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    Lipids for transfection of Eukaryotic cells
    1.
    发明授权
    Lipids for transfection of Eukaryotic cells 有权
    用于转染真核细胞的脂质

    公开(公告)号：US08759499B2

    公开(公告)日：2014-06-24

    申请号：US13073972

    申请日：2011-03-28

    申请人： Joel Jessee ,  Gulilat Gebeyehu

    发明人： Joel Jessee ,  Gulilat Gebeyehu

    IPC分类号： C07D487/00 ,  A61K48/00 ,  A61K51/00 ,  A61K36/14 ,  A01N63/00

    CPC分类号： C07K14/005 ,  A61K47/6455 ,  C12N15/87 ,  C12N2720/12022

    摘要： Compositions and methods for improved delivery of macromolecules into eukaryotic cells are provided. Fusogenic peptides from fusion proteins of non-enveloped viruses enhance the efficiency of transfection of eukaryotic cells mediated by transfection agents such as cationic lipids, polycationic polymers such as PEI and dendrimers. These fusogenic peptides are used as part of a transfection complex that efficiently delivers a macromolecule, for example, a nucleic acid, into a eukaryotic cell. Novel cationic lipids and compositions of cationic lipids also are provided that may be used for the introduction of macromolecules such as nucleic acids, proteins and peptides into a variety of cells and tissues. The lipids can be used alone, in combination with other lipids and/or in combination with fusogenic peptides to prepare transfection complexes.

    摘要翻译： 提供了将大分子转运到真核细胞中的组合物和方法。 来自非包膜病毒融合蛋白的融合肽提高了转染试剂如阳离子脂质，聚阳离子聚合物如PEI和树枝状大分子介导的真核细胞的转染效率。 这些融合肽用作转染复合物的一部分，其将大分子例如核酸有效地递送到真核细胞中。 还提供了新的阳离子脂质和阳离子脂质的组合物，其可用于将大分子如核酸，蛋白质和肽引入多种细胞和组织中。 脂质可以单独使用，与其他脂质组合使用和/或与融合肽组合使用以制备转染复合物。

    Transfection reagents
    3.
    发明授权
    Transfection reagents 有权
    转染试剂

    公开(公告)号：US07915450B2

    公开(公告)日：2011-03-29

    申请号：US12638336

    申请日：2009-12-15

    申请人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    发明人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    IPC分类号： C07C211/63 ,  C07C233/02 ,  C12N15/88 ,  A61K31/225 ,  A61K31/16 ,  A61K31/14

    CPC分类号： A61K47/48046 ,  A61K9/16 ,  A61K31/14 ,  A61K38/28 ,  A61K38/40 ,  A61K47/543 ,  C07C211/21 ,  C07C211/63 ,  C07C211/64 ,  C07C229/26 ,  C07D209/48 ,  C07D241/12 ,  C12N15/88 ,  Y10S435/81

    摘要： Disclosed are cationic lipid compounds and compositions of lipid aggregates for delivery of macromolecules and other compounds into cells. The compounds can be used alone or in combination with other compounds lo prepare liposomes and other lipid aggregates suitable for iransfcclion or delivery of compounds to target cells, cither in vitro or in vivo. The compounds are preferably polycationic and preferably form highly stable complexes with various anionic macromolecules. particularly polyanions such as nucleic acids. These compounds have the properly, when dispersed in water, of forming lipid aggregates which associate strongly, via their cationic portion, with polyanions. Also disclosed are intermediates for preparing the compound and compositions of the invention and methods of using the compounds to introduce other compounds into cells.

