4751
- Ohkubo, I. et al., 1999. Dipeptidyl peptidase III from rat liver cytosol: purification, molecular cloning and immunohistochemical localization. Biological chemistry, 380(12), pp.1421-1430.
- Patel, A., Smith, H.J. & Sewell, R.D., 1993. Inhibitors of enkephalin-degrading enzymes as potential therapeutic agents. Progress in medicinal chemistry, 30, pp.327-78.
- Perl A. 2013. Oxidative stress in the pathology and treatment of systemic lupus erythematosus. Nat. Rev. Rheumatol., 9(11), pp. 674-686.
- Perse M., 2013. Oxidative Stress in the Pathogenesis of Colorectal Cancer: Cause or Consequence?. BioMed Research International, Vol. 2013, Article ID 725710.
- Pham-Huy et al. 2008. Free Radicals, Antioxidants in Disease and Health. Int. J. Biomed. Sci., 4 (2), pp. 89-96.
- Pinheiro Da Silva, F. & Nizet, V., 2009. Cell death during sepsis: Integration of disintegration in the inflammatory response to overwhelming infection. Apoptosis, 14(4), pp.509-521.
- Pitocco et al. 2013. Oxidative Stress in Diabetes: Implications for Vascular and Other Complications. Int. J. Mol. Sci., 14, pp. 21525-21550.
- Pohanka M. 2013. Role of oxidative stress in infectious diseases. A review. Folia Microbiol., 58, pp. 503-513.
- Prajapati, S.C. & Chauhan, S.S., 2011. Dipeptidyl peptidase III: a multifaceted oligopeptide N-end cutter. FEBS Journal, 278(18), pp.3256-3276.
- Quiñonez-Flores C. M. 2016, Oxidative Stress Relevance in the Pathogenesis of the Rheumatoid Arthritis: A Systematic Review. BioMed Research International Volume 2016, Article ID 6097417.
- Raghupathi, R., 2004. Cell death mechanisms following traumatic brain injury. Brain pathology (Zurich, Switzerland), 14(2), pp.215-22.
- Rastija, V. et al., 2015. Synthesis, QSAR, and Molecular Dynamics Simulation of Amidino-substituted Benzimidazoles as Dipeptidyl Peptidase III Inhibitors. Acta Chimica Slovenica, 62, pp.867-878.
- Rittirsch, D., Huber-Lang, M., Flierl, M. Ward, P.: Immunodesign of experimental sepsis by cecal ligation and punc-ture,
Nature Protocols 4, — 31 — 36 (2009) - Rodrigo R. et al., 2013. Oxidative Stress and Pathophysiology of Ischemic Stroke: Novel Therapeutic Opportunities. CNS & Neurological Disorders – Drug Targets, DOI: 10.2174/1871527311312050015.
- Romanillos, G. et al. EP 2949332 A2
- Saed G. M. et al., 2017. Updates of the role of oxidative stress in the pathogenesis of ovarian cancer. Gynecologic Oncology, 145, pp. 595-602.
- Sanderink, G.J., Artur, Y. & Siest, G., 1988. Human aminopeptidases: a review of the literature. Journal of clinical chemistry and clinical biochemistry. Zeitschrift für klinische Chemie und klinische Biochemie, 26(12), pp.795-807.
- Sawicka E. et al. 2015. The role of oxidative stress in bladder cancer, Postepy Hig Med Dosw, 69, pp. 744-752.
- Schütte, M., Thullier, P., Pelat, T., Wezler, X., Rosenstock, P., Hinz, D., Kirsch, M.I.,Hasenberg, M., Frank, R., Schirrmann, T., Gunzer, M., Hust, M., Dübel, S., 2009. Identification of a putative Crf splice variant and generation of recombinant antibodies for the specific detection of Aspergillus fumigatus. PLoS One 4, e6625
- Schwarz K. B. 1996, Oxidative stress during viral infection. Free Radical Biology & Medicine, 21, No. 5, pp. 641-649.
- Shimamori, Y., Watanabe, Y. & Fujimoto, Y., 1986. Purification and Characterization of Dipeptidyl Aminopeptidase III from Human Placenta. Chem. Pharm. Bull., 34(8), pp.3333-3340.
