One-pot synthesis of multisubstituted propenylbenzenes from benzyl chlorides through relay catalysis of palladium.

A strategy for the synthesis of multisubstituted propenylbenzenes using benzyl chlorides as starting materials is described. The palladium-catalyzed allylative dearomatization and the subsequent Wagner-Meerwein rearrangement as well as the olefin isomerization proceeded smoothly under mild conditions to produce propenylation products in good yields with high regioselectivity. Control experiments and cyclic voltammetry analysis suggest that Bu3SnCl, a by-product generated in the first step of allylative dearomatization, plays an essential role in the third step of olefin isomerization in the presence of a Brønsted acid.

Palladium-catalyzed C-H dimethylamination of 1-chloromethyl naphthalenes with N,N-dimethylformamide as the dimethyl amino source.

Palladium-catalyzed remote C-H dimethylamination of 1-chloromethylnaphthalenes using N,N-dimethylformamide as the dimethylamino source is described for the first time. The dimethylamination took place exclusively at the 4-position of 1-chloromethylnaphthalenes in 2-methyltetrahydrofuran under mild conditions to afford 1-(N,N-dimethylamino)-4-alkylnaphthalenes in good to high yields. The halogen atom remained intact during the dimethylamination of 1-chloromethylnaphthalenes. A P,N bidentate ligand was conveniently synthesized and successfully utilized as the ligand in the Kumada-Corriu reaction.

Thermal Hysteresis Involving Reversible Self-Catalytic Reactions.

Hysteresis is ubiquitous in nature and biology. It appears in ferromagnetism, ferroelectrism, traffic congestion, river sedimentation, electronics, thermoresponses, cell division, differentiation, and apoptosis. Hysteresis phenomena are beyond equilibrium and involve nonlinear, bistable, time delay, and memory events, which are described in input/output profiles by different outputs during continuous decreases and increases in input intensity. Although hysteresis profiles in these phenomena appear similar, the mechanisms underlying them are complex, and their basic understanding is desired. In this Account, I describe thermal hysteresis caused by molecules dispersed in dilute solutions containing optically active helicene oligomers, which form homo- and heterodouble helices, the cooling and heating processes of which cause different structural changes with regard to their relative concentrations. Reversible self-catalytic reactions are involved in the formation of a double helix, which catalyzes its own formation. The reactions accelerate as they progress, in contrast to ordinary reactions, which exhibit monotonic retardation as they progress. Thermal hysteresis involving reversible self-catalytic reactions exhibits notable phenomena, when various cooling/heating inputs are applied during the reaction; these phenomena are shown herein with profiles of experimental results of Δε outputs obtained by circular dichroism (CD) plotted against temperature inputs. Thermal hysteresis is discussed in terms of (1) two states of the homodouble helix and a random coil involving one reversible self-catalytic reaction and (2) three states of enantiomeric heterodouble helices and a random coil involving two reversible self-catalytic reactions. Repeated cooling and heating processes provide the same stable thermal hysteresis loops, when the initial and final high-temperature states are under equilibrium, and nonloop and unstable thermal hysteresis appears when whole the systems are beyond equilibrium. Diverse thermal hysteresis loops are obtained under different temperature change conditions for different oligomers. The mechanism of thermal hysteresis involves different macroscopic mechanisms at a fixed temperature, when the relative concentrations of substrates/products and the reaction direction differ. Microscopic mechanisms, which are shown by energy diagrams, are fixed at a temperature irrespective of cooling or heating. A comparison of thermal hysteresis loops and equilibrium curves provides distances to the metastable states on the loops from equilibrium, and reactions occur from the metastable states toward equilibrium. Notable phenomena described herein include bistability, high sensitivity to small concentration changes, equilibrium crossing, three-state one-directional structural change caused by a single heating procedure, reaction shortcuts, the memory effect on thermal history, figure-eight thermal hysteresis, chemical oscillation, stable and unstable thermal hysteresis, double-helix formation only under heating, and chiral symmetry breaking.

Palladium-Catalyzed Para-C-H Bond Amination of 2-Aryl Chloromethylbenzenes.

