A reappraisal,based on (31)P NMR,of the direct coordination of a metal ion with the phosphoryl oxygen at the cleavage site of a hammerhead ribozyme

It has been generally accepted, on the basis of kinetic studies with phosphorothioate-containing substrates and analyses by NMR spectroscopy, that a divalent metal ion interacts directly with the pro-Rp oxygen at the cleavage site in reactions catalyzed by hammerhead ribozymes. However, results of our recent kinetic studies (Zhou, D.-M.; Kumar, P. K. R.; Zhang. L. H.; Taira, K. J. Am. Chem. Soc. 1996, 118, 8969-8970. Yoshinari, K.; Taira, K. Nucleic Acids Res. 2000, 28, 1730-1742) demonstrated that a Cd(2+) ion does not interact with the sulfur atom at the Rp position of the scissile phosphate (P1.1) in the ground state or in the transition state. Therefore, in the present study, we attempted to determine by (31)P NMR spectroscopy whether a Cd(2+) ion binds to the P1.1 phosphorothioate at the cleavage site in solution. In the case of the R32-S11S (ribozyme-substrate) complex, neither the Rp- nor the Sp-phosphorothioate signal from the S11S substrate at the cleavage site was perturbed (the change was less than 0.1 ppm) upon the addition of Cd(2+) ions (19 equiv) at pH 5.9 and 8.5. By contrast, we detected the significant perturbation of the P9 phosphorothioate signal from another known metal-binding site (the A9/G10.1 metal-binding motif). The Rp-phosphorothioate signal from A9/G10.1 was shifted by about 10 ppm in the higher field direction upon the addition of Cd(2+) ions. These observations support the results of our kinetic analysis and indicate that a Cd(2+) ion interacts with the sulfur atom of the phosphorothioate at the A9/G10.1 site (P9) but that a Cd(2+) ion does not interact with the sulfur atom at the Rp- or at the Sp-position of the scissile phosphate (P1.1) in the ground state.     

Self-assembly of cyclic hexamers of γ-cyclodextrin in a metallosupramolecular framework with d-penicillamine

Cyclodextrins are widely used cyclic oligosaccharides of d-glucose whose hydrophilic exterior is covered by hydroxyl groups and whose hydrophobic interior is surrounded by lipophilic moieties. Because of this structural feature, cyclodextrin molecules commonly aggregate into dimensional structures via intermolecular hydrogen bonds, and their aggregation into closed oligomeric architectures has been achieved only via the attachment of functional substituent groups to the cyclodextrin rings. Here, we report the first structurally characterized self-assembly of non-substituted γ-cyclodextrin molecules into cyclic hexamers, which was realized in a chiral coordination framework composed of complex-anions with d-penicillamine rather than l- or dl-penicillamine. The self-assembly is accompanied by the 3D-to-2D structural transformation of porous coordination frameworks to form helical hexagonal cavities that accommodate helical γ-cyclodextrin hexamers. This finding provides new insight into the development of cyclodextrin chemistry and host–guest chemistry based on chiral recognition and crystal engineering processes.

The complex anions with d-penicillamine are organized into a 3D porous framework that allows the inclusion of γ-CD. The inclusion is accompanied by the 3D-to-2D transformation of porous frameworks so as to accept cyclic hexamers of γ-CD.     

