YAKUGAKU ZASSHI
Online ISSN : 1347-5231
Print ISSN : 0031-6903
ISSN-L : 0031-6903
133 巻, 9 号
選択された号の論文の14件中1~14を表示しています
誌上シンポジウム
  • 角田 慎一
    2013 年 133 巻 9 号 p. 923-924
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
  • 鎌田 春彦
    2013 年 133 巻 9 号 p. 925-930
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Biopharmaceuticals represent new generation drugs that are more effective than conventional low molecular weight medicines for the treatment of cancer and other refractory diseases. In order to develop biopharmaceuticals it is critically important to explore suitable target molecules both for the diagnosis and treatment of disease. However, the choice of effective biomarker or drug target is often the most difficult part of the drug development process. Target discovery typically involves ‘omics’ techniques such as genomics, gene chip analysis and proteomics. Of these, proteomics is particularly important because proteins often comprise the final biomarker or drug target in the clinical sample (e.g., tissue or blood). Comprehensive proteomic analysis involves the identification of target proteins with different levels of expression between two states and is extremely useful at selecting promising candidates. Technology is then applied to further narrow down the list of relevant proteins. Here, I would like to introduce our novel procedure, termed “antibody proteomics technology”, which can generate monoclonal antibody from a minute amount (i.e., nanogram level) of protein. Antibody proteomics technology can be used to easily identify suitable target molecules in order to develop effective biomarkers and drug targets.
  • 加藤 和則
    2013 年 133 巻 9 号 p. 931-938
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Establishment of a system that allows selective drug delivery and gene silencing to a tumor is expected to enable targeted therapy. We constructed a genetically modified adenovirus incorporating an IgG Fc-binding motif from the Staphylococcus protein A, Z33 (Adv-FZ33). By cross-linking the Adv-FZ33 virus and the surface antigen molecules with the targeting monoclonal antibodies (mAbs), we attained highly enhanced gene deliveries into the respective antigen-positive cancer cells. Therefore, we aimed to establish a systematic screening method to search for antibody and cell surface target candidates that would provide highly selective anti-cancer reagents to malignant tumors. Using an Adv-FZ33, hybridoma libraries producing a variety of mAbs for human pancreatic, prostate, lung or ovarian carcinoma cells were screened, and we were able to selectively obtain several mAbs which had potent high affinity and recognized antigens of high structure. Within these mAbs, we have identified tumor cell target molecules including not only carcinoembryonic antigen (CEA), epithelial cell adhesion molecule (EpCAM), epidermal growth factor receptor (EGFR), prostate specific membrane antigen (PSMA) but also novel tumor surface target molecules such as phosphatidic acid phosphatase type 2a (PAP2a) and interleukin-13 receptor variant α2 (IL-13Rα2) as tumor antigens. Overall, these results indicate that this type of inductive method approach is a reliable strategy for screening in antibody therapy on par with antibody-dependent drug-delivery system.
  • 八木 秀樹, 益子 高
    2013 年 133 巻 9 号 p. 939-945
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Antibodies have greatly contributed to the development of medical science and pharmacology, because of their high specificity. The cell fusion method has developed monoclonal antibodies (mAb) technology, such that massive amounts of mAb with a uniform structure can be produced. Although mAb have been produced against many proteins so far, the production of mAb against multi-pass transmembrane proteins, such as G-protein coupled receptor (GPCR) and various transporter proteins has been extremely difficult. The complicated structures, poorly extracellular regions, and high hydrophobicity of multiple-transmembrane proteins make it difficult to produce mAb against them. Production of mAb that recognize the extracellular region of living cells is thought to be important in determining the ability of a protein. Based on these findings, we tried to produce mAb against a multi-pass transmembrane transporter using green fluorescent protein (GFP)-fused full-length target proteins as immunogens. Furthermore, the immunizing method has proved to be important in generating functional mAb. We succeeded in producing functional mAb that react against the extracellular region of a 12-pass transmembrane transporter in a living cell. Based on this success, we began to produce mAb against seven-transmembrane GPCR. In this symposium, we report on the results of producing mAb against S1P receptors, a type of GPCR.
