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1. 发明申请

US20060257997A1 High throughput synthesis system and synthesizer for automatically performing the system 审中-公开
标题翻译：高吞吐量合成系统和合成器，用于自动执行系统
公开(公告)号：US20060257997A1
公开(公告)日：2006-11-16
申请号：US10554434
申请日：2004-04-23
申请人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara , Riyo Morishita , Mihoro Saeki , Tomohisa Sato , Aya Kitamoto
发明人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara , Riyo Morishita , Mihoro Saeki , Tomohisa Sato , Aya Kitamoto
IPC分类号： C12M3/00
CPC分类号： C12P21/02 , C12P19/34
摘要： To develop system technology for a high throughput in vitro synthesis reaction method for biopolymers such as proteins, RNA, and the like. In detail, a cell-free synthesis system wherein a template substance is used as a raw material, and an automatic synthesizer using the system, includes the following systems, control systems therefor, or combinations thereof; 1) to make a template substance, a substrate, and a reaction solution into contact so as to lead to a synthetic reaction, 2) to put the reaction system outside of the synthetic reaction system to perform dilution treatment of the solution approximately around the decrease in the synthesis velocity, approximately around the stop of the synthetic reaction, or in the middle thereof, 3) to perform concentration treatment subsequent to the dilution treatment, and 4) to put the reaction system back into the synthetic reaction system, or 1) to make a template substance, a substrate, and a reaction solution into contact so as to lead to a synthetic reaction, 2) to put the reaction system outside of the synthetic reaction system to perform concentration treatment of the solution approximately around the decrease in the synthesis velocity, approximately around the stop of the synthetic reaction, or in the middle thereof, 3) to perform dilution treatment subsequent to the concentration treatment, and 4) to put the diluted reaction system back into the synthetic reaction system.
摘要翻译：开发用于生物聚合物如蛋白质，RNA等的高通量体外合成反应方法的系统技术。具体而言，其中使用模板物质作为原料的无细胞合成系统和使用该系统的自动合成器包括以下系统，其控制系统或其组合; 1）使模板物质，底物和反应溶液接触以导致合成反应，2）将反应体系置于合成反应体系外部，以进行大约减少的溶液的稀释处理在合成速度约合成反应停止或其中间，3）进行稀释处理后的浓缩处理，4）使反应体系回到合成反应体系中，或1）使模板物质，底物和反应溶液接触以导致合成反应，2）将反应体系置于合成反应体系外部，以在约合成速度大约在合成反应停止时，或在其中间，3）进行浓缩处理后的稀释处理，和4）将稀释的反应体系放回到合成反应体系中。

2. 发明授权

US07981617B2 Transcription template for cell-free protein synthesis and method using the same 有权
标题翻译：无细胞蛋白质合成的转录模板及其使用方法
公开(公告)号：US07981617B2
公开(公告)日：2011-07-19
申请号：US10363372
申请日：2001-08-28
申请人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara
发明人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara
IPC分类号： C12Q1/68
CPC分类号： C12P21/02 , C12Q1/686
摘要： Methods to construct a transcription template for cell-free protein synthesis that has a high translation template activity, using a 3′-side primer and a 5′-side primer for PCR are provided. The 5′-side primer for PCR has a sequence complementary to a base sequence containing at least a part of a promoter functional site from the 5′-end of a promoter and has a base sequence that does not contain a base sequence complementary to at least a part of a RNA polymerase-recognizing site of the 3′-end of the promoter. The other primer has a base sequence complementary to at least a part of the RNA polymerase-recognizing site of the 3′-end of the promoter and has a sequence that does not contain a base sequence complementary to at least a part of a promoter functional site from the 5′-end of the promoter.
摘要翻译：提供了使用3'侧引物和5'侧引物进行PCR构建具有高翻译模板活性的无细胞蛋白质合成转录模板的方法。用于PCR的5'侧引物具有与包含来自启动子的5'末端的至少一部分启动子功能位点的碱基序列互补的序列，并且其碱基序列不含有与启动子的3'末端的RNA聚合酶识别位点的至少一部分。另一个引物具有与启动子3'末端的RNA聚合酶识别位点的至少一部分互补的碱基序列，并且具有不包含与启动子功能的至少一部分互补的碱基序列的序列来自启动子的5'-末端的位点。

