Development of NanoFludarabine for Pediatric Leukemia using LCP Nanotechnology

利用 LCP 纳米技术开发治疗小儿白血病的纳米氟达拉滨

• 批准号: 8780357
• 负责人: Jun Li
• 金额: $ 15万
• 依托单位: QUALIBER, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-17 至 2015-06-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8780357
• 关键词: Adverse effects; Autoimmune Process; Biodistribution; Bone Marrow Suppression; CD44 gene; Calcium; Cancer Patient; Cell Culture Techniques; Cell Nucleus; Cells; Child; Childhood; Childhood Leukemia; Clinical Research; Cytoplasm; DNA Synthesis Inhibition; Development; Diagnosis; Drug Formulations; Effectiveness; Encapsulated; Endocytosis; Endosomes; Ensure; Fludarabine phosphate; Folate; Folic Acid; Future; Grant; Hematologic Neoplasms; Hyaluronan; Hyaluronic Acid; In Vitro; Incidence; Inhibitory Concentration 50; Kidney; Ligands; Lipid Bilayers; Lipids; Lung; Malignant Childhood Neoplasm; Malignant Neoplasms; Measurement; Metabolism; Modeling; Names; Nanotechnology; Neurologic; Normal Cell; Nucleic Acids; Nucleotides; Osmotic Pressure; Particle Size; Pharmaceutical Preparations; Phase; Plant Leaves; Precipitation; Process; Production; Reaction; Research; Reticuloendothelial System; Small Business Innovation Research Grant; Solid; Staging; Surface; Testing; Therapeutic Index; Toxic effect; Transfusion; Treatment Efficacy; base; calcium phosphate; cancer cell; chemotherapy; design; fludarabine; folate-binding protein; graft vs host disease; improved; in vitro activity; innovation; leukemia; leukemia/lymphoma; manufacturing process; nanoparticle; nanosized; novel; particle; phosphodiester; pre-clinical; public health relevance; receptor; scale up; tripolyphosphate; tumor; uptake; zeta potential

DESCRIPTION (provided by applicant): Development of NanoFludarabine for Pediatric Leukemia using LCP Nanotechnology NanoFludarabine is Qualiber's nanoparticle formulation of fludarabine phosphate (Fludara), a drug widely used for treating leukemia and other malignancies. While fludarabine phosphate is effective against tumors, it also has significant toxicities. Many of these side effects can potentially be minimized or avoided by modifying the biodistribution of fludarabine phosphate. Furthermore, fludarabine phosphate's therapeutic index can be improved if we can preferentially deliver the drug to cancer cells using nanoparticles which have been targeted to cancer cells. Our innovation is the encapsulation of fludarabine phosphate in a novel nanoparticle formulation, called Lipid-Calcium-Phosphate (LCP). The new formulation of fludarabine phosphate in LCP nanoparticles is named NanoFludarabine. The calcium phosphate core of LCP would encapsulate fludarabine phosphate with a high efficiency and rapidly dissolve in the acidic endosome releasing fludarabine phosphate in the cytoplasm where it is further phosphorylated to fludarabine triphosphate and transported to the nucleus for inhibition of DNA synthesis. In this Phase I, Qualiber will formulate LCP for the delivery of fludarabine phosphate (NanoFludarabine) and characterize them for in vitro activity and determine feasibility for development as pediatric leukemia treatment. Aim 1. Increase Current Production Scale of Encapsulated Fludarabine Phosphate in LCP Nanoparticles by 10 Fold with In-Process Controls (IPC) Qualiber aims to test the feasibility of scalable process to encapsulate fludarabine phosphate with solid analytical measurements. The preliminary results show that fludarabine phosphate can be encapsulated into LCP particles at lab scale with batch size of 1 mg of fludarabine phosphate encapsulated. Two challenges exist at this stage: (i) small scale of production and (ii) lack of process controls Aggregation of particles was observed partly due to a low surface zeta potential. Successful production to improve the scale of nanoparticles to 10x of current scale with batch size of >10 mg of fludarabine phosphate will be the criteria for progressing to Aim 2. Also, in-process controls such as drug encapsulation efficiency (>40%), particle size and poly-dispersity will be put in place to ensure future process designs for a scalable manufacturing process that would allow further pre-clinical development and clinical studies regulated by FDA. Aim 2. Select One Targeted Ligand and Make NanoFludarabine to Target Leukemia with Proof of Concept (POC) In Vitro Studies NanoFludarabine nanoparticles produced in the above scale-up process will be further targeted to leukemia cells. Qualiber is planning to make NanoFludarabine with at least two different ligands to target (i) CD44 receptor with Hyaluronan/Hyaluronic Acid (HA) and (ii) Folate receptor with PEGylated folic Acid (FA). Free fludarabine phosphate, NanoFludarabine without ligand, NanoFludarabine with HA or FA targeting ligands will be tested in in vitro cell culture models. The results will be used to determine the following: (a) Is NanoFludarabine more efficacious than free fludarabine phosphate and (b) Which targeting ligand provides highest cell uptake via endocytosis of the nanoparticles. One ligand will be chosen based on the targeting efficacy, availability and manufacturing feasibility to move forward. Successfully demonstrating that NanoFludarabine (with HA or FA targeting ligand) has a IC50 values at least 2-5 fold lower than free fludarabine phosphate will be a key milestone for progressing the product for further preclinical development and Phase II SBIR grant.