総説
  • 松尾(池田) 由理
    2013 年 133 巻 9 号 p. 947-954
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Although augmented prostaglandin E2 (PGE2) accumulation has been demonstrated at the lesion sites of rodent ischemia models, the role of postischemic PGE2 in neuronal survival has remained obscure. We recently identified the microsomal prostaglandin E synthase-1 (mPGES-1), an inducible terminal enzyme for prostaglandin E2 synthesis, as a critical factor in stroke-reperfusion injury. Co-induction of mPGES-1 and cyclooxygenase (COX)-2, an upstream enzyme for PGE2 production, was observed after brain ischemia. In mPGES-1 knockout (KO) mice, in which the postischemic PGE2 production in the cortex was completely absent, the ischemic injuries were less severe compared to those in wild-type (WT) mice. The ameliorated symptoms observed in KO mice after ischemia were reversed to almost the same severity as in the WT mice by intracerebroventricular injection of PGE2 into KO mice. The induction of mPGES-1 was also observed after glutamate exposure in cultured hippocampal slices. In mPGES-1 KO slices, glutamate-induced excitotoxicity was less severe compared to that in WT slices. Among the EP1-4 antagonists and agonists, only the EP3 antagonist attenuated and only the EP3 agonist augmented the glutamate-induced excitotoxicity. Furthermore, intraperitoneal injection of COX-2 inhibitor or EP3 antagonist reduced the ischemic injuries in WT mice, but not in mPGES-1 KO mice. In EP3 KO mice, the ischemic injuries were less severe compared to those in WT mice. These results suggest that mPGES-1 and COX-2 are co-induced by excessive glutamate in the ischemic brain and act together to exacerbate stroke injury through PGE2 production followed by activation of EP3 receptors.
  • 五十嵐 信智
    2013 年 133 巻 9 号 p. 955-961
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Aquaporins (AQPs) are membrane channels that transport water within the human body and are therefore important for the regulation of water homeostasis. However, little is known regarding the details of the physiological role of AQP3, which is predominantly expressed in the colon. Thus, we investigated the role of AQP3 in the colon using laxative agents (magnesium sulfate and bisacodyl). The results suggest that the laxative effect produced by magnesium sulfate, which is classified as an osmotic laxative, is not simply a result of the changes in osmotic pressure but is also associated with the increased expression of AQP3 in the mucosal epithelial cells of the colon. In addition, magnesium sulfate increased colonic AQP3 expression through adenylate cyclase activation, which is caused by an increase in the intracellular Mg2+ concentration. This effect may trigger CREB phosphorylation through PKA activation and promote AQP3 gene transcription. Meanwhile, bisacodyl, which is classified as a stimulant laxative, decreases the expression level of AQP3 in the mucosal epithelial cells of the colon, resulting in the inhibition of water transfer from the intestinal tract to the vascular side of the epithelium, eventually leading to the development of diarrhea. It was also observed that the direct activation of colon macrophages by bisacodyl increases the secretion of PGE2, which acts as a paracrine factor and decreases AQP3 expression in colon mucosal epithelial cells. Future studies of the enteric AQP3 expression level and water transport may aid in the development of new laxative and antidiarrheal agents that target AQP3.
  • 永野 恵司
    2013 年 133 巻 9 号 p. 963-974
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      The periodontal disease-associated bacterium Porphyromonas gingivalis primarily uses FimA fimbriae for adhesion to and colonization in the gingival tissues. The fimbriae show a filamentous structure that is composed of polymer of FimA encoded by the fimA gene. FimC, FimD and FimE are associated with the fimbriae as minor components. FimB anchors the fimbriae to the bacterial surface and regulates their length. The N terminus of FimA is digested in a maturation process, then mature FimA proteins are polymerized to form fimbriae in the outer membrane. Transcription of the fimA gene is regulated by the two-component regulatory system of FimS/R. In addition, expression of FimA is influenced by many environmental factors such as nutrients, environmental stresses, and other bacterial products. The fimA gene shows a genetic polymorphism and it is proposed that there are six genotypes (types I-V and Ib). Types II and IV are frequently isolated from severe periodontal patients. Therefore, they are predicted to be high virulent types, but the molecular mechanisms remain unclear. FimA fimbriae also exhibit a heterogenic antigenicity that is basically consistent with the fimA genotype. The fimbriae interact with many molecules such as surface molecules of host cells, extracellular matrix, salivary components, and bacterial components. Many reports argue binding of FimA residues in the fimbriae to the target molecules, but it is reported that accessory components of FimCDE critically function as an adhesin. Elucidation of adherent mechanism of P. gingivalis through the FimA fimbriae could lead to a development of prophylaxis against the bacterial infection.
