多功能雙性機丁聚醣-pyranine核殼奈米載體之設計合成及在藥物釋放,顯影, 細胞內酸鹼值測定之應用研究

Abstract

此研究計畫之目標在於利用本實驗室先前所開發之兩性幾丁聚醣並結合螢光分子曱氧 基苄腈，設計出一種新型多功能奈米粒子，其同時具有藥物傳遞、生物顯影以及細胞 内酸鹼值量測等功能。此新型奈米元件可經由兩分子間之靜電吸附力（帶正電荷之兩性 幾丁聚醣與負電荷之曱氧基苄腈）或是化學修飾法(利用酯化反應將曱氧基苄腈修飾於 幾丁聚醣上）組成核殼形態之奈米結構。兩性幾丁聚醣之自組織能力使其組成球形奈米 粒子，並且可裝載並傳遞如喜樹鹼及薑黃素之藥物。連接在兩性聚丁聚醣奈米粒子上 之曱氧基苄腈提供了細胞内與細胞外之螢光訊號以及酸鹼敏感偵測能力，前者可直接 量測細胞内酸鹼值並可即時監控，這是單獨的曱氧基苄腈分子無法達成地。目前初步 的成果提供了強力的證據證明此多功能奈米粒子開發成功之潛力。本研究計畫最重要 的是將此新型多功能生物材料提升到新的階段，經由在體外表徵與體内之表現，使此 新型多功能生物材料從技術性與實用性的觀點而有進一步的發展與突破，帶給此新型 多功能奈米粒子從早期診斷到後期階段性治療更完整與廣泛地應用。

Synthesis of a new type of multifunctional nanoparticle capable of integrating drug delivery, bio-imaging, and intracellular pH monitoring simultaneously via an assembly of an amphiphilic chitosan (termed as CHC, which was well-developed from this lab) and a fluorescence probe, known as pyranine, is proposed in this 2-year research project. This new type of nanoobject can be assembled into a core-shell-type nanostructure via either electrostatic attraction (between positively-charged CHC and negatively-charged pyranine) or chemical modification (pyranine-modified CHC via esterification interaction) between these two molecules. The self-assembly capability of the CHC ensures a spherical nanoparticle to be built, which also renders drugs (CPT and demethoxycurcumin, as model molecules) to be encapsulated and delivered. The presence of pyranine, associated with the CHC nanomatrix, ensures a fluorescence lightening and a pH-sensitive emission to be detected intracellularly and extracellularly, where for the former, enable a direct measurement of intracellular pH to be monitored on a real-time base (which has been realized to be impossible for pyranine alone). Preliminary outcomes gave strong evidences of potential success in the development of such multifunctional nanoparticle. It is more critical and important to bring this new multifunctional biomaterial to a stage through this

2-year project that enables a further exploitation and new discoveries from both technical and practical viewpoints through in-vitro characterization and in-vivo performance, making this new type of multifunctional nanoobject to be more robust in a wider variety of applications, ranging from early diagnosis to later-stage therapeutic treatment.