利索-全球专利检索

  1. 登录
  2. 注册

搜索结果

19 条记录 (耗时 0.038 秒)

    Staphylococcal nuclease fusion proteins for the production of recombinant peptides
    5.
    发明申请
    Staphylococcal nuclease fusion proteins for the production of recombinant peptides 审中-公开
    用于生产重组肽的葡萄球菌核酸酶融合蛋白

    公开(公告)号：US20090123972A1

    公开(公告)日：2009-05-14

    申请号：US12000370

    申请日：2007-12-12

    申请人： Zhengding Su ,  Feng Ni

    发明人： Zhengding Su ,  Feng Ni

    IPC分类号： C12P21/04 ,  C12N15/11 ,  C12N15/00 ,  C12N1/21 ,  C12N1/19

    CPC分类号： C12N9/22 ,  C07K2319/20 ,  C07K2319/21 ,  C07K2319/75 ,  C12N15/62

    摘要： Peptides are produced as fusions with a suitable carrier protein. The carrier protein disclosed herein are adapted from the N-terminal domain of staphylococcus nuclease. This novel carrier protein acts to promote the over-expression of the peptide-protein fusion in the form of inclusion bodies, which minimizes in-cell proteolysis of desired peptides. The fusion protein is readily purified by conventional procedures or His-tag affinity chromatography when His-tag is inserted into the fusion protein. The target peptide is released from the purified fusion protein by a simple cleavage step and separated from the librated carrier protein by use of a reverse-phase HPLC process or by repeating the same affinity purification method. A particular advantage of the disclosed method, in addition to the obvious advantage of high yields, is its use for producing isotopically labeled peptides for NMR characterization of bioactive peptides and their interactions with target proteins.

    摘要翻译： 肽与合适的载体蛋白融合产生。 本文公开的载体蛋白质从葡萄球菌核酸酶的N-末端结构域改编。 这种新的载体蛋白起着促进包涵体形式的肽 - 蛋白质融合物的过度表达的作用，其最小化所需肽的细胞内蛋白水解。 当His标签插入融合蛋白中时，融合蛋白可以通过常规方法或His-标签亲和层析容易地纯化。 目标肽通过简单的切割步骤从纯化的融合蛋白中释放，并通过使用反相HPLC方法或通过重复相同的亲和纯化方法与分离的载体蛋白分离。 所公开的方法的特别优点除了高产率的明显优点之外，其用于生产同位素标记的肽用于生物活性肽的NMR表征及其与靶蛋白的相互作用。

    Bivalent thrombin binding molecules comprising linkers
    7.
    发明授权
    Bivalent thrombin binding molecules comprising linkers 失效
    包含接头的二价凝血酶结合分子

    公开(公告)号：US08063018B2

    公开(公告)日：2011-11-22

    申请号：US11630462

    申请日：2005-06-20

    申请人： Feng Ni ,  Dmitri Tolkatchev ,  Zhengding Su

    发明人： Feng Ni ,  Dmitri Tolkatchev ,  Zhengding Su

    IPC分类号： A61K38/16 ,  C07K14/00

    CPC分类号： G01N33/542 ,  A61K38/00 ,  A61K47/66 ,  B82Y5/00 ,  C07K14/40 ,  G01N33/53 ,  G01N2500/02

    摘要： There is provided herein a multivalent binding molecule and uses thereof. The molecule is useful in binding a target under certain conditions and releasing it under other conditions. The molecule has the general formula (1) of BM1-L-(BM2)n (1) wherein, BM1 is a binding moiety 1 having an affinity for site 1 on the target, BM2 is a binding moiety 2 having an affinity for a site other than site 1 on the target, n is 1 or greater, and L is a linker joining BM1 and BM2, said linker being adapted to respond to a change in its environment with a change in conformation and/or flexibility, wherein BM1 and BM2 may be the same or different and are selected such that in use each of the BM1 and BM2 existing separately has a lower binding affinity then the complex of BM1 and BM2 does when they are linked to form the molecule. BM2 may have a single binding region or multiple binding regions with affinity for the target. The binding affinity of BM1 or BM2 to the target alone is no more than ½ the binding affinity of the molecule of formula (1). The molecule of formula (1) can be constructed using an oligomeric or polymeric linker, such as a polypeptide sequence. Such molecules can be useful in the delayed release of drugs, in screening assays, for stabilizing enzymes such as proteases, and for controlling reactions such as blood clotting.