  • 原田 慎吾
    2013 年 133 巻 9 号 p. 975-982
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Enantio- and diastereoselective one-pot synthesis of three- to seven-membered cis-azaheterocycles was achieved using a triggered asymmetric conjugate addition reaction of lithium amide with an enoate, followed by C- and N-alkylation of the resulting lithium 3-aminoenolate with α, ω-dihaloalkane, which can be regarded as a [1+2+n] cyclization. The usefulness of the developed methodology was clearly demonstrated by the short-step asymmetric syntheses of azirinomycin ester, nemonapride, and kopsinine.
  • 泉 安彦
    2013 年 133 巻 9 号 p. 983-988
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Parkinson disease is one of the most common neurodegenerative disorders and is characterized by the selective loss of dopaminergic neurons in the substantia nigra. Although a decrease in proteasome activity has been found in patients with sporadic Parkinson disease, the relationship between the ubiquitin-proteasome system and dopaminergic neuronal death remains to be elucidated. Here, we review a mechanism in which proteasome inhibition provides dopaminergic neuroprotection from oxidative stress. Treatment with lactacystin, a proteasome inhibitor, significantly suppressed 6-hydroxydopamine (6-OHDA)-induced toxicity and oxidative stress in PC12 cells. In addition, lactacystin enhanced glutathione synthesis via elevation of γ-glutamylcysteine synthetase (γ-GCS) mRNA levels. Expression of antioxidant enzymes, such as γ-GCS and hemeoxygenase-1 (HO-1), is regulated by the nuclear factor-erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway. Lactacystin induced Nrf2 accumulation and increased ARE activity. In mesencephalic cultures, lactacystin-induced upregulation of HO-1 in astrocytes contributed to dopaminergic neuroprotection against 6-OHDA-induced toxicity. These data suggest that proteasome inhibition provides cytoprotection against oxidative stress by activating the Nrf2-ARE pathway. Subsequently, we attempted to identify a novel Nrf2-ARE activator in dietary fruits and vegetables. Using bioactivity-guided fractionation, we identified 2′,3′-dihydroxy-4′,6′ -dimethoxychalcone (DDC) from green perilla leaves as the activator responsible for the increased activation of the ARE pathway. DDC upregulated γ-GCS and HO-1 and protected PC12 cells against 6-OHDA-induced toxicity. In conclusion, the activation of the Nrf2-ARE pathway may be an effective means to prevent dopaminergic neuronal death in patients with Parkinson disease.
  • 石原 慶一
    2013 年 133 巻 9 号 p. 989-994
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Down syndrome (DS), caused by triplication of human chromosome 21, is the most common aneuploidy. A mouse model of DS may be useful for the investigation of DS pathophysiology. Ts1Cje mouse, an established DS mouse model, is widely used in DS research. It carries a trisomic segment of mouse chromosome 16 that contains a syntenic region to human chromosome 21. The brain of the Ts1Cje mouse has been analyzed morphologically and biochemically to elucidate DS pathophysiology. We have also demonstrated some abnormal phenotypes of this mouse model, such as enlarged brain ventricles, reduced embryonic and adult neurogenesis, and increased lipid peroxidation. Elucidating the underlying molecular mechanisms in the Ts1Cje mouse may improve understanding of the etiology of the phenotypic abnormalities of DS, including cognitive impairment and developmental retardation, and aid in development of therapeutic strategy. High-throughput gene and protein expression analyses, such as transcriptomics and proteomics, are useful for identification of molecules associated with the development of DS symptoms. In this review, alteration of molecular expression in the brain of a DS mouse model is highlighted, and possible molecular mechanisms underlying DS phenotypic abnormalities such as cognitive impairment and developmental retardation are discussed.
