Novel Targeted Nanomedicine Delivering MicroRNA-30-5p ReplacementTherapy for Multi-drug Resistant Cancer Treatment

摘要： 在这个第二阶段的SBIR应用中，miRecule建议开发一种基于microRNA的miR- 30-5P(miRecule候选MC-30)，用于治疗多药耐药(MDR)癌症。头部和颈部 鳞癌(HNSCC)是第六种最常见的癌症。超过一半的患者 患有晚期III或IV期疾病，平均5年生存率~40%。HNSCC肿瘤有很高的 基因突变水平导致高度的肿瘤异质性和耐药性。MIR-30-5P表达为 MIR30基因在肿瘤组织中被广泛抑制，约30%的HNSCC中存在MIR30基因缺失。损失MIR-30- 口咽部HNSCC患者(OPSCC)中5P的表达与预后不良相关。 这代表了我们最初的临床人口。基于MIR的疗法提供了一种颠覆性方法 靶向原发致癌途径和内源性OR机制治疗多药耐药癌 后天抵抗力。EGFR靶向治疗通常通过过度表达生长因子来补偿 受体(GFRs)MET和IGF1R。然而，我们发现miR-30-5P同时靶向和 压制所有这三个GFR。MiR-30-5P替代疗法的基本原理是，它将在 治疗异质性晚期HNSCC的能力，因为它不仅能够调节EGFR，而且能够调节MET，IGF- 1R和其他20多个mRNAs在肿瘤组织中证实是去调控的，并与 增殖、黏附、迁移、细胞外基质重塑和分化。在我们的SBIR的第一阶段， 我们开发了一种经过化学修饰的miR-30-5P模拟物，其核酸酶稳定性和5倍活性提高了1000倍。 在HNSCC模型中与天然microRNA进行比较。我们还证明了同时抑制了十几种 HNSCC中的关键癌基因和我们的模拟TO可以克服顺铂和EGFR相关药物的证据 抵抗。我们还展示了我们的临床验证的LNP配方，它通过 一种针对转铁蛋白受体(TFR)的单链抗体，通过具有 小剂量1 mg/kg的体内活性。这一点在两个生物分布研究和四个 不同的HNSCC体内模型均对MC-30表现出较强的敏感性。在这项第二阶段SBIR研究中， 我们建议：1)表征MC-30的PK/PD和非GLP TOX研究。2)创建令人信服的数据 在同基因和PDX模型中展示MC-30剂量依赖效应的包。3)示范 MC-30与已批准的具有潜力的顺铂、西妥昔单抗和抗PD-1药物相比的竞争优势 用于联合使用。4)在小型体外临床试验中验证我们提出的OPSCC患者的临床群体 审判。5)优化MC-30的放大、制造和CMC释放测试。这些研究的总和将 支持提交孤儿药物申请，支持我们的IND前会议，指导我们的临床开发，以及 验证外部对MC-30的投资。

Abstract: In this phase 2 SBIR application, miRecule proposes to develop a microRNA-based therapeutic mimic of miR- 30-5p (miRecule candidate MC-30) for the treatment of multi-drug resistant (MDR) cancers. Head and Neck Squamous Cell Carcinoma (HNSCC) is the 6th most common form of cancer. Greater than half of patients present with late stage III or IV disease, with an average 5-year survival rate of ~40%. HNSCC tumors have high levels of genetic mutations leading to high tumor heterogeneity and drug resistance. miR-30-5p expression is widely repressed in tumor tissues and MIR30 gene deletion is observed in ~30% of HNSCCs. Loss of miR-30- 5p expression correlates with poor survival outcome in 100% of Oropharyngeal HNSCC patients (OPSCC), which represents our initial clinical population. miR-based therapeutics offer a disruptive approach for treatment of MDR cancer by targeting both primary oncogenic pathways and mechanisms of intrinsic or acquired resistance. EGFR targeted therapy is often compensated for by overexpression of growth factor receptors (GFRs) MET and IGF1R. However, we have discovered that miR-30-5p simultaneously targets and repress all three of these GFRs. The rationale for miR-30-5p replacement therapy is that it will be superior in its ability to treat heterogeneous late-stage HNSCC due to its ability to regulate not only EGFR, but also MET, IGF- 1R, and over two dozen other mRNAs confirmed to be deregulated in tumor tissue and associated with proliferation, adhesion, migration, extracellular matrix remodeling, and differentiation. In phase 1 of our SBIR, we developed a chemically-modified mimic of miR-30-5p with >1000x improved nuclease stability and 5X activity in HNSCC models compared to the natural microRNA. We also demonstrated simultaneous inhibition of a dozen critical oncogenes in HNSCC with evidence that our mimic to can overcome cisplatin and EGFR-related drug resistance. We also demonstrated that of our clinically validated LNP formulation, which targets solid tumors via an scFv against the transferrin receptor (TfR), overcomes the challenge of delivery to cancer cells by having activity at a low dose of 1 mg/kg in vivo. This was demonstrated in both biodistribution studies and four different in vivo models of HNSCC that all showed strong sensitivity to MC-30. In this Phase II SBIR study, we propose to: 1) Characterized PK/PD and non-GLP Tox studies of MC-30. 2) Create a compelling data package that demonstrates dose-dependent efficacy of MC-30 in syngeneic and PDX models. 3) Demonstrate the competitive advantage of MC-30 over approved cisplatin, cetuximab, and anti-PD-1 therapies with potential for combination use. 4) Validate our proposed clinical population of OPSCC patients in a mini ex vivo clinical trial. 5) Optimize scale-up, manufacturing, and CMC release tests for MC-30. The sum of these studies will support filing an orphan drug application, enable our pre-IND meeting, guide our clinical development, and validate outside investment in MC-30.