Discovery of Novel Chromenopyridine Derivatives as Readthrough-Inducing Drugs.

Hurler syndrome, a type of Mucopolysaccharidosis type I, is an inherited disorder caused by the accumulation of glycosaminoglycans (GAG) due to a deficiency in lysosomal α-L-iduronidase (IDUA), resulting in multiorgan dysfunction. In many patients with Hurler syndrome, IDUA proteins are not produced due to nonsense mutations in their genes; therefore, readthrough-inducing compounds, such as gentamycin, are expected to restore IDUA proteins by skipping the premature termination codon. In the present study, we synthesized a series of chromenopyridine derivatives to identify novel readthrough-inducing compounds. The readthrough-inducing activities of synthesized compounds were examined by measuring cellular IDUA activities and GAG concentrations in Hurler syndrome patient-derived cells. Compounds with a difluorophenyl group at the 2-position of chromenopyridine, a cyclobutyl group at the 3-position, and a basic side chain or basic fused ring exhibited excellent readthrough-inducing activities. KY-640, a chromenopyridine derivative with a tetrahydroisoquinoline sub-structure, increased the cellular IDUA activities of patient-derived cells by 3.2-fold at 0.3 µM and significantly reduced GAG concentrations, and also significantly increased enzyme activity in mouse models, suggesting its therapeutic potential in patients with Hurler syndrome.

Synthesis and Evaluation of Novel Triaryl Derivatives with Readthrough-Inducing Activity.

The readthrough mechanism, which skips the premature termination codon and restores the biosynthesis of the defective enzyme, is an emerging therapeutic tactic for nonsense mutation-related diseases, such as Hurler syndrome, a type of mucopolysaccharidosis. In the present study, novel triaryl derivatives were synthesized and their readthrough-inducing activities were evaluated by a luciferase reporter assay with a partial α-L-iduronidase (IDUA) DNA sequence containing the Q70X nonsense mutation found in Hurler syndrome and by measuring the enzyme activity of IDUA knockout cells transfected with the mutant IDUA gene. KY-516, a representative compound in which the meta position carboxyl group of the left ring of the clinically used ataluren was converted to the para position sulfamoylamino group, the central ring to triazole, and the right ring to cyanobenzene, exhibited the most potent readthrough-inducing activity in the Q70X/luciferase reporter assay. In Q70X mutant IDUA transgenic cells, KY-516 significantly increased enzyme activity at 0.1 µM. After the oral administration of KY-516 (10 mg/kg), the highest plasma concentration of KY-516 was above 5 µM in rats. These results indicate that KY-516, a novel triaryl derivative, exhibits potent readthrough-inducing activity and has potential as a therapeutic agent for Hurler syndrome.

Prevalence of Nosema species infections in Apis cerana japonica and Apis mellifera honeybees in the Tohoku region of Japan.

We investigated here, the prevalence of Nosema microsporidia infections in the honeybees, Apis cerana japonica and Apis mellifera, in the Tohoku region of Japan. We detected Nosema ceranae DNA in 14 (2.8%) of 509 A. cerana japonica and in 34 (21.9%) of 155 A. mellifera honeybees from Aomori, Iwate, Akita, Yamagata, and Fukushima prefectures. Nosema apis DNA was undetectable in A. cerana japonica and A. mellifera. The unidentifiable Nosema species that genetically differed from N. apis, N. ceranae, and N. neumanni in terms of small subunit (SSU) rDNA, large subunit rDNA, and internal transcribed spacer sequences was identified in 105 (20.6%) of 509 A. cerana japonica and in 1 (0.6%) of 155 A. mellifera honeybees, and from Iwate prefecture. A phylogenetic tree based on SSU rDNA sequences showed that the Nosema sp. belonged to the same clade as N. thomsoni detected in moth and solitary bees in North America and N. pieriae found in cabbage butterfly in Turkey, which have not hitherto been detected in honeybees. The morphological characteristics of the spores should be analyzed to enable species identification of the Nosema sp.

The prevalence and molecular characterization of Acarapis woodi and Varroa destructor mites in honeybees in the Tohoku region of Japan.

Honeybee acarapiosis and vorrosis were designated as Notifiable Infectious Diseases in the Act on Domestic Animal Infectious Diseases Control by the Minister of Agriculture, Forestry and Fisheries of Japan in 1997. However, the prevalences of A. woodi and V. destructor in Japan, especially in the Tohoku region, have not been sufficiently elucidated. This study was designed to clarify the prevalence of A. woodi and V. destructor mites in Apis cerana japonica and Apis mellifera in the Tohoku region and the characteristics of their mitochondrial cytochrome c oxidase I (COI) DNA. Acarapis woodi mites were detected from 13.5% of A. c. japonica and 0% of A. mellifera. Aomori prefecture, Japan is a new distribution locality for A. woodi. None of the honeybees examined showed infection by V. destructor mites. The COI sequences (1638 bp) of A. woodi were identical and phylogenetically closely related to those of A. woodi from Japan and the UK, suggesting that the mite would have been introduced into Japan with A. mellifera during the last 150 years and spread throughout the country.

Copper(II)/phenanthroline-mediated CD-enhancement and chiral memory effect on a meta-ethynylpyridine oligomer.

meta-Ethynylpyridine 18-mer associates with octyl ß-D-glucopyranoside to show an induced circular dichroism (CD) band at around 337 nm in CH(2)Cl(2). The addition of copper(II) triflate and o-phenanthroline enhanced the CD band. Even when an equimolar amount of ß-L-glucopyranoside was added to make the system racemic, the CD was memorized and remained for several weeks.

Copper(II)-mediated chiral helicity amplification and inversion of meta-ethynylpyridine polymers with metal coordination sites.

meta-Ethynylpyridine polymers bearing metal coordination sites associate with alkyl glycoside guests to show induced circular dichroism (ICD) bands. The addition of a Cu(II) ion changed the intensity or the sign of these ICD bands. The changes suggested chiral helicity amplification or inversion of the polymers by Cu(II)-mediated cross-linking at the coordinating side chains.

Azacrown-attached meta-ethynylpyridine polymer: saccharide recognition regulated by supramolecular device.

Polymeric synthetic host , azacrown-attached 2,6-pyridylene ethynylene polymer, was investigated for its saccharide recognition and the additive effect of triethylene tetramine-trifluoroacetic acid; heteroallosteric effects were observed on the basis of CD and UV/Vis analyses, which indicated saccharide-dependent stabilization and destabilization of helical complexes by the formation of pseudopolyrotaxanes.