Pharmacokinetics-Based DNA-Encoded Library Screening

项目摘要 这个K99/R 00提案寻求通过以下途径扩大可药物化蛋白质靶点的空间： 开发新的分析技术，将指导下一代，非 分子治疗学与诊断学。环肽，其不是药物样的（即，利平斯基- 例如），对于解决不可治疗的目标，特别是蛋白质-蛋白质相互作用（PPI），具有很大的希望。 然而，更大的分子量和更复杂的二级结构导致了不可预测的和几乎不可预测的结果。 总是低的细胞渗透性，显著削弱了它们作为候选药物甚至作为临床前工具的效用 细胞中的化合物。环肽很少会表现出异常高的渗透性。然而，数据不足 存在的目的是揭示渗透的规则，因为细胞渗透的高通量测量 根本不存在在这个建议中，一个高通量的体外渗透试验兼容的DNA编码的 将开发组合文库（DEL）筛选并将其小型化至微流体液滴规模。的 渗透试验将用于筛选DEL以鉴定有效渗透膜的物质 双层。渗透性大环命中物将在基于细胞的渗透测定中进一步验证，该测定还将 在项目中开发。大型筛选数据集将揭示渗透率和渗透率之间的关系。 结构，特别是对于超出5规则（bRo 5）的分子，将使用机器进一步分析 学习利用这些和其他经验推导的大环肽药代动力学性质的模型， 另一方面，我们可能最终能够将这些非Lipinski分子从实验室大规模转移到临床。这项建议 将大大提高PI的职业发展，并推动她朝着成为一名 在一家以研究为重点的机构担任独立调查员。拟议项目提供切割方面的培训- 边缘研究技能，包括基于细胞的检测开发和高内涵成像，DEL设计，合成 和筛选技术的发展，以及机器学习技术。加州大学欧文分校提供了一个理想的 学术培训环境，与世界知名的医学和计算化学专家， 化学生物学和微流体工程。此外，UCI提供了一个智力环境， 鼓励协作与合作，使候选人的成长为科学的一员 社区事实上，PI将参与旨在实现独立性的活动，包括实验室培训， 管理和培训，网络，以及为学术就业市场做准备。总之， 拟议的计划将使PI的科学和职业明智的增长和独立性，进一步定位她 以获得未来的R 01资金。

PROJECT ABSTRACT This K99/R00 proposal seeks to expand the space of druggable protein targets through the development of novel analytical technology that will guide the design of next-generation, non- Lipinski molecular therapeutics and diagnostics. Cyclic peptides, which are not drug-like (i.e., Lipinski- like), hold great promise for addressing undruggable targets, especially protein-protein interactions (PPIs). However, the larger molecular mass and more complex secondary structures result in unpredictable and almost always low cell permeation, significantly blunting their utility as drug candidates and even as preclinical tool compounds in cells. Rarely, a cyclic peptide will exhibit anomalously high permeation. However, insufficient data exist to uncover the rules dictating permeation because high-throughput measurements of cell permeation simply do not exist. In this proposal, a high-throughput in vitro permeation assay compatible with DNA-encoded combinatorial library (DEL) screening will be developed and miniaturized to the microfluidic droplet scale. The permeation assay will be applied to screen DELs to identify the species that efficiently permeate membrane bilayers. The permeable macrocycle hits will be further validated in a cell-based permeation assay that will also be developed in the project. The large screening data sets will reveal relationships between permeability and structure, especially for molecules of beyond the Rule of 5 (bRo5), which will be further analyzed using machine learning. With these and other empirically derived models of macrocyclic peptide pharmacokinetic properties in hand, we may finally be able to move such non-Lipinski molecules from the lab to the clinic at scale. This proposal will significantly enhance the PI’s career development and advance her toward her career goal of becoming an independent investigator at a research-focused institution. The proposed project provides training in cutting- edge research skills, including cell-based assay development and high-content imaging, DEL design, synthesis and screening technology development, and machine learning techniques. UC Irvine provides an ideal environment for academic training, with world-renowned experts in medicinal and computational chemistry, chemical biology and microfluidics engineering. In addition, UCI provides an intellectual environment that encourages collaboration and cooperation, enabling the candidate’s growth as a member of the scientific community. Indeed, the PI will engage in activities designed to achieve independence, including training in lab management and grantsmanship, networking, and preparation for the academic job market. In summary, the proposed plan will enable the PI’s scientific and career-wise growth and independence, further positioning her to attain future R01 funding.