Palladium-catalyzed para-C-H bond amination of 2-aryl chloromethylbenzenes is described for the first time. The reactions of 2-aryl chloromethylbenzenes with cyclic amines proceeded smoothly in the presence of Pd(acac)2, tri(2-furyl)phosphine, and NaH in tetrahydrofuran at 40 °C to provide para-C-H bond aminated products in satisfactory to high yields with acceptable regioselectivity in most cases. The electronic property of the substituents linked to the benzene rings did not significantly influence the reactivity of the 2-aryl chloromethylbenzene substrates and the reaction regioselectivity.

Spirocarbocycle Synthesis from Chloromethylarenes via Transition-Metal-Catalyzed Allylative Dearomatization and Ring Closure Metathesis.

A strategy for the synthesis of spirocarbocycles by using chloromethyl arenes as starting materials is described in this paper. The palladium-catalyzed allylative dearomatization and the subsequent ruthenium-catalyzed ring closure metathesis proceeded smoothly under mild conditions to produce the corresponding spirocarbocycle products with moderate to high yields. Benzene-ring-, naphthalene-ring-, and anthracene-ring-containing substrates can be easily transformed into spirocarbocycles by using the proposed method.

Synthesis of trans-stilbenes via phosphine-catalyzed coupling reactions of benzylic halides.

An efficient and practical phosphine-catalyzed homo-coupling reaction of benzyl chlorides is described. The reactions proceed smoothly in the presence of CsF/B(OMe)3 and NaH as the base, respectively, to provide trans-stilbenes in good yields with a broad scope. Unsymmetrical stilbenes are also generated from the reactions of benzyl chlorides with phosphonium salts. Several P-based key intermediates have been detected by NMR and HRMS analyses, which shed light on the postulated catalytic cycle. In the presence of different bases, the transformations involve two different pathways, in which phenylcarbene and phosphonium alkoxide are considered as key intermediates, respectively. The two pathways are complementary in synthesis but different in mechanisms. The synthetic utility, including gram-scale reactions and straightforward access to π-conjugated molecules, has been demonstrated as well.

Stable and Unstable Concentration Oscillations Induced by Temperature Oscillations on Reversible Nonequilibrium Chemical Reactions of Helicene Oligomers.

Temperature oscillations can affect behaviors of living things. In this article, we describe the effect of triangle temperature oscillations on reversible nonequilibrium chemical reactions detected as concentration oscillations. When amplification through self-catalytic reactions is involved in the chemical reactions, concentration oscillations exhibit diverse nonequilibrium phenomena, which include equilibrium intersecting, equilibrium noncontact, and equilibrium sliding. Both stable and unstable concentration oscillations occur, during which repeated cycles provide the same and different concentration oscillations, respectively. Concentration oscillations are classified according to their waveforms in concentration/time profiles, the shapes of hysteresis curves in concentration/temperature profiles, the nature of self-catalytic reactions, and their relationships with equilibrium. An unstable concentration oscillation may be transformed into a stable concentration oscillation, which is described on the basis of the classifications. Experimental examples are shown using reversible association and dissociation reactions of helicene oligomers.

Sequential self-catalytic reactions in the formation of hetero-double-helix and their self-assembled gels by pseudoenantiomer mixtures of ethynylhelicene oligomers.

A 1:1 mixture of pseudoenantiomer ethynylhelicene (P)-pentamer and (M)-tetramer in toluene formed hetero-double-helix and their self-assembled gels. Kinetic analysis under isothermal conditions showed a complex nonlinear nature with regard to temperature changes. At 60°C, sigmoidal kinetics were observed, which disappeared after seeding with the self-assembled gels. These findings indicate the involvement of self-catalytic reaction I, in which a hetero-double-helix catalyzes the reaction to form a hetero-double-helix from random-coils. At 20°C, stairwise biphasic kinetics were observed, which disappeared after seeding. This phenomenon was explained by the involvement of two reactions with sigmoidal kinetics, namely, the formation of self-assembled gel I from hetero-double-helix by self-catalytic reaction II and the formation of self-assembled gel II by self-catalytic reaction III. Constant-rate temperature change experiments between 90 and 5°C showed nonsigmoidal thermal hysteresis in accordance with the involvement of sequential self-catalytic reactions with different reaction rates.

Rhodium-Catalyzed Synthesis of Organosulfur Compounds Involving S-S Bond Cleavage of Disulfides and Sulfur.