3. 发明授权

US06869774B2 Methods of synthesizing cell-free protein 有权
标题翻译：合成无细胞蛋白的方法
公开(公告)号：US06869774B2
公开(公告)日：2005-03-22
申请号：US10344803
申请日：2001-08-28
申请人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara
发明人： Yaeta Endo , Tatsuya Sawasaki , Tomio Ogasawara
IPC分类号： C12P21/02 , C12P21/06
CPC分类号： C12P21/02
摘要： One embodiment of the present invention is a diffusion continuous batch cell-free protein-synthesis method characterized simultaneously by continuously supplying substrate and energy source molecules in the supply phase to the reaction phase by the free diffusion via interface between both phases and by transferring by-products formed in the reaction phase by enhancing the efficiency of the synthesis reaction by prolonging the reaction lifetime by directly contacting a synthesis reaction mixture (reaction phase) containing a biological extract with a substrate- and energy source-supplying solution (supply phase) without using barrier such as semi-permeable membrane or ultrafiltration membrane in a general cell-free protein-synthesis reaction means. Another embodiment of the present invention is a dilution batch cell-free protein synthesis method characterized by enhancing the efficiency of the protein synthesis by prolonging the reaction lifetime by adding a diluting solution to the reaction mixture after pre-incubating the reaction mixture in a cell-free protein-synthesis reaction means using a wheat-embryo extract. Another embodiment of the present invention is a method characterized by enhancing the efficiency of the synthesis reaction simultaneously by re-supplying substrate and energy sources necessary for the protein synthesis (e.g., amino acids, ATP, GTP, creatine phosphate) to the reaction mixture using a gel filtration column and/or semipermeable membrane and by discontinuously removing by-products formed during the reaction after the synthesis reaction stops in the batch cell-free protein synthesis method.
摘要翻译：本发明的一个实施方案是扩散连续分批无细胞蛋白质合成方法，其特征在于同时通过在两相之间的界面通过自由扩散连续供应反应相中的底物和能量源分子，通过使含有生物提取物的合成反应混合物（反应相）与底物和能量供应溶液（供应阶段）直接接触而延长反应寿命，通过提高合成反应的效率而在反应阶段形成的产物，而不使用一般无细胞蛋白合成反应手段的半透膜或超滤膜等障碍。本发明的另一个实施方案是一种稀释分批无细胞蛋白质合成方法，其特征在于通过在将反应混合物预先培养至细胞培养基中之后，通过向反应混合物中加入稀释溶液来延长反应寿命来提高蛋白质合成的效率，游离蛋白质合成反应意味着使用小麦胚提取物。本发明的另一个实施方案是一种方法，其特征在于通过向反应混合物中再次提供蛋白质合成所需的底物和能量源（例如氨基酸，ATP，GTP，肌酸磷酸），同时提高合成反应的效率，凝胶过滤柱和/或半透膜，并且在间歇无细胞蛋白质合成方法中，在合成反应停止后不连续地除去反应期间形成的副产物。

4. 发明申请

US20070141661A1 Cell extract for high-functioned cell-free protein synthesis and method of preparing the extract 审中-公开
标题翻译：用于高功能无细胞蛋白质合成的细胞提取物及其制备方法
公开(公告)号：US20070141661A1
公开(公告)日：2007-06-21
申请号：US10596538
申请日：2004-12-17
申请人： Yaeta Endo , Tomio Ogasawara
发明人： Yaeta Endo , Tomio Ogasawara
IPC分类号： C12P21/06 , C12N9/64
CPC分类号： C12P21/02
摘要： The subjects of the present invention are to prepare a highly functionalized cell extract for cell-free protein synthesis and to specify and eliminate inhibitors and unstable substances present in conventional and various cell extracts for cell-free protein synthesis. Also provided is a method for preparing the cell extract for use in a cell-free protein synthesis means, wherein ATP-mediated phosphorylation pathway of sugar present in the cell extract is controlled. In particular, the control is introducing at least one selected from the following: 1) removing monosaccharides, 2) removing phosphorylated sugars, 3) controlling production of monosaccharides from polysaccharides, and 4) controlling production of phosphorylated sugars from monosaccharides.
摘要翻译：本发明的目的是制备用于无细胞蛋白质合成的高度官能化的细胞提取物，并且规定和消除存在于常规和各种细胞提取物中的抑制剂和不稳定物质，用于无细胞蛋白质合成。还提供了一种制备用于无细胞蛋白质合成手段的细胞提取物的方法，其中控制细胞提取物中存在的糖的ATP介导的磷酸化途径。具体而言，该对照是引入选自以下的至少一种：1）除去单糖，2）去除磷酸化糖，3）控制多糖的单糖生产，4）控制单糖中磷酸化糖的产生。