- Šimaga, Š. et al., 1998. Dipeptidyl peptidase III in malignant and non-malignant gynaecological tissue. European Journal of Cancer, 34(3), pp.399-405.
- Šimaga, Š. et al., 2003. Tumor cytosol dipeptidyl peptidase III activity is increased with histological aggressiveness of ovarian primary carcinomas. Gynecologic Oncology, 91(1), pp.194-200.
- Singh, R. et al., 2014. Transcription factor C/EBP-beta mediates downregulation of dipeptidyl-peptidase III expression by interleukin-6 in human glioblastoma cells. FEBS Journal, 281, pp.1629-1641.
- Sobocanec et al., 2015. The role of 17β-estradiol in the regulation of antioxidant enzymes via the Nrf2-Keapl pathway in the livers of CBA/H mice. Life Sciences, 130, pp. 57-65
- Sobocanec et al., 2016. Prominent role of exopeptidase DPP III in estrogen-mediated protection against hyperoxia in vivo. Redox Biology, 8, pp. 49-159
- Sosa et al. 2013. Oxidative stress and cancer: An overview. Ageing Research Reviews, 12, pp. 376-390.
- Sureshbabu A. 2015. Oxidative stress and autophagy: Crucial modulators of kidney injury. Redox Biology, 4, pp. 208-214.
- Tian T. et al. 2017. Pathomechanisms of Oxidative Stress in Inflammatory Bowel Disease and Potential Antioxidant Therapies. Oxidative Medicine and Cellular Longevity, Vol. 2017, Article ID 4535194.
- Ullah A. et al. 2015. Diabetes mellitus and oxidative stress-A concise review. Saudi Pharmaceutical Journal, 24, pp. 547-553.
- Vairappan B. 2015. Endothelial dysfunction in cirrhosis: Role of inflammation and oxidative stress. World Journal of Hepatology, 7 (3), pp. 443-459.
- Valavanidis A. et al. 2013, Pulmonary Oxidative Stress, Inflammation and Cancer: Respirable Particulate Matter, Fibrous Dusts and Ozone as Major Causes of Lung Carcinogenesis through Reactive Oxygen Species Mechanisms. Int. J. Environ. Res. Public Health, 10, pp. 3886-3907.
- Vandenberg, I., King, F. & Kuchel, P., 1985. Enkephalin Degradation by Human Erythrocytes and Hemolysates Studied Using 1H NMR Spectroscopy. Archives of Biochemistry and Biophysics, 242(2), pp.515-522.
- Vanha-Perttula, T., 1988. Dipeptidyl peptidase III and alanyl aminopeptidase in the human seminal plasma: origin and biochemical properties. Clinica chimica acta; international journal of clinical chemistry, 177(2), pp.179-95.
- Volonte, D. et al., 2013. Inhibition of nuclear factor-erythoid 2-related factor (Nrf2) by caveolin-2 promotes stress-induced premature senescence. Molecular Biology of the Cell, 24, pp. 1852-1862.
- Wang Z. et al., 2016. Oxidative Stress and Liver Cancer: Etiology and Therapeutic Targets. Oxidative Medicine and Cellular Longevity, Vol. 2016, Article ID: 7891574.
- Wattiaux, R. et al., 2007. Lysosomes and Fas-mediated liver cell death. The Biochemical journal, 403(1), pp.89-95.
- Wild, David (2005). The Immunoassay Handbook, Elsevier LTD, Oxford; 3rd ed., ISBN-13: 978-0080445267
- Xu, Y., Yang, X. & Wang, E., 2010. Review: Aptamers in microfluidic chips. Analytica chimica acta, 683(1), pp.12-20.
- Yamamoto, Y. et al., 1998. Inhibitory action of spinorphin, an endogenous regulator of enkephalin-degrading enzymes, on carrageenan-induced polymorphonuclear neutrophil accumulation in mouse air-pouches. Life sciences, 62(19), pp.1767-73.
- Yamamoto, Y. et al., 2000. Characterization of tynorphin, a potent endogenous inhibitor of dipeptidyl peptidaseIII. Peptides, 21(4), pp.503-8.