Organosulfur compounds are widely used for the manufacture of drugs and materials, and their synthesis in general conventionally employs nucleophilic substitution reactions of thiolate anions formed from thiols and bases. To synthesize advanced functional organosulfur compounds, development of novel synthetic methods is an important task. We have been studying the synthesis of organosulfur compounds by transition-metal catalysis using disulfides and sulfur, which are easier to handle and less odiferous than thiols. In this article, we describe our development that rhodium complexes efficiently catalyze the cleavage of S-S bonds and transfer organothio groups to organic compounds, which provide diverse organosulfur compounds. The synthesis does not require use of bases or organometallic reagents; furthermore, it is reversible, involving chemical equilibria and interconversion reactions.

Elevation of sensitivity to anticancer agents of human lung adenocarcinoma A549 cells by knockdown of claudin-2 expression in monolayer and spheroid culture models.

Claudins, tight junctional proteins, regulate the paracellular permeability of ions and small molecules. Claudin-2 is highly expressed in human lung adenocarcinoma cells and is involved in the up-regulation of cell proliferation. However, the effect of claudin-2 on cellular sensitivity to anticancer agents has not been clarified. The cytotoxicity of anticancer agents such as cisplatin, gefitinib and doxorubicin (DXR) was increased by claudin-2 knockdown in A549 cells. Claudin-2 knockdown also significantly decreased the expression level of multidrug resistance-associated protein/ABCC2. The expression levels of other drug efflux transporters were unchanged. The intracellular accumulation of 5-chloromethylfluorescein diacetate (CMFDA) and DXR, substrates of ABCC2, was increased by claudin-2 knockdown, whereas the efflux was decreased. MK-571, an inhibitor of ABCC2, enhanced the cytotoxicity of anticancer agents. Claudin-2 knockdown decreased the levels of p-c-Jun and nuclear Sp1. SP600125, an inhibitor of c-Jun, and mithramycin, an inhibitor of Sp1, decreased the level of ABCC2. The promoter activity of ABCC2 was decreased by claudin-2 knockdown, SP600125 and mithramycin treatments, suggesting that claudin-2 is involved in the up-regulation of ABCC2 expression at the transcriptional level. Claudin-2 knockdown increased the paracellular permeability of DXR in a 2D monolayer culture model. In addition, the accumulation of DXR into spheroids was enhanced by claudin-2 knockdown, resulting in a reduction in cell viability. We suggest that claudin-2 may be a novel therapeutic target in lung adenocarcinoma, because claudin-2 knockdown increased the accumulation of anticancer agents in cancer cells and spheroids.

Ferroelectric Alkylamide-Substituted Helicene Derivative with Two-Dimensional Hydrogen-Bonding Lamellar Phase.

Alkylamide (-CONHC nH2 n+1)-substituted benzene and its pyrene derivatives have shown a discotic hexagonal columnar liquid-crystalline phase through a one-dimensional (1D) intermolecular N-H···O═ hydrogen-bonding interaction, the direction of which is inverted through the application of an alternate current voltage. The polar hydrogen-bonding chains and dipole inversion reveal a ferroelectric polarization-electric field ( P- E) hysteresis curve. Non-π-planar helicene derivatives bearing two -CONHC14H29 chains also indicate a ferroelectric response. The racemic helicene derivative shows a bilayer lamellar liquid-crystal phase within a temperature range of 330-420 K, whereas there is no liquid crystallinity for the optically active derivative because of the different molecular assembly structure. The racemic phase is constructed through a two-dimensional (2D) N-H···O═ hydrogen-bonding network, which shows ferroelectric P- E hysteresis curves at above 340 K. The collective dipole inversion in the 2D layer contributes to the ferroelectricity in the lamellar phase. The remanent polarization ( Pr) of 11.1 µC cm-2 is about 6 times higher than those of the π-planar benzene- and pyrene-based 1D ferroelectrics. Both the density of the hydrogen-bonding site and the domain orientation in the 2D system are higher than those of the 1D columnar system.

Long-Range Anisotropic Structural Films and Fibers Formed from Lyotropic Liquid Crystal Gels Containing Hetero-Double-Helices with C16 Terminal Groups.

