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    Chimeric antisense oligonucleotides and cell transfecting formulations thereof
    1.
    发明授权
    Chimeric antisense oligonucleotides and cell transfecting formulations thereof 失效
    嵌合反义寡核苷酸及其细胞转染制剂

    公开(公告)号：US06846921B2

    公开(公告)日：2005-01-25

    申请号：US09826519

    申请日：2001-04-03

    申请人： Michael A. Innis ,  Christoph J. Reinhard ,  Ronald N. Zuckermann

    发明人： Michael A. Innis ,  Christoph J. Reinhard ,  Ronald N. Zuckermann

    IPC分类号： A61K38/00 ,  A61K47/48 ,  C12N15/113 ,  C07H21/04

    CPC分类号： C12N15/113 ,  A61K38/00 ,  A61K47/54 ,  A61K47/554 ,  A61K47/6949 ,  B82Y5/00 ,  C12N2310/315 ,  C12N2310/317 ,  C12N2310/321 ,  C12N2310/346 ,  C12N2310/3515 ,  C12N2310/3521

    摘要： Chimeric oligonucleotide of the formula 5′-W—X1—Y—X2—Z-3′, where W represents a 5′-O-alkyl nucleotide, each of X1 and X2 represents a block of seven to twelve phosphodiester-linked 2′-O-alkyl ribonucleotides, Y represents a block of five to twelve phosphorothioate-linked deoxyribonucleotides, and Z represents a blocking group effective to block nuclease activity at the 3′ end of the oligonucleotide, are described. These compounds exhibit high resistance to endo- and exonucleases, high sequence specificity, and the ability to activate RNAse H, as evidenced by efficient and long-lasting suppression of target mRNA. The oligonucleotides are preferably transfected into cells in formulations containing a lipid-peptoid conjugate carrier molecule of the formula L-linker-[N(CH2CH2NH2)CH2(C═O)—N(CH2CH2R)CH2(C═O)—N(CH2CH2R)CH2(C═O)]3—NH2, where L is a lipid moiety, including a steroid, and each group R is independently selected from alkyl, aminoalkyl, and aralkyl.

    摘要翻译： 式5-WX 1 -YX 2 -Z-3'的嵌合寡核苷酸，其中W代表5'-O-烷基核苷酸，X 1和X 2各自表示 7至12个磷酸二酯连接的2'-O-烷基核糖核苷酸，Y表示5至12个硫代磷酸酯连接的脱氧核糖核苷酸的嵌段，Z表示有效阻断寡核苷酸3'末端核酸酶活性的封闭基团。 这些化合物表现出对内切核酸酶和外切核酸酶的高抗性，高序列特异性和激活RNA酶H的能力，如靶mRNA的有效和持久的抑制所证明的。 优选将寡核苷酸转染到含有脂质 - 拟肽结合物载体分子的制剂中，其中L是包含类固醇的脂质部分，每个基团R独立地选自烷基，氨基烷基和芳烷基。

    Chimeric antisense oligonucleotides and cell transfecting formulations thereof
    2.
    发明授权
    Chimeric antisense oligonucleotides and cell transfecting formulations thereof 有权
    嵌合反义寡核苷酸及其细胞转染制剂

    公开(公告)号：US06677445B1

    公开(公告)日：2004-01-13

    申请号：US09648254

    申请日：2000-08-25

    申请人： Michael A. Innis ,  Christoph J. Reinhard ,  Ronald N. Zuckermann

    发明人： Michael A. Innis ,  Christoph J. Reinhard ,  Ronald N. Zuckermann

    IPC分类号： C07H2104

    CPC分类号： C12N15/113 ,  A61K38/00 ,  A61K47/54 ,  A61K47/554 ,  A61K47/6949 ,  B82Y5/00 ,  C12N2310/315 ,  C12N2310/317 ,  C12N2310/321 ,  C12N2310/346 ,  C12N2310/3515 ,  C12N2310/3521

