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(申请人提供)：使用液晶聚合物纳米技术开发治疗儿童白血病的纳米氟达拉滨纳米氟达拉滨是Qualiber公司的磷酸氟达拉滨(Fludara)纳米颗粒配方，是一种广泛用于治疗白血病和其他恶性肿瘤的药物。虽然磷酸氟达拉滨对肿瘤有效，但它也有显著的毒性。通过改变磷酸氟达拉滨的生物分布，这些副作用中的许多可能被最小化或避免。此外，如果我们能够优先使用针对癌细胞的纳米粒将药物输送到癌细胞，则可以提高氟达拉滨的治疗指数。我们的创新是将磷酸氟达拉滨包裹在一种名为磷脂-钙-磷酸(LCP)的新型纳米制剂中。LCP纳米粒中磷酸氟达拉滨的新配方被命名为纳米氟达拉滨。LCP的磷酸钙核心可以高效地包裹磷酸氟达拉滨，并迅速溶解在酸性内体中，释放出胞浆中的磷酸氟达拉滨，在胞浆中进一步磷酸化为三磷酸氟达拉宾，并转运到细胞核中抑制DNA合成。在这一阶段，Qualiber将为磷酸氟达拉滨(纳米氟达拉滨)的输送制定LCP，并对它们的体外活性进行表征，并确定将其开发为治疗儿童白血病的可行性。目的1.采用过程中控制(IPC)技术将液晶微球囊化磷酸氟达拉滨的生产规模扩大10倍，目的是验证固体分析法可扩展工艺制备氟达拉宾磷酸盐的可行性 测量。初步结果表明，磷酸氟达拉滨可在实验室条件下以1 mg磷酸氟达拉滨为包封剂制备LCP微球。在这个阶段存在两个挑战：(1)生产规模小和(2)缺乏工艺控制，部分原因是由于表面Zeta电位低，观察到颗粒聚集。成功的生产将纳米粒的规模提高到当前规模的10倍，批量生产10毫克的氟达拉滨磷酸将是进展到目标2的标准。此外，还将实施药物包封率(&gt；40%)、颗粒大小和多分散性等过程控制，以确保未来可扩展制造工艺的工艺设计，使进一步的临床前开发和临床研究能够受到FDA的监管。目的：选择一个靶向配体，制备具有概念验证(POC)的纳米氟达拉滨靶向白血病体外研究以上放大过程中制备的纳米氟达拉滨纳米粒将进一步靶向白血病细胞。Qualiber计划制造具有至少两种不同配体的纳米氟达拉滨，以(I)以透明质酸/透明质酸(HA)为靶标的CD44受体和(Ii)以聚乙二醇化叶酸(FA)为靶标的叶酸受体。游离氟达拉滨磷酸、无配体的纳米氟达拉滨、带有HA或FA靶向配体的纳米氟达拉滨将在体外细胞培养模型中进行测试。结果将被用来确定：(A)纳米氟达拉滨是否比游离的氟达拉滨磷酸更有效，以及(B)靶向配体通过内吞纳米粒提供最高的细胞摄取。将根据靶向效力、可用性和制造可行性来选择一种配体以推动进展。成功证明纳米氟达拉滨(带有HA或FA靶向配体)的IC50值至少比游离的氟达拉滨磷酸低2-5倍，这将是推动该产品进一步临床前开发和第二阶段SBIR批准的关键里程碑。