- Zong, W.-X. & Thompson, C.B., 2006. Necrotic Cell Death as a Cell Fate. Genes & Development, 20, pp.1-5.
- Zuk et al., Enzyme Immunochromatography--A Quantitative Immunoassay Requiring No Instrumentation, Clinical Chemistry, 31 (7): 1144-1150 (1985)
- SEQ ID▫No. 1 – hDPP3 aa 1-737
-
MADTQYILPNDIGVSSLDCREAFRLLSPTERLYAYHLSRAAWYGGLAVLL QTSPEAPYIYALLSRLFRAQDPDQLRQHALAEGLTEEEYQAFLVYAAGVY SNMGNYKSFGDTKFVPNLPKEKLERVILGSEAAQQHPEEVRGLWQTCGEL MFSLEPRLRHLGLGKEGITTYFSGNCTMEDAKLAQDFLDSQNLSAYNTRL FKEVDGEGKPYYEVRLASVLGSEPSLDSEVTSKLKSYEFRGSPFQVTRGD YAPILQKVVEQLEKAKAYAANSHQGQMLAQYIESFTQGSIEAHKRGSRFW IQDKGPIVESYIGFIESYRDPFGSRGEFEGFVAVVNKAMSAKFERLVASA EQLLKELPWPPTFEKDKFLTPDFTSLDVLTFAGSGIPAGINIPNYDDLRQ TEGFKNVSLGNVLAVAYATQREKLTFLEEDDKDLYILWKGPSFDVQVGLH ELLGHGSGKLFVQDEKGAFNFDQETVINPETGEQIQSWYRSGETWDSKFS TIASSYEECRAESVGLYLCLHPQVLEIFGFEGADAEDVIYVNWLNMVRAG LLALEFYTPEAFNWRQAHMQARFVILRVLLEAGEGLVTITPTTGSDGRPD ARVRLDRSKIRSVGKPALERFLRRLQVLKSTGDVAGGRALYEGYATVTDA PPECFLTLRDTVLLRKESRKLIVQPNTRLEGSDVQLLEYEASAAGLIRSF SERFPEDGPELEEILTQLATADARFWKGPSEAPSGQA - SEQ ID No. 2 — hDPP3 aa 474-493 (N-Cys) — immunization peptide with additional N-terminal▫Cystein
-
CETVINPETGEQIQSWYRSGE - SEQ ID No. 3 — hDPP3 aa 477-482 — epitope of AK1967
-
INPETG - SEQ ID No. 4 – hDPP3 aa 480-483
-
ETGE - SEQ ID No. 5 – variable region of murine AK1967 in heavy chain
-
QVTLKESGPGILQPSQTLSLTCSFSGFSLSTSGMSVGWIRQPSGKGLEWL AHIWWNDNKSYNPALKSRLTISRDTSNNQVFLKIASVVTADTGTYFCARN YSYDYWGQGTTLTVSS - SEQ ID No. 6 – variable region of murine AK1967 in light chain
-
DVVVTQTPLSLSVSLGDPASISCRSSRSLVHSIGSTYLHWYLQKPGQSPK LLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVP WTFGGGTKLEIK - SEQ ID No. 7 – CDR1 of murine AK1967 in heavy chain
-
GFSLSTSGMS - SEQ ID No. 8 – CDR2 of murine AK1967 in heavy chain
-
IWWNDNK - SEQ ID No. 9 – CDR 3 of murine AK1967 in heavy chain
-
ARNYSYDY - SEQ ID No. 10 – CDR1 of murine AK1967 in light chain
-
RSLVHSIGSTY - CDR2 of murine AK1967 in light chain
-
KVS - SEQ ID No. 11 – CDR3 of murine AK1967 in light chain
-
SQSTHVPWT - SEQ ID No. 12 — humanized AK1967 — heavy chain sequence (IgG1κ: backbone)
-
MDPKGSLSWRILLFLSLAFELSYGQITLKESGPTLVKPTQTLTLTCTFSG FSLSTSGMSVGWIRQPPGKALEWLAHIWWNDNKSYNPALKSRLTITRDTS KNQVVLTMTNMDPVDTGTYYCARNYSYDYWGQGTLVTVSSASTKGPSVFP LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG - SEQ ID No. 13 — humanized AK1967 — light chain sequence (IgG1κ: backbone)
-
METDTLLLWVLLLWVPGSTGDIVMTQTPLSLSVTPGQPASISCKSSRSLV HSIGSTYLYWYLQKPGQSPQLLIYKVSNRFSGVPDRFSGSGSGTDFTLKI SRVEAEDVGVYYCSQSTHVPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC -
FIG. 1A illustrates Association- and dissociation curve of the AK1967-DPP3 binding analysis using Octet. AK1967 loaded biosensors were dipped into a dilution series of recombinant GST-tagged human DPP3 (100, 33.3, 11.1, 3.7 nM) and association and dissociation monitored. -
FIG. 1B is a Western Blot of dilutions of blood cell lysate and detection of DPP3 with AK1967 as primary antibody. -
FIG. 1C shows an Inhibition curve of native DPP3 from blood cells with inhibitory antibody AK1967. Inhibition of DPP3 by a specific antibody is concentration dependent, with an IC50 at ~15 ng/ml when analyzed against 15 ng/ml DPP3. -
FIG. 2A is an Experimental design of heart failure study of rats in septic shock. -
FIG. 2B illustrates Fluorescence images of DHE labelled myocardium of sham, CLP and CLP AK1967 animals. -
FIG. 2C illustrates Quantification of DHE stained areas and expression as percentage of area of interest (% of ROI).
Claims (11)
1-15. (canceled)
16. A dipeptidyl peptidase 3 (DPP3) binder wherein the binder is produced by a process comprising:
