ビニルエーテルの不斉オキシセレン化反応を用いる光学活性アセタール類の合成と天然物合成への応用に関する研究

取得学位：博士(薬学)，学位授与番号：博乙第234号，学位授与年月日：平成13年9月28日,学位授与年：200

固相担持型Mn(III)反応剤を用いる酸化的ラジカル環化反応の開発とその応用

金沢大学疾患モデル総合研究センター機器分析研究施設1.固相担持型Mn(III)反応剤の調製まず、Mn(III)反応剤としてMn(OAc)_3・2H_2Oを、固相担体としてCOOH基を交換基とするイオン交換樹脂であるAmberlite IRP-64 [100-400 mesh,〜10 meq/g(dry)]を用い、固相担持型Mn(III)反応剤の調製を試みた。すなわち、窒素雰囲気下、Mn(OAc)_3・2H_2O(1.07g,4mmol)のエタノール溶液にAmberliteIRP-64(1.0g)を加え、室温にて42時間撹拌した後、吸引ろ取した。次に、ろ取した樹脂をエタノール、つづいてヘキサンで洗浄した後、真空乾燥して赤褐色の樹脂を得た。原料として用いたMn(OAc)_3・2H_2Oはエタノールに溶解するが、この手法で処理した後の樹脂はエタノール中で撹拌してもMn(III)特有の赤色が全く溶出してくることがなかったことから、用いたMn(OAc)_3・2H_2Oは、ほぼ定量的にAmberlite IRP-64に担持されたものと推測された。2.固相担持型Mn(III)反応剤を用いた酸化的ラジカル環化反応の検討上記の方法で調製した樹脂を用い、実際にラジカル環化反応を試みた。研究実施計画に基づき、分子内にアルキン部を有しβ位にメチル基が1つ置換された鎖状β-ケトエステルを基質として用い、エタノール中、調製した樹脂とともに撹拌したところ、予想に反してラジカル環化はほとんど進行せず、5-exo環化体と6-endo環化体が合わせてもわずか6%の収率でしか得られず、その代わりに主生成物として基質の活性メチンがヒドロキシル化されたアルコール体が29%の収率で得られた。触媒量のMn(OAc)_2・4H_2Oを用い酸素雰囲気下で環状β-ケトエステルを反応させると同様の反応が起こることが既に知られているが、鎖状β-ケトエステルではうまくいかなかったと報告されていることから、本研究で得られた結果は興味深い。さらに、触媒量の樹脂を用い酸素雰囲気下で反応を行った場合にもこのヒドロキシル化反応は進行し、また反応後の後処理は、1)樹脂のろ去、2)ろ液の濃縮、3)カラム精製のみであることもこの樹脂を用いる大きな利点である。一方、本来の目的であった酸化的ラジカル環化反応に関しては、わずかながらではあるが環化反応が進行したことから、反応条件の更なる検討、用いる固相担体の修飾などによって改善される可能性が示唆された。研究課題/領域番号:15790006, 研究期間(年度):2003 – 2004出典：「固相担持型Mn(III)反応剤を用いる酸化的ラジカル環化反応の開発とその応用」研究成果報告書　課題番号15790006（KAKEN：科学研究費助成事業データベース（国立情報学研究所））（https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-15790006/)を加工して作

Novel Autologous Therapy for Long-Gap Peripheral Nerve Injury Using Human Sk-SCs

Losses in vital functions of the somatic motor and sensory nervous system are induced by severe long-gap peripheral nerve transection injury. In such cases, autologous nerve grafts are the gold standard treatment, despite the unavoidable sacrifice of other healthy functions, whereas the prognosis is not always favorable. Here, we use human skeletal muscle-derived stem cells (Sk-SCs) to reconstitute the function after long nerve-gap injury. Muscles samples were obtained from the amputated legs from 9 patients following unforeseen accidents. The Sk-SCs were isolated using conditioned collagenase solution, and sorted as CD34+/45- (Sk-34) and CD34-/45-/29+ (Sk-DN/29+) cells. Cells were separately cultured/expanded under optimal conditions for 2 weeks, then injected into the athymic nude mice sciatic nerve long-gap model (7-mm) bridging an acellular conduit. After 8-12 weeks, active cell engraftment was observed only in the Sk-34 cell transplanted group, showing preferential differentiation into Schwann cells and perineurial/endoneurial cells, as well as formation of the myelin sheath and perineurium/endoneurium surrounding regenerated axons, resulted in 87% of numerical recovery. Differentiation into vascular cell lineage (pericyte and endothelial cells) were also observed. A significant tetanic tension recovery (over 90%) of downstream muscles following electrical stimulation of the sciatic nerve (at upper portion of the gap) was also achieved. In contrast, Sk-DN/29+ cells were completely eliminated during the first 4 weeks, but relatively higher numerical (83% vs. 41% in axon) and functional (80% vs. 60% in tetanus) recovery than control were observed. Noteworthy, significant increase in the formation of vascular networks in the conduit during the early stage (first 2 weeks) of recovery was observed in both groups with the expression of key factors (mRNA and protein levels), suggesting the paracrine effects to angiogenesis. These results suggested that the human Sk-SCs may be a practical source for autologous stem cell therapy following severe peripheral nerve injury

Stereoselective synthesis of trans-3a-aryloctahydroindoles using cyclization of N-vinylic α-(methylthio)acetamides

金沢大学大学院自然科学研究科生理活性物質科学金沢大学薬学部Treatment of N-(2-arylcyclohex-l-enyl)-α-(methylthio)acetamide with NCS underwent cyclization to give 3a-arylhexahydroindol-2-one, which was stereoselectively converted into trans-3a-aryloctahydroindole. © 2006 The Japan Institute of Heterocyclic Chemistry

