Development of a Liposome Doxorubicin Product Drug Release Assay

脂质体阿霉素产品药物释放测定的开发

• 批准号: 8666586
• 负责人: Peter Working
• 金额: $ 46.78万
• 依托单位: ZONEONE PHARMA, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2014-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8666586
• 关键词: Development; Liposome; Doxorubicin; Product; Drug

Development of a Liposome Doxorubicin Product Drug Release Assay A barrier to the rational assessment for regulatory purposes of generic drug delivery system for parenterally administered drugs is the absence of validated mechanism-based drug release assays that reflect or predict in vivo drug release from the carrier. One such FDA licensed drug carrier is the sterically stabilized liposome product known as Doxil(R). Members of our team played important roles in the early development of Doxil(R) a liposome formulation that was specifically designed to be exceptionally stable at 37¿C in the presence of serum in order to provide a sustained release of drug over the course of three to four weeks. This inherent stability is a barrier when it comes to create in vitro doxorubicin release systems. Herein we propose a mechanism-based plan to devise, assess and validate in vitro release assays (also designated as dissolution assays) that distinguish liposome formulations with variations of composition and physical properties. Based upon the in vitro release profile of the innovator product Doxil(R), we will identify assays that better correlate with Doxil(R)'s in vivo drug release profile. In the background section, we provide a critical review of liposome content release assays for liposome formulations. We use this information to create a research plan with four specific aims. In the background section, we provide a critical review of content release assays for liposome formulations. In aim 1, we prepare and characterize a series of liposome formulations that span the range of compositions, source of materials and physical properties that can be encompassed by the product description of Doxil(R). The formulations are prepared using the technologies used in the manufacture of Doxil(R) and include modifications to the mean particle diameter, hydrogenated phosphatidylcholine lipid acyl chain composition, pegylated lipid mole percent, ammonium sulfate concentration, residual sulfate concentration in the liposome, internal pH, number of lamellae, suspending buffer composition and doxorubicin concentration. These liposome formulations will be compared in subsequent aims to the innovator product for drug retention and release characteristics. In aim 2, we will devise and validate multiple drug release assays that define release at multiple temperatures in the presence of whole blood, serum components or synthetic materials that absorb cholesterol, lipids or doxorubicin in a traditional flow through device where the liposomes are retained by dialysis membranes or in single unit systems that require separation of the liposomes from the released drug on size exclusion columns followed by assessment in the change in liposome chemical composition or physical properties as a function of time. In this aim, we also will introduce and validate a cell-based transwell assay that involves maintaining periphery blood mononuclear cells (PBMC) in one compartment and cancer cells lines that are sensitive to doxorubicin in a second compartment. The liposomes will be introduced to the PBMC compartment and the effect of released doxorubicin on cell viability/proliferation will be monitored on cancer cells in the second compartment. In aim 3, we will vary the drug release (dissolution) conditions and measure the extent of release and variability of release as a function of the principle components and operating conditions of the release assay. The innovator product Doxil(R) will serve as the gold standard for setting the operating conditions that lead to reproducible release of drug as a function of time until 66% of the doxorubicin content of the liposome is released. LipoDox(R) and the test formulations with the above noted variations will be assessed in the assays that provide the most consistent release and the greatest extent of release within a seven day period. In aim 4, we will identify those drug release assays which better correlate with published in vivo human data and identify the method or methods that provide better predictions of the in vivo release profile. Our goal is to create a standard doxorubicin release assay with precisely specified conditions, devices and components so that others can readily assess if follow-on liposome doxorubicin formulations have equivalent doxorubicin release in these relevant release assays to what is measured using Doxil(R).

阿霉素脂质体药物释放度检测方法的建立 对非专利药物非专利给药系统进行合理评估的障碍 给药是缺乏有效的、基于机制的药物释放分析，以反映或预测 体内药物从载体中释放。FDA许可的药物载体之一是空间稳定的脂质体 名为Doxil(R)的产品。我们团队的成员在Doxil(R)a的早期开发中发挥了重要作用 专门设计的脂质体配方，在37℃时，在 为了在三到四周的时间内提供药物的持续释放，给药人提供了一种新的血清。这与生俱来 稳定性是创建阿霉素体外释放系统的障碍。在此，我们提出一种 设计、评估和验证体外释放试验(也称为溶出度)的基于机制的计划 分析)，区分不同成分和物理性质的脂质体制剂。基座 根据创新者产品Doxil(R)的体外释放情况，我们将确定更好地关联的分析方法 S的体内药物释放概况。在背景部分，我们对脂质体进行了评论 脂质体制剂的含量释放分析。我们使用这些信息来创建一个研究计划，其中包括四个 明确的目标。在背景部分，我们对脂质体的内容释放试验进行了综述。 配方。在目标1中，我们制备了一系列脂质体制剂，并对其进行了表征 产品描述中可包含的成分、材料来源和物理性能 Doxil(R)。该制剂使用在生产Doxil(R)中使用的技术和 包括对平均粒径、氢化磷脂酰胆碱脂酰链的修饰 组成，聚乙二醇化脂肪摩尔百分比，硫酸铵浓度，残留硫酸盐浓度 脂质体、内pH、片层数、悬浮缓冲液组成和阿霉素浓度。 这些脂质体制剂将在随后的目标中与创新者产品进行药物保留的比较 和释放特性。在目标2中，我们将设计和验证多种药物释放试验，以确定 在全血、血清成分或合成材料存在的情况下在多种温度下释放 在传统的流过装置中吸收胆固醇、脂类或阿霉素，其中脂质体被保留 通过透析膜或在需要从释放的脂质体中分离的单一单元系统中 药物大小排除柱，随后评估脂质体化学成分的变化或 物理性质作为时间的函数。在这个目标中，我们还将介绍和验证一种基于细胞的Transwell 外周血单核细胞(PBMC)维持在一个隔室的检测和癌症 对阿霉素敏感的细胞系在第二个隔间。脂质体将被引入到 将监测PBMC隔室和释放的阿霉素对细胞存活/增殖的影响 癌细胞在第二个隔间。在目标3中，我们将改变药物释放(释放)条件和 作为主要组成部分的函数来衡量释放的程度和可变性 释放试验的操作条件。创新产品Doxil(R)将成为 将导致药物可重复释放的操作条件设置为时间的函数，直到66% 释放脂质体中的阿霉素含量。Lipodox(R)和上述试验制剂 将在提供最一致的释放和最大程度的 在七天内释放。在目标4中，我们将确定那些更好地关联的药物释放分析 与已发表的活体人类数据进行比较，并确定能够更好地预测 体内释放情况。我们的目标是创建一种标准的阿霉素释放试验， 条件、设备和成分，以便其他人可以容易地评估后续脂质体阿霉素 在这些相关释放试验中，制剂的阿霉素释放量与使用 Doxil(R)。