title: Fully human antibodies against the Protease-Activated Receptor-2 (PAR-2) with anti-inflammatory activity.
abstract: PAR-2 belongs to a family of G-protein coupled Protease-Activated Receptors (PAR) which are activated by specific proteolytic cleavage in the extracellular N-terminal region. PAR-2 is activated by proteases such as trypsin, tryptase, proteinase 3, factor VIIa, factor Xa and is thought to be a mediator of inflammation and tissue injury, where elevated levels of proteases are found. Utilizing the HuCAL GOLD® phage display library we generated fully human antibodies specifically blocking the protease cleavage site in the N-terminal domain. In vitro affinity optimization resulted in antibodies with up to 1000-fold improved affinities relative to the original parental antibodies with dissociation constants as low as 100 pM. Corresponding increases in potency were observed in a mechanistic protease cleavage assay. The antibodies effectively inhibited PAR-2 mediated intracellular calcium release and cytokine secretion in various cell types stimulated with trypsin. In addition, the antibodies demonstrated potent inhibition of trypsin induced relaxation of isolated rat aortic rings ex vivo. In a short term mouse model of inflammation, the trans vivo DTH model, anti-PAR-2 antibodies showed inhibition of the inflammatory swelling response. In summary, potent inhibitors of PAR-2 were generated which allow further assessment of the role of this receptor in inflammation and evaluation of their potential as therapeutic agents.
authors: Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, USA.
authors: Patricia
authors: P
authors: Giblin
authors: Rainer
authors: R
authors: Boxhammer
authors: Sudha
authors: S
authors: Desai
authors: Rachel
authors: R
authors: Kroe-Barrett
authors: Gale
authors: G
authors: Hansen
authors: John
authors: J
authors: Ksiazek
authors: Maret
authors: M
authors: Panzenbeck
authors: Kerry
authors: K
authors: Ralph
authors: Racheline
authors: R
authors: Schwartz
authors: Clare
authors: C
authors: Zimmitti
authors: Catrin
authors: C
authors: Pracht
authors: Sandra
authors: S
authors: Miller
authors: Jeanne
authors: J
authors: Magram
authors: Tobias
authors: T
authors: Litzenburger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D019138:Antibodies, Blocking / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D001011:Aorta / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D006968:Hypersensitivity, Delayed / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D007249:Inflammation / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D007700:Kinetics; D008252:Macaca fascicularis; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids; D051381:Rats; D044464:Receptor, PAR-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014162:Transfection; D014357:Trypsin / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000893:Anti-Inflammatory Agents; D019138:Antibodies, Blocking; D019151:Peptide Library; D044464:Receptor, PAR-2; D014357:Trypsin; D002118:Calcium