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    ONE STEP IMAC (MCAC) PURIFICATION OF PROTEINS
    1.
    发明申请
    ONE STEP IMAC (MCAC) PURIFICATION OF PROTEINS 审中-公开
    一步IMAC（MCAC）蛋白质的纯化

    公开(公告)号：WO2008006899A1

    公开(公告)日：2008-01-17

    申请号：PCT/EP2007/057239

    申请日：2007-07-13

    Applicant: LEK PHARMACEUTICALS D.D. ,  MENART, Viktor ,  GABERC POREKAR, Vladka ,  PRERADOV VASLE, Tatjana ,  JESENKO, Ana ,  PODOBNIK, Barbara

    Inventor： MENART, Viktor ,  GABERC POREKAR, Vladka ,  PRERADOV VASLE, Tatjana ,  JESENKO, Ana ,  PODOBNIK, Barbara

    IPC: C07K1/22

    CPC classification number: C07K1/22

    Abstract: This invention relates to a process for isolation and/or purification of a protein of interest and to the use of IMAC as a capture and the main separation step. In one embodiment, the invention relates to a process for the isolation and/or purification of a protein of interest by use of immobilized metal affinity chromatography (IMAC), comprising the steps of (a) applying a mixture or solution comprising said protein to an IMAC matrix, (b) separating contaminant(s) by use of metal ion affinity of said protein to said IMAC matrix, under conditions suitable to keep said protein bound to the matrix, (c) separating the protein of interest from further contaminant(s) by use of both a non-metal ion affinity interaction and a metal ion affinity interaction between said protein and said IMAC matrix.

    Abstract translation: 本发明涉及用于分离和/或纯化目的蛋白质的方法以及IMAC作为捕获和主要分离步骤的用途。 在一个实施方案中，本发明涉及通过使用固定化金属亲和层析（IMAC）分离和/或纯化目的蛋白的方法，其包括以下步骤：（a）将包含所述蛋白质的混合物或溶液施用于 IMAC基质，（b）在适合于保持所述蛋白质结合到基质上的条件下，（c）将所述蛋白质与另外的污染物（s）分离，将所述蛋白质的金属离子亲和力与所述IMAC基质分离出来 ）通过使用所述蛋白质和所述IMAC基质之间的非金属离子亲和相互作用和金属离子亲和力相互作用两者。

    MONOCLONAL ANTIBODIES AGAINST SOLUBLE TNF-'alpha' RECEPTORS p55 and p75 AS WELL AS AGAINST TNF-'alpha' AND ITS ANALOGUES
    2.
    发明申请
    MONOCLONAL ANTIBODIES AGAINST SOLUBLE TNF-'alpha' RECEPTORS p55 and p75 AS WELL AS AGAINST TNF-'alpha' AND ITS ANALOGUES 审中-公开
    抗肿瘤坏死因子-α及其类似物可溶性TNF-α受体p55和p75的单克隆抗体

    公开(公告)号：WO1997006251A1

    公开(公告)日：1997-02-20

    申请号：PCT/SI1996000018

    申请日：1996-07-31

    Applicant: LEK, TOVARNA FARMACEVTSKIH IN KEMIC¿NIH IZDELKOV, D.D. ,  ZAVOD REPUBLIKE SLOVENIJE ZA TRANSFUZIJO KRVI ,  C¿URIN S¿ERBEC, Vladka ,  SCHUURBIERS, Annette ,  MENART, Viktor ,  GADERC POREKAR, Vladimira ,  S¿TALC, Anton ,  Z¿UNEC, Petra ,  OCVIRK, Janja ,  S¿TABUC, Borut

    Inventor： LEK, TOVARNA FARMACEVTSKIH IN KEMIC¿NIH IZDELKOV, D.D. ,  ZAVOD REPUBLIKE SLOVENIJE ZA TRANSFUZIJO KRVI

    IPC: C12N15/06

    CPC classification number: C07K14/7151 ,  A61K38/00 ,  C07K14/525 ,  C07K16/241 ,  C07K16/2866

    Abstract: The present invention belongs to the field of pharmaceutical industry and relates to novel monoclonal antibodies and fragments thereof which are bound to the available surface of a human recombinant TNF- alpha , TNF- alpha analogues and TNF- alpha soluble receptors p55 and p75. Further, the invention relates to stable hybridoma cell lines, which lines are capable of producing such monoclonal antibodies, to a process for the preparation thereof and to pharmaceutical preparations containing them. The invention also relates to a method for the quantitative determination of human TNF- alpha , free or complexed with soluble TNF- alpha receptors p55 and p75, and to a kit used therefor. By hitherto disclosed monoclonal antibodies and reagents developed therefrom, it has not been possible to simultaneously determine free TNF- alpha and its soluble receptors p55 and p75 as well as complexes thereof. Another object of the invention is also the TNF- alpha analogue TNF- alpha Cys95His107His108Cys148, which in a complex with TNF- alpha soluble receptors, is used as an antigen for immunization of mice.