    摘要翻译： 公开了阳离子脂质化合物和用于将大分子和其它化合物递送到细胞中的脂质聚集体的组合物。 化合物可以单独使用或与其它化合物组合使用，以制备适合于将体外或体内化合物转运至靶细胞的脂质体和其他脂质聚集体。 该化合物优选是阳离子型的，优选与各种阴离子大分子形成高度稳定的络合物。 特别是聚阴离子如核酸。 当分散在水中时，这些化合物可以适当地形成通过其阳离子部分与聚阴离子强烈缔合的脂质聚集体。 还公开了制备本发明化合物和组合物的中间体以及使用该化合物将其它化合物引入细胞的方法。

    Novel reagents for transfection of eukaryotic cells
    4.
    发明申请
    Novel reagents for transfection of eukaryotic cells 有权
    用于转染真核细胞的新型试剂

    公开(公告)号：US20090023215A1

    公开(公告)日：2009-01-22

    申请号：US11434765

    申请日：2006-05-17

    申请人： Joel Jessee ,  Gulilat Gebeyehu

    发明人： Joel Jessee ,  Gulilat Gebeyehu

    IPC分类号： C12N15/00 ,  C07C211/09 ,  C07K2/00 ,  C07C211/13 ,  C12N15/63 ,  C07K7/00 ,  C07K4/00 ,  C07C53/00 ,  C07K14/00

    CPC分类号： C07K14/005 ,  A61K47/6455 ,  C12N15/87 ,  C12N2720/12022

    摘要： Compositions and methods for improved delivery of macromolecules into eukaryotic cells are provided. Fusogenic peptides from fusion proteins of non-enveloped viruses enhance the efficiency of transfection of eukaryotic cells mediated by transfection agents such as cationic lipids, polycationic polymers such as PEI and dendrimers. These fusogenic peptides are used as part of a transfection complex that efficiently delivers a macromolecule, for example, a nucleic acid, into a eukaryotic cell. Novel cationic lipids and compositions of cationic lipids also are provided that may be used for the introduction of macromolecules such as nucleic acids, proteins and peptides into a variety of cells and tissues. The lipids can be used alone, in combination with other lipids and/or in combination with fusogenic peptides to prepare transfection complexes.

    摘要翻译： 提供了将大分子转运到真核细胞中的组合物和方法。 来自非包膜病毒融合蛋白的融合肽提高了转染试剂如阳离子脂质，聚阳离子聚合物如PEI和树枝状大分子介导的真核细胞的转染效率。 这些融合肽用作转染复合物的一部分，其将大分子例如核酸有效地递送到真核细胞中。 还提供了新的阳离子脂质和阳离子脂质的组合物，其可用于将大分子如核酸，蛋白质和肽引入多种细胞和组织中。 脂质可以单独使用，与其他脂质组合使用和/或与融合肽组合使用以制备转染复合物。

    Transfection reagents
    5.
    发明授权
    Transfection reagents 有权

    公开(公告)号：US07470817B2

    公开(公告)日：2008-12-30

    申请号：US11617625

    申请日：2006-12-28

    申请人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    发明人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    IPC分类号： C07C233/00 ,  C07C235/00 ,  C07C237/00 ,  C07C239/00 ,  C07C277/00 ,  C07C211/00 ,  A61K31/13 ,  A61K9/127 ,  A61K9/50 ,  C12N15/86

    CPC分类号： A61K47/48046 ,  A61K9/16 ,  A61K31/14 ,  A61K38/28 ,  A61K38/40 ,  A61K47/543 ,  C07C211/21 ,  C07C211/63 ,  C07C211/64 ,  C07C229/26 ,  C07D209/48 ,  C07D241/12 ,  C12N15/88 ,  Y10S435/81

