No abstract

No abstract

A mild and selective C(sp3)−H aerobic oxidation enabled by decatungstate photocatalysis has been developed. The reaction can be significantly improved in a microflow reactor enabling the safe use of oxygen and enhanced irradiation of the reaction mixture. Our method allows for the oxidation of both activated and unactivated C−H bonds (30 examples). The ability to selectively oxidize natural scaffolds, such as (−)‐ambroxide, pregnenolone acetate, (+)‐sclareolide, and artemisinin, exemplifies the utility of this new method.

Participation of p38 mitogen-activated protein kinase (p38) in insulin-induced glucose uptake was suggested using pyridinylimidazole p38 inhibitors (e.g. SB203580). However, the role of p38 in insulin action remains controversial. We further test p38 participation in glucose uptake using a dominant-negative p38 mutant and two novel pharmacological p38 inhibitors related to but different from SB203580. We present the structures and activities of the azaazulene pharmacophores A291077 and A304000. p38 kinase activity was inhibited in vitro by A291077 and A304000 (IC 50 ‫؍‬ 0.6 and 4.7 M). At higher concentrations A291077 but not A304000 inhibited JNK2␣ (IC 50 ‫؍‬ 3.5 M). Pretreatment of 3T3-L1 adipocytes and L6 myotubes expressing GLUT4myc (L6-GLUT4myc myotubes) with A291077, A304000, SB202190, or SB203580 reduced insulin-stimulated glucose uptake by 50 -60%, whereas chemical analogues inert toward p38 were ineffective. Expression of an inducible, dominant-negative p38 mutant in 3T3-L1 adipocytes reduced insulin-stimulated glucose uptake. GLUT4 translocation to the cell surface, immunodetected on plasma membrane lawns of 3T3-L1 adipocytes or on intact L6-GLUT4myc myotubes, was not altered by chemical or molecular inhibition of p38. We propose that p38 contributes to enhancing GLUT4 activity, thereby increasing glucose uptake. In addition, the azaazulene class of inhibitors described will be useful to decipher cellular actions of p38 and JNK.The p38 mitogen-activated protein kinases (p38), also referred to as stress-activated protein kinases-2, are a family of proline-directed serine/threonine kinases (1, 2). At least four isoforms, the products of different genes, have been cloned and are 60 -70% identical in their amino acid sequence. The most commonly used nomenclature of these isoforms are p38␣ (3, 4), p38␤ (5, 6), p38␥ (7,8), and p38␦ (9, 10). A splice variant of the p38␤, referred to as p38␤2, has also been described (11). Northern blot analysis has shown a wide tissue distribution of these isoforms, although p38␤ and p38␥ are preferentially expressed in skeletal muscle (5, 9). In addition to stressors, members of this family of protein kinases can also be activated by growth factors (12-15).Full activation of p38 by pro-inflammatory cytokines requires phosphorylation of Thr-180 and Tyr-182 found within a TGY tripeptide motif in the activation loop of the kinase (16). This double phosphorylation is catalyzed by the dual-specific MAPK 1 kinases MKK3 and MKK6 and possibly via auto-phosphorylation (17). It is remarkable that stimuli that increase p38 phosphorylation such as insulin-like growth factor-1 (18), muscle contraction (19 -21), lipoic acid (22), 5-aminoimidazole-4-carboxamide ribonucleoside (23), pro-inflammatory cytokines (18), protein synthesis inhibitors (24, 25), hyperosmolar stress (26), and preconditioning (ischemia/reperfusion) (27) also elevate glucose uptake. Importantly, the pyridinylimidazole inhibitor of p38, SB203580, reduced the stimulation of glucose uptake by all of the above stimuli incl...

