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    Modification of DNA and oligonucleotides using metal complexes of polyaza ligands
    1.
    发明授权
    Modification of DNA and oligonucleotides using metal complexes of polyaza ligands 失效
    使用多加配体的金属络合物修饰DNA和寡核苷酸

    公开(公告)号：US5504075A

    公开(公告)日：1996-04-02

    申请号：US128308

    申请日：1993-09-28

    申请人： Cynthia J. Burrows ,  Steven E. Rokita ,  Xiaoying Chen

    发明人： Cynthia J. Burrows ,  Steven E. Rokita ,  Xiaoying Chen

    IPC分类号： C12Q1/68 ,  A01N55/02

    CPC分类号： C12Q1/68

    摘要： A process is provided for oxidatively modifying nucleic acids containing a target nucleotide. The nucleic acid is contacted with a polyaza metal complex in the presence of an oxidizing agent so that the nucleic acid is modified at or near the target nucleotide. Also provided are a kit for carrying out the process and a method for treating neoplastic growth by administering to a subject having neoplastic growth, an effective amount of a polyaza metal complex which is capable of modifying DNA.

    摘要翻译： 提供了用于氧化修饰含有靶核苷酸的核酸的方法。 核酸在氧化剂存在下与多金属络合物接触，使核酸在靶核苷酸处或其附近被修饰。 还提供了用于实施该方法的试剂盒和通过向具有肿瘤生长的受试者施用能够修饰DNA的有效量的多金属络合物来治疗肿瘤生长的方法。

    Modification of DNA and oligonucleotides using metal complexes of polyaza ligands
    3.
    发明授权
    Modification of DNA and oligonucleotides using metal complexes of polyaza ligands 失效
    使用多加配体的金属络合物修饰DNA和寡核苷酸

    公开(公告)号：US5272056A

    公开(公告)日：1993-12-21

    申请号：US637163

    申请日：1991-01-03

    申请人： Cynthia J. Burrows ,  Steven E. Rokita ,  Xiaoying Chen

    发明人： Cynthia J. Burrows ,  Steven E. Rokita ,  Xiaoying Chen

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/68

    摘要： A process is provided for oxidatively modifying nucleic acids containing a target nucleotide. The nucleic acid is contacted with a polyaza metal comlex in the presence of an oxidizing agent so that the nucleic acid is modified at or near the target nucleotide. Also provided are a kit for carrying out the process and a method for treating neoplastic growth by administering to a subject having neoplastic growth, an effective amount of a polyaza metal complex which is capable of modifying DNA.

    摘要翻译： 提供了用于氧化修饰含有靶核苷酸的核酸的方法。 核酸在氧化剂的存在下与多金属金属comlex接触，使得核酸在靶核苷酸处或其附近被修饰。 还提供了用于实施该方法的试剂盒和通过向具有肿瘤生长的受试者施用能够修饰DNA的有效量的多金属络合物来治疗肿瘤生长的方法。

    Ion triggered alkylation of biological targets by silyloxy aromatic agents
    5.
    发明授权
    Ion triggered alkylation of biological targets by silyloxy aromatic agents 失效
    离子触发由甲硅烷氧基芳香剂引发生物靶标的烷基化