injecting an animal with a DPP3-peptide or a conjugate comprising a DPP3-peptide,
fusing activated B-cells from the animal after injection with a myeloma cell line and culturing the resulting cells,
selecting and culturing cells expressing the binder,
wherein said binder is defined as that which binds to an epitope according to SEQ ID NO.: 2, and binds to at least three amino acids of SEQ ID NO.: 2, wherein said epitope according to SEQ ID NO.: 2 is comprised in SEQ ID NO.: 1, and wherein said binder exhibits an affinity towards DPP3 at an affinity constant of at least 107 M-1.
17. The dipeptidyl peptidase 3 binder of claim 16 , wherein said binder is binds to an epitope according to SEQ ID NO.: 3, and wherein said binder binds to at least three amino acids of SEQ ID NO.: 3.
18. The dipeptidyl peptidase 3 binder of claim 16 , wherein said binder binds to an epitope according to SEQ ID NO.: 4, and wherein said DPP3 binder binds to at least three amino acids of SEQ ID NO.: 4.
19. The dipeptidyl peptidase 3 binder of claim 16 , wherein said binder is selected from a group consisting of an antibody or antigen binding fragment or non-Ig scaffold.
20. The dipeptidyl peptidase 3 binder of claim 16 , wherein said binder is selected from a group consisting of a monospecific antibody or a monospecific antigen binding fragment or a monospecific non-Ig scaffold.
21. A method of treatment comprising:
administering the binder of claim 16 to a patient suffering from a disease or acute condition associated with oxidative stress.
22. The method of claim 21 wherein said disease or acute condition that is associated with oxidative stress is selected from a group consisting of neurodegenerative diseases, metabolic syndrome, cardiovascular disorders, autoimmune diseases, inflammatory lung diseases, kidney diseases, liver diseases, digestive diseases, viral infectious diseases, cancer, inflammation, sepsis, septic shock and SIRS.
23. The method of claim 22 wherein
said neurodegenerative disease is selected from a group consisting of Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS)),
said metabolic syndrome is selected from a group consisting of insulin resistance, obesity, hyperglycemia, dyslipidemia, hypertension and diabetes,
said cardiovascular disorder is selected from a group consisting of aterosclerosis, hypertension, heart failure, cardiovascular ischemia, cerebral ischemic injury, stroke and myocardial infarction,
said autoimmune disease is selected from a group consisting of rheumatoid arthritis, systemic lupus erythematosus,
said inflammatory lung disease is selected from a group consisting of COPD, asthma,
said kidney disease is selected from a group consisting of acute kidney injury (AKI), chronic kidney disease (CKD), diabetic nephropathy, end-stage renal disease (ESRD),
said liver disease is selected from a group consisting of viral hepatitis, and cirrhosis,
said digestive disease is selected from a group consisting of inflammatory bowel disease e.g. Ulcerative colitis, Crohn’s disease, gastritis, pancreatitis and peptic ulcer,
said viral infectious disease may be selected from a group consisting of blood-borne hepatitis viruses (B, C, and D), human immunodeficiency virus (HIV), influenza A, Epstein-Barr virus, respiratory syncytial virus,
said cancer is selected from a group consisting of prostate cancer, breast cancer, lung cancer, colorectal cancer, bladder cancer, ovarian cancer, skin cancer, stomach cancer, liver cancer,
said inflammation is selected, and
said sepsis, septic shock or SIRS is selected.
