SBIR Phase I: Cell-penetrating monobodies targeting oncogenic KRAS

这个小企业创新研究（SBIR）第一阶段项目的更广泛影响在于一种称为细胞穿透单体（CPM）的新药物干预的技术开发。这种新的CPM干预的验证将通过设计针对某些癌症的有效和选择性的CPM抑制剂来证明。该项目的成功将建立一种强大的制药技术，用于设计个性化疗法，以治疗致癌性肿瘤驱动的肺癌、胰腺癌和结直肠癌，这些癌症的治疗选择非常有限。该项目还将加强迈阿密与工业界的伙伴关系，加强布法罗地区蓬勃发展的生命科学生态系统，并为西纽约地区培养具有全球竞争力的STEM劳动力。拟议的项目解决了单抗体临床转化的关键障碍-这是一类强大的工具生物制剂，尽管体积小，但不能渗透细胞。CPM技术通过结合正交交联技术克服了这一障碍，正交交联技术是一种专有方法，通过特定位点的链间交联来硬化单体结构，并具有单体表面增压功能。因此，CPM技术潜在地具有几个创新特征：1）遗传修饰促进CPM在细菌中的重组生产，无论是在研究规模还是用于生产; 2）使用成熟的展示技术可以容易地获得针对细胞内癌基因靶点的高结合亲和力和特异性;和3）由于刚性支架和可调的表面电荷，可以获得稳健的胞质转运效率。该项目旨在通过鉴定有效和选择性的抑制剂致癌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实现的。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KRAS突变体携带细胞系中鉴定出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知识价值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.