title: The generation of rat dorsal root ganglion cell lines to identify the target of KW-7158, a novel treatment for overactive bladder.
abstract: KW-7158 is a drug candidate for the treatment of overactive bladder. Although pharmacological studies have suggested that it suppresses afferent nerve conduction, its molecular target is unknown. We herein report the establishment of dorsal root ganglion (DRG) cell lines useful for identification of the target of this compound. First, we confirmed that the target exists in rat primary DRG by [(3)H]KW-7158 binding. To establish DRG cell lines, we used DRG from transgenic rats harboring the temperature-sensitive large T-antigen. The immortalized cells were initially screened for their expression of neuronal markers, and 72 positive clones were obtained (designated as TRD cells). Next, in order to select TRD cells expressing the target of KW-7158, we measured the binding affinity and amount of the binding sites present in each clone. Most clones expressed two binding sites, one with low affinity and one with high affinity. Differential binding of KW-7158 derivatives to each site revealed that the high affinity site is pharmacologically relevant. Therefore, we successfully identified "TRD-10" which express the largest amount of the high affinity site. These cell lines will therefore be useful tools to identify the target of KW-7158.
authors.affiliation: Biologics Research Laboratories, Kyowa Hakko Kirin Co., Ltd., 3-6-6 Asahi-machi, Machida-City, Tokyo 194-8533, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Nishiya
authors.forename: Sachiko
authors.initials: S
authors.lastname: Yokokawa
authors.forename: Ayumu
authors.initials: A
authors.lastname: Fukuda
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Yamagata
authors.forename: Atsushi
authors.initials: A
authors.lastname: Inayoshi
authors.forename: Masuo
authors.initials: M
authors.lastname: Obinata
authors.forename: Kenji
authors.initials: K
authors.lastname: Shibata
mesh_terms: D000818:Animals; D001582:Benzothiepins / Q000378:metabolism / Q000493:pharmacokinetics*; D018929:Cell Culture Techniques / Q000379:methods*; D002461:Cell Line, Transformed; D005727:Ganglia, Spinal / Q000187:drug effects* / Q000473:pathology / Q000502:physiology; D008297:Male; D061251:Primary Cell Culture; D051381:Rats; D055647:Rats, Transgenic; D017208:Rats, Wistar; D011984:Sensory Receptor Cells / Q000187:drug effects* / Q000473:pathology / Q000502:physiology; D053201:Urinary Bladder, Overactive / Q000188:drug therapy* / Q000473:pathology / Q000503:physiopathology
chemical_list: D001582:Benzothiepins; C466985:KW 7158