title: Single cycle structure-based humanization of an anti-nerve growth factor therapeutic antibody.
abstract: Most forms of chronic pain are inadequately treated by present therapeutic options. Compelling evidence has accumulated, demonstrating that Nerve Growth Factor (NGF) is a key modulator of inflammatory and nociceptive responses, and is a promising target for the treatment of human pathologies linked to chronic and inflammatory pain. There is therefore a growing interest in the development of therapeutic molecules antagonising the NGF pathway and its nociceptor sensitization actions, among which function-blocking anti-NGF antibodies are particularly relevant candidates.In this respect, the rat anti-NGF αD11 monoclonal antibody (mAb) is a potent antagonist, able to effectively antagonize rodent and human NGF in a variety of in vitro and in vivo systems. Here we show that mAb αD11 displays a significant analgesic effect in two different models of persistent pain in mice, with a remarkable long-lasting activity. In order to advance αD11 mAb towards its clinical application in man, anti-NGF αD11 mAb was humanized by applying a novel single cycle strategy based on the a priori experimental determination of the crystal and molecular structure of the parental Fragment antigen-binding (Fab). The humanized antibody (hum-αD11) was tested in vitro and in vivo, showing that the binding mode and the NGF neutralizing biological activities of the parental antibody are fully preserved, with even a significant affinity improvement. The results firmly establish hum-αD11 as a lead candidate for clinical applications in a therapeutic area with a severe unmet medical need. More generally, the single-cycle structure-based humanization method represents a considerable improvement over the standard humanization methods, which are intrinsically empirical and require several refinement cycles.
authors.affiliation: Lay Line Genomics, SpA, Roma, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Covaceuszach
authors.forename: Sara
authors.initials: S
authors.lastname: Marinelli
authors.forename: Ivet
authors.initials: I
authors.lastname: Krastanova
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ugolini
authors.forename: Flaminia
authors.initials: F
authors.lastname: Pavone
authors.forename: Doriano
authors.initials: D
authors.lastname: Lamba
authors.forename: Antonino
authors.initials: A
authors.lastname: Cattaneo
mesh_terms: D000595:Amino Acid Sequence; D000700:Analgesics / Q000737:chemistry* / Q000276:immunology* / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry* / Q000276:immunology* / Q000494:pharmacology / Q000627:therapeutic use; D000918:Antibody Specificity; D018360:Crystallography, X-Ray; D015195:Drug Design; D005557:Formaldehyde / Q000009:adverse effects; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000737:chemistry / Q000276:immunology*; D009437:Neuralgia / Q000139:chemically induced / Q000188:drug therapy; D011487:Protein Conformation; D051381:Rats
chemical_list: D000700:Analgesics; D061067:Antibodies, Monoclonal, Humanized; D005557:Formaldehyde; D020932:Nerve Growth Factor