Anti-IL-6 Receptor Antibody Causes Less Promotion of Tuberculosis Infection than Anti-TNF-α Antibody in Mice

Objective. Our aim was to investigate the effects of IL-6 blockade on the progression of Mycobacterium tuberculosis (TB) and compare them with those of TNF-α blockade in mice. Methods. Mice were intravenously infected with TB and injected with antibodies. Survival was monitored and histological and immunological studies were carried out. Results. All anti-IL-6R Ab-treated mice and 8 of 10 control mice survived until sacrificed 224 days after TB challenge, whereas anti-TNF-α Ab-treated mice all died between 120 and 181 days. Anti-IL-6R Ab-treated mice exhibited no significant differences in TB CFU in organs, including the lungs, and no deterioration in histopathology compared to control mice at 4 weeks. In contrast, anti-TNF-α Ab-treated mice exhibited increased TB CFU and greater progression of histopathological findings in organs than control mice. Spleen cells from anti-TNF-α Ab-treated mice had decreased antigen-specific response in IFN-γ release and proliferation assays. The results in anti-IL-6R Ab-treated mice suggest that spleen cell responses were decreased to a lesser degree. Similar results were obtained in IL-6 knockout (KO) mice, compared with TNF receptor 1 (TNFR1) KO and TNFR1/IL-6 double KO (DKO) mice. Conclusion. IL-6R blockade promotes the progression of TB infection in mice far less than TNF-α blockade

Construction of a genetic AND gate under a new standard for assembly of genetic parts

<p>Abstract</p> <p>Background</p> <p>Appropriate regulation of respective gene expressions is a bottleneck for the realization of artificial biological systems inside living cells. The modification of several promoter sequences is required to achieve appropriate regulation of the systems. However, a time-consuming process is required for the insertion of an operator, a binding site of a protein for gene expression, to the gene regulatory region of a plasmid. Thus, a standardized method for integrating operator sequences to the regulatory region of a plasmid is required.</p> <p>Results</p> <p>We developed a standardized method for integrating operator sequences to the regulatory region of a plasmid and constructed a synthetic promoter that functions as a genetic AND gate. By standardizing the regulatory region of a plasmid and the operator parts, we established a platform for modular assembly of the operator parts. Moreover, by assembling two different operator parts on the regulatory region, we constructed a regulatory device with an AND gate function.</p> <p>Conclusions</p> <p>We implemented a new standard to assemble operator parts for construction of functional genetic logic gates. The logic gates at the molecular scale have important implications for reprogramming cellular behavior.</p

Assessment of the Efficacy of ReoGo-J Robotic Training Against Other Rehabilitation Therapies for Upper-Limb Hemiplegia After Stroke: Protocol for a Randomized Controlled Trial

Background: Stroke patients experience chronic hemiparesis in their upper extremities leaving negative effects on quality of life. Robotic therapy is one method to recover arm function, but its research is still in its infancy. Research questions of this study is to investigate how to maximize the benefit of robotic therapy using ReoGo-J for arm hemiplegia in chronic stroke patients.Methods: Design of this study is a multi-center parallel group trial following the prospective, randomized, open-label, blinded endpoint (PROBE) study model. Participants and setting will be 120 chronic stroke patients (over 6 months post-stroke) will be randomly allocated to three different rehabilitation protocols. In this study, the control group will receive 20 min of standard rehabilitation (conventional occupational therapy) and 40 min of self-training (i.e., sanding, placing and stretching). The robotic therapy group will receive 20 min of standard rehabilitation and 40 min of robotic therapy using ReoGo®-J device. The combined therapy group will receive 40 min of robotic therapy and 20 min of constraint-induced movement therapy (protocol to improve upper-limb use in ADL suggests). This study employs the Fugl-Meyer Assessment upper-limb score (primary outcome), other arm function measures and the Stroke Impact Scale score will be measured at baseline, 5 and 10 weeks of the treatment phase. In analysis of this study, we use the mixed effects model for repeated measures to compare changes in outcomes between groups at 5 and 10 Weeks. The registration number of this study is UMIN000022509.Conclusions: This study is a feasible, multi-site randomized controlled trial to examine our hypothesis that combined training protocol could maximize the benefit of robotic therapy and best effective therapeutic strategy for patients with upper-limb hemiparesis

Lubiprostone Decreases the Small Bowel Transit Time by Capsule Endoscopy: An Exploratory, Randomised, Double-Blind, Placebo-Controlled 3-Way Crossover Study

The aim of this study was to investigate the usefulness of lubiprostone for bowel preparation and as a propulsive agent in small bowel endoscopy. Six healthy male volunteers participated in this randomized, 3-way crossover study. The subjects received a 24 μg tablet of lubiprostone 60 minutes prior to the capsule ingestion for capsule endoscopy (CE) and a placebo tablet 30 minutes before the capsule ingestion (L-P regimen), a placebo tablet 60 minutes prior to CE and a 24 μg tablet of lubiprostone 30 minutes prior to CE (P-L regimen), or a placebo tablet 60 minutes prior to r CE and a placebo tablet again 30 minutes prior to CE (P-P regimen). The quality of the capsule endoscopic images and the amount of water in the small bowel were assessed on 5-point scale. The median SBTT was 178.5 (117–407) minutes in the P-P regimen, 122.5 (27–282) minutes in the L-P regimen, and 110.5 (11–331) minutes in the P-L regimen (P=0.042). This study showed that the use of lubiprostone significantly decreased the SBTT. We also confirmed that lubiprostone was effective for inducing water secretion into the small bowel during CE