公开(公告)号：US07928236B2

公开(公告)日：2011-04-19

申请号：US11916061

申请日：2006-06-08

申请人： Masumi Taki ,  Masahiko Sisido

发明人： Masumi Taki ,  Masahiko Sisido

IPC分类号： C07D221/18 ,  A61K38/02

CPC分类号： C09B15/00 ,  C07K14/005 ,  C09B5/36 ,  C12N2740/16322 ,  Y02P20/55

摘要： The subject of the present invention is to provide a fluorescent substance excitable under visible light, having higher photostability and a long fluorescence lifetime. Another subject is to provide a fluorescent substance consists of a non-natural amino acid applicable to peptide synthesis systems.Searching fluorescent substances, which are fluorescent amino acids and excitable under visible light, having the lowest possible molecular weight for a high photostability resulted in forming a condensed polycyclic aromatic compound by subjecting a compound having an acridone structure to substitution with an amino acid and further condensation with a benzene ring. Thus, the subject is achieved by the fluorescent substance consisting of amino acid-substituted benzoacridone derivative excitable under visible light.

摘要翻译： 本发明的主题是提供在可见光下可激发的荧光物质，具有较高的光稳定性和较长的荧光寿命。 另一个主题是提供由适用于肽合成系统的非天然氨基酸组成的荧光物质。 在具有高光稳定性的情况下，以具有最低可能分子量的荧光氨基酸和可见光激发的荧光物质搜索导致通过使具有吖啶酮结构的化合物用氨基酸取代并进一步缩合形成稠合多环芳族化合物 与苯环。 因此，通过由可见光可激发的氨基酸取代的苯并吖啶酮衍生物构成的荧光物质来实现。

公开(公告)号：US07847098B2

公开(公告)日：2010-12-07

申请号：US12306784

申请日：2007-07-03

申请人： Masumi Taki ,  Masahiko Sisido

发明人： Masumi Taki ,  Masahiko Sisido

IPC分类号： C07D213/00 ,  A61K38/00

CPC分类号： C07D219/06 ,  C09K11/06 ,  C09K2211/1029

摘要： The present invention provides a fluorescent amino acid derivative which can be synthesized by simple steps, can be excited particularly by a blue laser ray region of visible light, and has an improved light stability. These objects can be achieved by a fluorescent amino acid derivative which is an acridone derivative substituted with an amino acid to comprise an electrophilic substituent group between the amino acid and the acridone derivative. Instead of a conventional strategy that aminophenylalanine is used as a starting material to form a fluorescent group through a coupling reaction and an intramolecular cyclization reaction, a fluorescent acridone derivative is used as a starting material to furnish the material to a reactive group by a position-specific electrophilic substitution reaction, and then the acridone derivative having the reactive group is allowed to couple with an amino acid derivative.

摘要翻译： 本发明提供了可以通过简单的步骤合成的荧光氨基酸衍生物，特别可以通过可见光的蓝色激光射线区域被激发，并且具有改善的光稳定性。 这些目的可以通过荧光氨基酸衍生物来实现，其是由氨基酸取代的吖啶酮衍生物，其在氨基酸和吖啶酮衍生物之间包含亲电取代基。 代替使用氨基苯丙氨酸作为通过偶联反应和分子内环化反应形成荧光基团的起始材料的常规策略，使用荧光吖啶酮衍生物作为原料，通过位置 - 取代基将该材料提供给反应性基团， 特异性亲电取代反应，然后使具有反应性基团的吖啶酮衍生物与氨基酸衍生物偶联。

公开(公告)号：US20100152417A1

公开(公告)日：2010-06-17

申请号：US11916061

申请日：2006-06-08

申请人： Masumi Taki ,  Masahiko Sisido

发明人： Masumi Taki ,  Masahiko Sisido

IPC分类号： C07K2/00 ,  C07D221/18 ,  C07K14/00

CPC分类号： C09B15/00 ,  C07K14/005 ,  C09B5/36 ,  C12N2740/16322 ,  Y02P20/55

摘要： The subject of the present invention is to provide a fluorescent substance excitable under visible light, having higher photostability and a long fluorescence lifetime. Another subject is to provide a fluorescent substance consists of a non-natural amino acid applicable to peptide synthesis systems.Searching fluorescent substances, which are fluorescent amino acids and excitable under visible light, having the lowest possible molecular weight for a high photostability resulted in forming a condensed polycyclic aromatic compound by subjecting a compound having an acridone structure to substitution with an amino acid and further condensation with a benzene ring. Thus, the subject is achieved by the fluorescent substance consisting of amino acid-substituted benzoacridone derivative excitable under visible light.

