2014-05-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659714摘要：為加速發展與完成台灣生物科技產業鏈，著眼於亞洲特有疾病是最省時及有效率的策略之一，可以增加台灣於藥物及體外試劑發展的競爭力。腸病毒 71型首先在 1969年於加州發現，尤其是在泛亞太地區，從 1997年後即成為一種致命性的新興病原體。幾乎所有的病毒皆傾向演化出不同的抗原變異株與抗藥性，這種特性造成了發展有效地病毒治療策略的阻礙。至今，對於腸病毒 71型的感染尚無有效的治療方式或是疫苗問世。第一型干擾素可以提供抗病毒的第一線防線，隨後更會進一步的導致後天免疫的建立。於病毒早期感染時，施予干擾素治療可以限制病毒的擴散，但是，病毒卻可以透過干擾或破壞干擾素訊息傳遞相關的分子，逃脫干擾素的攻擊。最近，我們發現的一個新的機制，其中，腸病毒會誘發宿主微核醣核酸-146a(miR-146a)的表現，進而抑制干擾素產生與逃脫免疫。因此，我們希望發展出一個以微核醣核酸為基礎的抗腸病毒 71型策略。在這個計畫中，我們以體外的報導基因分析法與腸病毒 71型小鼠感染模式從一群抗微核醣核酸-146a的衍生物中，挑選出活性最好的兩、三個衍生物。再以報導基因分析法偵測抗微核醣核酸-146a的 IC50以及核子影像收集早期藥物動力學/藥效學的資料，隨後將建立以 MALDI-TOF或 ELISA直接測量微核醣核酸-146a量的方法學，並用於偵測微核醣核酸-146a 的組織分布、半生期與藥物動力學/藥效學的資料。我們亦會定性分析微核醣核酸-146a的化學與物化特性。此外，ADME、單一劑量急性毒性試驗、重複劑量毒性試驗以及新血管、呼吸與中樞神經等三大系統的藥物安全性都將會被研究。MALDI-TOF/ELISA的微核醣核酸-146a測量法亦會用於預配方研究與建立微核醣核酸-146a製程的品質控制。最後，微核醣核酸-146a對於受腸病毒 71型感染小鼠的細胞性與體液性免疫影響也將列入評估。經過這計劃的努力，我們希望抗腸病毒 71型核酸藥物的開發應可以從先導藥物的階段進入候選藥物階段，甚至是臨床前期，為台灣生技產業升級貢獻一己之力。 <br> Abstract: To develop and complete the value chain of Bio-Technology industry focusing on Asia-unique diseases is one of the most time- and cost-effective means to increase competitiveness of Taiwan in pharmaceutics and development of in vitro diagnosis (IVD). Enterovirus 71 (EV71), which was first identified in California in 1969, has become a newly emerging life-threatening pathogen, particularly in the Asia-Pacific region, since 1997. All of viruses are predisposed to evolve new antigenic variations and drug resistances that impede the development of effective antiviral therapies. Up to date, there is no effective therapy or vaccine against EV71 infection. Type I IFNs provide a first-line of defense in antiviral responses and further lead to the establishment of adaptive immunity. Administration of IFNs can limit a virus from spreading at an early phase during virus infections but virus can escape IFN attacks by influencing or disrupting IFN signalling-associated molecules by viral components. Recently we discovered a new strategy, by which Enterovirus induces cellular miR-146a expression to inhibit IFN production and escape host immune attacks. Hence, we would like to develop a new microRNA-based therapeutic strategy against EV71 infection. In this proposal, we will determine IC50 of anti-miR-146a by an in vitro reporter assay and the early Pharmacokinetics/Pharmacodynamics will use molecular image platform for anti-miR-146a modification, radio-labeling, SPECT/PET/CT/MRI image, bio-distribution and organ pharmacokinetic study. We then will establish direct measurement of anti-miR-146a by MALDI-TOF or ELISA for determining bio-distribution and half-life as well as Pharmacokinetics/Pharmacodynamics of anti-miR-146a. The chemistry and physicochemical properties of anti-miR-146a will be determined. In vitro ADME, toxicology tests for acute single dose/repeated dose toxicity studies will be performed and pharmacological safety will be evaluated in three major organs, cardiovascular, respiratory and central nervous systems. MALDI-TOF or ELISA anti-miR-146a detection is also used to evaluate the efficiency of different pre-formulations and establish the quality control of anti-miR-146a manufacture. Finally the inference of anti-miR-146a on the cellular and humoral responses of EV71-infected and mock mice will be determined. Through this study the development of anti-EV71 oligonucleotide should be progressed from Lead to "Lead to Candidate" and "Candidate to IND enabling" and contributes to the upgrade of Taiwan Biotechnology industries.腸病毒 71型干擾素微核醣核酸抗病毒藥EV71IFNmicroRNAanti-virus drug