Showing papers by "Taisho Pharmaceutical Co. published in 2004"
TL;DR: The H-InvDB as discussed by the authors is a database of 41,118 full-length cDNAs that capture the gene transcripts as complete functional cassettes, providing an unequivocal report of structural and functional diversity at the gene level.
Abstract: The human genome sequence defines our inherent biological potential; the realization of the biology encoded therein requires knowledge of the function of each gene. Currently, our knowledge in this area is still limited. Several lines of investigation have been used to elucidate the structure and function of the genes in the human genome. Even so, gene prediction remains a difficult task, as the varieties of transcripts of a gene may vary to a great extent. We thus performed an exhaustive integrative characterization of 41,118 full-length cDNAs that capture the gene transcripts as complete functional cassettes, providing an unequivocal report of structural and functional diversity at the gene level. Our international collaboration has validated 21,037 human gene candidates by analysis of high-quality full-length cDNA clones through curation using unified criteria. This led to the identification of 5,155 new gene candidates. It also manifested the most reliable way to control the quality of the cDNA clones. We have developed a human gene database, called the H-Invitational Database (H-InvDB; http://www.h-invitational.jp/). It provides the following: integrative annotation of human genes, description of gene structures, details of novel alternative splicing isoforms, non-protein-coding RNAs, functional domains, subcellular localizations, metabolic pathways, predictions of protein three-dimensional structure, mapping of known single nucleotide polymorphisms (SNPs), identification of polymorphic microsatellite repeats within human genes, and comparative results with mouse full-length cDNAs. The H-InvDB analysis has shown that up to 4% of the human genome sequence (National Center for Biotechnology Information build 34 assembly) may contain misassembled or missing regions. We found that 6.5% of the human gene candidates (1,377 loci) did not have a good protein-coding open reading frame, of which 296 loci are strong candidates for non-protein-coding RNA genes. In addition, among 72,027 uniquely mapped SNPs and insertions/deletions localized within human genes, 13,215 nonsynonymous SNPs, 315 nonsense SNPs, and 452 indels occurred in coding regions. Together with 25 polymorphic microsatellite repeats present in coding regions, they may alter protein structure, causing phenotypic effects or resulting in disease. The H-InvDB platform represents a substantial contribution to resources needed for the exploration of human biology and pathology.
365 citations
TL;DR: It is suggested that perinatal brain masculinization requires AR function and that expression of male-typical behaviors in adults is mediated by both AR-dependent and -independent androgen signaling.
Abstract: Testicular testosterone produced during a critical perinatal period is thought to masculinize and defeminize the male brain from the inherent feminization program and induce male-typical behaviors in the adult. These actions of testosterone appear to be exerted not through its androgenic activity, but rather through its conversion by brain aromatase into estrogen, with the consequent activation of estrogen receptor (ER)-mediated signaling. Thus, the role of androgen receptor (AR) in perinatal brain masculinization underlying the expression of male-typical behaviors remains unclear because of the conversion of testosterone into estrogen in the brain. Here, we report a null AR mutation in mice generated by the Cre-loxP system. The AR-null mutation in males (ARL-/Y) resulted in the ablation of male-typical sexual and aggressive behaviors, whereas female AR-null homozygote (ARL-/L-) mice exhibited normal female sexual behaviors. Treatment with nonaromatizable androgen (5α-dihydrotestosterone, DHT) was ineffective in restoring the impaired male sexual behaviors, but it partially rescued impaired male aggressive behaviors in ARL-/Y mice. Impaired male-typical behaviors in ERα-/- mice were restored on DHT treatment. The role of AR function in brain masculinization at a limited perinatal stage was studied in ARL-/L- mice. Perinatal DHT treatment of females led to adult females sensitive to both 17β-estradiol and DHT in the induction of male-typical behaviors. However, this female brain masculinization was abolished by AR inactivation. Our results suggested that perinatal brain masculinization requires AR function and that expression of male-typical behaviors in adults is mediated by both AR-dependent and -independent androgen signaling.
306 citations
TL;DR: The aryloxyanilide derivatives, which have been derived by opening the diazepine ring of 1, are a novel class as PBR ligands and have exhibited high and selective affinity for peripheral benzodiazepine receptors (PBRs).