  • 三宅 正晃
    2013 年 133 巻 9 号 p. 995-1006
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      The suppository preparation, which can improve the absorption of poorly absorbable drugs safer than commercially available suppositories, was developed by utilizing sodium laurate and taurine. Additionally, the novel oral absorption-improving system was also established by utilizing polyamines and bile acids. Furthermore, to evaluate the efficacy of these new formulations and estimate the absorbability of new drug candidates in humans, the in vitro prediction system utilizing an isolated human intestinal tissues was developed and successfully predicted the fraction of dose absorbed for several model drugs. These findings would contribute to the development of new dosage forms and new drugs for oral administration.
  • 伊藤 卓也
    2013 年 133 巻 9 号 p. 1007-1015
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      The aminocyclitol family is a relatively new class of natural products such as gentamicin, kanamycin, and streptomycin, which have been used clinically for decades as potent antimicrobial agents. These secondary metabolites are chiefly produced by microorganisms, especially Actinomycetes. Their chemical structures most commonly contain a C7N unit, 2-epi-5-epi-valiolone or 3-amino-5-hydroxybenzoic acid (3,5-AHBA) which are known to be responsible for their biological activities. In the course of current study, the biosynthesis of the C7N-containing metabolites, validamycin and acarbose, pactamycin, have been evaluated. We studied N-formamide salicylic acid (FSA) moiety which is a C7N unit synthesized from tryptophan by microorganisms. A strong antifungal agent antimycin, isolated from several Streptomyces sp., contains an FSA moiety, and constitutes a unique nine-membered dilactone ring with L-threonine, short-chain fatty acid, and an amide linkage connecting it to an FSA moiety. Also, an antitumor antibiotic asukamycin, produced by Streptomyces nodosus subsp. asukaensis ATCC 29757, consists of both 3,4-AHBA and C5N, cyclohexane ring linked to trans-triens. To improve the efficacy and reduce the toxicity of these metabolites, further structural modification is needed. Total chemical synthesis of these complex compounds is difficult. Therefore, alternative approaches are required, e.g., biosynthetic or genetic modification methods. This review presents the biosynthetic study on these compounds for creating new analogs using mutasyntheis.
ノート
  • 森野 博文, 小泉 朋子, 三浦 孝典, 福田 俊昭, 柴田 高
    2013 年 133 巻 9 号 p. 1017-1022
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      Noroviruses are one of the most important causes of acute gastroenteritis throughout the world. The aim of this study is to evaluate the efficacy of a chlorine dioxide gas-generating gel (ClO2 gel, 60 g) against feline calicivirus (FCV), a norovirus surrogate, in the wet state on glass dishes in a test sink (43 cm long, 75 cm wide, and 29 cm deep). The ClO2 gel permits sustained release of gaseous ClO2 (1.7 mg/h at 25°C), and was placed in one corner of the test sink. The glass dishes containing FCV suspension were placed at three positions in the test sink. We demonstrated that FCV was inactivated within 5h (>2 or >3 log10 reductions at three positions, n=20) in the test sink where the ClO2 gel was placed. These small quantities of ClO2 gel might be a useful tool for reducing the risk of infection by norovirus in wet environments such as kitchens and bathrooms under optimal condition.
  • 栗本 蕗, 堀 里子, 佐藤 宏樹, 三木 晶子, 澤田 康文
    2013 年 133 巻 9 号 p. 1023-1034
    発行日: 2013/09/01
    公開日: 2013/09/01
    ジャーナル フリー
      For drug fostering and evolution, it is important to collect information directly from patients on the efficacy and safety of drugs as well as patient needs. At present, however, information gathered by healthcare professionals, pharmaceutical companies, or governments is not sufficient. There is concern that patients may fail to recognize the importance of providing information voluntarily. The present study was conducted to provide drug information to patients/consumers, to enlighten them on the importance of providing drug information by themselves, and to develop an Internet website, called “Minkusu,” for collecting drug information from patients. This website is based on a registration system (free of charge). It is designed to provide information on proper drug use, and to collect opinions about drugs. As of May 31, 2012, a total of 1149 people had been registered. The male/female ratio of registered members was approximately 1:1, and patients/consumers accounted for 23%. According to the results of a questionnaire survey, several patient/consumer members appreciated the usefulness of the information service, and they took an opportunity to know of the concepts of drug development and evolution (Ikuyaku, in Japanese) through the information services provided by this site. In conclusion, the developed information system would contribute to the proper use of drugs by patients/consumers and to the promotion of drug development and evolution.
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