title: Discovery of the improved antagonistic prolactin variants by library screening.
abstract: Prolactin (PRL), a potent growth stimulator of the mammary epithelium, has been suggested to be a factor contributing to the development and progression of breast and prostate cancer. Several PRL receptor (PRLR) antagonists have been identified in the past decades, but their in vivo growth inhibitory potency was restricted by low receptor affinity, rendering them pharmacologically unattractive for clinical treatment. Thus, higher receptor affinity is essential for the development of improved PRLR antagonistic variants with improved in vivo potency. In this study, we generated Site 1 focused protein libraries of human G129R-PRL mutants and screened for those with increased affinity to the human PRLR. By combining the mutations with enhanced affinities for PRLR, we identified a novel G129R-PRL variant with mutations at Site 1 that render nearly 50-fold increase in the antagonistic potency in vitro.
authors.affiliation: Novo Nordisk China R&D, 29 Life Science Park Road, Beijing, China. yunl@novonordisk.com
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Gong
authors.forename: Jens
authors.initials: J
authors.lastname: Breinholt
authors.forename: Leif
authors.initials: L
authors.lastname: Nørskov-Lauritsen
authors.forename: Jinchao
authors.initials: J
authors.lastname: Zhang
authors.forename: Qinhong
authors.initials: Q
authors.lastname: Ma
authors.forename: Jianhe
authors.initials: J
authors.lastname: Chen
authors.forename: Svetlana
authors.initials: S
authors.lastname: Panina
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.forename: Tengkun
authors.initials: T
authors.lastname: Li
authors.forename: Jingyuan
authors.initials: J
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Kong
authors.forename: Zibing
authors.initials: Z
authors.lastname: Liu
authors.forename: Jingjing
authors.initials: J
authors.lastname: Mao
authors.forename: Leif
authors.initials: L
authors.lastname: Christensen
authors.forename: Sean
authors.initials: S
authors.lastname: Hu
authors.forename: Lingyun
authors.initials: L
authors.lastname: Wang
mesh_terms: D001943:Breast Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D019382:Human Growth Hormone / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D008297:Male; D009154:Mutation; D011388:Prolactin / Q000235:genetics / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D011981:Receptors, Prolactin / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D011981:Receptors, Prolactin; D019382:Human Growth Hormone; D011388:Prolactin