163 citations
TL;DR: Results indicate that R278995/CRA0450 is an orally active CRF (1) and sigma(1) receptor antagonist with potent anxiolytic-like and antidepressant-like activities.
142 citations
TL;DR: Findings suggest that NCI formation is accelerated by the progression of the disease process, and that in MSA, NNI formation is an earlier phenomenon thanNCI formation.
Abstract: Multiple system atrophy (MSA) is a sporadic neurodegenerative disease characterized by the presence of neuronal and oligodendroglial alpha-synuclein aggregates. To investigate the relationship between the occurrence of neuronal cytoplasmic and intranuclear inclusions (NCIs and NNIs, respectively) and the progression of neuronal degeneration, we performed a quantitative analysis of the pontine and inferior olivary nuclei based on 14 cases of MSA. alpha-Synuclein immunohistochemistry revealed that NCIs and NNIs were present in both brain nuclei in all the cases. The average incidence of NCIs in the pontine and inferior olivary nuclei was 9.1% and 25.8%, respectively, and that of NNIs was 9.2% and 9.0%, respectively. The number of NNIs was strongly correlated with that of neurones in the pontine and inferior olivary nuclei. Although the number of NCIs was not correlated with the neuronal population in both nuclei, the NCI count in patients with moderate MSA was higher than in patients with mild MSA. The NNI count was much higher than the NCI count in the pontine nucleus in four patients, and was the same in the olivary nucleus in three of the four patients. Moreover, the neuronal population in the NNI-predominant cases was significantly higher than in the NCI-predominant cases. These findings suggest that NCI formation is accelerated by the progression of the disease process, and that in MSA, NNI formation is an earlier phenomenon than NCI formation.
122 citations
TL;DR: The widespread accumulation of phosphorylated α-synuclein in both glial and neuronal cells is a pathological feature in patients suffering from MSA.
Abstract: We immunohistochemically examined the brain and peripheral sympathetic ganglia from eight patients with multiple system atrophy (MSA), using an antibody specific for phosphorylated alpha-synuclein (anti-PSer129). Phosphorylated alpha-synuclein was deposited in five cellular locations: oligodendroglial cytoplasm and nucleus, and neuronal cytoplasm, processes and nucleus. Many neuronal cytoplasmic inclusions (NCIs) were found in the pontine and inferior olivary nuclei and, to a lesser extent, in the substantia nigra, locus ceruleus, and neocortical and hippocampal neurons. NCIs were also found in the sympathetic ganglia in two out of the eight cases. Moreover, anti-PSer129 immunohistochemistry revealed extensive neuropil pathology; swollen neurites were abundant in the pontine nucleus, delicate neurites were observed in the deeper layers of the cerebral cortex and thalamus, and neuropil threads and dot-like structures were distributed in the basal ganglia and brainstem. Diffuse neuronal cytoplasmic staining (pre-NCI) was frequently found in the pontine and inferior olivary nuclei. Thus, the widespread accumulation of phosphorylated alpha-synuclein in both glial and neuronal cells is a pathological feature in patients suffering from MSA.
114 citations
TL;DR: Scratching up the skin with toenails seemed to be the most important factor leading to dermatitis in NC mice.
80 citations
TL;DR: Some derivatives of imidazole 1 which had an amino group on the side chain showed potent and selective inhibitory activity, and found that a dimethylaminohexyloxy derivative showed potency and selectivity for cytochrome P450s.
76 citations
TL;DR: It is shown that intranasal administration of eotaxin reconstituted pulmonary eosinophilia but not AHR and mucus hypersecretion in OVA-inhalated STAT6-/- mice was induced, suggesting that STAT6 also plays a critical role at late phase of Th2-dependent allergy induction.