Long-range anisotropic structural materials exhibit notable optical and mechanical properties, and an efficient method for synthesizing such materials involving self-assembly of well-defined monodispersed organic molecules is described here. Hetero-double-helices are formed in toluene using a pseudoenantiomeric mixture of an ethynylhelicene ( M)-tetramer with C16 terminal groups and a ( P)-pentamer. When the concentration of the mixture was increased, the hetero-double-helices self-assembled to form lyotropic liquid crystal gels. On evaporating the solvent by drop casting, a long-range anisotropic structural film with a single domain and a size of up to centimeter order was spontaneously formed. Kinetics analysis of the film formation indicated the generation of perpendicularly aligned liquid crystal domains at the interface of the liquid and solid phases. When the lyotropic liquid crystal gel was extruded into methanol, a long-range anisotropic structural fiber with a single domain was formed. Different shapes of long-range anisotropic structural materials were obtained by different mechanical treatments of lyotropic liquid crystal gels.

Continued exposure of anti-cancer drugs to human iPS cell-derived cardiomyocytes can unmask their cardiotoxic effects.

Although the cardiotoxicity of anti-cancer drugs is an important issue, the underlying mechanisms remain unknown. To develop a sensitive assay system for cardiotoxicity, we examined effects of anti-cancer drugs on contractile functions of human iPS cell-derived cardiomyocytes by using non-invasive motion field imaging analysis with extended drug exposure time. We succeeded in continuously measuring stable contractile function. The continued exposure revealed that the difference in cardiotoxicity between cardiotoxic doxorubicin and less toxic erlotinib was more evident after 8 days of treatment than with 3 days of treatment, suggesting that continued exposure improved the predictive power for cardiotoxicity of anti-cancer drugs.

Proximate stochastic chiral symmetry breaking is mechanically tunable: formation of enantiomeric hetero-double-helices and aggregates from racemic oxymethylenehelicene oligomers.

A mixture of oxymethylenehelicene (P)-hexamer and (M)-hexamer in solution exhibited chiral symmetry breaking, which was induced by mechanical stirring, during the formation of enantiomeric hetero-double-helices and their aggregates. A racemic 50 : 50 mixture was heated to 90 °C to form a dissociated state, and then cooled to 25 °C. Mechanical stirring with a magnetic stirrer at 2000 rpm for 100 h resulted in the exhibition of a strong negative Cotton effect at 322 nm. Repeated experiments provided negative Cotton effects, which is due to the deterministic chiral symmetry breaking. No change in the Cotton effect occurred in the absence of stirring. The (P)-hexamer to (M)-hexamer mixing molar fraction was varied, and a positive Cotton effect appeared at molar fractions between 40 : 60 and 46 : 54 and a negative Cotton effect at molar fractions between 48 : 52 and 60 : 40, which was reversed at 47 : 53. The slight deviation of symmetry from that at 50 : 50 was termed proximate stochastic chiral symmetry breaking. The process of chiral symmetry breaking could be tuned by varying the procedures of mechanical stirring and mixing procedures for solutions of (P)-hexamer and/or (M)-hexamer.

The RING finger- and PDZ domain-containing protein PDZRN3 controls localization of the Mg2+ regulator claudin-16 in renal tube epithelial cells.

Ion exchange in the renal tubules is fundamental to the maintenance of physiological ion levels. Claudin-16 (CLDN16) regulates the paracellular reabsorption of Mg2+ in the thick ascending limb of Henle's loop in the kidney, with dephosphorylation of CLDN16 increasing its intracellular distribution and decreasing paracellular Mg2+ permeability. CLDN16 is located in the tight junctions, but the mechanism regulating its localization is unclear. Using yeast two-hybrid systems, we found that CLDN16 binds to PDZRN3, a protein containing both RING-finger and PDZ domains. We also observed that the carboxyl terminus of the cytoplasmic CLDN16 region was required for PDZRN3 binding. PZDRN3 was mainly distributed in the cytosol of rat kidney cells and upon cell treatment with the protein kinase A inhibitor H-89, colocalized with CLDN16. H-89 also increased mono-ubiquitination and the association of CLDN16 with PDZRN3. Mono-ubiquitination levels of a K275A mutant were lower, and its association with PDZRN3 was reduced compared with wild-type (WT) CLDN16 and a K261A mutant, indicating that Lys-275 is the major ubiquitination site. An S217A mutant, a dephosphorylated form of CLDN16, localized to the cytosol along with PDZRN3 and the endosomal marker Rab7. PDZRN3 siRNA increased cell-surface localization of WT CLDN16 in H-89-treated cells or containing the S217A mutant and also suppressed CLDN16 endocytosis. Of note, H-89 decreased paracellular Mg2+ flux in WT CLDN16 cells, and PDZRN3 siRNA increased Mg2+ flux in the H-89-treated WT CLDN16 and S217A mutant cells. These results suggest that PDZRN3 mediates endocytosis of dephosphorylated CLDN16 and represents an important component of the CLDN16-trafficking machinery in the kidney.

