Takatoshi Kawai

The rationale for E2020 as a potent acetylcholinesterase inhibitor.

The phase III drug-candidate, E2020, developed for treatment of Alzheimers disease, and possibly other demenitas, and its analogues have been the focus of extensive molecular pharmacological and structural studies. The potency and selectivity of E2020 as an inhibitor of acetylcholinesterase, AChE, in the brain is established. A combination of molecular modeling and QSAR studies have been used throughout the evolution of the AChE inhibitor program leading to the benzylpiperidine series, and, ultimately, E2020. QSAR studies have identified requirements of optimize inhibition activity as a function of substituent choice on both the indanone and benzyl rings in the E2020 class of inhibitors. A combination of X-ray crystal structure studies of E2020 isomers and the molecular shape analysis, MSA, of E2020 and its analogues has led to a postulated active conformation, and molecular shape, for these AChE inhibitors. The active molecular shape corresponds to a high degree of shape similarity between the two E2020 isomers which, in turn, is consistent with the observed high inhibition potencies of both of these compounds. Intermolecular docking studies were carried out for E2020 and some analogues with the crystal structure of AChE when it became available. The docking simulations involving E2020 analogues suggest these inhibitors do not bind at the acetylcholine, ACh, active site, but rather at the most narrow location of the long channel leading to the active site. Intermolecular binding geometries are consistent with the postulated active conformations derived from structure-activity (receptor geometry independent) information.

Reduced neuropeptide Y mRNA levels in the frontal cortex of people with schizophrenia and bipolar disorder.

To study the change of gene expression in the brain tissues of schizophrenia, we used the gene expression monitoring technology and compared two sets of pools each containing four RNA samples of frontal cortex that were randomly selected from the control or schizophrenia group. We found that the expression of two genes were commonly altered in four pairwise comparisons; the expression of DEAD-box protein p72 (p72) gene was increased and neuropeptide Y (NPY) gene expression was decreased in the schizophrenia group compared with the control group. To substantiate these results, we estimated their mRNA levels by the real time TaqMan method in the 15 samples of each frontal or temporal cortex of four matched groups of schizophrenia, bipolar disorder, major depression and normal controls. A statistically significant decrease was observed for NPY in the frontal, but not in the temporal cortex, in the schizophrenia group (P=0.003). A decrease was also observed in the frontal cortex of the bipolar disorder group (P=0.031). In contrast, p72 gene expression showed no significant difference among the four groups. In conclusion, by novel technology of DNA array and TaqMan PCR analyses, we found that neuropeptide Y mRNA levels were significantly reduced in the frontal cortex in both schizophrenia and bipolar disorder.

Chronic Intracerebroventricular Administration of Relaxin-3 Increases Body Weight in Rats

Bolus-administered intracerebroventricular (ICV) relaxin-3 has been reported to increase feeding. In this study, to examine the role of relaxin-3 signaling in energy homeostasis, we studied the effects of chronically administered ICV relaxin-3 on body weight gain and locomotor activity in rats. Two groups of animals received vehicle or relaxin-3 at 600 pmol/head/day, delivered with Alzet osmotic minipumps. In animals receiving relaxin-3, food consumption and weight gain were statistically significantly higher than those in the vehicle group during the 14-day infusion. During the light phase on days 2 and 7 and the dark phase on days 3 and 8, there was no difference in locomotor activity between the two groups. Plasma concentrations of leptin and insulin in rats chronically injected with relaxin-3 were significantly higher than in the vehicle-injected controls. These results indicate that relaxin-3 up-regulates food intake, leading to an increase of body weight and that relaxin-3 antagonists might be candidate antiobesity agents.

Characterization of mRNA species that are associated with postsynaptic density fraction by gene chip microarray analysis

We previously reported the partial identification by random sequencing of mRNA species that are associated with the postsynaptic density (PSD) fraction prepared from the rat forebrain [Tian et al., 1999. Mol. Brain Res. 72, 147-157]. We report here further characterization by gene chip analysis of the PSD fraction-associated mRNAs, which were prepared in the presence of RNase inhibitor. We found that mRNAs encoding various postsynaptic proteins, such as channels, receptors for neurotransmitters and neuromodulators, proteins involved in signaling, scaffold and adaptor proteins and cytoskeletal proteins, were highly concentrated in the PSD fraction, whereas those encoding housekeeping proteins, such as enzymes in the glycolytic pathway, were not. We extracted approximately 1900 mRNA species that were highly concentrated in the PSD fraction. mRNAs related to certain neuronal diseases were also enriched in the PSD fraction. We also constructed a cDNA library using the PSD fraction-associated mRNAs as templates, and identified 1152 randomly selected clones by sequencing. Our data suggested that the PSD fraction-associated mRNAs are a very useful resource, in which a number of as yet uncharacterized mRNAs are concentrated. Identification and functional characterization of them are essential for complete understanding of synaptic function.

Synthetic studies on (−)-tetrodotoxin (3) nitrogenation through overman rearrangement and guanidine ring formation

Abstract The guanidinium part of tetrodotoxin was synthesized from a sugar derivative, levoglucosenone, in optically active form via Overman rearrangement as key step.

Formation of 1,2-dioxolane in the singlet oxygenation of a silicon-silicon σ-bond: peroxonium ion intermediate

Abstract Singlet oxygenation of 1,1,2,2-tetramesityl-1,2-disilirane 1a afforded the corresponding cyclic peroxide 3a . Trapping experiments and theoretical studies of the initially formed peroxidic intermediate were also carried out. Peroxonium ion type intermediate is required to rationalize the results.

SYNTHESIS AND BIOLOGICAL EVALUATION OF BOTH ENANTIOMERS OF DYNEMICIN A MODEL COMPOUND

Abstract A novel 1,4-asymmetric induction was developed for the synthesis of chiral dynemicin A model compound. By using this reaction, both enantiomers were synthesized from chiral alcohol prepared by lipase catalyzed resolution. The biological activities of these compounds were examined.

1,3-Dipolar cycloaddition of di-1-menthyl benzylidenemalonate to (z)n,α-diphenylnitrone: Explanation for diastereoselectivity

Abstract 1,3-Dipolar cycloaddition of di-1-menthyl benzylidenemalonate to (Z)-N,α-diphenylnitrone proceeds by Si-face preference to give the endo-adduct as the major product among all four possible adducts. An explanation for this diastereofacial selectivity (Si-face preference) is proposed based on X-ray crystallographic analysis of the malonate as well as effects of high-pressure to the ratio of the adducts.

Discovery of a potent, metabolically stabilized resorcylic lactone as an anti-inflammatory lead

With bioactivity-guided phenotype screenings, a potent anti-inflammatory compound f152A1 has been isolated, characterized and identified as the known natural product LL-Z1640-2. Metabolic instability precluded its use for the study on animal disease models. Via total synthesis, a potent, metabolically stabilized analog ER-803064 has been created; addition of the (S)-Me group at C4 onto f152A1 has resulted in a dramatic improvement on its metabolic stability, while preserving the anti-inflammatory activities.

Picrotoxane terpenoids from Picrodendron baccatum

Abstract Three new picrotoxane type terpenoids, picrodendrins E, F and I, were isolated from the bark of Picrodendron baccatum . The structures were determined by spectroscopic evidence and X-ray crystallographic analysis.