Abstract: When wild-type BALB/c mice were transferred with OVA-specific Th2 cells followed by OVA inhalation, a severe eosinophilia, mucus hypersecretion and airway hyper-responsiveness (AHR) was induced in parallel with a marked elevation of IL-4, IL-5 and IL-13 levels in bronchoalveolar lavage fluid (BALF). However, neither eosinophilia, AHR nor mucus hypersecretion was induced in Th2 cell-transferred STAT6-/- mice. The failure of eosinophilia was not due to the defect of Th2 cytokine production in BALF of STAT6-/- mice transferred with Th2 cells, but because of the defect of STAT6-dependent eotaxin production. Indeed, intranasal administration of eotaxin reconstituted pulmonary eosinophilia but not AHR and mucus hypersecretion in OVA-inhalated STAT6-/- mice. These results initially provided direct evidence that STAT6-dependent eotaxin production is essential for pulmonary eosinophilia. We also dissociated the role of STAT6 for eosinophilia from that for AHR and mucus hypersecretion. Thus, STAT6 also plays a critical role at late phase of Th2-dependent allergy induction.
75 citations
TL;DR: It is suggested that blocking the vasoconstrictor actions of 20-HETE may be useful to prevent the acute fall in cerebral blood flow following subarachnoid hemorrhage and to increase cerebral vascular tone both in vivo and in vitro.
71 citations
TL;DR: Prostaglandin D2 plays a physiological role in inhibiting pruritus of NC/Nga mice via their specific prostanoid DP1 receptors, and that prostagland in D2 and/or a prostanoids DP1 receptor agonist may have therapeutic effects for cases of consecutive skin inflammation.
TL;DR: In this article, the authors examined some derivatives of imidazole 1 which had an amino group on the side chain, and found that a dimethylaminohexyloxy derivative (3g; IC(50) value of 8.8 nM) showed potent and selective inhibitory activity.
Abstract: In a previous paper, we reported that an imidazole derivative 1 exhibited a potent inhibitory activity of 20-HETE synthase (1; IC(50) value of 5.7 nM), but this compound also exhibited little selectivity for cytochrome P450s (CYPs). We examined some derivatives of imidazole 1 which had an amino group on the side chain, and found that a dimethylaminohexyloxy derivative (3g; IC(50) value of 8.8 nM) showed potent and selective inhibitory activity.
TL;DR: The results suggest that this method shows a good correlation with the effectiveness of drugs prescribed for itching in humans with atopic dermatitis, and histamine and serotonin do not play an important role in causing the scratching behavior seen by NC/Nga mice.
TL;DR: The 4-fluoro derivative was found to exhibit better DPP-IV inhibitory activity and higher plasma drug concentrations after oral administration to rats than the 4-unsubstituted derivative.
TL;DR: It was suggested that activation of each of the ERK1/2, p38, and JNK MAPK cascades in addition to that of PKC and cPLA2 played an important role in the Cd-stimulated biosynthesis of PGE2 in mouse osteoblastic cells.
TL;DR: The requirements of calcium, magnesium and phosphorus in Japanese young adults, ranging from 18 to 28 y old, took part in mineral balance studies after written informed consent was obtained, finding that there was not much of a correlation for Ca.
Abstract: This study was conducted to estimate the requirements of calcium (Ca), magnesium (Mg) and phosphorus (P) in Japanese young adults. From 1986 to 2000, 109 volunteers 123 males, 86 females), ranging from 18 to 28 y old, took part in mineral balance studies after written informed consent was obtained. The duration of the study periods ranged from 5 to 12 d, with 2-4 d of adaptation. Foodstuffs used in each study were selected from those commercially available. The minerals present in diet, feces, urine and sweat were measured by atomic absorption spectrophotometer (Ca, Mg) or spectrophotometer (P). The dietary intakes of Ca, Mg and P ranged from 4.83-23.58, 2.44-7.83 and 13.46-45.69 mg/ kg BW/d, respectively. Dietary intake (Intake) of Ca was positively correlated to apparent absorption (A.A.) (r2 = 0.425), which was also correlated with urine excretion (Urine) (r2 = 0.327) and balance (Bal) (r2 = 0.382). Intake of Ca was slightly but significantly correlated with Bal (r2 = 0.036, p = 0.048). The mean value and upper limit of the 95% confidence interval for the regression equation between Intake and Bal when balance is equal to zero (Mean and upper limit) for Ca were 11.752 and 12.555 mg/kg BW/d, respectively. Intake of Mg was positively correlated to A.A. (r2 = 0.451), which was also correlated with Urine (r2 = 0.486) and Bal (r2 = 0.349). However, Intake of Mg was not correlated with Bal. Intake of P was positively correlated with A.A. (r2 = (0.959), which was also correlated with both Urine (r2 = 0.908) and Bal (r2 = 0.135). Intake of P was slightly but significantly correlated with Bal (r2 = 0.103, p = 0.0013). Mean and upper limits for P were 22.584 and 24.059 mg/kg BW/d, respectively. Intakes of Mg and P correlated negatively with their respective A.A. rates (%) (r2 = 0.120 for Mg, r2 = 0.109 for P). However, there was not much of a correlation for Ca. Balance of Ca was positively correlated with that of Mg (r2 = 0.541), but not with that of P.