24. The method of claim 21 wherein said disease or acute condition that is associated with oxidative stress is selected from a group consisting of sepsis, septic shock, and SIRS.
25. The method of claim 21 wherein said disease or acute condition that is associated with oxidative stress is myocard.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/984,611 US20230295344A1 (en) | 2017-10-25 | 2022-11-10 | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP17198420 | 2017-10-25 | ||
EP17198420.6 | 2017-10-25 | ||
PCT/EP2018/079197 WO2019081595A2 (en) | 2017-10-25 | 2018-10-24 | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress |
US202016758881A | 2020-04-24 | 2020-04-24 | |
US17/984,611 US20230295344A1 (en) | 2017-10-25 | 2022-11-10 | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress |
Related Parent Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2018/079197 Continuation WO2019081595A2 (en) | 2017-10-25 | 2018-10-24 | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress |
US16/758,881 Continuation US11530276B2 (en) | 2017-10-25 | 2018-10-24 | DPP3 binder directed to and binding to specific DPP3-epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress |
Publications (1)
Publication Number | Publication Date |
---|---|
US20230295344A1 true US20230295344A1 (en) | 2023-09-21 |
Family
ID=60186108
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/758,881 Active 2039-02-13 US11530276B2 (en) | 2017-10-25 | 2018-10-24 | DPP3 binder directed to and binding to specific DPP3-epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress |
US17/984,611 Pending US20230295344A1 (en) | 2017-10-25 | 2022-11-10 | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/758,881 Active 2039-02-13 US11530276B2 (en) | 2017-10-25 | 2018-10-24 | DPP3 binder directed to and binding to specific DPP3-epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress |
Country Status (12)
Country | Link |
---|---|
US (2) | US11530276B2 (en) |
EP (1) | EP3700937A2 (en) |
JP (2) | JP7424972B2 (en) |
KR (1) | KR20200083509A (en) |
CN (1) | CN111542548A (en) |
AU (1) | AU2018356441A1 (en) |
BR (1) | BR112020006563A2 (en) |
CA (1) | CA3080251A1 (en) |
IL (1) | IL274009A (en) |
MX (1) | MX2020004271A (en) |
SG (1) | SG11202002594PA (en) |
WO (1) | WO2019081595A2 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115769076A (en) | 2020-03-16 | 2023-03-07 | 4Teen4制药有限公司 | DPP3 in patients infected with coronaviruses |
EP3922993A1 (en) | 2020-06-12 | 2021-12-15 | 4TEEN4 Pharmaceuticals GmbH | Dpp3 in patients infected with coronavirus |
WO2024126793A1 (en) | 2022-12-15 | 2024-06-20 | 4TEEN4 Pharmaceuticals GmbH | Dpp3 inhibitor for improvement of pulmonary function in critically ill patients |
Family Cites Families (30)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5807715A (en) | 1984-08-27 | 1998-09-15 | The Board Of Trustees Of The Leland Stanford Junior University | Methods and transformed mammalian lymphocyte cells for producing functional antigen-binding protein including chimeric immunoglobulin |
JPS6188884A (en) | 1984-10-04 | 1986-05-07 | Sankyo Co Ltd | Encephalinase b-inhibiting substance and its preparation |
AU634716B2 (en) | 1988-08-01 | 1993-03-04 | Ciba Corning Diagnostics Corp. | Method for detection of an analyte using acridinium esters and liposomes |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