5. 发明申请

US20050153390A1 Lyophilized preparation for synthesis of cell-free protein 审中-公开
标题翻译：用于合成无细胞蛋白的冻干制剂
公开(公告)号：US20050153390A1
公开(公告)日：2005-07-14
申请号：US10514855
申请日：2003-05-06
申请人： Yaeta Endo , Tomio Ogasawara
发明人： Yaeta Endo , Tomio Ogasawara
IPC分类号： C12P21/00 , C12P21/06 , C12N11/04
CPC分类号： C12P21/00
摘要： Lyophilized preparations from cell extract for use in cell-free protein, which exhibit an activity for protein synthesis being in no way inferior to that exhibited in low-temperature storage; and a method of utilizing the same. In particular, preparations are formed by performing lyophilization after reducing the concentration of deliquescent substances in a solution containing cell extract for synthesis of cell-free protein to a level not detrimental to the quality of preparations after lyophilization.
摘要翻译：来自细胞提取物的冻干制剂用于无细胞蛋白质，其表现出蛋白质合成的活性绝不逊于在低温储存中显示的活性; 及其利用方法。特别地，通过在将含有细胞提取物的溶液的溶液中减少潮解物质浓度以合成无细胞蛋白质之后进行冷冻干燥来形成制剂，使其在冻干后不会对制剂质量产生不利影响。

6. 发明授权

US5037981A Intermediates for synthesizing BH.sub.4 and its derivatives 失效
标题翻译：用于合成BH4及其衍生物的中间体
公开(公告)号：US5037981A
公开(公告)日：1991-08-06
申请号：US515962
申请日：1990-04-27
申请人： Masayasu Kurono , Takehiko Suzuki , Tomio Ogasawara , Nobuko Ohishi , Kunio Yagi
发明人： Masayasu Kurono , Takehiko Suzuki , Tomio Ogasawara , Nobuko Ohishi , Kunio Yagi
IPC分类号： C07D405/12 , C07D475/04 , C12P25/00
CPC分类号： C07D405/12 , C07D475/04 , C12P25/00
摘要： The present invention relates to intermediates for synthesizing BH.sub.4 and derivatives thereof. The intermediates are shown as follows; ##STR1## wherein R.sub.1 is a hydrogen atom, alkyl, aralkyl, or aryl group; R.sub.2 is an alkyl, hydroxyalkyl, or polyhydroxyalkyl group; R.sub.3 and R.sub.4 are the same or different and represent alkyl, aralkyl, or aryl group; R.sub.5, R.sub.6, R.sub.7, and R.sub.8 are the same or different and represent a hydrogen atom or acyl group; R.sub.9 is an alkyl, aralkyl, or aryl group; R.sub.10 and R.sub.11 are the same or different and represent a hydrogen atom or acyl group; n is an integer of 5 or less; and HX is an acid. The invention also relates to a process for the preparation of L-biopterin.

7. 发明授权

US4528184A Polymer-metal complexes containing quaternary nitrogen atoms used in the treatment of hypercholesteremia 失效
标题翻译：含有用于治疗高胆固醇血症的季氮原子的聚合物 - 金属络合物
公开(公告)号：US4528184A
公开(公告)日：1985-07-09
申请号：US438487
申请日：1982-11-02
申请人： Masayasu Kurono , Osamu Nakagawa , Takafumi Iida , Yoshiro Ishiwata , Tomio Ogasawara , Meiji Kuwayama
发明人： Masayasu Kurono , Osamu Nakagawa , Takafumi Iida , Yoshiro Ishiwata , Tomio Ogasawara , Meiji Kuwayama
IPC分类号： C08G59/00 , A61K31/785 , A61K31/80 , A61P9/10 , C08G73/00 , C08G73/02 , C08G79/00 , C08L63/00 , A61K31/74
CPC分类号： A61K31/80 , A61K31/785 , C08G73/0226
摘要： A novel polymer-metal complex containing quaternary nitrogen atoms, which is obtained by subjecting polyethylenepolyamine or polyethyleneimine to a reaction with a bifunctional compound represented by the formulaY--A--Zwherein Y and Z are same or different and are --CH.sub.2 X, --CH(OH)CH.sub.2 X or ##STR1## group, respectively, A is a single bond, --(CH.sub.2).sub.m --, --CH.sub.2 --O--CH.sub.2 -- or --CH.sub.2 --O--(CH.sub.2).sub.k --O--CH.sub.2 -- group, X is chlorine, bromine or iodine atom, m is an integral of 1 to 3, and k is an integral of 1 to 4, and then subjecting the resulting cross-linked polymer to a coordination of a polyvalent metal ion, a salt thereof, a process for the preparation thereof and a pharmaceutical agent comprising the same.
摘要翻译：通过使聚亚乙基多胺或聚乙烯亚胺与式YAZ表示的双官能化合物进行反应，其中Y和Z相同或不同，为-CH 2 X，-CH（OH）CH 2 X，得到含有季氮原子的新型聚合物 - 金属配合物或基团，A是单键， - （CH 2）m - ， - CH 2 -O-CH 2 - 或-CH 2 -O-（CH 2）k O-CH 2 - 基，X是氯，溴或碘原子，m为1〜3的整数，k为1〜4的整数，然后使所得交联聚合物与多价金属离子，其盐，其制备方法和包含该药剂的药剂。