    摘要： Disclosed are compounds capable of facilitating transport of biologically active agents or substances into cells having the general structure: wherein Q is selected from the group consisting of N, O and S; L is any bivalent organic radical capable of linking each Q, such as C, CH, (CH2)I, or {(CH2)i-Y—(CH2)j}k, wherein Y is selected from the group consisting of CH2, an ether, a polyether, an amide, a polyamide, an ester, a sulfide, a urea, a thiourea, a guanidyl, a carbamoyl, a carbonate, a phosphate, a sulfate, a sulfoxide, an imine, a carbonyl, and a secondary amino group and wherein Y is optionally substituted by —X1-L′-X2-Z or -Z; R1-R6, independently of one another, are selected from the group consisting of H, —(CH2)p-D-Z, an alkyl, an alkenyl, an aryl, and an alkyl or alkyl ether optionally substituted by one or more of an alcohol, an aminoalcohol, an amine, an amide, an ether, a polyether, a polyamide, an ester, a mercaptan, an alkylthio, a urea, a thiourea, a guanidyl, or a carbamoyl group, and wherein at least one of R1, R3, R4 and R6 is a straight chain or branched, cyclic, alkyl, alkenyl, alkynyl or aryl group; and anyone of R1, R3, R4 and/or R6 may optionally be covalently linked with each other, with Y or with L when L is C or CH to form a cyclic moiety; Z is selected from the group consisting of amine, spermiyl, carboxyspermiyl, guanidyl, spermidinyl, putricinyl, diaminoalkyl, pyridyl, piperidinyl, pyrrolidinyl, polyamine, amino acid, peptide, and protein; X1 and X2, independently of one another, are selected from the group consisting of NH, O, S, alkylene, and arylene; L′ is selected from the group consisting of alkylene, alkenylene, alkynylene, arylene, alkylene ether, and polyether; D is Q or a bond; A1 and A2, independently of one another, are selected from the group consisting of CH2O, CH2S, CH2NH, C(O), C{NH), C(S) and (CH2)t; X is a physiologically acceptable anion; m, n, r, s, u, v, w and y are 0 or 1, with the proviso that when both m and n are 0 at least one of r, s, u and y is other than 0; i, j, k, l, p and are integers from 0 to about 100; q is an integer from 1 to about 1000; and a is the number of positive charge divided by the valence of the anion.

    Methods for producing polypeptides through enhanced synthesis of encoding nucleic acid molecules
    6.
    发明授权
    Methods for producing polypeptides through enhanced synthesis of encoding nucleic acid molecules 有权
    通过增强编码核酸分子的合成产生多肽的方法

    公开(公告)号：US07344863B2

    公开(公告)日：2008-03-18

    申请号：US10899139

    申请日：2004-07-27

    申请人： Wu-Bo Li ,  Joel A. Jessee ,  David Schuster ,  Jiulin Xia ,  Gulilat Gebeyehu

    发明人： Wu-Bo Li ,  Joel A. Jessee ,  David Schuster ,  Jiulin Xia ,  Gulilat Gebeyehu

    IPC分类号： C12P21/00 ,  C12P19/34

    CPC分类号： C12P19/34 ,  C07H21/00 ,  C12Q1/6848 ,  C12Q2527/125

    摘要： The present invention is directed to compositions and methods for enhancing synthesis of nucleic acid molecules, particularly GC-rich nucleic acid molecules. Specifically, the invention provides compositions comprising one or more nitrogen-containing organic compounds having a formula selected from the group consisting of formula I and formula II (or salts or derivatives thereof), preferably 4-methylmorpholine N-oxide or betaine (carboxymethyltrimethylammonium), and further comprising one or more compounds selected from the group consisting of proline and an N-alkylimidazole compound, and more preferably proline, 1-methylimidazole or 4-methylimidazole. The invention further relates to methods for enhanced, high-fidelity synthesis of nucleic acid molecules, including via amplification (particularly PCR), reverse transcription, and sequencing methods. The invention also relates to nucleic acid molecules synthesized by these methods, to fragments or derivatives thereof, and to vectors and host cells comprising such nucleic acid molecules, fragments, or derivatives. The invention also relates to kits for synthesizing, amplifying, reverse transcribing or sequencing nucleic acid molecules comprising one or more of the compositions of the invention.

    摘要翻译： 本发明涉及用于增强核酸分子，特别是富含GC的核酸分子的合成的组合物和方法。 具体地说，本发明提供包含一种或多种具有选自式I和式II（或其盐或衍生物），优选4-甲基吗啉N-氧化物或甜菜碱（羧甲基三甲基铵）的式的含氮有机化合物的组合物， 并且还包含一种或多种选自脯氨酸和N-烷基咪唑化合物，更优选脯氨酸，1-甲基咪唑或4-甲基咪唑的化合物。 本发明还涉及用于核酸分子增强，高保真合成的方法，包括经由扩增（特别是PCR），逆转录和测序方法。 本发明还涉及通过这些方法合成的核酸分子，其片段或衍生物以及包含该核酸分子，片段或衍生物的载体和宿主细胞。 本发明还涉及用于合成，扩增，逆转录或测序包含一种或多种本发明组合物的核酸分子的试剂盒。

