FP Assay for Isolation of Hsp90 Inhibitors (RMI)

描述（由申请人提供）：本项目的长期目标是鉴定用于癌症治疗和神经退行性疾病治疗的新型Hsp90抑制剂。目前努力的目标是将我们的Hsp90荧光偏振分析转化为MLSCN中心。该实验探讨了红移cy3b标记的格尔达霉素与细胞裂解物Hsp90的竞争结合。该检测的开发资金部分由R03 NS050838-01提供；Chiosis, G （PI）Hsp90是一种伴侣蛋白，在维持癌细胞的转化和提高癌细胞的生存和生长潜能方面发挥着重要作用。最近的证据表明Hsp90抑制剂在神经退行性疾病、神经损伤、炎症和感染中的其他应用。几种灭活Hsp90功能的天然产物在体外和体内癌症模型中具有抗肿瘤作用。然而，由于Hsp90在正常细胞稳态中的作用，目前尚不清楚Hsp90抑制剂是否具有足够的特异性，可以用作治疗剂。第一个进入临床试验的Hsp90抑制剂17AAG的早期临床结果表明，这些担忧可能是没有根据的。这些研究证实了Hsp90是一种有希望的新型癌症治疗靶点，并为临床引入Hsp90抑制剂铺平了道路。由于现有抑制剂的局限性（即17AAG不能穿过血脑屏障，BBB）， Hsp90抑制剂作为其他疾病治疗药物的潜力尚未得到充分探索。因此，目前人们对开发这种蛋白质的新型抑制剂越来越感兴趣。我们在该领域进行了开创性的研究，并设计了第一类合成的Hsp90抑制剂，嘌呤支架类（pu类）。我们还设计并开发了几种检测Hsp90的生化和细胞抑制的方法。本文提出了一种用于MLSCN的检测方法。发现基于不同化学骨架的新型Hsp90抑制剂是充分挖掘伴侣蛋白治疗潜力的必要条件，我们建议通过筛选针对该靶点的大型化合物文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.