    摘要翻译： 本文提供了多价结合分子及其用途。 该分子在某些条件下可用于结合靶并在其它条件下释放。 该分子具有BM1-L-（BM2）n（1）的通式（1），其中，BM1是对靶标上的位点1具有亲和性的结合部分1，BM2是具有与 位于靶上的位点1之外的位置，n为1或更大，L为连接BM1和BM2的接头，所述接头适于响应其环境变化与构象和/或柔性的变化，其中BM1和 BM2可以相同或不同，并且被选择使得在使用中分别存在的BM1和BM2中的每一个具有较低的结合亲和力，而当BM1和BM2的复合物连接形成分子时，BM2和BM2的复合物才会起作用。 BM2可以具有对靶标具有亲和性的单一结合区域或多个结合区域。 BM1或BM2对靶单独的结合亲和力不超过式（1）分子的结合亲和力的1/2。 式（1）的分子可以使用寡聚或聚合的接头，如多肽序列构建。 这样的分子可用于药物的延迟释放，筛选测定中，用于稳定酶如蛋白酶，以及用于控制诸如血液凝固的反应。

    Peptide inhibitors of thrombin as potent anticoagulants
    8.
    发明申请
    Peptide inhibitors of thrombin as potent anticoagulants 审中-公开
    凝血酶肽抑制剂作为有效的抗凝剂

    公开(公告)号：US20090137779A1

    公开(公告)日：2009-05-28

    申请号：US12219031

    申请日：2008-07-15

    申请人： Feng Ni ,  Dmitri Tolkatchev ,  Anna Natapova ,  Anatol Koutychenko

    发明人： Feng Ni ,  Dmitri Tolkatchev ,  Anna Natapova ,  Anatol Koutychenko

    IPC分类号： C07K14/00

    CPC分类号： C07K14/815 ,  A61K38/00

    摘要： The tetrapeptide Phe-Asn-Pro-Arg (SEQ ID NO: 3) is a structurally-optimized sequence for binding to the active site of thrombin. By conjugating this tetrapeptide or variants thereof to a C-terminal fragment of hirudin, we were able to generate a series of new multivalent inhibitors of thrombin containing only genetically encodable natural amino acids. We found that synergistic binding to both the active site and an exosite of thrombin can be enhanced through substitutions of amino acid residues at the P4, P3 and P3′ sites of the active-site directed sequence, Xaa (P4)-Yaa (P3)-Pro (P2)-Arg (P1)-Pro(P1′)-Gln(P2′)-Zaa(P3′). Complementary to rational design, a phage library was constructed to explore further the residue requirements at the P4, P3 and P3′ sites for multivalent and optimized bridge-binding. Panning of the phage library has led to thrombin-inhibitory peptides possessing strong anti-clotting activities in the low nanomolar range and yet interfering only partially with the catalytic active site of thrombin. In all, the availability of potent and genetically-encodable polypeptide inhibitors of thrombin opens the door for much wider applications of this clinically-successful class of anticoagulants, e.g. through more cost-effective recombinant peptide production, in areas such as gene therapy as well as to improve clinical efficacy/safety through the incorporation of homing peptides for targeted delivery.

    摘要翻译： 四肽Phe-Asn-Pro-Arg（SEQ ID NO：3）是结合到凝血酶活性位点的结构优化的序列。 通过将这种四肽或其变体与水蛭素的C末端片段缀合，我们能够产生一系列含有仅基因可编码天然氨基酸的凝血酶的新型多价抑制剂。 我们发现通过在活性位点定向序列Xaa（P4）-Yaa（P3）的P4，P3和P3'位点处的氨基酸残基的取代可以增强与凝血酶的活性位点和外部位点的协同结合， -Pro（P2）-Arg（P1）-Pro（P1'）-G1n（P2'）-Zaa（P3'）。 与理性设计相辅相成，构建了噬菌体文库，进一步探索P4，P3和P3位点的残留要求，进行多价和优化的桥接。 噬菌体文库的淘选导致凝血酶抑制肽在低纳摩尔范围内具有强烈的抗凝血活性，但仅部分地与凝血酶的催化活性位点相互干扰。 总之，有效的和可遗传编码的凝血酶多肽抑制剂的可用性为这种临床上成功的一类抗凝剂的应用开辟了广泛的应用。 通过在基因治疗等领域通过更具成本效益的重组肽生产，以及通过并入用于靶向递送的归巢肽来提高临床疗效/安全性。