Doublecortin marks a new population of transiently amplifying muscle progenitor cells and is required for myofiber maturation during skeletal muscle regeneration.

Muscle satellite cells are indispensable for muscle regeneration, but the functional diversity of their daughter cells is unknown. Here, we show that many Pax7(+)MyoD(-) cells locate both beneath and outside the basal lamina during myofiber maturation. A large majority of these Pax7(+)MyoD(-) cells are not self-renewed satellite cells, but have different potentials for both proliferation and differentiation from Pax7(+)MyoD(+) myoblasts (classical daughter cells), and are specifically marked by expression of the doublecortin (Dcx) gene. Transplantation and lineage-tracing experiments demonstrated that Dcx-expressing cells originate from quiescent satellite cells and that the microenvironment induces Dcx in myoblasts. Expression of Dcx seems to be necessary for myofiber maturation because Dcx-deficient mice exhibited impaired myofiber maturation resulting from a decrease in the number of myonuclei. Furthermore, in vitro and in vivo studies suggest that one function of Dcx in myogenic cells is acceleration of cell motility. These results indicate that Dcx is a new marker for the Pax7(+)MyoD(-) subpopulation, which contributes to myofiber maturation during muscle regeneration.

Synthesis and Self-Assembly of Chiral Cylindrical Molecular Complexes: Functional Heterogeneous Liquid-Solid Materials Formed by Helicene Oligomers.

Chiral cylindrical molecular complexes of homo- and hetero-double-helices derived from helicene oligomers self-assemble in solution, providing functional heterogeneous liquid-solid materials. Gels and liotropic liquid crystals are formed by fibril self-assembly in solution; molecular monolayers and fibril films are formed by self-assembly on solid surfaces; gels containing gold nanoparticles emit light; silica nanoparticles aggregate and adsorb double-helices. Notable dynamics appears during self-assembly, including multistep self-assembly, solid surface catalyzed double-helix formation, sigmoidal and stairwise kinetics, molecular recognition of nanoparticles, discontinuous self-assembly, materials clocking, chiral symmetry breaking and homogeneous-heterogeneous transitions. These phenomena are derived from strong intercomplex interactions of chiral cylindrical molecular complexes.

Hypotonic Stress-induced Down-regulation of Claudin-1 and -2 Mediated by Dephosphorylation and Clathrin-dependent Endocytosis in Renal Tubular Epithelial Cells.

Hypotonic stress decreased claudin-1 and -2 expression levels in renal tubular epithelial HK-2 and Madin-Darby canine kidney cells. Here, we examined the regulatory mechanism involved in this decrease. The hypotonicity-induced decrease in claudin expression was inhibited by the following: SB202190, a p38 MAPK inhibitor, but not by U0126, a MEK inhibitor; Go6983, a protein kinase C inhibitor; or SP600125, a Jun N-terminal protein kinase inhibitor. Hypotonic stress increased transepithelial electrical resistance, which was inhibited by SB202190. The mRNA expression level of claudin-1 was decreased by hypotonic stress but that of claudin-2 was not. Hypotonic stress decreased the protein stability of claudin-1 and -2. The hypotonicity-induced decrease in claudin expression was inhibited by the following: chloroquine, a lysosome inhibitor; dynasore and monodansylcadaverine, clathrin-dependent endocytosis inhibitors; and siRNA against clathrin heavy chain. Claudin-1 and -2 were mainly distributed in the cytosol and tight junctions (TJs) in the chloroquine- and monodansylcadaverine-treated cells, respectively. Hypotonic stress decreased the phosphorylation levels of claudin-1 and -2, which were inhibited by the protein phosphatase inhibitors okadaic acid and cantharidin. Dephosphorylated mutants of claudin-1 and -2 were mainly distributed in the cytosol, which disappeared in response to hypotonic stress. In contrast, mimicking phosphorylation mutants were distributed in the TJs, which were not decreased by hypotonic stress. We suggest that hypotonic stress induces dephosphorylation, clathrin-dependent endocytosis, and degradation of claudin-1 and -2 in lysosomes, resulting in disruption of the TJ barrier in renal tubular epithelial cells.

