公开(公告)号：EP1270746A1

公开(公告)日：2003-01-02

申请号：EP02076171.4

申请日：1999-07-16

申请人： Inoxell A/S

发明人： Jensen, Allan ,  Halkier, Torben ,  Jespersen, Lene

IPC分类号： C12Q1/68 ,  C12N15/10

CPC分类号： C12N15/1044 ,  C12N15/1079 ,  C12N15/1086

摘要： The invention provides for methods for identification of biologically active biomolecules. In one aspect, a biologically active biomolecule such as RNA or a peptide is identified by incorporating random nucleotide sequences in a scaffold constituted by an enzyme activity modulator, transforming substantially identical host cells with the construct obtained thereby and screening the transformed cells to identify those where a preselected phenotypic trait has been altered. The randomized DNA is subsequently isolated from the phenotypically altered cells and the peptide and/or RNA encoded by the random sequence is determined. In turn, interaction partners which are putative drug targets are identified and isolated by use of the peptide and/or RNA as part of affinity reagents. A preferred scaffold is derived from the potato inhibitor I family of protease inhibitors and exemplified is the barley chymotrypsin inhibitor 2A (CI-2A). Another aspect relates to the identification of novel enzyme inhibitors by using substantially the same approach, but screening specifically for changes in target enzyme activity. Also disclosed are methods of producing the relevant transformation and expression vectors as well as methods for identifying lead compounds and drug targets for use in drug development. Finally, the invention also includes within its scope a method for the preparation of a medicinal product.

摘要翻译： 本发明提供了鉴定生物活性生物分子的方法。 在一个方面，生物活性生物分子如RNA或肽通过将随机核苷酸序列并入由酶活性调节剂构成的支架中，通过将由此获得的构建体转化基本相同的宿主细胞并筛选转化细胞来鉴定那些 预选的表型性状已被改变。 随后的DNA随后从表型改变的细胞中分离，并确定随机序列编码的肽和/或RNA。 反过来，通过使用肽和/或RNA作为亲和试剂的一部分来鉴定和分离作为推定药物靶标的相互作用配偶体。 优选的支架衍生自蛋白酶抑制剂的马铃薯抑制剂I家族，并且例示的是大麦胰凝乳蛋白酶抑制剂2A（CI-2A）。 另一方面涉及通过使用基本上相同的方法鉴定新型酶抑制剂，但专门筛选靶酶活性的变化。 还公开了生产相关转化和表达载体的方法以及用于鉴定用于药物开发的铅化合物和药物靶标的方法。 最后，本发明在其范围内还包括制备药用产品的方法。