A series of 3-substituted aminocyclopentanes has been identified as highly potent and selective NR2B receptor antagonists. Incorporation of a 1,2,4-oxadiazole linker and substitution of the pendant phenyl ring led to the discovery of orally bioavailable analogues that showed efficient NR2B receptor occupancy in rats. Unlike nonselective NMDA antagonists, the NR2B-selective antagonist 22 showed no adverse affects on motor coordination in the rotarod assay at high dose. Compound 22 was efficacious following oral administration in a spinal nerve ligation model of neuropathic pain and in an acute model of Parkinson's disease in a dose dependent manner.

A novel series of annulated tricyclic compounds was synthesized and evaluated as NMDA/NR2B antagonists. Structure-activity development was directed towards in vitro optimization of NR2B activity and selectivity over the hERG K(+) channel. Preferred compounds were subsequently evaluated for selectivity in an alpha(1)-adrenergic receptor binding counter-screen and a cell-based assay of NR2B activity.

The discovery of a novel series of NR2B subtype selective N-methyl-d-aspartate (NMDA) antagonists is reported. Initial optimization of a high-throughput screening lead afforded an aminopyridine derivative 13 with significant NR2B antagonist potency but limited selectivity over hERG-channel and other off-target activities. Further structure-activity studies on the aminoheterocycle moiety and optimization of the carbamate led to the highly potent 2-aminopyrimidine derivative 20j with a significantly improved off-target activity profile and oral bioavailability in multiple species coupled with good brain penetration. Compound 20j demonstrated efficacy in in vivo rodent models of antinociception, allodynia, and Parkinson's disease.

Retina-specific nuclear receptor (RNR), also known as PNR and NR2E3, is an orphan nuclear receptor expressed exclusively in photoreceptor cells of the retina. Here we describe homogeneous cell-based resonance energy transfer assay for identification of RNR agonists using beta-lactamase as the reporter gene. Bacterial beta-lactamase reporter construct containing GAL4 response elements was randomly integrated into the genome with subsequent selection of responsive cell pools by fluorescence-activated cell sorting. Chimeric RNR (RNR hinge and ligand-binding domains fused to GAL4 DNA-binding domain) was stably transfected into mammalian Flp-In Chinese hamster ovary cells using Flp-mediated recombination into a single pre-integrated Flp recombination target site. Since no RNR ligand could be used as a control for monitoring the development of the RNR assay, we developed a parallel cell line with the functionally related well-characterized thyroid hormone nuclear receptor. This parallel thyroid hormone nuclear receptor system was used as a "guide" in optimizing the RNR assay for ultra-high-throughput screening in 3,456-well nanoplate format. The assay was successfully used to screen a large compound collection for RNR agonists. In this study we demonstrated the feasibility of developing and optimization of the high-throughput screening-compatible assay for the orphan nuclear receptor in the absence of its cognitive ligand.

We described skeletal deformities in Willamette River fishes from larval and juvenile specimens collected in 2002 and 2003. Deformities were found in most taxa examined but were more frequent in native broadcast spawners, especially minnows and suckers, than in native or exotic nest builders. Caudal deformities were uniformly distributed throughout the river, but precaudal deformities were more localized near the towns of Newberg and Salem (Wheatland Ferry). In northern pikeminnow, Ptychocheilus oregonensis, deformities were dependent on relative hatch date, with early season fish having about three times the deformity load as late season fish. In a subsample of northern pikeminnow and chiselmouth, Acrocheilus alutaceus, number of deformities was directly related to number of trematode metacercariae and precaudal deformities were twice as likely as caudal deformities to be associated with metacercariae. Based on a logistic regression, the probability of a precaudal deformity was dependent on number of metacercariae and geographic area with the area effect disappearing as the number of cysts increased. A separate analysis showed that some types of deformities were unlikely to be associated with metacercariae. However, even in cases where metacercariae were unlikely to be associated with deformities, metacercariae were usually present elsewhere in the fish and an indirect effect could not be dismissed. The taxonomic, spatial and temporal patterns of skeletal deformities in Willamette River fishes may be due to differences in intermediate host (snail or fish) resistance or susceptibility to parasites, to differences in life history ecology, or to anthropogenic effects that are manifest in increased precaudal deformities near Newberg and Wheatland Ferry or decreased rates elsewhere.

ABSTRACT The Willamette River, one of only 14 American Heritage Rivers, flows through the most densely