Supramolecular Tandem Membrane Assays

超分子串联膜测定

• 批准号: 333358947
• 负责人: Professor Dr. Andreas Hennig
• 金额: --
• 依托单位: Institut für Chemie neuer Materialien
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/333358947?language=en
• 关键词: Supramolecular; Tandem; Membrane; Assays

Membrane transport of organic molecules plays a crucial role in the discovery and delivery of small-molecule drugs as well as in the mechanism of action of antimicrobial peptides and cell-penetrating peptides (CPPs). The measurement of the actual transport of these compounds across the lipid bilayer membrane remains nevertheless a major experimental challenge. We have recently shown that liposome-encapsulated supramolecular reporter pairs (consisting of a fluorescent dye and a macrocyclic receptor molecule) qualify to unambiguously detect membrane transport of organic molecules and peptides by a simple and fluorescence-based method. Within this proposal, we will comprehensively establish the utility and applicability of this method.In detail, we will encapsulate supramolecular reporter pairs in liposomes of varying size and membrane composition in order to investigate membrane transport of selected organic molecules and peptides by absorption and fluorescence spectroscopic methods. This includes spectroscopic characterizations, time-resolved measurements, including stopped-flow techniques for rapid kinetics, as well as fluorescence microscopy with giant unilamellar vesicles (GUVs) as well as live cells.The supramolecular tandem membrane assays to be developed herein will be utilized to follow membrane transport of small, unlabeled molecules and CPPs. This will afford membrane permeability coefficients and activation energies for membrane transport of small, organic molecules as experimental benchmarks, for example, for computational studies. The adaptation to parallel artificial membrane transport assays (PAMPA) will demonstrate potential future applications in high-throughput screening in the pharmaceutical industry. We will also establish liposome-encapsulated supramolecular reporter pairs as key components in advanced supramolecular sensing systems, in which the membrane transport process improves the detection limit and selectivity of supramolecular sensor systems, thereby enabling orders of magnitude sensitivity enhancements and the discrimination of peptide sequences.

有机分子的膜转运在小分子药物的发现和递送以及抗菌肽和细胞穿透肽（CPP）的作用机制中起着至关重要的作用。然而，测量这些化合物穿过脂质双层膜的实际运输仍然是一个主要的实验挑战。我们最近已经表明，脂质体封装的超分子报告对（由荧光染料和大环受体分子组成）有资格通过简单的基于荧光的方法明确检测有机分子和肽的膜转运。在这个提议中，我们将全面建立这种方法的实用性和适用性。详细地说，我们将在不同大小和膜组成的脂质体中封装超分子报告对，以便通过吸收和荧光光谱方法研究选定的有机分子和肽的膜转运。这包括光谱表征，时间分辨的测量，包括停流技术的快速动力学，以及荧光显微镜与巨大的单层囊泡（GUV）以及活celles.The超分子串联膜测定将在此开发将被用来遵循膜运输的小，未标记的分子和CPP。这将提供膜渗透系数和活化能的膜运输的小，有机分子作为实验基准，例如，计算研究。适应平行人工膜转运试验（PAMPA）将展示潜在的未来应用在制药工业的高通量筛选。我们还将建立脂质体封装的超分子报告对作为先进的超分子传感系统的关键组成部分，其中膜传输过程提高了超分子传感器系统的检测限和选择性，从而使数量级的灵敏度增强和肽序列的歧视。