A practical procedure for the synthesis of esonarimod, (R,S)-2-acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid,an antirheumatic agent (part 1)

An efficient and practical procedure for the synthesis of esonarimod, (R,S)-2-acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (1), a new antirheumatic drug, has been developed. The intermediate, 2-methylene-4-(4-methylphenyl)-4-oxobutanoic acid (2), was prepared by Friedel-Crafts acylation of toluene with itaconic anhydride (3) in the presence of aluminum trichloride and nitrobenzene in 63% yield without silica gel column purification. Compound 1 was prepared by Michael addition of 2 with thioacetic acid (4) in 74% yield. Overall, 1 was obtained in 47% yield from 3. The structures and synthetic mechanisms of by-products (five compounds) of 2 were also clarified.     

Determination of domoic acid in Japanese mussels by enzyme immunoassay

Ten samples of commercial blue mussels (Mytilus edulis) from Japan were analyzed for domoic acid by an indirect competitive enzyme immunoassay (idc-EIA) based on an anti-domoic acid monoclonal antibody. Domoic acid was found in all samples at low concentrations (0.11-1.81 ng/g mussel tissue). The presence of domoic acid was confirmed by liquid chromatography coupled with immunoaffinity chromatography using an anti-domoic acid monoclonal antibody as ligand. To our knowledge, this is the first reported detection of domoic acid, a causative agent of amnesic shellfish poisoning, in Japanese mussels.     

Efficient and widely applicable method of constructing neo-proteoglycan utilizing copper(I) catalyzed 1,3-dipolar cycloaddition

Efficient constructions of two types of neo-proteoglycan are described. Enzymatically prepared alkyne containing chondroitin 6-sulfate chains and chemically synthesized azido group having compounds are linked by utilizing the copper(I) catalyzed 1,3-dipolar cycloaddition.     

Thio-sugars. IV. Synthesis of carbohydrate 1,3-glycol (six-membered) thionocarbonates and their attempted O-S rearrangement

Four six-membered cyclic thionocarbonates (two sec-sec and two prim-sec) were prepared and their radical-promoted O-S rearrangement reaction was examined. The results revealed that the reaction in six-membered rings is difficult compared with the simple rearrangement in five-membered rings. In the course of syntheses of the substrates, interesting acyl migration during the stannylation process occurred.     

Product analyses and crosslink mechanism of plasma-polymerized methyl methacrylate

Plasma-polymerized methyl methacrylate (PPMMA) film was produced by a radio-frequency glow discharge of methyl methacrylate (MMA) gas with argon. PPMMA deposited under a plasma of mild energy was separated into a soluble fraction (sol-PPMMA) and an insoluble matrix (insol-PPMMA), which is highly crosslinked, by solvent-extraction and filtration. The chemical structure of the sol-PPMMA was determined by pyrolysis/gas chromatography, IR spectroscopy, ¹H-NMR, ¹³C-NMR, and elemental analysis techniques. On the other hand, the structure of the insol-PPMMA was examined with IR spectroscopy, ¹³C-NMR with CP/MAS method, and elemental analysis. The crosslinking mechanism in the plasma polymerization of MMA is discussed on the basis of product analyses of the sol-PPMMA and the insol-PPMMA.     

Catalytic flow-injection determination of copper at nanogram levels by using color formation of N-phenyl-p-phenylenediamine with m-phenylenediamine in the presence of pyridine and ammonia as activators

A spectrophotometric flow-injection method for determining copper(II) has been developed. It is based on the catalytic effect of copper(II) on the oxidative coupling of N-phenyl-p-phenylenediamine with m-phenylenediamine in the presence of hydrogen peroxide. Pyridine and ammonia as activators increased the absorbance for the copper(II)-catalyzed coloration, and the dye formed was stabilized by adding a non-ionic surfactant. The working range of the method was 0.1-2.0 ng ml(-1) of cooper(II) with a relative standard deviation 2.4% at a sampling rate of 30 h(-1). Interference from iron(III) was effectively suppressed by citric acid. Copper in natural water samples can be determined easily.     