AU633698B2 (en) | 1990-01-12 | 1993-02-04 | Amgen Fremont Inc. | Generation of xenogeneic antibodies |
US5427908A (en) | 1990-05-01 | 1995-06-27 | Affymax Technologies N.V. | Recombinant library screening methods |
WO1992020791A1 (en) | 1990-07-10 | 1992-11-26 | Cambridge Antibody Technology Limited | Methods for producing members of specific binding pairs |
GB9015198D0 (en) | 1990-07-10 | 1990-08-29 | Brien Caroline J O | Binding substance |
EP0746609A4 (en) | 1991-12-17 | 1997-12-17 | Genpharm Int | Transgenic non-human animals capable of producing heterologous antibodies |
US6818418B1 (en) | 1998-12-10 | 2004-11-16 | Compound Therapeutics, Inc. | Protein scaffolds for antibody mimics and other binding proteins |
WO2001016334A2 (en) | 1999-09-01 | 2001-03-08 | Incyte Genomics, Inc. | Human hydrolytic enzymes |
IL160289A0 (en) | 2001-08-30 | 2004-07-25 | Biorexis Pharmaceutical Corp | Modified transferrin fusion proteins |
EP2298278B1 (en) | 2002-06-07 | 2015-11-11 | Dyax Corp. | Prevention and reduction of blood loss and inflammatory response |
US20050164301A1 (en) | 2003-10-24 | 2005-07-28 | Avidia Research Institute | LDL receptor class A and EGF domain monomers and multimers |
US20100028995A1 (en) | 2004-02-23 | 2010-02-04 | Anaphore, Inc. | Tetranectin Trimerizing Polypeptides |
WO2005106486A2 (en) | 2004-04-28 | 2005-11-10 | Bayer Healthcare Ag | Diagnostic and therapeutics for diseases associated with dipeptidyl-peptidase 3(dpp3) |
AU2005287557B2 (en) | 2004-09-21 | 2011-10-13 | Biontech Ag | Use of microproteins as tryptase inhibitors |
JP2006230318A (en) | 2005-02-25 | 2006-09-07 | Maruishi Pharmaceutical Co Ltd | Method for diagnosing rheumatoid arthritis |
WO2007117444A2 (en) | 2006-03-31 | 2007-10-18 | Yinghe Hu | Protein detection by aptamers |
EP2030025A2 (en) | 2006-06-07 | 2009-03-04 | Tethys Bioscience, Inc. | Markers associated with arteriovascular events and methods of use thereof |
ATE527353T1 (en) | 2007-12-19 | 2011-10-15 | Affibody Ab | PDGF-BINDING POLYPEPTIDE FROM PROTEIN A |
NZ592591A (en) | 2008-11-03 | 2012-04-27 | Molecular Partners Ag | Binding proteins comprising ankyrin repeast domains that inhibit the vegf-a receptor interaction |
US8715928B2 (en) * | 2009-02-13 | 2014-05-06 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Molecular-based method of cancer diagnosis and prognosis |
WO2010113096A1 (en) * | 2009-03-30 | 2010-10-07 | Tel Hashomer Medical Research Infrastructure And Services Ltd. | Methods of predicting clinical course and treating multiple sclerosis |
BR112012004314A2 (en) | 2009-08-27 | 2016-11-29 | Covagen Ag | il-17 binding compounds and medicinal uses of these compounds |
CA2778871C (en) | 2009-12-14 | 2017-08-01 | Scil Proteins Gmbh | Modified ubiquitin proteins having a specific binding activity for the extradomain b of fibronectin |
WO2011154420A2 (en) | 2010-06-08 | 2011-12-15 | Pieris Ag | Tear lipocalin muteins binding il-4 r alpha |
CN105228633A (en) | 2013-01-28 | 2016-01-06 | 新天然替代品公司 | For the compositions of the whole body therapeutic of the pathological state by oxidative stress and/or the unbalance generation of oxidoreduction |
CN104946646B (en) * | 2014-03-25 | 2020-03-31 | 中国医学科学院基础医学研究所 | Small nucleic acid molecule, DNA molecule, protein and application for preventing and/or treating Ebola viral hemorrhagic fever |
JP2019520550A (en) * | 2016-04-21 | 2019-07-18 | スフィンゴテック セラピューティクス ゲゼルシャフト ミット ベシュレンクテル ハフツング | Method of determination and treatment of DPP3 |