8. 发明授权

US5061691A Enkephalin analogs 失效
标题翻译： ENKEPHALIN ANALOGS
公开(公告)号：US5061691A
公开(公告)日：1991-10-29
申请号：US235867
申请日：1988-08-23
申请人： Kunio Yagi , Yasuyuki Shimohigashi , Hiroaki Kodama , Tomio Ogasawara , Takuya Koshizaka , Masayasu Kurono
发明人： Kunio Yagi , Yasuyuki Shimohigashi , Hiroaki Kodama , Tomio Ogasawara , Takuya Koshizaka , Masayasu Kurono
IPC分类号： A61K38/00 , C07K14/70
CPC分类号： C07K14/702 , A61K38/00
摘要： The invention relates to enkephalin analogs of the formula ##STR1## wherein in case of X is D-Val, D-Phe, Pro, D-Met, D-Met(O), D-Leu, D-Glu, D-Glu(Obzl), D-Lys, D-Lys(Z), or D-Arg,Y is Gly or Phe; andR isa) a direct bond,b) an alkylene group having from 1 to 6 carbon atoms,c) o-, m-, or p-phenylene group,d) a cycloalkane; orwherein in case ofX is D-Ala,Y is Phe; andR isa) a direct bond,b) an alkylene group having from 1 to 6 carbon atoms,c) o-, m-, or p-phenylene group, ord) a cycloalkane,its salts and hydrates, as well as a therapeutic composition containing at least one of the enkephalin analogs, as an effective component.

9. 发明授权

US4937342A Intermediates for synthesizing BH.sub.4 and its derivatives 失效
标题翻译：用于合成BH4及其衍生物的中间体
公开(公告)号：US4937342A
公开(公告)日：1990-06-26
申请号：US277109
申请日：1988-11-29
申请人： Masayasu Kurono , Takehiko Susuki , Tomio Ogasawara , Nobuko Ohishi , Kunio Yagi
发明人： Masayasu Kurono , Takehiko Susuki , Tomio Ogasawara , Nobuko Ohishi , Kunio Yagi
IPC分类号： C07D405/12 , C07D475/04 , C12P25/00
CPC分类号： C07D405/12 , C07D475/04 , C12P25/00
摘要： The present invention relates to intermediates for synthesizing BH.sub.4 and derivatives thereof. The intermediates are shown as follows: ##STR1## wherein R.sub.1 is a hydrogen atom, alkyl, aralkyl, or aryl group; R.sub.2 is an alkyl, hydroxyalkyl, or polyhydroxyalkyl group; R.sub.3 and R.sub.4 are the same or different and represent alkyl, aralkyl, or aryl group; R.sub.5, R.sub.6, R.sub.7, and R.sub.8 are the same or different and represent a hydrogen atom or acyl group; R.sub.9 is an alkyl, aralkyl, or aryl group; R.sub.10 and R.sub.11 are the same or different and represent a hydrogen atom or acyl group; n is an integer of 5 or less; and HX is an acid. The invention also relates to a process for the preparation of L-biopterin.

10. 发明授权

US4906477A Antineoplastic agent-entrapping liposomes 失效
标题翻译：抗肿瘤药物包埋脂质体
公开(公告)号：US4906477A
公开(公告)日：1990-03-06
申请号：US153302
申请日：1988-02-08
申请人： Masayasu Kurono , Hitoshi Noda , Tomio Ogasawara , Hidefumi Yamakawa , Takafumi Iida , Kunio Yagi
发明人： Masayasu Kurono , Hitoshi Noda , Tomio Ogasawara , Hidefumi Yamakawa , Takafumi Iida , Kunio Yagi
IPC分类号： A61K9/10 , A61K9/127 , A61K31/70 , A61P35/00
CPC分类号： A61K9/1272 , Y10T428/2984
摘要： Some of antineoplastic agents, for instance, adriamycin and daunorubicin, have a positive charge at physiological pH. Such drugs possess high affinity for binding to steroidal sulfate by means of electrostatic force. The invention utilizes this property to prepare steroidal sulfate liposomes having a net negative charge and encapsulating the antineoplastic agent.

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