    Quantitative ranking of transient ligand binding to target biomolecules
    9.
    发明申请
    Quantitative ranking of transient ligand binding to target biomolecules 审中-公开
    瞬态配体与靶生物分子结合的定量分级

    公开(公告)号：US20050287527A1

    公开(公告)日：2005-12-29

    申请号：US10500878

    申请日：2003-01-10

    申请人： Feng Ni ,  Zhengding Su ,  Ping Xu ,  Dmitri Tolkatchev ,  Michael Osborne ,  Anatol Koutychenko

    发明人： Feng Ni ,  Zhengding Su ,  Ping Xu ,  Dmitri Tolkatchev ,  Michael Osborne ,  Anatol Koutychenko

    IPC分类号： C12Q1/68 ,  G01N24/08 ,  G01N33/00 ,  G01N33/542 ,  G01R33/46 ,  G01R33/465

    CPC分类号： G01N33/542 ,  G01N24/08 ,  G01R33/4625 ,  G01R33/465

    摘要： There is provided a method of quantitatively ranking transient ligand binding to target biomolecules by means of NMR relaxation dispersion profiles. The present invention also relates to a method to identify ligand site obeying two-state and more complex binding behavior in a transient complex of a ligand with a target molecule, still with the use of NMR. There is also provided an efficient method to quantitate fast dissociation rates of ligands containing at least one magnetic nuclei by performing NMR relaxation dispersion experiments at different protein concentrations, enabling the evaluation of populations and exchange rates, and extending the practical applicability of the NMR relaxation dispersion experiments.

    摘要翻译： 提供了一种通过NMR松弛分散谱定量分析瞬时配体与靶生物分子结合的方法。 本发明还涉及在使用NMR的情况下，鉴定配体与靶分子的瞬时复合物中符合双态和更复杂结合行为的配体位点的方法。 还提供了通过在不同蛋白质浓度下进行NMR松弛分散实验来定量含有至少一个磁核的配体的快速解离速率的有效方法，使得能够评价群体和交换速率，并且扩展了NMR松弛分散体的实际适用性 实验。

    Locally-active anticoagulants and anti-inflammatory agents
    10.
    发明授权
    Locally-active anticoagulants and anti-inflammatory agents 有权

    公开(公告)号：US10155802B2

    公开(公告)日：2018-12-18

    申请号：US14112565

    申请日：2012-04-13

    申请人： Feng Ni ,  Ping Xu ,  Sazzard Hossain ,  Dmitri Tolkatchev ,  Kenji Tonan

    发明人： Feng Ni ,  Ping Xu ,  Sazzard Hossain ,  Dmitri Tolkatchev ,  Kenji Tonan

    IPC分类号： C07K14/745 ,  C07K14/81 ,  C12N9/74 ,  C07K5/107 ,  C07K16/36 ,  A61K38/00

    摘要： A locally-activatable bivalent thrombin binding agent is provided having two thrombin binding moieties for non-overlapping sites on a surface of thrombin linked together by a linker. The linker is a polypeptide having 5 to 30 amino acid residues existing in a folded state under an environmental condition where the binding agent is inactive. The linker changes conformation from the folded state to an unfolded state in response to a change in bulk temperature and/or to the presence of hyper-mobile water thereby activating the binding agent. Such locally-activatable thrombin binding agents can be administered systemically while only targeting specific sites of coagulation or inflammation since the thrombin binding agent will only activate at the site where the existence of atherosclerotic plaques has changed the local bulk temperature and/or created hyper-mobile water sufficiently to unfold the linker and activate the binding agent. Such binding agents are useful as site-specific anti-coagulant, anti-thrombotic and/or antiinflammatory agents.