    Abstract translation: 本发明属于制药工业领域，涉及与人重组TNF-α，TNF-α类似物和TNF-α可溶性受体p55和p75的可用表面结合的新型单克隆抗体及其片段。 此外，本发明涉及能够产生这种单克隆抗体的稳定的杂交瘤细胞系，其制备方法和含有它们的药物制剂。 本发明还涉及用于定量测定与可溶性TNF-α受体p55和p75无游离或复合的人TNF-α的方法，以及用于其的试剂盒。 迄今为止公开了从其开发的单克隆抗体和试剂，不可能同时测定游离TNF-α及其可溶性受体p55和p75及其复合物。 本发明的另一个目的还是TNF-α类似物TNF-αCys95His107His108Cys148，其在与TNF-α可溶性受体的复合物中用作小鼠免疫的抗原。

    ORGANIC COMPOUNDS
    3.
    发明申请
    ORGANIC COMPOUNDS 审中-公开
    有机化合物

    公开(公告)号：WO2008074724A1

    公开(公告)日：2008-06-26

    申请号：PCT/EP2007/063900

    申请日：2007-12-13

    Applicant: LEK PHARMACEUTICALS d.d. ,  MENART, Viktor ,  JESENKO, Ana ,  PRERADOV VASLE, Tatjana

    Inventor： MENART, Viktor ,  JESENKO, Ana ,  PRERADOV VASLE, Tatjana

    IPC: C07K1/14 ,  C07K1/36 ,  C12N1/06 ,  C12P21/00

    CPC classification number: C07K1/36 ,  C07K14/535 ,  C12P21/02

    Abstract: The present invention relates to a new process for the production of a polypeptide of interest, or of a precursor thereof, wherein the protein or the precursor are expressed in a host cell comprising a cell wall and at least one cell membrane, followed by extraction from the intact host cell by partial permeabilization of the cell membrane under non-denaturing conditions. During extraction the polypeptide of interest, or the precursor thereof, which is expressed in at least partially correctly folded form, passes through the cell wall and the cell membrane.

    Abstract translation: 本发明涉及生产目的多肽或其前体的新方法，其中蛋白质或前体在包含细胞壁和至少一个细胞膜的宿主细胞中表达，随后从 完整的宿主细胞通过在非变性条件下细胞膜的部分透化。 在提取期间，以至少部分正确折叠的形式表达的目标多肽或其前体通过细胞壁和细胞膜。

    PROCESS FOR THE ISOLATION AND/OR PURIFICATION OF PROTEINS
    4.
    发明申请
    PROCESS FOR THE ISOLATION AND/OR PURIFICATION OF PROTEINS 审中-公开
    分离和/或纯化蛋白质的方法

    公开(公告)号：WO2005112982A1

    公开(公告)日：2005-12-01

    申请号：PCT/EP2005/005507

    申请日：2005-05-20

    Applicant: LEK PHARMACEUTICALS d.d. ,  MENART, Viktor ,  KENIG, Maja ,  GABERC POREKAR, Vladka ,  FONDA, Irena

    Inventor： MENART, Viktor ,  KENIG, Maja ,  GABERC POREKAR, Vladka ,  FONDA, Irena

    IPC: A61K38/18

    CPC classification number: C07K1/22 ,  C07K14/525

    Abstract: The invention relates to the process for the isolation and / or purification of biologically active proteins, preferably TNF-alpha or TNF-alpha analogues. The process of the present invention results in the production of high yields of proteins, preferably TNF-alpha or TNF­alpha analogues with a purity of greater than 98%. The described process is particularly suitable for the industrial production of proteins, preferably TNF-alpha or TNF-alpha analogues.