摘要翻译： 本发明的主题是提供在可见光下可激发的荧光物质，具有较高的光稳定性和较长的荧光寿命。 另一个主题是提供由适用于肽合成系统的非天然氨基酸组成的荧光物质。 在具有高光稳定性的情况下，以具有最低可能分子量的荧光氨基酸和可见光激发的荧光物质搜索导致通过使具有吖啶酮结构的化合物用氨基酸取代并进一步缩合形成稠合多环芳族化合物 与苯环。 因此，通过由可见光可激发的氨基酸取代的苯并吖啶酮衍生物构成的荧光物质来实现。

公开(公告)号：US20090240026A1

公开(公告)日：2009-09-24

申请号：US12306784

申请日：2007-07-03

申请人： Masumi Taki ,  Masahiko Sisido

发明人： Masumi Taki ,  Masahiko Sisido

IPC分类号： C07K2/00 ,  C07D219/00

CPC分类号： C07D219/06 ,  C09K11/06 ,  C09K2211/1029

摘要： An object of the present invention is to provide a novel fluorescent amino acid derivative which can be synthesized by simple steps, can be excited particularly by a blue laser ray region of visible light, and has an improved light stability. Another object is to provide a production method of the fluorescent amino acid derivative. These objects can be achieved by a fluorescent amino acid derivative which is an acridone derivative substituted with an amino acid to comprise an electrophilic substituent group between the amino acid and the acridone derivative. Instead of a conventional strategy that aminophenylalanine is used as a starting material to form a fluorescent group through a coupling reaction and an intramolecular cyclization reaction, a fluorescent acridone derivative is used as a starting material to furnish the material to a reactive group by a position-specific electrophilic substitution reaction, and then the acridone derivative having the reactive group is allowed to couple with an amino acid derivative.

摘要翻译： 本发明的目的是提供可以通过简单的步骤合成的新的荧光氨基酸衍生物，特别可以通过可见光的蓝色激光射线区域被激发，并且具有改善的光稳定性。 另一个目的是提供荧光氨基酸衍生物的制备方法。 这些目的可以通过荧光氨基酸衍生物来实现，其是由氨基酸取代的吖啶酮衍生物，其在氨基酸和吖啶酮衍生物之间包含亲电取代基。 代替使用氨基苯丙氨酸作为通过偶联反应和分子内环化反应形成荧光基团的起始材料的常规策略，使用荧光吖啶酮衍生物作为原料，通过位置 - 取代基将该材料提供给反应性基团， 特异性亲电取代反应，然后使具有反应性基团的吖啶酮衍生物与氨基酸衍生物偶联。

公开(公告)号：US07972816B2

公开(公告)日：2011-07-05

申请号：US10567168

申请日：2004-08-09

申请人： Yasuomi Takagi ,  Kazunari Taira ,  Masumi Taki ,  Yoshio Kato ,  Makoto Miyagishi

发明人： Yasuomi Takagi ,  Kazunari Taira ,  Masumi Taki ,  Yoshio Kato ,  Makoto Miyagishi

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/04

CPC分类号： C12N15/66 ,  A61K31/711 ,  A61K31/713 ,  C12N15/10 ,  C12N15/111 ,  C12N2310/11 ,  C12N2310/53 ,  C12N2330/30

摘要： The present invention provides a simple method for producing a dumbbell-shaped DNA.A method for producing a dumbbell-shaped DNA, wherein each of sense and antisense strands is connected at both the 5′ and 3′ ends of a linear-shaped double stranded DNA by a single stranded DNA of loop structure, comprising the steps of; 1) amplifying a target DNA in a template DNA by PCR using sense and antisense primers, wherein each of the sense and antisense primers contains the following sequence (a) at the 5′ end and also contains the following sequences (b), (c), and (d) in order from the 5′ end to the 3′ end, (a) a part of a sense sequence of a nickase recognition sequence, comprising the sequence of a region between the site where a nick is introduced by the action of a nickase and the 3′ end, (b) a sequence capable of forming a loop structure from a single strand, (c) the entire antisense sequence of the nickase recognition sequence (a), (d) a sequence complementary to all or part of the sequence of the target DNA; 2) treating the amplified DNA product of step 1) with a nickase of (a); 3) heating and then annealing the nickase treated amplified DNA product of step 2); and 4) treating the heated and annealed amplified DNA product of step 3) with DNA ligase, wherein the sense and antisense primers used in step 1) are phosphorylated at the 5′ end, or the amplified DNA product is phosphorylated at the 5′ end after step 1) but before step 4).

