Ketorolac and Related NSAIDs for Targeting Rho-family GTPases in Ovarian Cancer

项目摘要 Rho家族GT酶新活性物质R-酮咯酸的化学信息学鉴定 卵巢癌的抑制活性激发了两层验证策略。临床前和体外试验 将评估FDA批准的临床使用的[R，S]-酮咯酸用于卵巢癌的新用途， R-酮咯酸用于研究性新药备案。Rho家族GTP酶（Rac、Rho和Cdc 42）统称 控制细胞增殖、粘附和迁移，并作为功能性治疗靶点， 许多上皮癌。一个虚拟的药物筛选含羧酸的药物首先预测的R- 批准的药物[R，S]-酮咯酸的对映体作为高价值的临床相关GT3抑制剂。之前 在我们的工作中，S-对映体被认为是外消旋药物制剂中的唯一活性组分， 对环加氧酶（考克斯）的选择性活性，用于减轻炎症和急性疼痛。gtp酶活性 测量结果表明，R-酮咯酸是鸟嘌呤核苷酸结合的变构抑制剂，而 酮咯酸的S-对映异构体对GTP酶几乎没有活性。在基于细胞的测定中，R-酮咯酸阻断 Rac 1和Cdc 42激活、肌动蛋白重塑与下游细胞粘附、迁移和侵袭 卵巢癌细胞系。我们的回顾性研究表明， 显著的生存获益和我们的前瞻性0期试验。[R，S]-酮咯酸给药， 药代动力学分布，降低GT3活性，并抑制与侵袭相关的行为， 患者肿瘤细胞的转移。根据我们全面的已发表数据，我们假设， 重新利用[R，S]-酮咯酸并将R-酮咯酸开发为研究性新药将最大限度地减少 卵巢癌复发通过抑制Rac 1和Cdc 42依赖的肿瘤转移， 专门的壁龛，以增加对化疗的脆弱性。一系列的三个目标将验证1）好处 使用肿瘤复发的动物异种移植模型，2）酮咯酸在减少肿瘤复发中的影响， 酮咯酸通过离体器官型和生物反应器培养最大限度地减少肿瘤细胞-生态位相互作用，以及 3)酮咯酸/卵巢癌作为预测的治疗/适应症对。这些研究将告知“去与不去” 在卵巢癌患者中实施[R，S]酮咯酸和R-酮咯酸单药治疗的2a期临床试验的决策 癌症患者。研究的另一个结果是提高提交FDA申请的准备程度。 R-ketorolac作为卵巢癌新疗法的研究性新药申请，以克服毒性 和与外消旋药物相关的局限性。

PROJECT SUMMARY Cheminformatics identification of R-ketorolac as a novel pharmacologic entity with Rho-family GTPase inhibitory activity for ovarian cancer motivates a two-tiered validation strategy. Preclinical and ex vivo testing will evaluate repurposed new use of the FDA approved, clinically used [R,S]-ketorolac for ovarian cancer and R-ketorolac for an Investigational New Drug filing. Rho family GTPases (Rac, Rho and Cdc42) collectively control cell proliferation, adhesion and migration and are of interest as functional therapeutic targets in numerous epithelial cancers. A virtual drug screen of -carboxylate containing drugs first predicted the R- enantiomer of the approved drug [R,S]-ketorolac as a high value, clinically relevant GTPase inhibitor. Prior to our work, the S-enantiomer was considered the sole active component in the racemic drug formulations, with selective activity against cyclooxygenases (COX) for mitigating inflammation and acute pain. GTPase activity measurements demonstrate that R-ketorolac is an allosteric inhibitor of guanine nucleotide binding, while the S-enantiomer of ketorolac shows little to no activity against GTPases. In cell-based assays, R-ketorolac blocks Rac1 and Cdc42 activation, actin remodeling and downstream cell adhesion, migration and invasion of human ovarian cancer cell lines. Human efficacy is supported by both our retrospective study demonstrating significant survival benefit and our prospective Phase 0 trial. [R,S]-ketorolac administration, has favorable pharmacokinetic distribution, reduces GTPase activities, and inhibits behaviors associated with invasion and metastasis in patient tumor cells. Based on our comprehensive published data, we hypothesize that repurposing [R,S]-ketorolac and development of R-ketorolac as an Investigational New Drug will minimize ovarian cancer relapse by inhibiting Rac1 and Cdc42 dependent tumor metastasis, and reducing protection in specialized niches to increase vulnerability to chemotherapy. A series of three aims will validate 1) the benefit of ketorolac in reducing tumor relapse using an animal xenograft model of tumor recurrence, 2) the impact of ketorolac on minimizing tumor cell-niche interactions through ex vivo organotypic and bioreactor cultures, and 3) ketorolac/ovarian cancer as a predicted therapeutic/ indication pair. These studies will inform `Go vs. No Go' decision-making for Phase 2a clinical trial implementation of [R,S] ketorolac and R-ketorolac alone in ovarian cancer patients. An additional outcome of the studies will be to enhance readiness to submit an FDA Investigational New Drug filing for R-ketorolac as a new therapy for ovarian cancer to overcome the toxicities and limitations associated with racemic drug.