Claudin-18 inhibits cell proliferation and motility mediated by inhibition of phosphorylation of PDK1 and Akt in human lung adenocarcinoma A549 cells.

Abnormal expression of claudin subtypes has been reported in various cancers. However, the pathological role of each claudin has not been clarified in detail. Claudin-18 was absent in human non-small cell and small cell lung cancers, although it is expressed in normal lung tissues. Here, we examined the effect of claudin-18 expression on the expression of junctional proteins, cell proliferation, and cell motility using human lung adenocarcinoma A549 cells. Real-time PCR and western blotting showed that exogenous expression of claudin-18 had no effect on the expression of junctional proteins including claudin-1, zonula occludens-1 (ZO-1), occludin, and E-cadherin. Claudin-18 was mainly distributed in cell-cell contact areas concomitant with ZO-1. Cell proliferation was significantly decreased at 48 and 72h after seeding of claudin 18-expressing cells. Claudin-18 suppressed cell motility, whereas it increased cell death in anoikis. Claudin-18 decreased phosphorylated (p)-3-phosphoinositide-dependent protein kinase-1 (PDK1) and p-Akt levels without affecting p-epidermal growth factor receptor and p-phosphatidylinositol-3 kinase (PI3K) levels. Furthermore, claudin-18 was bound with PDK1 and suppressed the nuclear localization of PDK1. We suggest that claudin-18 suppresses the abnormal proliferation and motility of lung epithelial cells mediated by inhibition of the PI3K/PDK1/Akt signaling pathway.

Up-Regulation of Transient Receptor Potential Melastatin 6 Channel Expression by Tumor Necrosis Factor-α in the Presence of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor.

Anti-epidermal growth factor receptor (EGFR) drugs such as erlotinib and gefitinib cause a side effect of hypomagnesemia, but chemotherapy to treat this has not yet been developed. The transient receptor potential melastatin 6 (TRPM6) channel is involved in the reabsorption of Mg2+ in the renal tubule. We reported previously that the expression of TRPM6 is up-regulated by epidermal growth factor (EGF) in renal tubular epithelial NRK-52E and HEK293 cells. EGF-induced elevation of TRPM6 expression was inhibited by erlotinib, gefitinib, and lapatinib. We found that tumor necrosis factor-α (TNF-α) increases TRPM6 expression in the presence of erlotinib. Therefore, we investigated what molecules are involved in the up-regulation of TRPM6 expression by TNF-α. EGF increased the levels of phosphorylated extracellular signal-regulated kinase 1 and 2 (p-ERK1/2), which were inhibited by erlotinib. TNF-α did not change p-ERK1/2 levels, but increased the phosphorylation and nuclear localization of nuclear factor-κB (NF-κB), which were blocked by the NF-κB inhibitors BAY 11-7082 and pyrrolidinedithiocarbamate ammonium. Similarly, luciferase reporter activity of human TRPM6 was increased by TNF-α, which was blocked by NF-κB inhibitors, and was inhibited by a mutation in the κB-binding site in the proximal region of the TRPM6 promoter. A chromatin immunoprecipitation assay revealed that NF-κB binds to the κB-binding site, which was blocked by NF-κB inhibitors. In the presence of erlotinib, TNF-α increased Mg2+ influx, which was blocked by NF-κB inhibitors. These results suggest that TNF-α reverses the reduction in Mg2+ reabsorption caused by anti-EGFR drugs. J. Cell. Physiol. 232: 2841-2850, 2017. © 2016 Wiley Periodicals, Inc.