Department of Pathology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA, 94305, USA.
Department of Chemical and System Biology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA, 94305, USA.
Angew Chem Int Ed Engl. 2020 Oct 19;59(43):19143-19152. doi: 10.1002/anie.202006719. Epub 2020 Aug 26.
Fluorescently quenched probes that are specifically activated in the cancer microenvironment have great potential application for diagnosis, early detection, and surgical guidance. These probes are often designed to target specific enzymes associated with diseases by direct optimization using single purified enzymes. However, this can result in painstaking chemistry efforts to produce a probe with suboptimal performance when applied in vivo. We describe here an alternate, unbiased activity-profiling approach in which whole tissue extracts are used to directly identify optimal peptide sequences for probe design. Screening of tumor extracts with a hybrid combinatorial substrate library (HyCoSuL) identified a combination of natural and non-natural amino-acid residues that was used to generate highly efficient tumor-specific probes. This new strategy simplifies and enhances the process of probe optimization without any a priori knowledge of enzyme targets and has the potential to be applied to diverse disease states using clinical or animal-model tissue samples.
荧光猝灭探针在癌症微环境中特异性激活，具有很大的应用潜力，可用于诊断、早期检测和手术指导。这些探针通常通过直接使用单一纯化酶进行优化来靶向与疾病相关的特定酶。然而，当在体内应用时，这可能会导致产生性能不佳的探针的艰苦化学努力。我们在这里描述了一种替代的、无偏见的活性分析方法，其中使用整个组织提取物来直接确定用于探针设计的最佳肽序列。用杂交组合底物文库（HyCoSuL）筛选肿瘤提取物，鉴定出一组天然和非天然氨基酸残基，用于生成高效的肿瘤特异性探针。这种新策略简化并增强了探针优化过程，而无需预先了解酶靶标，并且有可能使用临床或动物模型组织样本应用于多种疾病状态。