    公开(公告)号：US5493012A

    公开(公告)日：1996-02-20

    申请号：US71116

    申请日：1993-06-02

    申请人： Steven E. Rokita ,  Tianhu Li ,  Qingping Zeng

    发明人： Steven E. Rokita ,  Tianhu Li ,  Qingping Zeng

    IPC分类号： C07D207/404 ,  C07F7/18 ,  C07H19/04 ,  C07H21/00 ,  C07H23/00 ,  C07H21/04

    CPC分类号： C07D207/404 ,  C07F7/1852 ,  C07F7/1856 ,  C07H21/00 ,  C07H23/00

    摘要： A silyloxy aromatic derivative capable of alkylating a target biological molecule when activated by ionic strength. A sequence directed reagent may be constructed by conjugating a methyl silyloxy aromatic derivative to a hexamethyiamino linker attached to either the 5' or 3' terminus of an oligonucleotide. Annealing this modified fragment of DNA to its complementary sequence allows for target modification subsequent to ionic activation. The product of this reaction is a covalent crosslink between the reagent and target strands resulting from an alkylation of DNA by the activated silyloxy aromatic derivative. In a preferred embodiment, a nitrophenyl or bromo group is attached to a methyl group of the silyloxy aromatic derivative. This reagent may be similarly linked to an oligonucleotide probe. Activation of the alkylating agent by an ionic signal (X) which may naturally occur, or may be introduced into the media containing the target molecule, such as by the introduction of a salt (MX).

    摘要翻译： 当通过离子强度活化时，能够将靶生物分子烷基化的甲硅烷氧基芳族衍生物。 序列导向试剂可以通过将甲基甲硅烷氧基芳族衍生物与连接于寡核苷酸的5'或3'末端的六甲基氨基接头缀合来构建。 将该修饰的DNA片段退火至其互补序列允许在离子活化后进行靶修饰。 该反应的产物是由活化的甲硅烷氧基芳族衍生物的DNA烷基化产生的试剂和靶链之间的共价交联。 在优选的实施方案中，硝基苯基或溴基连接到甲硅烷氧基芳族衍生物的甲基上。 该试剂可以类似地连接到寡核苷酸探针。 通过离子信号（X）活化烷基化剂，其可以天然存在，或者可以通过引入盐（MX）引入含有靶分子的介质中。

    Ion triggered alkylation of biological targets by silyloxy aromatic agents
    6.
    发明授权
    Ion triggered alkylation of biological targets by silyloxy aromatic agents 失效
    离子触发由甲硅烷氧基芳香剂引发生物靶标的烷基化

    公开(公告)号：US5296350A

    公开(公告)日：1994-03-22

    申请号：US606463

    申请日：1990-10-31

    申请人： Steven E. Rokita ,  Tianhu Li

    发明人： Steven E. Rokita ,  Tianhu Li

    IPC分类号： C07F7/18 ,  C07H21/00 ,  C07H23/00 ,  C12Q1/68

    CPC分类号： C07H21/00 ,  C07F7/1852 ,  C07F7/1856 ,  C07H23/00

    摘要： A sequence directed reagent is constructed by conjugating a methyl silyloxy aromatic derivative to a hexamethylamino linker attached to either the 5' or 3' terminus of an oligonucleotide. Annealing this modified fragment of DNA to its complementary sequence allows for target modification subsequent to ionic activation. The product of this reaction is a covalent crosslink between the reagent and target strands resulting from an alkylation of DNA by the activated silyloxy aromatic derivative. In a preferred embodiment, a nitrophenyl group is attached to the methyl group of the silyloxy aromatic derivative. This reagent is similarly linked to an oligonucleotide probe. Activation of this probe linked alkylating agent by an ionic signal, (X) which may naturally occur, or may be introduced into the media containing the target molecule, such as by the introduction of a salt (MX).

    摘要翻译： 通过将甲基甲硅烷氧基芳族衍生物与连接到寡核苷酸的5'或3'末端的六甲基氨基接头缀合来构建顺序导向的试剂。 将该修饰的DNA片段退火至其互补序列允许在离子活化后进行靶修饰。 该反应的产物是由活化的甲硅烷氧基芳族衍生物的DNA烷基化产生的试剂和靶链之间的共价交联。 在优选的实施方案中，硝基苯基与甲硅烷氧基芳族衍生物的甲基连接。 该试剂类似地与寡核苷酸探针连接。 通过离子信号（X）活化该探针连接的烷基化剂，例如通过引入盐（MX），可天然存在或可以引入含有靶分子的介质中。