    Abstract translation: 本发明涉及分离和/或纯化生物活性蛋白质，优选TNF-α或TNF-α类似物的方法。 本发明的方法导致产生高产率的蛋白质，优选纯度大于98％的TNF-α或TNFα类似物。 所述方法特别适用于工业生产蛋白质，优选TNF-α或TNF-α类似物。

    PROCESS FOR THE PURIFICATION AND/OR ISOLATION OF BIOLOGICALLY ACTIVE GRANULOCYTE COLONY STIMULATING FACTOR
    7.
    发明申请
    PROCESS FOR THE PURIFICATION AND/OR ISOLATION OF BIOLOGICALLY ACTIVE GRANULOCYTE COLONY STIMULATING FACTOR 审中-公开
    净化和/或分离生物活性粒细胞集落刺激因子的方法

    公开(公告)号：WO2003051922A1

    公开(公告)日：2003-06-26

    申请号：PCT/EP2002/013810

    申请日：2002-12-05

    Applicant: LEK PHARMACEUTICALS D.D. ,  GABERC POREKAR, Vladka ,  MENART, Viktor

    Inventor： GABERC POREKAR, Vladka ,  MENART, Viktor

    IPC: C07K14/535

    CPC classification number: C07K14/535 ,  A61K38/193

    Abstract: The invention relates to the process for the isolation of biologically active granulocyte colony stimulating factor (G-CSF), which enables the separation of correctly folded biologically active monomeric molecules of G-CSF from the incorrectly folded, biologically inactive monomeric, oligo- or polymeric and also from aggregated molecules of G-CSF by using immobilised metal affinity chromatography. The process of the invention, if desired the whole process, can be advantageously performed under native conditions. The biologically active G-CSF with a purity of greater than 95% is thus obtained. Only two additional chromatographic steps, cationic exchange chromatography and gel filtration, are then preferably applied to remove the traces of impurities. The entire process results in the production of higher yields of G-CSF with a purity of greater than 99%. The described process is particularly suitable for the industrial production of G-CSF.

    Abstract translation: 本发明涉及分离生物活性粒细胞集落刺激因子G-CSF的方法，其能够将正确折叠的G-CSF的生物活性单体分子与错误折叠的生物活性单体，寡聚体或聚合物分离，并且还 通过使用固定化金属亲和层析从G-CSF的聚集分子。 如果需要，本发明的方法可以有利地在天然条件下进行整个过程。 从而得到纯度大于95％的生物活性G-CSF。 然后优选仅进行两次另外的色谱步骤，阳离子交换层析和凝胶过滤以除去痕迹的杂质。 整个过程导致产生更高产率的G-CSF，其纯度大于99％。 所述方法特别适用于G-CSF的工业生产。

    SELENIUM CONTAINING MODIFYING AGENTS AND CONJUGATES
    8.
    发明申请
    SELENIUM CONTAINING MODIFYING AGENTS AND CONJUGATES 审中-公开
    含有修饰剂和硒化物的硒

    公开(公告)号：WO2009106239A1

    公开(公告)日：2009-09-03

    申请号：PCT/EP2009/001039

    申请日：2009-02-13

    Applicant: LEK PHARMACEUTICALS D.D. ,  KUNSTELJ, Menci ,  MENART, Viktor ,  AMBROZIC, Gabriela ,  GABERC POREKAR, Vladka

    Inventor： KUNSTELJ, Menci ,  MENART, Viktor ,  AMBROZIC, Gabriela ,  GABERC POREKAR, Vladka

    IPC: A61K47/48 ,  C07K14/535 ,  C07C391/00

    CPC classification number: C07K14/535 ,  A61K47/60 ,  A61K49/0423 ,  B82Y5/00 ,  C07C391/00

    Abstract: The invention relates to a modifying agent comprising a water soluble polymer, wherein the water soluble polymer comprises at least one reactive selenium group, said reactive selenium group being capable of reacting with a thiol group thereby forming an -Se-S- bond. Furthermore, the invention relates to a method for producing said modifying agents and their use in the modification of pharmaceutically active agents, e.g. G-CSF. Additionally, the invention concerns conjugates comprising a water-soluble polymer and a pharmaceutically active agent, wherein the water-soluble polymer is linked via a S-Se-bond to agent and a method for their production and their use as medicaments. Finally, the invention concerns a pharmaceutical composition comprising the inventive conjugates.

    Abstract translation: 本发明涉及包含水溶性聚合物的改性剂，其中所述水溶性聚合物包含至少一种反应性硒基团，所述反应性硒基团能够与硫醇基团反应从而形成-Se-S-键。 此外，本发明涉及一种生产所述改性剂的方法及其在药学活性剂的改性中的用途。 G-CSF。 此外，本发明涉及包含水溶性聚合物和药物活性剂的缀合物，其中水溶性聚合物通过S-Se键与药剂连接，其制备方法及其用作药物。 最后，本发明涉及包含本发明缀合物的药物组合物。

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