Translational Development of a Targeted and Stroma-penetrating Nanoparticle Drug for Pancreatic Cancer Therapy

项目摘要 药物递送的低效率和侵袭性肿瘤生物学是肿瘤细胞对治疗耐药的主要原因。 胰腺癌致密的纤维性肿瘤间质的存在产生了药物递送屏障，并促进了肿瘤的生长。 胰腺癌细胞的侵袭性生物学和耐药性。我们开发了一种靶向基质 携带伊立替康活性代谢物SN 38的透明质酸纳米粒。临床前结果 研究表明，显著增强的纳米颗粒/药物递送到导管癌细胞中， 在耐药胰腺癌患者来源的异种移植物（PDX）模型中具有强的治疗效果。 重要的是，基质穿透纳米药物具有调节肿瘤基质的独特性质， 纳米药物可以穿过基质屏障，但保留保护性基质结构。用于临床翻译 对于这种新的纳米药物，我们的SBIR I期研究集中在重新设计无标签的基质穿透配体， ATFmmp 14（R2），其由尿激酶纤溶酶原激活物受体（uPAR）结合结构域和 MMP 14的催化结构域，遵循FDA Pre-IND团队的建议。我们开发了 具有在无内毒素细菌中大规模生产能力的无标签ATFmmp 14（R2）表达载体， 系统我们还证明了ATFmmp 14（R2）缀合的HANP/SN 38具有相当的肿瘤靶向性， 第一代His标记的ATFmmp 14（R1）-HANP/SN 38在胰腺癌中的能力和治疗功效 癌症PDX模型。拟议的第二阶段项目旨在通过开展 重要的研究性新药（IND）-使临床前研究。在目标1中，我们将优化并最终确定 MIGRA-NP 01纳米制剂用于GLP/GMP条件下的大规模生产。然后我们将 表征MIGRA-NP 01的体外和体内特性，包括生物物理和生物化学参数 体外和全身毒性、药代动力学/药效学、生物分布和正常 和荷瘤小鼠（目的2）。在这些研究期间，我们将制定展台操作规程（SOP） 和MIGRA-NP 01用于药物开发的质量控制（QC）指标（目标2）。接下来，我们将确定一个 MIGRA-NP 01在具有不同病理和遗传的胰腺PDX模型中的优化剂量和方案 特征和对化疗的敏感性（目的3）。最后，胰腺PDX肿瘤的治疗反应将 与基因表达特征相关，以鉴定MIGRA的潜在预测性生物标志物- NP 01治疗（目标3）。该转化研究项目的成果包括：1）获得最终的 MIGRA-NP 01的优化纳米制剂，具有MIGRA-NP 01的GLP/GMP生产的既定SOP 并生产两批GLP等级的MIGRA-NP 01;和2）测定全身毒性，生物分布， 图10示出了MIGRA-NP 01在正常小鼠和胰腺PDX荷瘤小鼠中的治疗剂量/时间表。结果 SBIR第二阶段项目的实施将对未来提交IND申请产生重大影响， FDA批准在晚期胰腺癌患者中进行1期临床试验。

Project Summary Low efficiency in drug delivery and aggressive tumor biology are the major causes of resistance to therapy in pancreatic cancer. The presence of a dense fibrous tumor stroma creates a drug delivery barrier and promotes aggressive biology and drug resistance in pancreatic cancer cells. We have developed a targeted stroma penetrating hyaluronic acid nanoparticle carrying SN38, an active metabolite of irinotecan. Results of preclinical studies demonstrated significantly enhanced nanoparticle/drug delivery into ductal carcinoma cells to produce strong therapeutic effects in drug resistant pancreatic cancer patient derived xenograft (PDX) models. Importantly, the stroma-penetrating nanodrug has unique properties to modulate tumor stroma that allows the nanodrug to pass through the stromal barrier but retain a protective stromal structure. For clinical translation of this novel nanodrug, our SBIR phase I study focused on the re-engineering a tagless stroma penetrating ligand, ATFmmp14 (R2), that consists of a urokinase plasminogen activator receptor (uPAR) binding domain and a catalytic domain of MMP14, following the recommendation of the FDA Pre-IND team. We have developed the tagless ATFmmp14(R2) with the capacity of large scale production in an endotoxin-free bacterial expressing system. We also demonstrated that ATFmmp14 (R2) conjugated HANP/SN38 has comparable tumor targeting ability and therapeutic efficacy to the first generation of His-tagged ATFmmp14(R1)-HANP/SN38 in pancreatic cancer PDX models. The proposed phase II project aims to accelerate the translational process by conducting important Investigational New Drug (IND)-enabling preclinical studies. In Aim 1, we will optimize and finalize nanoformulation of MIGRA-NP01 for large-scale production under GLP/GMP conditions. We will then characterize in vitro and in vivo properties of MIGRA-NP01, including biophysical and biochemical parameters in vitro, and systemic toxicity, pharmacokinetics/pharmacodynamics, biodistribution, and clearance in normal and tumor bearing mice (Aim 2). During those studies, we will establish the Stand Operating Protocols (SOPs) and metrics for quality control (QC) of MIGRA-NP01 for drug development (Aim 2). Next, we will determine an optimized dose and schedule for MIGRA-NP01 in pancreatic PDX models with different pathological and genetic features, and sensitivity to chemotherapy (Aim 3). Finally, therapeutic responses of pancreatic PDX tumors will be correlated with the gene expression signatures to identify the potential predicative biomarkers for MIGRA- NP01 treatment (Aim 3). The outcomes of this translational research project include: 1) obtaining the final optimized nanoformulation of MIGRA-NP01 with established SOPs for the GLP/GMP production of MIGRA-NP01 and producing two lots of the GLP grades of MIGRA-NP01; and 2) determining systemic toxicity, biodistribution, therapeutic dose/schedule of the MIGRA-NP01 in normal mice and pancreatic PDX tumor bearing mice. Results of the SBIR phase II project should have a great impact on the future submission of an IND application for the FDA approval of a phase 1 clinical trial in advanced pancreatic cancer patients.