    摘要： Chimeric oligonucleotide of the formula 5′-W-X1-Y-X2-Z-3′, where W represents a 5′-O-alkyl nucleotide, each of X1 and X2 represents a block of seven to twelve phosphodiester-linked 2′-O-alkyl ribonucleotides, Y represents a block of five to twelve phosphorothioate-linked deoxyribonucleotides, and Z represents a blocking group effective to block nuclease activity at the 3′ end of the oligonucleotide, are described. These compounds exhibit high resistance to endo- and exonucleases, high sequence specificity, and the ability to activate RNAse H, as evidenced by efficient and long-lasting suppression of target mRNA. The oligonucleotides are preferably transfected into cells in formulations containing a lipid-peptoid conjugate carrier molecule of the formula L-linker-[N(CH2CH2NH2)CH2(C═O)—N(CH2CH2R)CH2(C═O)—N(CH2CH2R)CH2(C═O)]3—NH2, where L is a lipid moiety, including a steroid, and each group R is independently selected from alkyl, aminoalkyl, and aralkyl.

    摘要翻译： 式5-WX 1 -YX 2 -Z-3'的嵌合寡核苷酸，其中W代表5'-O-烷基核苷酸，X 1和X 2各自表示 7至12个磷酸二酯连接的2'-O-烷基核糖核苷酸，Y表示5至12个硫代磷酸酯连接的脱氧核糖核苷酸的嵌段，Z表示有效阻断寡核苷酸3'端的核酸酶活性的封闭基团。 这些化合物表现出对内切核酸酶和外切核酸酶的高抗性，高序列特异性和激活RNA酶H的能力，如靶mRNA的有效和持久的抑制所证明的。 优选将寡核苷酸转染到含有脂质 - 拟肽结合物载体分子的制剂中，其中L是包含类固醇的脂质部分，每个基团R独立地选自烷基，氨基烷基和芳烷基。

    DNA expression vector and use thereof
    6.
    发明授权
    DNA expression vector and use thereof 失效
    DNA表达载体及其用途

    公开(公告)号：US5045463A

    公开(公告)日：1991-09-03

    申请号：US287688

    申请日：1988-12-19

    申请人： Michael A. Innis ,  David H. Gelfand ,  James H. Meade

    发明人： Michael A. Innis ,  David H. Gelfand ,  James H. Meade

    IPC分类号： C12N9/34 ,  C12N15/10 ,  C12N15/62 ,  C12N15/70 ,  C12N15/81 ,  C12P7/06

    CPC分类号： C12N9/2428 ,  C12N15/10 ,  C12N15/625 ,  C12N15/70 ,  C12N15/81 ,  C12P7/06 ,  C07K2319/02 ,  C07K2319/036 ,  Y02E50/17

    摘要： A gene having a DNA sequence complementary to that of the glucoamylase polypeptide mRNA from a fungal species, preferably Aspergillus awamori, is prepared. The mRNA is an approximately 2.2 kilobase poly A RNA obtained from fungal cells grown under conditions of glucoamylase induction. Reverse transcription of the mRNA provides a glucoamylase probe used to identify genomic digest fragments containing glucoamylase gene regions, which are sequenced to locate the introns and exons. The genomic fragments are spliced together to form a gene having a DNA sequence with altered or deleted introns which codes for fungal glucoamylase protein and is capable, when correctly combined with a cleaved DNA expression vector, of expressing a non-native protein having glucoamylase enzyme activity upon transformation of a host organism by the vector. The host is preferably bacteria or yeast. The transformed yeast host may be used to produce ethanol.

    摘要翻译： 制备具有与来自真菌种，优选泡盛曲霉的葡糖淀粉酶多肽mRNA的DNA序列互补的DNA序列的基因。 mRNA是从葡糖淀粉酶诱导条件下生长的真菌细胞获得的约2.2千碱基聚A RNA。 mRNA的逆转录提供了用于鉴定含有葡糖淀粉酶基因区域的基因组消化片段的葡糖淀粉酶探针，其被测序以定位内含子和外显子。 将基因组片段拼接在一起以形成具有改变或缺失的内含子的DNA序列的基因，其编码真菌葡糖淀粉酶蛋白，并且当与切割的DNA表达载体正确组合时，能够表达具有葡糖淀粉酶酶活性的非天然蛋白质 通过载体转化宿主生物体。 宿主优选是细菌或酵母。 转化的酵母宿主可用于生产乙醇。