Journal Article•
TL;DR: The findings suggest that VGA1155 inhibits endothelial cell growth and angiogenesis by inhibiting VEGF function but not non-specific cytotoxicity, and exhibits promise as an antiangiogenic or anti-tumor agent with fewer side-effects.
Abstract: The process of angiogenesis involves the formation of new blood vessels from established vasculature and is essential for progressive tumor growth and metastasis. Since vascular endothelial growth factor (VEGF) plays a pivotal role in tumor angiogenesis, it is reasonable to expect that antagonizing VEGF binding to its receptor may be effective in cancer therapy. Our previous study found that a novel low molecular weight compound, VGA1155, inhibited binding between radioisotope-labelled VEGF and cells overexpressing its two receptors, Fit-1 and KDR/Flk-1, that is, NIH3T3-Flt-1 and NIH3T3-KDR, respectively. In the present study, we investigated the anti-angiogenic effects of VGA1155 based on VEGF inhibition. VGA1155 inhibited VEGF-induced DNA synthesis of human umbilical vein endothelial cells (HUVEC) and human retinal endothelial cells (HREC) in a concentration-dependent manner. VGA1155 also inhibited VEGF-induced tube formation of HUVEC in vitro and tumor angiogenesis toward B16-BL6 melanoma after orthotopic implantation into the skin of the back. On the other hand, VGA1155 did not affect the proliferation of human epidermoid carcinoma (KB) cells and mouse mammary carcinoma (MM2) cells. It also had no effect on the activity of several cytosolic kinases such as p55 fyn and p56 lck . These findings suggest that VGA1155 inhibits endothelial cell growth and angiogenesis by inhibiting VEGF function but not non-specific cytotoxicity. VGA1155 thus exhibits promise as an anti-angiogenic or anti-tumor agent with fewer side-effects.
TL;DR: Observations show that NNK predominantly induces DNA adducts leading to A:T to T:A and/or A-T to C:G mutations in the transgene.
Abstract: 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) found in chewing tobacco, snuff, cigarettes, and cigars is a tobacco-specific nitrosamine and classified as a possible human carcinogen (Class 2B) by the International Agency for Research on Cancer (IARC). NNK given intraperitoneally was seen to induce lung and liver adenomas. To evaluate the genotoxicity of NNK in vivo, NNK was intraperitoneally administered to Muta Mouse at two concentrations (125 and 250 mg/kg, once a week for 4 weeks) followed by the measurement of mutant frequencies in the lacZ and cII genes from lung and liver in the same mice. Characterization of the types of the mutation was determined by sequencing the cII genes from mutant plaques. The mutant frequencies in both target genes from both organs dose-dependently increased up to 10 times compared to those of the control group. For the types of mutations, the ratio of the G:C to A:T mutation in the total number of mutants was less than the ratio of A:T to T:A and A:T to C:G transversion, contrary to a previous report. The A:T to T:A transversion was the most highly induced mutation both in the lung and liver cII genes. The increasing rate of mutant frequencies in lung and liver over the vehicle control was 55 and 56 times, respectively, while the increasing rate of G:C to A:T transition was only 1.9 and 2.8 times, respectively. These observations show that NNK predominantly induces DNA adducts leading to A:T to T:A and/or A:T to C:G mutations in the transgene.
TL;DR: From the ease of the technique, automation of the microtitre plate assay and application of the single concentration method, the method might be useful for inhibitory assessment of cytochromes P450 more than that of current conventional methods.
