Discovery of N-(4-Fluoro-3-methoxybenzyl)-6-(2-(((2S,5R)-5-(hydroxymethyl)-1,4-dioxan-2-yl)methyl)-2H-tetrazol-5-yl)-2-methylpyrimidine-4-carboxamide. A Highly Selective and Orally Bioavailable Matrix Metalloproteinase-13 Inhibitor for the Potential Treatment of Osteoarthritis.

Matrix metalloproteinase-13 (MMP-13) is a zinc-dependent protease responsible for the cleavage of type II collagen, the major structural protein of articular cartilage. Degradation of this cartilage matrix leads to the development of osteoarthritis. We previously have described highly potent and selective carboxylic acid containing MMP-13 inhibitors; however, nephrotoxicity in preclinical toxicology species precluded development. The accumulation of compound in the kidneys mediated by human organic anion transporter 3 (hOAT3) was hypothesized as a contributing factor for the finding. Herein we report our efforts to optimize the MMP-13 potency and pharmacokinetic properties of non-carboxylic acid leads resulting in the identification of compound 43a lacking the previously observed preclinical toxicology at comparable exposures.

Environmental fate of pyrethroids in urban and suburban stream sediments and the appropriateness of Hyalella azteca model in determining ecological risk.

According to several recent studies using standard acute Hyalella azteca sediment bioassays, increased pyrethroid use in urban and suburban regions in California has resulted in the accumulation of toxic concentrations of pyrethroids in sediments of area streams and estuaries. However, a critical review of the literature indicates that this is likely an overestimation of environmental risk. Hyalella azteca is consistently the most susceptible organism to both aqueous and sediment-associated pyrethroid exposures when compared to a suite of other aquatic taxa. In some cases, H. azteca LC50 values are less than the community HC10 values, suggesting that the amphipod is an overly conservative model for community- or ecosystem-level impacts of sediment-associated pyrethroids. Further, as a model for responses of field populations of H. azteca, the laboratory bioassays considerably overestimate exposure, because the amphipod is more appropriately characterized as an epibenthic organism, not a true sediment dweller; H. azteca preferentially inhabit aquatic macrophytes, periphyton mats, and leaf litter, which drastically reduces their exposure to contaminated sediments. Sediment-bound pyrethroids are transported via downstream washing of fine particulates resulting in longer range transport but also more efficient sequestration of the chemical. In addition, site-specific variables such as sediment organic carbon content, grain size, temperature, and microbial activity alter pyrethroid bioavailability, degradation, and toxicity on a microhabitat scale. The type and source of the carbon in particular, influences the pyrethroid sequestering ability of sediments. The resulting irregular distribution of pyrethroids in stream sediments suggests that sufficient nonimpacted habitat may exist as refugia for resident sediment-dwelling organisms for rapid recolonization to occur. Given these factors, we argue that the amphipod model provides, at best, a screening level assessment of pyrethroid impacts and can correctly identify those sediments not toxic to benthic organisms but cannot accurately predict where sediments will be toxic.

Discovery of (pyridin-4-yl)-2H-tetrazole as a novel scaffold to identify highly selective matrix metalloproteinase-13 inhibitors for the treatment of osteoarthritis.

Potent, highly selective and orally-bioavailable MMP-13 inhibitors have been identified based upon a (pyridin-4-yl)-2H-tetrazole scaffold. Co-crystal structure analysis revealed that the inhibitors bind at the S(1)(') active site pocket and are not ligands for the catalytic zinc atom. Compound 29b demonstrated reduction of cartilage degradation biomarker (TIINE) levels associated with cartilage protection in a preclinical rat osteoarthritis model.

Clutch morphology and the timing of exposure impact the susceptibility of aquatic insect eggs to esfenvalerate.