    Compositions and methods for enhanced synthesis of nucleic acid molecules
    7.
    发明授权
    Compositions and methods for enhanced synthesis of nucleic acid molecules 有权
    用于增强核酸分子合成的组合物和方法

    公开(公告)号：US06787305B1

    公开(公告)日：2004-09-07

    申请号：US09266935

    申请日：1999-03-12

    申请人： Wu-Bo Li ,  Joel A. Jessee ,  David Schuster ,  Jiulin Xia ,  Gulilat Gebeyehu

    发明人： Wu-Bo Li ,  Joel A. Jessee ,  David Schuster ,  Jiulin Xia ,  Gulilat Gebeyehu

    IPC分类号： C12Q168

    CPC分类号： C12P19/34 ,  C07H21/00 ,  C12Q1/6848 ,  C12Q2527/125

    摘要： The present invention is directed to compositions and methods for enhancing synthesis of nucleic acid molecules, particularly GC-rich nucleic acid molecules. Specifically, the invention provides compositions comprising one or more nitrogen-containing organic compounds having a formula selected from the group consisting of formula I and formula II (or salts or derivatives thereof), preferably 4-methylmorpholine N-oxide or betaine (carboxymethyltrimethylammonium), and further comprising one or more compounds selected from the group consisting of proline and an N-alkylimidazole compound, and more preferably proline, 1-methyliimidazole or 4-methylimidazole. The invention further relates to methods for enhanced, high-fidelity synthesis of nucleic acid molecules, including via amplification (particularly PCR), reverse transcription, and sequencing methods. The invention also relates to nucleic acid molecules synthesized by these methods, to fragments or derivatives thereof, and to vectors and host cells comprising such nucleic acid molecules, fragments, or derivatives. The invention also relates to kits for synthesizing, amplifying, reverse transcribing or sequencing nucleic acid molecules comprising one or more of the compositions of the invention.

    摘要翻译： 本发明涉及用于增强核酸分子，特别是富含GC的核酸分子的合成的组合物和方法。 具体地说，本发明提供包含一种或多种具有选自式I和式II（或其盐或衍生物），优选4-甲基吗啉N-氧化物或甜菜碱（羧甲基三甲基铵）的式的含氮有机化合物的组合物， 并且还包含一种或多种选自脯氨酸和N-烷基咪唑化合物的化合物，更优选脯氨酸，1-甲基咪唑或4-甲基咪唑。 本发明还涉及用于核酸分子增强，高保真合成的方法，包括经由扩增（特别是PCR），逆转录和测序方法。 本发明还涉及通过这些方法合成的核酸分子，其片段或衍生物以及包含该核酸分子，片段或衍生物的载体和宿主细胞。 本发明还涉及用于合成，扩增，逆转录或测序包含一种或多种本发明组合物的核酸分子的试剂盒。

    Transfection reagents
    9.
    发明授权
    Transfection reagents 有权
    转染试剂

    公开(公告)号：US08785200B2

    公开(公告)日：2014-07-22

    申请号：US13420091

    申请日：2012-03-14

    申请人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    发明人： Yongliang Chu ,  Malek Masoud ,  Gulilat Gebeyehu

    IPC分类号： C12N15/88

    CPC分类号： A61K47/48046 ,  A61K9/16 ,  A61K31/14 ,  A61K38/28 ,  A61K38/40 ,  A61K47/543 ,  C07C211/21 ,  C07C211/63 ,  C07C211/64 ,  C07C229/26 ,  C07D209/48 ,  C07D241/12 ,  C12N15/88 ,  Y10S435/81

    摘要： Disclosed are cationic lipid compounds and compositions of lipid aggregates for delivery of macromolecules and other compounds into cells. The compounds can be used alone or in combination with other compounds to prepare liposomes and other lipid aggregates suitable for transfection or delivery of compounds to target cells, either in vitro or in vivo. The compounds are preferably polycationic and preferably form highly stable complexes with various anionic macromolecules, particularly polyanions such as nucleic acids. These compounds have the property, when dispersed in water, of forming lipid aggregates which associate strongly, via their cationic portion, with polyanions. Also disclosed are intermediates for preparing the compound and compositions of the invention and methods of using the compounds to introduce other compounds into cells.