    Interferon-.alpha.61
    7.
    发明授权
    Interferon-.alpha.61 失效
    干扰素α61

    公开(公告)号：US4973479A

    公开(公告)日：1990-11-27

    申请号：US414054

    申请日：1982-09-02

    申请人： Michael A. Innis

    发明人： Michael A. Innis

    IPC分类号： A61K38/00 ,  C07K14/56

    CPC分类号： C07K14/56 ,  A61K38/00 ,  Y10S435/811

    摘要： A new polypeptide, called IFN-.alpha.61, produced by E. coli transformed with a newly isolated and characterized human IFN-.alpha. gene is described. The polypeptide exhibits interferon activities such as antiviral activity, cell growth regulation, and regulation of production of cell-produced substances.

    摘要翻译： 描述了用新分离和表征的人IFN-α基因转化的大肠杆菌产生的称为IFN-α61的新多肽。 多肽表现出干扰素活性，例如抗病毒活性，细胞生长调节和细胞产生物质的生产调节。

    Expression of interferon genes in Chinese hamster ovary cells
    8.
    发明授权
    Expression of interferon genes in Chinese hamster ovary cells 失效
    干扰素基因在中国仓鼠卵巢细胞中的表达

    公开(公告)号：US4966843A

    公开(公告)日：1990-10-30

    申请号：US761180

    申请日：1985-07-31

    申请人： Francis P. McCormick ,  Michael A. Innis ,  Gordon M. Ringold

    发明人： Francis P. McCormick ,  Michael A. Innis ,  Gordon M. Ringold

    IPC分类号： A61K38/00 ,  C07K14/555 ,  C07K14/56 ,  C07K14/565 ,  C12N15/20 ,  C12N15/22

    CPC分类号： C07K14/565 ,  C07K14/555 ,  C07K14/56 ,  A61K38/00 ,  Y10S435/811

    摘要： DNA constructs are prepared which operably link human interferon genes, selective, eukaryotic marker genes, and promoter and expression control sequences for the expression of human interferon in Chinese hamster ovary (CHO) cells or progeny thereof. The human recombinant interferon so produced contains glycans which are a subset of the population of glycans which are contained in the native counterpart, and may be used in therapeutic formulations. The CHO cells yield high levels of human interferon with no detectable amounts of host IFN, either constitutive or inductive.

    摘要翻译： 制备DNA构建体，其可操作地连接人干扰素基因，选择性真核标记基因，以及用于在中国仓鼠卵巢（CHO）细胞或其后代中表达人干扰素的启动子和表达控制序列。 如此制备的人重组干扰素含有聚糖，其是包含在天然对应物中的聚糖群体的子集，并且可用于治疗制剂中。 CHO细胞产生高水平的人干扰素，没有可检测量的宿主IFN，组成型或归因的。

    Chimeric proteins
    10.
    发明授权
    Chimeric proteins 失效
    嵌合蛋白

    公开(公告)号：US06783960B2

    公开(公告)日：2004-08-31

    申请号：US09741106

    申请日：2000-12-21

    申请人： Michael A. Innis ,  Abla A. Creasey

    发明人： Michael A. Innis ,  Abla A. Creasey

    IPC分类号： C07K1400

    CPC分类号： C07K14/8114 ,  A61K38/00 ,  C07K2319/00

    摘要： Chimeric proteins possessing Kunitz-type domain 1 of TFPI-2 and Kunitz-type domain 2 of TFPI are disclosed, as are muteins of TFPI and TFPI-2. Nucleic acid sequences, expression vectors and transformed host cells encoding and capable of producing the disclosed chimeric proteins and muteins are also disclosed. Finally, methods for prevention and treatment of septic shock using the chimeric proteins and muteins are disclosed.

    摘要翻译： 公开了具有TFPI-2的Kunitz型结构域1和TFPI的Kunitz型结构域2的嵌合蛋白质，以及TFPI和TFPI-2的突变蛋白。 还公开了核酸序列，表达载体和编码并能够产生所公开的嵌合蛋白和突变蛋白的转化宿主细胞。 最后，公开了使用嵌合蛋白质和突变蛋白质预防和治疗败血性休克的方法。