摘要翻译： 本发明提供了一种制备哑铃型DNA的简单方法。 一种制备哑铃型DNA的方法，其中有义链和反义链中的每一个通过环结构的单链DNA连接在线形双链DNA的5'和3'末端，包括以下步骤： 1）使用正义和反义引物通过PCR扩增模板DNA中的靶DNA，其中有义和反义引物中的每一个在5'末端包含以下序列（a），并且还包含以下序列（b），（c ）和（d）从5'末端到3'末端的顺序，（a）切口酶识别序列的有义序列的一部分，其包含在切口被引入的位点之间的区域的序列 切口酶和3'端的作用，（b）能够从单链形成环结构的序列，（c）切口酶识别序列（a）的整个反义序列，（d）与所有的互补序列互补的序列 或靶DNA的序列的一部分; 2）用（a）的切口酶处理步骤1）的扩增的DNA产物; 3）加热然后对步骤2）的经切割处理的扩增的DNA产物进行退火; 和4）用DNA连接酶处理步骤3）的加热和退火的扩增DNA产物，其中步骤1）中使用的有义和反义引物在5'末端被磷酸化，或者扩增的DNA产物在5'末端被磷酸化 在步骤1之后），但在步骤4之前）。

公开(公告)号：US20080153763A1

公开(公告)日：2008-06-26

申请号：US10567168

申请日：2004-08-09

申请人： Yasuomi Takagi ,  Kazunari Taira ,  Masumi Taki ,  Yoshio Kato ,  Makoto Miyagishi

发明人： Yasuomi Takagi ,  Kazunari Taira ,  Masumi Taki ,  Yoshio Kato ,  Makoto Miyagishi

IPC分类号： A61K31/711 ,  C12P19/34 ,  C12N15/64 ,  C07H21/04

CPC分类号： C12N15/66 ,  A61K31/711 ,  A61K31/713 ,  C12N15/10 ,  C12N15/111 ,  C12N2310/11 ,  C12N2310/53 ,  C12N2330/30

摘要： The present invention provides a simple method for producing a dumbbell-shaped DNA.A method for producing a dumbbell-shaped DNA, wherein each of sense and antisense strands is connected at both the 5′ and 3′ ends of a linear-shaped double stranded DNA by a single stranded DNA of loop structure, comprising the steps of; 1) amplifying a target DNA in a template DNA by PCR using sense and antisense primers, wherein each of the sense and antisense primers contains the following sequence (a) at the 5′ end and also contains the following sequences (b), (c), and (d) in order from the 5′ end to the 3′ end, (a) a part of a sense sequence of a nickase recognition sequence, comprising the sequence of a region between the site where a nick is introduced by the action of a nickase and the 3′ end, (b) a sequence capable of forming a loop structure from a single strand, (c) the entire antisense sequence of the nickase recognition sequence (a), (d) a sequence complementary to all or part of the sequence of the target DNA; 2) treating the amplified DNA product of step 1) with a nickase of (a); 3) heating and then annealing the nickase treated amplified DNA product of step 2); and 4) treating the heated and annealed amplified DNA product of step 3) with DNA ligase, wherein the sense and antisense primers used in step 1) are phosphorylated at the 5′ end, or the amplified DNA product is phosphorylated at the 5′ end after step 1) but before step 4).

摘要翻译： 本发明提供了一种制备哑铃型DNA的简单方法。 一种制备哑铃型DNA的方法，其中有义链和反义链中的每一个通过环结构的单链DNA连接在线形双链DNA的5'和3'末端，包括以下步骤： 1）使用正义和反义引物通过PCR扩增模板DNA中的靶DNA，其中有义和反义引物中的每一个在5'末端包含以下序列（a），并且还包含以下序列（b），（c ）和（d）从5'末端到3'末端的顺序，（a）切口酶识别序列的有义序列的一部分，其包含在切口被引入的位点之间的区域的序列 切口酶和3'端的作用，（b）能够从单链形成环结构的序列，（c）切口酶识别序列（a）的整个反义序列，（d）与所有的互补序列互补的序列 或靶DNA的序列的一部分; 2）用（a）的切口酶处理步骤1）的扩增的DNA产物; 3）加热然后对步骤2）的经切割处理的扩增的DNA产物进行退火; 和4）用DNA连接酶处理步骤3）的加热和退火的扩增DNA产物，其中步骤1）中使用的有义和反义引物在5'末端被磷酸化，或者扩增的DNA产物在5'末端被磷酸化 在步骤1之后），但在步骤4之前）。