設計並合成具標靶傳送與增進癌細胞內累積量之長效型艾黴素製劑

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研究生：	盧泉潓 Chuan-Hui Lu
論文名稱：	設計並合成具標靶傳送與增進癌細胞內累積量之長效型艾黴素製劑 Design and Synthesis of Long-Acting Doxorubicin Derivatives Aiming Targeted Delivery and Enhanced Drug Translocation
指導教授：	阮若屈 Ruoh-Chyu Ruaan
口試委員:
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程與材料工程學系 Department of Chemical & Materials Engineering
論文出版年：	2013
畢業學年度：	101
語文別：	英文
論文頁數：	69
中文關鍵詞：	抗生胜肽、藥物載體
外文關鍵詞：	Cell-penetrating peptide
相關次數：	點閱：15 下載：0
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在此項研究中，我們設計一個具有專一性傳遞與增加藥物在細胞內累積量的長效型艾黴素(DOX)載體。從先前的研究中發現，IL類似物可攜帶小分子螢光物質FITC進到細胞裡; 根據這樣的發現，我們進一步想利用此條胜肽與小分子藥物-艾黴素進行共價接合增加攜帶藥物進入癌細胞中的累積量。更進一步，我們將一具有生物相容性的高分子聚乙二醇(polyethylene glycol, PEG) 藉以保護胜肽，避免酵素將胜肽切割進而達到延長體內循環的時間。此外我們也設計一段蛋白酶分解片段，使載體能在特定細胞周圍釋放。研究結果中發現，在聚乙二醇保護下，切割一定量的胜肽從十分鐘延長至四小時，可證明聚乙二醇可有效的保護胜肽。同時，在蛋白酶切割後，我們觀察到藥物在細胞內累積量上升。因此，我們認為此一藥物載體設計可應用於實際藥物傳遞，並且能增進藥物的累積量。

In this study, we designed and synthesized the novel doxorubicin (DOX) formulation for enhanced drug delivery and cancer cell targeting. An Indolicidin analogue was covalently conjugated to DOX and acted as a transmembrane carrier for enhancing drug translocation into HepG2 cells. The bioinert polymer, polyethylene glycol (PEG), was conjugated with peptide for prolonging in vivo drug circulation. The protease cleavable sequences were designed for cancer cell recogition. The PEG segment could be detached from DOX-peptide in the presence of protease. It was found that the half-life in trypsin after PEG protection could be increased from 10 minutes to 4 hours. And the enhanced penetration could be observed after protease cleavage. Thus, this DOX formulation is able to be a potential to apply in pharmaceutical use.

Abstract (English)    I
Abstract (Chinese)    II
List of Figures    V
List of Tables    VII
Chapter 1    Introduction    1
1    Research motivation    1
2    Research purpose    3
Chapter 2    Literature survey    4
1    Importance of drugs or biologics delivery    4
1.1    Small molecule drugs    4
1.2    Peptide and protein drug    6
1.3    Genetic drugs    7
2    Carriers in drug delivery    7
2.1    Metal-based Nanoparticles    9
2.2    Polymer-based carriers    11
2.3    Liposomal-based carriers    12
2.3.1    Enhanced permeability and retention (EPR) effect    13
2.4    Peptide-based carriers    14
2.4.1    Antimicrobial peptides (AMPs)    14
2.4.2    Cell-penetrating peptides (CPPs)    15
3    DOX delivery    16
3.1    Structure of Doxil®    17
3.2    Improved PEGylated liposomal doxorubicin    18
4    Targeting design for drug delivery    20
4.1    Antibody    20
4.2    Matrix metalloproteinase-2/9    21
5    Biostability in drug delivery system    23
6    Molecular design for DOX delivery    24
Chapter 3    Materials and Methods    25
1    Chemical compounds and biomolecules    25
2    Experiment framework    26
2.1    PEGylation process    27
2.2    Protease digestion    28
2.3    Analysis of high-performance liquid chromatography    29
2.4    Peptide conjugated with DOX using sulfo-SMPB    29
2.5    Complete formulation synthesis process    29
2.6    Cytotoxicity assay    30
Chapter 4    Delivery System Design    31
1    Role of CPP for DOX intracellular translocation    31
1.1    The role of free CPP on DOX intracellular translocation    32
1.2    Synthesis of CPP and DOX conjugates    34
1.3    The role of conjugated CPP on DOX intracellular translocation    36
2    Shortest MMP-2 targeting sequence design    38
3    Enzyme digestion of peptides and its PEGylated form    42
3.1    Peptide PEGylation and purification    43
3.2    Effect of PEG length on the protease stability of CPP    46
3.3    MMP-2 digestion of PEGylated MMP-2 recognizable peptide    48
4    Novel formulation of PEG-GPLGI-P17-DOX conjugate    49
4.1    Synthesis of PEG-GPLGI-P17-DOX conjugate    49
4.2    Cell toxicity of PEG-GPLGI-P17-DOX conjugate    50
Chapter 5    Conclusions    52
Reference    53


                                

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