    摘要翻译： 公开了阳离子脂质化合物和用于将大分子和其它化合物递送到细胞中的脂质聚集体的组合物。 化合物可以单独使用或与其他化合物组合使用以制备适合于在体外或体内转染或递送化合物到靶细胞的脂质体和其他脂质聚集体。 该化合物优选是阳离子型的，优选与各种阴离子大分子，特别是聚阴离子如核酸形成高度稳定的络合物。 这些化合物在分散于水中时具有形成通过其阳离子部分强力缔合聚阴离子的脂质聚集体的性质。 还公开了制备本发明化合物和组合物的中间体以及使用该化合物将其它化合物引入细胞的方法。

    TRANSFECTION REAGENTS
    10.
    发明申请
    TRANSFECTION REAGENTS 有权

    公开(公告)号：US20120238747A1

    公开(公告)日：2012-09-20

    申请号：US13420091

    申请日：2012-03-14

    申请人： Yongliang CHU ,  Malek Masoud ,  Gulilat Gebeyehu

    发明人： Yongliang CHU ,  Malek Masoud ,  Gulilat Gebeyehu

    IPC分类号： C07C211/63 ,  C07D273/08 ,  C07C215/18 ,  C07C237/10 ,  C07J1/00

    CPC分类号： A61K47/48046 ,  A61K9/16 ,  A61K31/14 ,  A61K38/28 ,  A61K38/40 ,  A61K47/543 ,  C07C211/21 ,  C07C211/63 ,  C07C211/64 ,  C07C229/26 ,  C07D209/48 ,  C07D241/12 ,  C12N15/88 ,  Y10S435/81

    摘要： Disclosed are compounds capable of facilitating transport of biologically active agents or substances into cells having the general structure: wherein Q is selected from the group consisting of N, O and S; L is any bivalent organic radical capable of linking each Q, such as C, CH, (CH2)l, or {(CH2)i-Y—(CH2)j}k, wherein Y is selected from the group consisting of CH2, an ether, a polyether, an amide, a polyamide, an ester, a sulfide, a urea, a thiourea, a guanidyl, a carbamoyl, a carbonate, a phosphate, a sulfate, a sulfoxide, an imine, a carbonyl, and a secondary amino group and wherein Y is optionally substituted by —X1-L′-X2—Z or —Z; R1-R6, independently of one another, are selected from the group consisting of H, —(CH2)p-D-Z, an alkyl, an alkenyl, an aryl, and an alkyl or alkyl ether optionally substituted by one or more of an alcohol, an aminoalcohol, an amine, an amide, an ether, a polyether, a polyamide, an ester, a mercaptan, an alkylthio, a urea, a thiourea, a guanidyl, or a carbamoyl group, and wherein at least one of R1, R3, R4 and R6 is a straight chain or branched, cyclic, alkyl, alkenyl, alkynyl or aryl group; and anyone of R1, R3, R4 and/or R6 may optionally be covalently linked with each other, with Y or with L when L is C or CH to form a cyclic moiety; Z is selected from the group consisting of amine, spermiyl, carboxyspermiyl, guanidyl, spermidinyl, putricinyl, diaminoalkyl, pyridyl, piperidinyl, pyrrolidinyl, polyamine, amino acid, peptide, and protein; X1 and X2, independently of one another, are selected from the group consisting of NH, O, S, alkylene, and arylene; U is selected from the group consisting of alkylene, alkenylene, alkynylene, arylene, alkylene ether, and polyether; D is Q or a bond; A1 and A2, independently of one another, are selected from the group consisting of CH2O, CH2S, CH2NH, C(O), C{NH), C(S) and (CH2)t; X is a physiologically acceptable anion; m, n, r, s, u, v, w and y are 0 or 1, with the proviso that when both m and n are 0 at least one of r, s, u and y is other than 0; i, j, k, l, p and are integers from 0 to about 100; q is an integer from 1 to about 1000; and a is the number of positive charge divided by the valence of the anion.