EFFECTIVENESS OF PHOTOFRIN II IN ACIWATION OF MACROPHAGES AND in vitro KILLING OF RETINOBLASTOMA CELLS

Abstract Administration of a small dose (300 ng/mouse) of photofrin II (PII) to mice, followed by 4 days of exposure to only ambient fluorescent light in animal quarters, induced Fc-receptor-mediated phagocytic and superoxide-generating capacities of peritoneal macrophages by five- and seven-fold, respectively. When these mice were kept in the dark for 4 days, no activation of macrophages was observed. These results suggest that macrophage activation is a consequence of photodynamic activation. Much higher doses (> 3000 ng/mouse) suppressed macrophage activity. However, 2 months after administration of 3000 ng PII/mouse, greatly enhanced phagocytic and superoxide-generating capacities of peritoneal macrophages were observed.
In vitro photodynamic activation of macrophages was analyzed after white or red fluorescent light exposure of mouse peritoneal cells (mixture of macrophages and B and T lymphocytes) in media containing PII. A short (10 s) white fluorescent light treatment of peritoneal cells in a medium containing 0.03 ng PII/mL produced the maximal level of phagocytic activity of macrophages. Illumination with the same total fluence of red fluorescent light requires a threefold higher concentration of PII to achieve the same extent of enhanced phagocytic activity of macrophages. Thus, photodynamic activation of macrophages with PII by white fluorescent light was more efficient than by red fluorescent light. Similarly, photodynamic killing of retinoblastoma cells was more efficient with white than red fluorescent light. The concentration of hematoporphyrin (HP) or PII required for direct photodynamic killing of retinoblastoma cells was roughly four orders of magnitude greater than that required for activation of macrophages. These results suggest that effective photodynamic therapy may be achieved with milder treatments that stimulate macrophage activity, an important component of immunopotentiation.     

Unexpected highly chemoselective deprotection of the acetals from aldehydes and not ketones: TESOTf-2,6-lutidine combination

Acetal functions are recognized as good protecting groups of carbonyl groups. Although many deprotecting methods of acetals to carbonyl functions have already been developed, there is no methodology which can deprotect acetals in the presence of ketals because the usual acidic or radical reactions occur more easily via the more stable cationic or radical intermediates from the ketals. On the other hand, this new method can proceed in a reverse manner to that described in previous reports. That is, the method can deprotect aliphatic acetals in the presence of ketals. The reaction condition is common for silylation, i.e., the TESOTf-2,6-lutidine combinations. Although the TMSOTf-2,6-lutidine combination can also deprotect acetals, it lacks chemoselectivity in deprotection of the acetals from aldehydes and ketones. The treatment of acetals with TESOTf and 2,6-lutidine in CH2Cl2 followed by a H2O workup gave the corresponding aldehydes. Of course, the compounds, which have both acetal and hydroxyl functions afforded the compounds obtained by the usual silylation of an alcohol and deprotection of an acetal without any problem. However, deprotection of the ketals from ketones was not observed during the conversion reaction of acetals from aldehydes. This chemoselectivity was confirmed in the reactions of the compounds that have the acetal and ketal in the same molecule. In both cases, the acetal functions were deprotected to give aldehydes with intact ketals. Furthermore, under the conditions described here, many functional groups such as methoxy, acetoxy, allyl alcohol, and silyloxy ether are intact. This method is very mild and available for many compounds.     

TUMORICIDAL CAPACITIES OF MACROPHAGES PHOTODYNAMICALLY ACTIVATED WITH HEMATOPORPHYRIN DERIVATIVE

Four days after administration to mice of small amounts (30-600 ng/mouse) of hematoporphyrin derivative (HPD), peritoneal macrophages exhibited a greatly enhanced Fc-receptor mediated phagocytic capacity as assayed by ingestion activity of IgG-coated sheep erythrocytes. Much higher doses (greater than 3000 ng/mouse) did not have this effect. The peritoneal macrophages activated by administration of HPD have tumoricidal capacity for IgG-coated retinoblastoma cells. We then studied in vitro photodynamic activation of macrophages by white and red fluorescent light irradiation of mouse peritoneal cells (mixture of macrophages and B and T lymphocytes) in media containing very low concentrations of HPD. A short (5 s) white fluorescent light exposure (1Wm-2) of peritoneal cells in a medium containing 0.03 ng HPD/mL produced the maximal level of ingestion activity of macrophages. A 15 s red fluorescent light exposure (1Wm-2) of peritoneal cells in a medium containing 0.1 ng HPD/mL produced the maximal level of ingestion activity of macrophages. Thus, photodynamic activation of macrophages with white fluorescent light is more efficient than that with red fluorescent light. This can be explained by the fact that HPD has a large absorption peak at about 364 nm which extends into the visible range, and decreasingly smaller absorption bands at 500, 535, 570 and 630 nm. In vitro photodynamically activated macrophages showed efficient tumoricidal activity regardless of the type (white or red) of light used. These results suggest that a low level of HPD promotes therapeutic immunopotentiation.     