-
2018
- 2018-10-24 CN CN201880069013.0A patent/CN111542548A/en active Pending
- 2018-10-24 KR KR1020207014841A patent/KR20200083509A/en not_active Application Discontinuation
- 2018-10-24 SG SG11202002594PA patent/SG11202002594PA/en unknown
- 2018-10-24 MX MX2020004271A patent/MX2020004271A/en unknown
- 2018-10-24 JP JP2020523261A patent/JP7424972B2/en active Active
- 2018-10-24 WO PCT/EP2018/079197 patent/WO2019081595A2/en unknown
- 2018-10-24 US US16/758,881 patent/US11530276B2/en active Active
- 2018-10-24 CA CA3080251A patent/CA3080251A1/en active Pending
- 2018-10-24 BR BR112020006563-3A patent/BR112020006563A2/en unknown
- 2018-10-24 EP EP18795970.5A patent/EP3700937A2/en active Pending
- 2018-10-24 AU AU2018356441A patent/AU2018356441A1/en active Pending
-
2020
- 2020-04-16 IL IL274009A patent/IL274009A/en unknown
-
2022
- 2022-11-10 US US17/984,611 patent/US20230295344A1/en active Pending
-
2024
- 2024-01-18 JP JP2024006108A patent/JP2024041975A/en active Pending
Also Published As
Publication number | Publication date |
---|---|
WO2019081595A2 (en) | 2019-05-02 |
JP2024041975A (en) | 2024-03-27 |
AU2018356441A1 (en) | 2020-04-16 |
IL274009A (en) | 2020-05-31 |
CN111542548A (en) | 2020-08-14 |
US11530276B2 (en) | 2022-12-20 |
US20210206876A1 (en) | 2021-07-08 |
JP2021500376A (en) | 2021-01-07 |
MX2020004271A (en) | 2020-07-29 |
RU2020116450A3 (en) | 2021-12-21 |
BR112020006563A2 (en) | 2020-10-13 |
JP7424972B2 (en) | 2024-01-30 |
RU2020116450A (en) | 2021-11-25 |
CA3080251A1 (en) | 2019-05-02 |
KR20200083509A (en) | 2020-07-08 |
EP3700937A2 (en) | 2020-09-02 |
SG11202002594PA (en) | 2020-04-29 |
WO2019081595A4 (en) | 2019-09-06 |
WO2019081595A3 (en) | 2019-06-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20230295344A1 (en) | Dpp3 binder directed to and binding to specific dpp3-epitopes and its use in the prevention or treatment of diseases/acute conditions that are associated with oxidative stress | |
US11726094B2 (en) | Methods for determining DPP3 and therapeutic methods | |
WO2017194499A1 (en) | Tissue-specific exosomes as biomarkers | |
RU2787033C2 (en) | Dpp3-binding molecule aimed at specific dpp3 epitopes and binding to them and its use for prevention or treatment of diseases/acute conditions, which are related to oxidative stress | |
WO2021185786A1 (en) | Dpp3 in patients infected with coronavirus | |
KR20220145866A (en) | Anti-ADM-antibody binding to free N-terminus and combination with vitamin C for accelerated conversion of ADM-Gly to bio-ADM in patients with ADM-Gly/bio-ADM ratio above threshold | |
US20160327558A1 (en) | Means and methods for detecting activated malt1 | |
RU2771824C2 (en) | Methods for determining dpp3 and therapeutic methods | |
WO2011107586A1 (en) | Smoc1, tenascin-c and brain cancers | |
US20230250166A1 (en) | Anti-adrenomedullin (adm) binder for use in therapy of patients in shock | |
WO2024126793A1 (en) | Dpp3 inhibitor for improvement of pulmonary function in critically ill patients | |
WO2024023368A1 (en) | Prediction of an increase of dpp3 in a patient with septic shock | |
CA3207969A1 (en) | Anti-adrenomedullin (adm) antibody or anti-adm antibody fragment or anti-adm non-ig scaffold for use in therapy or prevention of shock and prediction of an increase of dpp3 in a critically ill patient |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |