Targeted chemotherapeutic drug discovery enabled by direct screening of peptide-drug conjugate libraries

通过直接筛选肽-药物缀合物库实现靶向化疗药物的发现

• 批准号: 413623402
• 负责人: Dr. Yen-Chun Lee
• 金额: --
• 依托单位: Department of Chemistry
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/413623402?language=en
• 关键词: Targeted; chemotherapeutic; drug; discovery; enabled

The scientific and medical community has been devoted to investigating different approaches for the treatment of cancer for over 100 years. In the mid-20th century, chemotherapeutic agents were introduced as adjuvant therapy to eliminate remaining cancer cells after surgery and radiation. Such therapies, for many cancer patients, have been able to slow tumor growth or even cure the cancer outright. However, conventional chemotherapy drugs are not able to differentiate between healthy cells and cancer cells, leading to several undesired side effects. In this regard, targeted chemotherapies with drugs that carry ligands for selective binding to specific receptors on cancer cells were developed to address this problem. Conjugating known receptor-targeting peptides to marketed drugs is a reliable method to generate bioactive PDCs as a targeted-delivery strategy. Because the number of known bioactive peptide sequences is limited, cell permeability often decreases after cargo attachment, and peptide-based drugs usually face short plasma half-life time, a streamlined approach for PDC development is needed. In this project, such a strategy is described by direct screening of an unnatural peptide-drug conjugate library in solution, which provides a straight forward approach to revealing de novo peptide sequences of PDC with protease resistance. A peptide library will be generated by SPPS assisted split-and-mix synthesis, followed by sequential Pt(IV) prodrug conjugation and PDC cleavage from solid support to yield a library of PDCs. The transmembrane protein, HER2, is overexpressed on various cancer cell lines and is a tested biological target for cancer treatment. After pull-down experiments for PDC binding to HER2, the mixture will be subjected to nLC-MS/MS for high throughput decoding, finally yielding potential hit PDCs. These PDCs will be prepared with an AFPS and PDC-HER2 binding will be confirmed by BioLayer Interferometry. Additionally, fluorescence microscopy experiments can be used to evaluate PDC internalization and MTT assays can assess their anticancer activity. The validation of hit PDCs offers opportunities to elucidate receptor-mediated internalization mechanisms. Thus, I would like to demonstrate an approach for the direct screening of PDC libraries to discover new, targeted cancer treatments as well as to develop molecular tools for elucidating new biology.

100多年来，科学和医学界一直致力于研究治疗癌症的不同方法。在世纪中期，化疗剂被引入作为辅助治疗，以消除手术和放射后残留的癌细胞。对于许多癌症患者来说，这种疗法已经能够减缓肿瘤生长，甚至彻底治愈癌症。然而，传统的化疗药物不能区分健康细胞和癌细胞，导致一些不希望的副作用。在这方面，开发了具有携带用于选择性结合癌细胞上的特异性受体的配体的药物的靶向化疗以解决该问题。将已知的受体靶向肽缀合至市售药物是产生生物活性PDCs作为靶向递送策略的可靠方法。由于已知的生物活性肽序列的数量有限，细胞渗透性通常在货物附着后降低，并且基于肽的药物通常面临短的血浆半衰期，因此需要用于PDC开发的简化方法。在该项目中，通过在溶液中直接筛选非天然肽-药物缀合物文库描述了这样的策略，其提供了一种直接的方法来揭示具有蛋白酶抗性的PDC的从头肽序列。肽文库将通过SPPS辅助的分裂-混合合成产生，然后依次进行Pt（IV）前药缀合和PDC从固体支持物裂解以产生PDC文库。跨膜蛋白HER 2在各种癌细胞系上过表达，并且是用于癌症治疗的经测试的生物靶标。在PDC与HER 2结合的下拉实验后，将混合物进行nLC-MS/MS以进行高通量解码，最终产生潜在的命中PDC。将使用AFPS制备这些PDC，并通过BioLayer干涉法确认PDC-HER 2结合。此外，荧光显微镜实验可用于评估PDC内化，MTT测定可评估其抗癌活性。命中PDCs的验证为阐明受体介导的内化机制提供了机会。因此，我想展示一种直接筛选PDC文库的方法，以发现新的靶向癌症治疗方法，并开发用于阐明新生物学的分子工具。