    Nucleic acids labeled with naphthoquinone probe
    7.
    发明授权
    Nucleic acids labeled with naphthoquinone probe 失效
    用萘醌探针标记的核酸

    公开(公告)号：US5292873A

    公开(公告)日：1994-03-08

    申请号：US442947

    申请日：1989-11-29

    申请人： Steven E. Rokita ,  Moneesh Chatterjee

    发明人： Steven E. Rokita ,  Moneesh Chatterjee

    IPC分类号： C07H21/00

    CPC分类号： C07H21/00

    摘要： A sequence directed reagent is constructed by conjugating a methyl naphthoquinone derivative to a hexamethylamino linker attached to the 5' terminus of an oligonucleotide. Annealing this modified fragment of DNA to its complementary sequence allows for target modification subsequent to photochemical activation. The product of this reaction is a covalent crosslink between the reagent and target strands resulting from an alkylation of DNA by the photoexcited quinone. The modified sequence is not labile to acid, base or reductants, and blocks the exonuclease activity of the Klenow fragment of DNA polymerase I. In another embodiment, a highly reactive moiety, such as Br is attached to the methyl group of the naphthoquinone derivative. This reagent is similarly linked to an oligonucleotide probe. Activation of this probe linked alkylating agent is by a reductive signal which may either naturally occur within the cell, such as an enzyme, or introduced into the media containing the target molecule.

    摘要翻译： 通过将甲基萘醌衍生物与连接到寡核苷酸的5'末端的六甲基氨基接头缀合来构建顺序导向的试剂。 将该修饰的DNA片段退火至其互补序列允许在光化学活化之后的靶修饰。 该反应的产物是由光激发的醌对DNA产生的试剂和靶链之间的共价交联。 修饰的序列对于酸，碱或还原剂不是不稳定的，并且阻断DNA聚合酶I的Klenow片段的核酸外切酶活性。在另一个实施方案中，诸如Br的高反应性部分连接到萘醌衍生物的甲基上。 该试剂类似地与寡核苷酸探针连接。 该探针连接的烷基化剂的活化是通过还原信号，其可以天然存在于细胞内，例如酶，或引入含有靶分子的培养基中。

    Reactive anthraquinone derivatives and conjugates thereof
    8.
    发明授权
    Reactive anthraquinone derivatives and conjugates thereof 失效
    活性蒽醌衍生物及其偶联物

    公开(公告)号：US5650399A

    公开(公告)日：1997-07-22

    申请号：US420326

    申请日：1995-04-11

    申请人： Steven E. Rokita ,  Hyunmin Kang

    发明人： Steven E. Rokita ,  Hyunmin Kang

    IPC分类号： C07H21/00 ,  C07H21/02 ,  C07H21/04

    CPC分类号： C07H21/00

    摘要： Anthraquinone derivatives are constructed for alkylation of target molecules. The anthraquinone derivative includes an anthraquinone ring system which has been modified by the addition of inducibly reactive substituents at positions on the ring system. The anthraquinone ring system can be selectively activated for alkylation of a target molecule. The substituents may be from 1 to 4 methyl groups, whereupon activation is achieved by a photochemical signal. In an additional alternative embodiment, the inducible reactive substituents may include both methyl groups and substituted methylene groups. In this embodiment, the activation of the anthraquinone derivative is achieved by photochemical and/or reductive signals. Alternatively, the substituents may be from 1 to 8 methyl groups modified to include a reactive group, whereupon activation is achieved by a chemical or enzymatic reductive signal. The anthraquinone derivatives of the invention may be used for non-specific modification of target molecules. Alternatively, the anthraquinone derivative may be further derivatized to permit specific modification of target molecules. The further derivatization of the anthraquinone includes attaching a probe to the anthraquinone ring system by way of a linking group. The probe is chosen to selectively localize to a target molecule, whereupon the anthraquinone derivative may be activated to react with and aklylate the target molecule. Also the process of employing such anthraquinone derivatives for use in vitro or in vivo for alkylating, specifically or non-specifically alkylating, target molecules.