Engagement of the T cell antigen receptor (TcR)1 with the antigen-major histocompatibility complex on antigen-presenting cells triggers a complex TcR signaling cascade that leads to T cell activation and cytokine secretion (1). During this process, T cells express the autocrine growth factor interleukin 2 (IL-2), which promotes T cell proliferation by interacting with the IL-2 receptor, which is also up-regulated on activated T cells. The transcriptional regulation of the IL-2 gene has been extensively analyzed at the IL-2 promoter, a 275-bp region located upstream of the transcriptional start site of the gene (2, 3). Several transcription factors have been identified to bind elements within this regulatory region, including AP-1, NF-B, and the nuclear factor of activated T cells (NFAT) (2).The transcription factor NFAT plays an essential role in IL-2 expression. Binding sites for NFATs have also been found within the promoter regions of several other cytokine genes, including IL-3, IL-4, IL-5, IL-8, IL-13, tumor necrosis factor ␣, granulocyte-macrophage colony-stimulating factor, and ␥-IFN (4, 5). NFAT is a complex composed of a cytoplasmic subunit and an inducible nuclear component comprised of AP-1 (Fos/ Jun) family members. At least four structurally related NFAT cytoplasmic subunit members, NFATp/NFAT1, NFATc/ NFAT2, NFAT3, and NFATX/NFATc3/NFAT4, have been identified (5). NFAT proteins share a conserved domain located toward the C terminus (6) that binds DNA and also participates in cooperative protein-protein interactions with AP-1 transcription factors (7,8). Immediately N-terminal to the DNA-binding domain is a second conserved module of ϳ300 residues known as the NFAT homology (NFAT-h) region. The N terminus of NFAT, including the NFAT-h region, regulates nuclear/cytoplasm trafficking in response to changes in intracellular Ca 2ϩ concentrations. In resting T cells, the protein is retained in the cytoplasm and its NFAT-h domain is heavily phosphorylated. Engagement of the TcR or treatment of cells with the Ca 2ϩ ionophore activates the Ca 2ϩ /calmodulin-dependent Ser/Thr phosphatase, calcineurin. CaN dephosphorylates the NFAT-h domain, resulting in translocation of NFAT to the nucleus (9).

No abstract

A series of bis(trifluoromethyl)pyrazoles (BTPs) has been found to be a novel inhibitor of cytokine production. Identified initially as inhibitors of IL-2 synthesis, the BTPs have been optimized in this regard and even inhibit IL-2 production with a 10-fold enhancement over cyclosporine in an ex vivo assay. Additionally, the BTPs show inhibition of IL-4, IL-5, IL-8, and eotaxin production. Unlike the IL-2 inhibitors, cyclosporine and FK506, the BTPs do not directly inhibit the dephosphorylation of NFAT by calcineurin.

Technologies that enable rapid screening of diverse reaction conditions are of critical importance to methodology development and reaction optimization, especially when molecules of high complexity and scarcity are involved. The lackofageneral solid dispensing method for chemical reagents on micro-and nanomole scale prevents the full utilization of reaction screening technologies.W eherein report the development of at echnology in whichg lass beads coated with solid chemical reagents (ChemBeads) enable the delivery of nanomole quantities of solid chemical reagents efficiently.B y exploring the concept of preferred screening sets,the flexibility and generality of this technology for high-throughput reaction screening was validated.High-throughput reaction screening is at ool that enables the investigation of large numbers of reaction conditions in parallel for ap articular chemical transformation. Reaction screening is an essential component for any new synthetic methodology development and is often utilized in complex natural product total synthesis.Whenconducted in parallel, it has the potential to offer high speed and efficiency in identifying an ovel or optimal reaction condition. Importantly,itisideal to miniaturize the reaction scale so that only milligram quantities of high-value intermediates are consumed when running arrays of reaction conditions. [1] Historically,the advancement and broad impact of high-throughput reaction screening has been hindered by al ack of suitable technologies to effectively handle reaction miniaturization. Only in the last few years have transformative advancements in this field started to emerge.S eminal papers from Merck and Pfizer have demonstrated the use of bioassay equipment and flow instrumentation to enable nanomole scale reaction screening in ah igh-throughput fashion. [2] However, ac ritical field-wide challenge that has yet to be addressed is how to effectively dispense diverse solid chemical reagents on nanomole (submilligram) scale accurately and efficiently. [3] Our search for ar obotic platform capable of dispensing av ariety of solids reagents in small quantities (< 1mg) was unsuccessful. [4] Each reagent would require individualized protocol development for accurate robotic dispensing, making it unrealistic to have as ingle platform for all solids. [2b,3b] As ar esult, tedious manual weighing has been the only reliable method. With nanomole quantities of material, this becomes unfeasible.T hus,n anomole scale reaction screening relies almost exclusively on the use of stock solutions made from reagents which are soluble in the reaction solvent. [2,5] Ideally, any combination of reagents,i nitially soluble or not, need to be incorporated in as creening set in order to have an unbiased assessment of reactivity.I nl ight of this field-wide challenge,w ee ndeavored to develop au niversal method for dispensing solid chemical reagents on nanomole scale with accuracy and efficiencyi nt he context of high throughput reaction screening.During our research, we became awa...