2-(4-Boc-piperazinyl)-2-phenylacetic acid

Nuclear factor erythroid 2-related factor 2 (NRF2) is a pleiotropic transcription factor which regulates the constitutive and inducible transcription of a wide array of genes and confers protection against a variety of pathologies. Directly disrupting Kelch-like ECH-associated protein 1 (KEAP1)-NRF2 protein-protein interaction (PPI) has been explored as a promising strategy to activate NRF2. We reported here the first identification of a series of 2-oxy-2-phenylacetic acid substituted naphthalene sulfonamide derivatives as potent KEAP1-NRF2 inhibitors. Our efforts led to the potent small molecule KEAP1-NRF2 inhibitor, 20c, which exhibited a Kd of 24 nM to KEAP1 and an IC50 of 75 nM in disrupting KEAP1-NRF2 interaction. Subsequent biological studies provided consistent evidence across mouse macrophage cell-based and in vivo models that 20c induced NRF2 target gene expression and enhanced downstream antioxidant and anti-inflammatory activities. Our study not only demonstrated that small molecule KEAP1-NRF2 PPI inhibitors can be potential preventive and therapeutic agents for diseases and conditions involving oxidative stress and inflammation but also enriched the chemical diversity of the KEAP1-NRF2 inhibitors.

Introduction: Auxins are hormones that regulate plant growth and development. To accurately quantify the low levels of auxins present in plant and soil samples, sensitive detection methods are needed. In this study, the design and construction of two different whole cell auxin bioassays is illustrated. Both use the auxin responsive element HpaA as an input module but differ in output module. The first bioassay incorporates the gfp gene to produce a fluorescent bioassay. Whereas the second one utilizes the genes phzM and phzS to produce a pyocyanin producing bioassay whose product can be measured electrochemically. Results: The fluorescent bioassay is able to detect 4-hydroxyphenylacetic acid (4-HPA) and 2-phenylacetic acid (PAA) concentrations from 60 μM to 3 mM in a dose-responsive manner. The pyocyanin producing bioassay can detect 4-HPA concentrations from 1.9 to 15.625 μM and PAA concentrations from 15.625 to 125 μM, both in a dose-responsive manner. Conclusion: A fluorescent whole cell auxin bioassay and an electrochemical whole cell auxin bioassay were constructed and tested. Both are able to detect 4-HPA and PAA at concentrations that are environmentally relevant to plant growth.

Metamitron (1) does not undergo hydrolysis at pH 1-8 and up to 5 M H(2)SO(4). The product of its two-electron reduction, 1, 6-dihydrometamitron (2), on the other hand, undergoes at pH <3 relatively fast hydrolysis. The dependence of the measured rate constant on acidity indicates that the completely protonated form (AH(2)(2+)) predominating in strongly acidic media undergoes hydrolysis slower than the species bearing one less proton (AH(+)). The latter most reactive species is present in highest concentration in solutions of pH between 0 and 2. This species is protonated on the 2,3-azomethine bond and yields as final products 2-hydrazino-2-phenylacetic acid (4) and acethydrazide (5). Kinetic, polarographic, and spectrophotometric measurements indicated for the first dissociation an average value pK(a) = -0.8, for the second pK(a) = 0.95. These observations together with the easy reduction of the 1,6-bond in metamitron (1) indicate that in nature the cleavage of metamitron may be preceded by its reduction to 1, 6-dihydrometamitron (2), which is then hydrolyzed. Thus, anaerobic, reductive conditions are likely preferable for the total microbial degradation of metamitron.