    摘要翻译： 构建蒽醌衍生物用于靶分子的烷基化。 蒽醌衍生物包括通过在环系上的位置添加可诱导的反应性取代基而被修饰的蒽醌环体系。 可以选择性地活化蒽醌环体系用于靶分子的烷基化。 取代基可以是1至4个甲基，因此通过光化学信号实现活化。 在另外的替代实施方案中，可诱导的反应性取代基可以包括甲基和取代的亚甲基。 在该实施方案中，通过光化学和/或还原信号实现蒽醌衍生物的活化。 或者，取代基可以是经修饰以包括反应性基团的1至8个甲基，因此通过化学或酶还原信号实现活化。 本发明的蒽醌衍生物可用于靶分子的非特异性修饰。 或者，可以将蒽醌衍生物进一步衍生化，以允许目标分子的特异性修饰。 蒽醌的进一步衍生化包括通过连接基团将探针附着到蒽醌环体系上。 选择该探针以选择性地定位于靶分子，于是该蒽醌衍生物可以被活化以与靶分子反应并使其成为碱基。 也是使用这种蒽醌衍生物在体外或体内用于烷基化，特异性或非特异性烷基化靶分子的方法。

    Ion triggered alkylation of biological targets by silyloxy aromatic agents
    10.
    发明授权
    Ion triggered alkylation of biological targets by silyloxy aromatic agents 失效
    离子触发由甲硅烷氧基芳香剂引发生物靶标的烷基化

    公开(公告)号：US5831073A

    公开(公告)日：1998-11-03

    申请号：US603221

    申请日：1996-02-20

    申请人： Steven E. Rokita ,  Tianhu Li ,  Qingping Zeng

    发明人： Steven E. Rokita ,  Tianhu Li ,  Qingping Zeng

    IPC分类号： C07D207/404 ,  C07F7/18 ,  C07H19/04 ,  C07H21/00 ,  C07H23/00

    CPC分类号： C07F7/1852 ,  C07D207/404 ,  C07F7/1856 ,  C07H21/00 ,  C07H23/00

    摘要： A silyloxy aromatic derivative capable of alkylating a target biological molecule when activated by ionic strength. A sequence directed reagent may be constructed by conjugating a methyl silyloxy aromatic derivative to a hexamethylamino linker attached to either the 5' or 3' terminus of an oligonucleotide. Annealing this modified fragment of DNA to its complementary sequence allows for target modification subsequent to ionic activation. The product of this reaction is a covalent crosslink between the reagent and target strands resulting from an alkylation of DNA by the activated silyloxy aromatic derivative. In a preferred embodiment, a nitrophenyl or bromo group is attached to a methyl group of the silyloxy aromatic derivative. This reagent may be similarly linked to an oligonucleotide probe. Activation of the alkylating agent by an ionic signal (X) which may naturally occur, or may be introduced into the media containing the target molecule, such as by the introduction of a salt (MX).

    摘要翻译： 当通过离子强度活化时，能够将靶生物分子烷基化的甲硅烷氧基芳族衍生物。 序列导向的试剂可以通过将甲基甲硅烷氧基芳族衍生物与连接到寡核苷酸的5'或3'末端的六甲基氨基接头共轭来构建。 将该修饰的DNA片段退火至其互补序列允许在离子活化后进行靶修饰。 该反应的产物是由活化的甲硅烷氧基芳族衍生物的DNA烷基化产生的试剂和靶链之间的共价交联。 在优选的实施方案中，硝基苯基或溴基连接到甲硅烷氧基芳族衍生物的甲基上。 该试剂可以类似地连接到寡核苷酸探针。 通过离子信号（X）活化烷基化剂，其可以天然存在，或者可以通过引入盐（MX）引入含有靶分子的介质中。