FP Assay for Isolation of Hsp90 Inhibitors (RMI)

描述(由申请人提供)：该计划的长期目标是确定用于癌症治疗和神经退行性疾病治疗的新型Hsp90抑制剂。目前工作的目标是将我们的Hsp90荧光偏振分析转换到MLSCN中心。本实验从细胞裂解产物中检测红移的cy3B标记的格尔达霉素与Hsp90的竞争结合。R03 NS050838-01；Chisis，G(PI)提供了开发该分析的部分资金。HSP90是一种伴侣蛋白，在维持癌细胞转化和提高癌细胞的存活和生长潜能方面具有重要作用。最近的证据表明，Hsp90抑制剂在神经退行性疾病、神经损伤、炎症和感染中有更多的应用。在体外和体内癌症模型中，几种使Hsp90功能失活的天然产物具有抗肿瘤作用。然而，由于Hsp90在正常细胞内稳态中的作用，目前尚不清楚Hsp90抑制剂是否具有足够的特异性用于治疗。17AAG是第一个进入临床试验的Hsp90抑制剂，其早期临床结果表明，这些担忧可能是没有根据的。这些研究证实了Hsp90是一种很有前途的新型癌症治疗靶点，并为Hsp90抑制剂在临床上的引入铺平了道路。由于目前抑制剂的局限性(即17AAG不能越过血脑屏障，BBB)，Hsp90抑制剂作为其他疾病的治疗药物的潜力被较少地探索。因此，目前人们对开发这种蛋白质的新型抑制剂的兴趣越来越大。我们在该领域进行了开创性的研究，并设计了第一个合成的Hsp90抑制剂类，即嘌呤-支架类(PU-类)。我们还设计和开发了几种检测Hsp90的生化和细胞抑制的方法。这里提出了一种用于MLSCN的这样的分析方法。发现基于不同化学骨架的新型Hsp90抑制剂对于充分挖掘伴侣的治疗潜力是必要的，我们在这里提出，可以通过筛选针对这一靶点的大量化合物库来获得这种多样性。

DESCRIPTION (provided by applicant): The long-term goal of this initiative is to identify novel Hsp90 inhibitors with use in cancer therapy and in the treatment of neurodegenerative diseases. The goal of the current effort is to translate our Hsp90 fluorescence polarization assay to MLSCN centers. This assay probes the competitive binding of red-shifted cy3B-labeled geldanamycin to Hsp90 from cell lysates. Funding for the development of the assay has been provided in part by R03 NS050838-01; Chiosis, G (PI). Hsp90 is a chaperone with important roles in maintaining transformation and in elevating the survival and growth potential of cancer cells. Recent evidence suggests additional applications of Hsp90 inhibitors in neurodegenerative diseases, nerve injuries, inflammation and infection. Several natural products that inactivate Hsp90 function have anti-tumor effects in in vitro and in vivo models of cancer. However, due to the role of Hsp90 in normal cellular homeostasis, it remained unclear whether Hsp90 inhibitors will be sufficiently specific for use as therapeutic agents. Early clinical results with 17AAG, the first Hsp90 inhibitor to enter clinical trials, suggest that these fears may be unfounded. These studies confirm that Hsp90 is a promising target for novel cancer therapeutics and pave the road for the introduction of Hsp90 inhibitors in the clinic. The potential of Hsp90 inhibitors as therapeutics in other diseases has been less explored due to limitations with the current inhibitors (i.e. 17AAG is not crossing the blood-brain barrier, BBB). Currently there is thus, increasing interest in developing novel inhibitors of this protein. We have conducted pioneering research in the area and designed the first synthetic class of Hsp90 inhibitors, the purine-scaffold class (PU-class). We have also designed and developed several assays that probe biochemical and cellular inhibition of Hsp90. One such assay is proposed here for use in MLSCN. Discovery of novel Hsp90 inhibitors based on diverse chemical skeletons is necessary to fully harvest the therapeutic potential of the chaperone, and we propose here that such diversity may be obtained by screening large libraries of compounds against this target.