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標題: | 以裝載Vacuolar ATPase 阻斷劑之奈米顆粒用於抗流
感病毒感染之效果 Antiviral Efficacy of the Nanoparticulate Vacuolar ATPase Blockers against Influenza Virus Infection |
作者: | You-Ting Chen 陳宥廷 |
指導教授: | 陳慧文 |
關鍵字: | 流感病毒,Vacuolar ATPase 阻斷劑,Diphyllin,Bafilomycin,奈米顆粒, Influenza virus,Vacuolar ATPase inhibitor,Diphyllin,Bafilomycin,Nanoparticles, |
出版年 : | 2018 |
學位: | 碩士 |
摘要: | 流感病毒每年在全球造成許多發病、重症及死亡,是全球相當關心的公共衛生議題,傳統的治療策略為使用拮抗病毒蛋白質的藥物,然而帶有抗藥性突變的流感病毒毒株的出現,成為臨床治療上一大挑戰。Diphyllin與Bafilomycin是強效的Vacuolar ATPase阻斷劑,先前被證實對流感病毒具有抑制效果,然而,它們極差的水溶性和可能的副作用限制了它們的臨床應用。此研究的核心目標即是設計新穎的抗流感病毒治療策略,利用奈米顆粒來裝載與遞送抗病毒藥物,以提高藥物的安全性與效性。本研究中,我們分別製備了裝載Diphyllin和Bafilomycin的奈米顆粒,以動態光散射儀與穿透性電子顯微鏡觀察奈米顆粒的型態、粒徑與表面電位,並以高效液相色譜分析藥物在奈米顆粒中的包覆率。實驗結果顯示,我們所製備的奈米顆粒能有效地遞送進入Fcwf-4、ARPE-19、MH-S等多種細胞內部,並緩慢的釋放。在MDCK及MH-S細胞中顯示,以奈米顆粒包覆遞送之藥物比未包覆之藥物具有較低的細胞毒性,與顯著較高的抗流感病毒活性。在活體的安全性研究中,分別由靜脈內及鼻腔內給予小鼠Diphyllin奈米顆粒,觀測其體重變化及血液生化參數,發現小鼠對於Diphyllin奈米顆粒有良好的耐受性,並在流感病毒攻毒之後,有給予Diphyllin奈米顆粒之組別與未給藥組相比,有較少的體重減輕。另一方面,我們也製備了外覆紅血球膜的奈米顆粒,透過紅血球膜與流感病毒的交互作用,此奈米顆粒能夠專一地被流感病毒感染後的細胞攝入,有機會發展為標靶治療藥物之平台。總結來說,本研究顯示以奈米載體遞送抗病毒藥物為一新穎、有潛力之治療策略,能夠對病毒傳染病之藥物發展帶來更多希望。 Influenza virus infections are a major public health concern and cause significant morbidity and mortality worldwide. Conventional treatments against the disease are designed to target viral proteins. Nevertheless, the emergence of new influenza viral strains carrying drug-resistant mutations that can outpace the development of pathogen-targeting antivirals presents a major clinical challenge. Diphyllin and bafilomycin are potent vacuolar ATPase inhibitors, and previously known to have anti-influenza virus activity. However, their poor water solubility and potential off-target effect may limit the clinical application. The central objective of this project is to generate a novel anti-influenza therapeutic strategy, integrating nanoparticle technology to enhance host-targeting antiviral delivery towards improved drug safety and efficacy. In this study, we successfully constructed diphyllin-loaded nanoparticles and bafilomycin-loaded nanoparticles. The shape, size, and zeta potential of these nanoparticles were measured by dynamic light scattering and transmission electron microscopy, and the drug encapsulation was analyzed by high-performance liquid chromatography. The nanoparticles can be efficiently delivered intracellularly to multiple cell lines including Fcwf-4, ARPE-19, and MH-S cells. Moreover, the drug-loaded nanoparticles have a sustained drug release profile. The nanoformulated drugs exhibited lower cytotoxicity as compared to the free drug formulation in MDCK and MH-S cells. Furthermore, drug-loaded nanoparticles demonstrated prominent broad-spectrum anti-influenza activity in vitro. In an in vivo safety study that evaluated body weight and blood chemistry parameters following intravenous or intranasal administration, diphyllin nanoparticles were well-tolerated in mice. Furthermore, after infection, the mice treated with diphyllin-loaded nanoparticles had less body weight loss as compared to the control group. In addition, to further improve targeting delivery of nanoparticles to virus-infected cells, fluorescent nanoparticles were coated with the red blood cell membrane to validate the viral infection-specific targeting of nanoparticles. Collectively, this work highlights nanoformulated vacuolar ATPase inhibitors as potential host-targeted treatments against influenza. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/77769 |
DOI: | 10.6342/NTU201801911 |
全文授權: | 有償授權 |
顯示於系所單位: | 獸醫學系 |
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