Abstract: 1. The inhibitory effects of various test compounds on recombinant human CYP3A4 activity assayed by fluorescent metabolite formation from 7-benzyloxyquinoline (7-BQ) and the effect of pre-incubation on inhibition were evaluated using the microtitre plate assay with multiple concentrations of test compounds (multiple concentration method). 2. Among the test compounds studied, ketoconazole inhibited CYP3A4 activity most extensively, followed by miconazole, troleandomycin, terfenazine and midazolam. The IC(50) values of other compounds exceeded 10 microM, but those of many compounds decreased after pre-incubation. The inhibitory effects of verapamil, amiodarone and diltiazem after pre-incubation were 205, 154 and 833 times greater than those in the case of co-incubation, respectively. 3. To assess the inhibitory effects more readily, the validity of the microtitre plate assay with a single concentration of the test compound (single concentration method) was studied. The accuracy of the automated dispensation and the coefficient of variation on enzyme activity were approximately 3%. 4. The IC(50) values estimated using the per cent of residual activity from the single concentration method matched closely those from the multiple concentration method. When the IC(50) value as inhibitor concentration was used for a single concentration method, the method enabled easy estimation of inhibitory patterns (such as competitive or time-dependent inhibition) on cytochromes P450. Therefore, from the ease of the technique, automation of the microtitre plate assay and application of the single concentration method might be useful for inhibitory assessment of cytochromes P450 more than that of current conventional methods.
Patent•
15 Mar 2004TL;DR: In this article, a monoclonal antibody specifically recognizing human epiregulin (hEPR), a hybridoma producing the same and a highly sensitive method of detecting hEPR using the antibody was presented.
Abstract: It is intended to establish a convenient and highly sensitive method of detecting hEPR and provide a novel method of detecting a human tumor expressing hEPR. More specifically speaking, a monoclonal antibody specifically recognizing human epiregulin (hEPR), a hybridoma producing the same and a highly sensitive method of detecting hEPR using the antibody.
TL;DR: Synthetic studies of FD-891 noticed the synthesized fragments including the C12-C15 conjugated double bond moiety showed a quite different signal pattern in 1H NMR, especially the chemical shifts of the double bonds compared to those reported in natural FD- 891 andFD-892.
TL;DR: The therapeutic efficacies of TKs against systemic infections and respiratory tract infection (RTI) caused by gram-positive bacteria in mice are superior to those of CAM and AZM, and the pharmacokinetics of TE-802 are similar to Those of AZM in mice and monkeys, suggesting the potential for once-daily administration in humans.
Abstract: The in vitro and in vivo antibacterial activities of tricyclic ketolides (TKs: TE-802, TE-806, TE-935, TE-943) have been compared with those of clarithromycin (CAM), azithromycin (AZM) and rokitamycin (RKM). TKs were active against not only erythromycin (EM)-susceptible organisms; aerobic gram-positive bacteria, some gram-negative bacteria, anaerobic bacteria and Mycoplasma pneumoniae, but also EM-resistant Staphylococcus aureus (inducible macrolide-resistant strains) as well as EM-resistant Streptococcus pneumoniae (efflux-resistant strains). The therapeutic efficacies of TKs against systemic infections and respiratory tract infection (RTI) caused by gram-positive bacteria in mice are superior to those of CAM and AZM. The peak plasma levels (Cmax, p.o.) of TE-802 in mice were equal to that of CAM, but the plasma area under the concentration-time curve (AUC(24 hours)) was 4.7 times that for CAM. The plasma Cmax (p.o.) value for TE-802 in monkey was equal to that of CAM, whereas the AUC(8 hours) value was three-fourths that of CAM. The pharmacokinetics of TE-802 are similar to those of AZM in mice and monkeys, suggesting the potential for once-daily administration in humans.
TL;DR: The results suggest that M cells exhibit features of not only hepatocytes but also myofibroblasts, and TGF-beta1 plays an important role in the formation of collagen fiber networks in this culture system.
TL;DR: The results suggest that the Na+/Ca2+ exchanger is involved in the stunned myocardium of dogs after reperfusion, and that SEA0400 has a protective effect against myocardial stunning in dogs.
TL;DR: The established method was validated and demonstrated to be applicable in the determination of total amount of CS in a commercial ophthalmic solution and application of the method to disaccharide compositional analysis was also performed.