The flameless atomic absorption spectrometric determination of aluminum with a carbon atomization system

The atomic absorption spectrometry of aluminum with a carbon tube atomizer is described, with particular reference to the heating cycle, argon flow rate and amplifier time constant. Interferences are eliminated by pre-atomization heating or with a hydrogen-argon-entrained air flame. Under optimal conditions, the sensitivity (1 % absorbance) is 5·10^?11g, and the relative standard deviation is 2.3 for 2.5 ng Al.     

Functionalization of chitosan with 3-nitro-4-amino benzoic acid moiety and its application to the collection/concentration of molybdenum in environmental water samples

A chitosan resin functionalized with 3-nitro-4-amino benzoic acid moiety (CCTS-NABA resin) was newly synthesized for the collection/concentration of trace molybdenum by using cross-linked chitosan (CCTS) as base material. The carboxyl group of the moiety was chemically attached to amino group of cross-linked chitosan through amide bond formation. The adsorption behavior of molybdenum as well as other 60 elements on the resin was examined by passing the sample solutions through a mini-column packed with the resin. After the elution of the elements collected on the resin with 1 M HNO₃, the eluates were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) and atomic emission spectrometry (ICP-AES).

The CCTS-NABA resin can adsorb several metal ions, such as vanadium, gallium, arsenic, selenium, silver, bismuth, thorium, tungsten, tin, tellurium, copper, and molybdenum at appropriate pHs. Among these metal ions, only molybdenum could be adsorbed almost completely on the resin at acidic regions. An excellent selectivity toward molybdenum could be obtained at pH 3–4. The adsorption capacity of CCTS-NABA resin for Mo(VI) was 380 mg g⁻¹ resin. Through the column pretreatment, alkali and alkaline earth metals in river water and seawater samples were successfully removed.

The CCTS-NABA resin was applied to the adsorption/collection of molybdenum in river water and seawater samples. The concentrations of molybdenum in river water samples were found in the range of 0.84 and 0.95 ppb (ng g⁻¹), whereas molybdenum in seawater was about 9 ppb. The validation of the proposed method was carried out by determining molybdenum in the certified reference materials of SLRS-4, CASS-4, and NASS-5 after passing through the CCTS-NABA resin; the results showed good agreement with the certified values.     

Neutron activation analysis of mercury contents in head hair of dentists in Japan

Mercury contents in head hair of 58 dentists employed at the NUSD hospital and 50 dentists employed at the private hospitals or clinics were determined using neutron activation analysis. The arithmetic means were 5.8 ppm and 5.2 ppm, and geometric means were 5.4 ppm and 4.8 ppm, respectively. They were much lower than the values reported in the past year, and agreed well with those of normal Japanese men of the same age. Therefore, it was concluded that the mercury pollution in the working environment of dentists might be practically non-existent in Japan today.     

Engineered biosynthesis of plant polyketides: chain length control in an octaketide-producing plant type III polyketide synthase

The chalcone synthase (CHS) superfamily of type III polyketide synthases (PKSs) produces a variety of plant secondary metabolites with remarkable structural diversity and biological activities (e.g., chalcones, stilbenes, benzophenones, acrydones, phloroglucinols, resorcinols, pyrones, and chromones). Here we describe an octaketide-producing novel plant-specific type III PKS from aloe (Aloe arborescens) sharing 50-60% amino acid sequence identity with other plant CHS-superfamily enzymes. A recombinant enzyme expressed in Escherichia coli catalyzed seven successive decarboxylative condensations of malonyl-CoA to yield aromatic octaketides SEK4 and SEK4b, the longest polyketides known to be synthesized by the structurally simple type III PKS. Surprisingly, site-directed mutagenesis revealed that a single residue Gly207 (corresponding to the CHS's active site Thr197) determines the polyketide chain length and product specificity. Small-to-large substitutions (G207A, G207T, G207M, G207L, G207F, and G207W) resulted in loss of the octaketide-forming activity and concomitant formation of shorter chain length polyketides (from triketide to heptaketide) including a pentaketide chromone, 2,7-dihydroxy-5-methylchromone, and a hexaketide pyrone, 6-(2,4-dihydroxy-6-methylphenyl)-4-hydroxy-2-pyrone, depending on the size of the side chain. Notably, the functional diversity of the type III PKS was shown to evolve from simple steric modulation of the chemically inert single residue lining the active-site cavity accompanied by conservation of the Cys-His-Asn catalytic triad. This provided novel strategies for the engineered biosynthesis of pharmaceutically important plant polyketides.