Pro-Drug Enolase Inhibitors in Precision Oncology

摘要。主要肿瘤抑制基因的基因组缺失是癌症中的常见事件，但目前 在精确肿瘤学的治疗上仍然不可操作。我们的实验室开创了一种创新的 治疗范例称为附带致死性，其中邻近TSG基因座的基因编码一个关键的 管家酶被巧合地删除。我们发现， 通过抑制酶的冗余同种型，可以在治疗上利用这种附带缺失。 这一框架的标志是携带糖酵解酶烯醇化酶1纯合缺失的癌症 （ENO1）。由于糖酵解是一个重要的生物能量过程，因此ENO1纯合子缺失的癌症完全是由糖酵解引起的。 依赖于ENO2进行糖酵解并确保细胞活力。抑制ENO2选择性杀死 ENO1纯合子删除了癌症，而正常组织未受影响。对…采取行动 针对这一漏洞，我们实验室开发了一种ENO2首选抑制剂HEX。作为强者的证明 我们已经证明HEX能够完全根除ENO1， 在小鼠中的胶质母细胞瘤的同卵缺失颅内原位模型中， 非人类灵长类动物这种强大的抗肿瘤作用在胶质母细胞瘤的背景下是前所未有的， 间接杀伤方法的威力对附带杀伤力的关注点的一种批评是其范围： 在ENO1缺失的情况下，只有一小部分患者能够受益。拓宽 治疗范围的附带致死性，该提案将集中于靶向ENO1杂合缺失的癌症， 占所有人类癌症的约20%。这将通过添加肿瘤亚型特异性前体蛋白来实现。 药物部分到HEX上以改善其递送。虽然ENO1杂合缺失的癌症在总体上是缺乏的， 烯醇化酶，它们几乎不像ENO1纯合缺失型癌症那样耗尽。因为HEX是一个负 荷电分子，肿瘤亚型特异性前药附着到HEX上不仅会增强其细胞增殖， 渗透性，但也将提高其肿瘤特异性。总之，这两个特性将使药物剂量在较低的 浓度，以提供足够大的治疗窗口来治疗ENO1杂合缺失的癌症 而不影响正常组织总的来说，该提案利用了合理的前药设计概念， 提高我们的核心ENO2抑制剂的特异性，以便我们可以扩大侧支致死的治疗范围。

ABSTRACT. Genomic deletions of major tumor suppressor genes are frequent events in cancer yet presently remain therapeutically unactionable for the purpose of precision oncology. Our lab has pioneered an innovative therapeutic paradigm known as collateral lethality, whereby genes neighboring a TSG locus encoding a key housekeeping enzyme are coincidentally deleted. We discovered that the metabolic vulnerabilities arising from such collateral deletions may be therapeutically exploited through inhibition of the enzyme’s redundant isoform. Emblematic of this framework are cancers harboring homozygous deletion of the glycolytic enzyme Enolase 1 (ENO1). As glycolysis is an essential bioenergetic process, ENO1-homozygous deleted cancers are entirely reliant on the ENO2 to perform glycolysis and ensure the cellular viability. Inhibition of ENO2 selectively kills ENO1-homozygous deleted the cancers while leaving normal tissues unperturbed. To pharmacologically act on this vulnerability, our lab has developed an ENO2-preferred inhibitor, HEX. As testament to the strong therapeutic viability of collateral lethality, we have shown that HEX is capable of completely eradicating ENO1- homzoygous deleted intracranial orthotopic models of glioblastoma in mice at concentrations well-tolerated in non-human primates. Such robust anti-neoplastic effects are unprecedented in the context of glioblastoma and speak to the power of the collateral lethality approach. One critique of the focus of collateral lethality is its scope: in the case of ENO1-deletions, only a small percentage of patients would be able to benefit. To broaden the therapeutic reach of collateral lethality, this proposal will focus on targeting ENO1-heterozygous deleted cancers, which comprise ~20% of all human cancers. This will be accomplished by adding tumor subtype-specific pro- drug moieties onto HEX to improve its delivery. While ENO1-heterozygous deleted cancers are deficient in total Enolase, they are not nearly as depleted as ENO1-homozygous deleted cancers are. As HEX is a negatively charged molecule, tumor-subtype specific pro-drug attachment onto HEX will not only enhance its cell permeability but will also improve its tumor specificity. Together, these two traits will enable drug dosing at lower concentrations to afford a therapeutic window sufficiently large to treat ENO1-heterozygous deleted cancers without the perturbing normal tissues. Overall, this proposal leverages the concept of rational pro-drug design to improve the specific of our core ENO2 inhibitor so that we may widen the therapeutic reach of collateral lethality.