Characterisation of nanomedicine heterogeneity with potential for impact on product performance

目的：众所周知，纳米药物制剂在制剂中的单个纳米颗粒之间表现出显著的多分散性。这些异质性没有得到很好的研究，也很少有人知道这些异质性对产品性能的影响。表征制剂的总体平均性质，而这在纳米颗粒亚组分内可能显著变化。我们的目标是更好地了解异质性对纳米医学特性的影响，以指导该领域未来的产品设计优化。 1.使用不对称流场流分级（AF 4）表征囊泡和立方体纳米药物制剂的异质性，可能对产品性能产生负面影响。 2.通过具有耗散的石英晶体微天平（QCM-D）使用CBM 40-GM 3相互作用表征纳米药物的超选择性靶向，与某些癌症的活性药物靶向相关。3.测定样品异质性对nanomedicine.Methodology的超选择性靶向疗效的影响：用于分馏多分散纳米药物样品的主要技术将是AF 4，这是一种色谱技术，分离颗粒的大小和形状。通过多角度光散射和动态光散射（以及UV/维斯和RI）进行的在线表征将提供有关从这些制剂中分离的亚组分的大小和形状的信息。我们将开发工作流程来研究药物包封和药物释放曲线如何在这些亚组分中变化，并对这些组分进行进一步的离线表征，例如通过TEM。将使用低亲和力、高选择性CBM-40/GM 3相互作用研究超选择性靶向，这是Ralf Richter组中线性聚合物的既定靶向系统。该项目将探索如何通过QCM-D在球形自组装纳米颗粒制剂中调节超选择性靶向的性质，其中CBM-40功能化纳米药物靶向GM 3功能化膜表面。这也将由TIRF显微镜进行补充。我们将表征纳米药物在不同GM 3浓度和不同CBM-40功能化颗粒上的膜上的开和关速率以及Kd和停留时间。最后，这两条链将结合在一起，以确定纳米药物异质性（AF 4分馏）对纳米药物超选择性靶向（QCM-D）的影响。影响：纳米医学在通过增强治疗剂在体内的空间和时间定位来增强药物的药理学特性方面具有很大的前景。尽管他们的承诺，在一些早期的成功，药物纳米制剂一直缓慢转化为临床。由于所有颗粒不相同（如单独的小分子药物的情况），也存在重大的监管挑战。纳米药物制剂中的异质性可能是进行体内研究时纳米药物性能降低或意外的一个重要原因。需要更好地理解这些异质性及其对这些影响功能的纳米颗粒的相关性质的影响，以促进改进产品设计并促进该领域的产品开发管道。预期合格品：1。两种模型纳米药物制剂中尺寸、形状、组成、载药量和释放曲线的异质性表征。2.了解超选择性靶向原理如何转化为柔软的球形纳米颗粒制剂。3.了解产品异质性将如何影响异质纳米药物制剂亚群的靶向性能。

Aims:Nanomedicine formulations are well known to exhibit significant polydispersity in properties between individual nanoparticles in a formulation. These heterogeneities are not well studied and little is known about the impact of these heterogeneities on product performance. The ensemble average properties of a formulation are characterised, while this may vary significantly within nanoparticle sub-fractions. We aim to get a better understanding of the impact of heterogeneities on nanomedicine properties to guide future optimisation of product design in this field. 1. Characterisation of heterogeneity in vesicle and cubosome nanomedicine formulations using asymmetric flow field flow fractionations (AF4) with potential for negative impact on product performance. 2. Characterisation of superselective targeting of nanomedicines by Quartz Crystal Microbalance with Dissipation (QCM-D) using the CBM40-GM3 interaction, relevant in active drug targeting to some cancers. 3. Determination of the impact of sample heterogeneity on superselective targeting efficacy of a nanomedicine.Methodology:The primary technique used for fractionating polydisperse nanomedicine samples will be AF4, which is a chromatography technique that separates particles by size and shape. Inline characterisation by multi-angle light scattering and dynamic light scattering (as well as UV/vis and RI) will give information on the size and shape of sub-fractions separated from within these formulations. We will develop workflows to investigate how drug encapsulation and drug release profiles vary across these sub-fractions and also conduct further offline characterisation of these fractions, such as by TEM. Superselective targeting will be investigated using low affinity, high selectivity CBM-40/GM3 interactions, which is an established targeting system in the group of Ralf Richter for linear polymers. This project will explore how the properties of superselective targeting may be modulated in spherical self-assembled nanoparticle formulations by QCM-D with CBM-40 functionalised nanomedicines targeting GM3 functionalised membrane surfaces. This will also be complemented by TIRF microscopy. We will characterisation on and off rates as well as Kd and residence times of nanomedicines on the membrane with varying GM3 concentration and varied CBM-40 functionalisation on the particles. Finally these two strands will be brought together to characterise the impact of nanomedicine heterogeneity (fractionation by AF4) on superselective targeting of nanomedicines (QCM-D). Impact:Nanomedicines hold great promise in enhancing the pharmacological properties of drugs through enhancing the spatial and temporal localisation of the therapeutic in the body. Despite their promise, following some early successes, drug nanoformulations have been slow to translate to the clinic. There are also significant regulatory challenges due to all particles not being identical (as is the case for a small molecule drug alone). Heterogeneous properties in nanomedicine formulations are likely to be one significant cause of reduced or unexpected performance of nanomedicines when conducting in vivo studies. A better understanding of these heterogeneities and their impact on relevant properties of these nanoparticles that impact function is required to facilitate improved product design and facilitate the product development pipeline in this field. Expected Deliverables:1. Characterisation of heterogeneities in size, shape, composition, drug loading and release profiles in two model nanomedicine formulations. 2. An understanding of how principles of superselective targeting translate to soft, spherical nanoparticle formulations. 3. An understanding of how product heterogeneity will impact the targeting performance of sub-populations of heterogeneous nanomedicine formulations.