TL;DR: Results of the baculovirus expression system showed that lornoxicam 5'-hydroxylation via CYP2C9 was markedly decreased by the substitution of Ile359Leu (CYP2C 9.3), whereas the effect of the substituted Arg144Cys was nonexistent or negligible, and additional in vivo studies are required to confirm that individuals with homologous CYP1C9*3 allele exhibit impaired lORNoxicam clearance.
Abstract: The effects of allelic variants of CYP2C9 ( CYP2C9*2 and CYP2C9*3 ) on lornoxicam 5′-hydroxylation were studied using the corresponding variant protein expressed in baculovirus-infected insect cells and human liver microsomes of known genotypes of CYP2C9. The results of the baculovirus expression system showed that CYP2C9.3 gives higher K m and lower V max values for lornoxicam 5′-hydroxylation than does CYP2C9.1. In contrast, K m and V max values of CYP2C9.1 and CYP2C9.2 for the reaction were comparable. Lornoxicam 5′-hydroxylation was also determined in liver microsomes of 12 humans genotyped for the CYP2C9 gene ( \*1/\*1, n = 7; \*1/\*2, n = 2; \*1/\*3, n = 2; \*3/\*3, n = 1). A sample genotyped as \*3/\*3 exhibited 8- to 50-fold lower intrinsic clearance for lornoxicam 5′-hydroxylation than did samples genotyped as \*1/\*1 . However, the values for intrinsic clearance for \*1/\*3 were within the range of values exhibited by samples of \*1/\*1 . In addition, no appreciable differences were observed in kinetic parameters for lornoxicam 5′-hydroxylation between \*1/\*1 and \*1/\*2 . In conclusion, this study showed that lornoxicam 5′-hydroxylation via CYP2C9 was markedly decreased by the substitution of Ile359Leu (CYP2C9.3), whereas the effect of the substitution of Arg144Cys (CYP2C9.2) was nonexistent or negligible. Additional in vivo studies are required to confirm that individuals with homologous CYP2C9*3 allele exhibit impaired lornoxicam clearance.
TL;DR: The findings of the present suggest that histamine produced by enhanced HDC activity is involved in the induction of short scratchings in NC mice.
TL;DR: The data support the notion that high levels of circadian variations of endogenous glucocorticoid lead to a lack of protection against bacteria and a persistence of S. aureus colonization on the skin in NC/Nga mice.
TL;DR: Improvement of the physical properties of pyrazole derivative 1, which was reported previously as a potent and selective 20-HETE synthase inhibitor, is described, and introduction of a sufficient substituted-amino group on the side chain enhanced the water-solubility of 1.
TL;DR: It is suggested that CYP2D6 have the highest catalytic activity of DMAE 5′-O-demethylation in human liver microsomes, followed by CYP1A2 to a small extent.
Abstract: The human cytochrome P450 (CYP) isoforms catalyzing the oxidation metabolism of desglymidodrine (DMAE), an active metabolite of midodrine, were studied. Recombinant human CYP2D6, 1A2 and 2C19 exhibited appreciable catalytic activity with respect to the 5′-O-demethylation of DMAE. TheO-demethylase activity by the recombinant CYP2D6 was much higher than that of other CYP isoforms. Quinidine (a selective inhibitor of CYP2D6) inhibited the O-demethylation of DMAE in pooled human microsomes by 86%, while selective inhibitors for other forms of CYP did not show any appreciable effect. Although the activity of CYP2D6 was almost negligible in the PM microsomes, the O-demethylase activity of DMAE was found to be maintained by about 25% of the pooled microsomes. Furafylline (a selective inhibitor of CYP1A2) inhibited the M-2 formation in the PM microsomes by 57%. The treatment of pooled microsomes with an antibody against CYP2D6 inhibited the formation of M-2 by about 75%, whereas that of the PM microsomes did not show drastic inhibition. In contrast, the antibody against CYP1A2 suppressed the activity by 40 to 50% in the PM microsomes. These findings suggest that CYP2D6 have the highest catalytic activity of DMAE 5′-O-demethylation in human liver microsomes, followed by CYP1A2 to a small extent.