描述（由申请人提供）：使用LCP纳米技术开发用于儿科白血病的NanoFludarabine NanoFludarabine是一种广泛用于治疗白血病和其他恶性肿瘤的药物磷酸氟达拉滨（Fludara）的纳米颗粒制剂。虽然磷酸氟达拉滨对肿瘤有效，但它也具有显著的毒性。通过改变磷酸氟达拉滨的生物分布，可以最大限度地减少或避免许多这些副作用。此外，如果我们可以使用靶向癌细胞的纳米颗粒优先将药物递送到癌细胞，则可以提高磷酸氟达拉滨的治疗指数。我们的创新是将磷酸氟达拉滨封装在一种称为脂质磷酸钙（LCP）的新型纳米颗粒制剂中。LCP纳米颗粒中磷酸氟达拉滨的新制剂被命名为NanoFludarabine。LCP的磷酸钙核心将高效地包封磷酸氟达拉滨，并迅速溶解在酸性内体中，在细胞质中释放磷酸氟达拉滨，在细胞质中磷酸氟达拉滨进一步磷酸化为三磷酸氟达拉滨并转运到细胞核以抑制DNA合成。在该I期研究中，Amber将配制用于输送磷酸氟达拉滨（NanoFludarabine）的LCP，并对其进行体外活性表征，并确定开发为儿科白血病治疗的可行性。目标1.通过过程中控制（IPC）将LCP纳米颗粒中包封的磷酸氟达拉滨的当前生产规模增加10倍，该公司旨在测试用固体分析物包封磷酸氟达拉滨的可扩展工艺的可行性。 测量.初步结果表明，磷酸氟达拉滨可以在实验室规模下包封到LCP颗粒中，包封的磷酸氟达拉滨的批量为1 mg。在该阶段存在两个挑战：（i）小规模生产和（ii）缺乏工艺控制，观察到颗粒聚集部分是由于低表面ζ电位。成功生产以将纳米颗粒的规模提高到当前规模的10倍，批量>10 mg磷酸氟达拉滨将是进展到目标2的标准。此外，还将实施过程中控制，如药物包封率（>40%）、粒度和多分散性，以确保未来的工艺设计能够实现可扩展的生产工艺，从而允许FDA监管的进一步临床前开发和临床研究。目标2.选择一种靶向配体并通过概念验证（POC）制备靶向白血病的纳米氟达拉滨体外研究在上述放大过程中产生的纳米氟达拉滨纳米颗粒将进一步靶向白血病细胞。生物制药公司正计划用至少两种不同的配体来制造NanoFludarabine，以靶向（i）用Hyaluronan/Hyaluronic Acid（HA）靶向CD 44受体，（ii）用PEG化叶酸（FA）靶向叶酸受体。将在体外细胞培养模型中检测游离磷酸氟达拉滨、不含配体的NanoFludarabine、含HA或FA靶向配体的NanoFludarabine。结果将用于确定以下内容：（a）纳米氟达拉滨是否比游离磷酸氟达拉滨更有效，以及（B）哪种靶向配体通过纳米颗粒的内吞作用提供最高的细胞摄取。将根据靶向功效、可用性和制造可行性选择一种配体以向前推进。成功证明NanoFludarabine（具有HA或FA靶向配体）的IC 50值比游离磷酸氟达拉滨低至少2-5倍，将是推进产品进一步临床前开发和II期SBIR资助的关键里程碑。