We investigated Baetis spp. (mayfly), Hesperoperla pacifica (stonefly), and Brachycentrus americanus (caddisfly) susceptibility at the egg stage to esfenvalerate, a synthetic pyrethroid insecticide. Eggs were obtained from the field or from field-collected gravid females at sites near Corvallis (OR, USA) and the Metolius River at Camp Sherman (OR, USA) for static exposures under controlled conditions for temperature and light. Eggs were exposed to esfenvalerate for 48 h at concentrations ranging from 0.025 to 4.0 microg/L. No effect on mortality or posthatch growth was detected in H. pacifica eggs exposed to esfenvalerate concentrations up to 1.0 microg/L. Exposure to 0.07 microg/L of esfenvalerate, however, caused a significant increase in Baetis spp. egg mortality, and exposure of near-eclosion eggs to lower concentrations (0.025 and 0.05 microg/L) resulted in behavioral effects and reduced survivorship in newly hatched Baetis nymphs. Early stage B. americanus eggs were 10-fold more sensitive to esfenvalerate when removed from the gelatinous clutch before exposure, an indication that the gelatin affords protection from toxicant exposure. Exposures of near-hatch B. americanus clutches to esfenvalerate concentrations ranging between 0.035 and 0.2 microg/L, however, resulted in significant clutch death within clutches resulting from behavioral aberrations of first-instar larvae. The results of the present study suggest that aquatic insect egg clutch morphology can be a strong influence on susceptibility of embryos to esfenvalerate exposure.

Effects of dietary esfenvalerate exposures on three aquatic insect species representing different functional feeding groups.

Given the chemical properties of synthetic pyrethroids, it is probable that compounds, including esfenvalerate, that enter surface waters may become incorporated into aquatic insect food sources. We examined the effect of dietary esfenvalerate uptake in aquatic insects representing different functional feeding groups. We used three field-collected aquatic insect species: A grazing scraper, Cinygmula reticulata McDunnough (Ephemeroptera: Heptageniidae); an omnivorous filter feeder, Brachycentrus americanus Banks (Trichoptera: Brachycentridae); and a predator, Hesperoperla pacifica Banks (Plecoptera: Perlidae). Laboratory-cultured algae were preexposed for 24 h to esfenvalerate concentrations of 0, 0.025, 0.05, and 0.1 microg/L and provided to two C. reticulata age classes (small and final-instar nymphs). Reduction in small nymph growth was observed following three weeks of feeding on algae exposed to 0.05 and 0.1 microg/L of esfenvalerate, and the highest dietary exposure reduced egg production in final-instar nymphs. The diet for B. americanus and H. pacifica consisted of dead third-instar Chironomus tentans larvae preexposed for 24 h to esfenvalerate concentrations ranging between 0.1 and 1.0 microg/L. Consumption of larvae exposed to 0.5 to 1.0 microg/L of esfenvalerate caused case abandonment and mortality in B. americanus caddisfly larvae. Although H. pacifica nymphs readily consumed esfenvalerate-exposed larvae, no adverse effects were observed during the present study. Furthermore, no evidence of esfenvalerate-induced feeding deterrence was found in any of the species tested, suggesting that aquatic insects may not be able to distinguish between pyrethroid-contaminated and uncontaminated food sources. These findings indicate that feeding deterrence is not a factor in regulating aquatic insect dietary exposures to synthetic pyrethroids.

Impact of aquatic insect life stage and emergence strategy on sensitivity to esfenvalerate exposure.

We investigated the impact of aquatic insect life stage and emergence strategy on sensitivity to esfenvalerate, a synthetic pyrethroid insecticide, using field-collected Brachycentrus americanus Banks (Trichoptera: Brachycentridae) and Cinygmula reticulata McDunnough (Ephemeroptera: Heptageniidae) insects. Final-instar C. reticulata emergence was observed for one week following three environmentally relevant, 48-h esfenvalerate exposures (0.005, 0.01, and 0.015 microg/L). Emergence was significantly depressed following exposure to esfenvalerate and resulted from an increase in nymph mortality during the emergence process. This experiment was duplicated for late-instar C. reticulata nymphs, which were similar in size to the final-instar nymphs but were not near emergence. Late-instar C. reticulata mayflies were approximately fivefold less sensitive to esfenvalerate exposures as gauged by one-week mortality rates. Brachycentrus americanus pupal mortality was significantly increased over that in controls following 48-h esfenvalerate exposures of 0.1 and 0.2 microg/L. These response concentrations correlated closely with those for case-abandonment rates of fourth-instar B. americanus larvae (a sublethal effect of esfenvalerate exposure). Pupal mortality rates were approximately 16-fold higher than those observed in larvae. Adult female egg weight as a percentage of total body weight was significantly decreased following pupal esfenvalerate exposures of 0.05, 0.1, and 0.2 microg/L. These findings suggest that exposure to esfenvalerate may impair hemimetabolous insect emergence behaviors and may decrease fecundity in holometabolous aquatic insects.