SBIR Phase I: Cell-penetrating monobodies targeting oncogenic KRAS

小型企业创新研究(SBIR)第一阶段项目的更广泛影响在于一种名为细胞穿透单体(CPM)的新药物干预的技术开发。这种新的CPM干预措施的有效性将通过设计针对某些癌症的基于CPM的有效和选择性抑制剂来证明。该项目的成功将为设计个性化疗法建立强大的制药技术，以治疗由癌基因突变驱动的肺癌、胰腺癌和结直肠癌，这些癌症的治疗选择极其有限。该项目还将加强学术界和产业界的合作伙伴关系，加强布法罗地区蓬勃发展的生命科学生态系统，并为纽约西部地区发展一支具有全球竞争力的STEM劳动力队伍。拟议的项目解决了单体临床翻译中的一个关键障碍-单体是一类强大的工具生物制品，尽管其体积很小，但不能渗透细胞。CPM技术通过结合正交交联剂克服了这一障碍，这是一种专利方法，通过特定部位的链间交联剂与单体体表面增压来硬化单体体结构。因此，CPM技术具有几个潜在的创新特征：1)基因修饰有助于在研究规模和生产中在细菌中重组CPM；2)利用成熟的展示技术可以很容易地获得与细胞内癌基因靶标的高结合亲和力和特异性；以及3)由于坚硬的支架和可调的表面电荷，可以获得强大的胞浆转运效率。该项目旨在通过鉴定有效和选择性的抑制物致癌KRAS突变体来释放CPM技术的商业价值，这些突变体被证明是小分子方法难以捉摸的。将使用已报道的基于单体的KRAS抑制剂作为模板，对电荷分布和物理化学性质进行广泛的优化，目标是在含有KRAS突变的细胞系中确定一种具有亚微摩尔抑制活性的CPM。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase I project lies in technological development of a new pharmaceutical intervention called cell-penetrating monobodies (CPM). The validation of this new CPM intervention will be demonstrated by designing potent and selective CPM-based inhibitors targeting certain cancers. The success of this project will establish a powerful pharmaceutical technology for designing personalized therapies to treat the oncogenic mutant-driven lung, pancreatic, and colorectal cancers, where treatment options are extremely limited. This project will also enhance the academia-industry partnership, strengthen a burgeoning life science ecosystem in the Buffalo area, and develop a globally competitive STEM workforce for the Western New York region.The proposed project addresses a critical barrier in the clinical translation of monobodies - a class of powerful tool biologics that are not cell-permeable despite their small size. The CPM technology overcomes this barrier by combining orthogonal crosslinking  a proprietary method to rigidify monobody structure through site-specific inter-strand crosslinking - with monobody surface supercharging. As a result, the CPM technology potentially possesses several innovative features: 1) genetic modifications facilitate recombinant production of CPM in bacteria both at research scale and for manufacturing; 2) high binding affinity and specificity toward intracellular oncogene targets can be readily obtained using well-established display technologies; and 3) robust cytosolic transport efficiency can be obtained owing to the rigid scaffold and tunable surface charge. This project aims to unlock the commercial value of CPM technology by identifying potent and selective inhibitors oncogenic KRAS mutants that proved to be elusive with the small-molecule approach. Extensive optimizations of charge distribution and physicochemical properties will be performed using a reported monobody-based KRAS inhibitor as a template, with a goal to identify one CPM with sub-micromolar inhibitory activity in the KRAS mutant-harboring cell lines.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.