Versatile Oxidation Methods for Organic and Inorganic Substrates Catalyzed by Platinum‐Group Metals on Carbons

Platinum‐group metals on activated carbon catalysts, represented by Pd/C, Ru/C, Rh/C, etc., are widely utilized to accomplish green and sustainable organic reactions due to their favorable features, such as easy handling, recoverability, and reusability. The efficient oxidation methods of various organic compounds using heterogeneous platinum‐group metals on carbons with or without added oxidants are summarized in this Personal Account. The oxidation of internal alkynes into diketones was effectively catalyzed by Pd/C in the presence of dimethyl sulfoxide and molecular oxygen or pyridine N‐oxide. The Pd/C‐catalyzed mild combustion of gaseous hydrogen with molecular oxygen provided hydrogen peroxide, which could be directly utilized for the oxidation of sulfide derivatives into sulfoxides. Furthermore, the Ru/C‐catalyzed aerobic oxidation of primary and secondary alcohols gave the corresponding aldehydes and ketones, respectively. On the other hand, the dehydrogenative oxidation of secondary alcohols into ketones was achieved using Rh/C in water, and primary alcohols were effectively dehydrogenated by Pd/C in water under mildly reduced pressure to produce carboxylic acids.

     

An Extremely Porous Hydrogen‐Bonded Framework Composed of d‐Penicillaminato CoIII2AuI3 Complex Anions and Aqua Cobalt(II) Cations: Formation and Stepwise Structural Transformation

A unique example of a hydrogen‐bonded ionic solid with a porosity of 80 %, [Co(H₂O)₆]₃[Co₂Au₃(d ‐pen‐N,S)₆]₂ ( 1 ; d ‐H₂pen=d ‐penicillamine), composed of [Co(H₂O)₆]²⁺ cations and [Co₂Au₃(d ‐pen‐N,S)₆]^3? anions, is reported. Solid 1 was kinetically produced and was then transformed stepwise into two more thermodynamically stable solids with lower porosities, [Co(H₂O)₄][Co(H₂O)₆]₂[Co₂Au₃(d ‐pen‐N,S)₆]₂ ( 2 ) and [Co(H₂O)₄]₃[Co₂Au₃(d ‐pen‐N,S)₆]₂ ( 3 ), through the coordination of the free carboxylate groups in [Co₂Au₃(d ‐pen‐N,S)₆]^3? to Co^II centers. Solids 1 – 3 were structurally characterized, and the selective adsorption of small molecules into their pores was investigated.     

Structure elucidation of 21,22-dihydroxyonnamides A1-A4 from the marine sponge Theonella swinhoei: an empirical rule to assign the relative stereochemistry of linear 1,5-diols

A polar cytotoxic fraction of the marine sponge Theonella swinhoei was analyzed to be a mixture of four isomeric compounds, 21,22-dihydroxyonnamides A₁, A₂, A₃, and A₄. They were separated after conversion to the isopropyridene derivatives. The structures of the 21S,22S- and 21R,22R-isomers were determined by comparison of their spectral data with those prepared from onnamide A by asymmetric dihydroxylation. During the analysis of NMR data of these derivatives, an empirical rule was implied to assign the relative stereochemistry of linear 1,5-diol. This rule was applied to assign the stereochemistry of the remaining congeners.     

Synthesis of the indole core structures of conophylline

Conophylline is a bisindole alkaloid of unique structure that shows anti‐cancer and anti‐diabetic activities. Two indole core structures of conophylline, namely 6,7‐dimethoxy‐5‐methoxymethoxy‐1H‐indole ( 1 ), and N‐benzoyl‐N‐[2‐(6‐methoxymethoxy‐2‐vinyl‐1H‐indol‐3‐yl)ethyl]amine ( 2 ) has been synthesized starting with substituted benzene derivatives.