2015-10-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/646871摘要：本計畫目標在開發可抑制無譯密碼之新型藥物。鑒於約30%的人類遺傳疾病均由無譯密碼突變所引起，由於本計畫所成功研發之新型藥物具有治療所有由無譯密碼突變所引起的遺傳疾病患者的潛力，因此極具開發價值。我們將以罕見疾病Dravet症候群，也就是由SCN1A基因突變所引起的小兒嚴重型癲癇作為疾病模式。大部分的Dravet症候群病人為異型合子，且>50%是無譯密碼(或終止密碼)突變，因此開發抑制無譯密碼突變而回復全長蛋白質表現的新型藥物，對臨床治療將有極大助益。本實驗室先前利用knock-in技術已成功產製具Scn1a基因無譯密碼突變的基因置入小鼠，且完成分析後顯示此小鼠具有Dravet病人的表現型(Tsai et al., 2015)，因此為臨床前測試的極佳活體藥物篩選平台。本團隊並利用已建立的具無譯密碼的細胞平台及國家型科技計畫(NRPB)的高通量小分子藥物篩選平台，自12萬個化合物資料庫中得到200個初選化合物(衛福部計畫2012~1014年)。綜合以上成果，本團隊提出本計畫，主要目標是完成具抑制無譯密碼的候選藥物篩選，實行目標如下:目標一: 測試初選化合物抑制無譯密碼之活性。(1)依據結構、溶解度、生物可利用性，EC50等分析及歸類初選化合物。 (2)利用細胞平台與小鼠測試至少5~10個化合物的抑制無譯密碼的能力。目標二: 以Dravet小鼠模式測試前導化合物之藥效。(1)利用Dravet症候群小鼠確認前導藥物的藥效，包括癲癇控制，過動控制，改善自閉症光譜障礙等。(2)確認前導藥物安全性與毒性。<br> Abstract: The goal of this project is to develop new therapeutic compounds with potential of nonsense suppression. Given that nearly 30% of human diseases are caused by nonsense mutation, the novel compounds that will be developed in this project will potentially benefit all patients suffering from disasters of nonsense mutations. We will use Dravet syndrome, the severe myoclonic epilepsy caused by mutations in SCN1A gene, as a disease model. The Dravet syndrome patients are heterozygous with a mutation allele and > 50% patients are nonsense (or stop-codon) mutations. Therefore, developing nonsense suppression drugs to allow re-expression of the full-length protein will be of great clinical implications. Our group has generated a knock-in mouse line containing a nonsense mutation in the Scn1a gene, and completed analyses to show that this mouse model displays phenotypes mimicking the Dravet syndrome patients (Tsai, et al., 2015). The mice will be suitable for in vivo drug screen. With the support from the Ministry of Health and Welfare (2012~2014), we have also established and used a cell-based screening platform, and the (NRPB) core facility of ultra High-throughput screening service to obtain 200 hit compounds from approximately 120,000 compounds. With all the achievement, we propose this project with the goal to identify candidate compounds with nonsense suppression potentials. The specific aims are the following. Specific Aim 1: validation of hit compounds with nonsense suppression activity. (1)Analyzing and categorizing the hit compounds according to structure, solubility, bioavailability, EC50 etc.(2)Validating 5~10 best hit compounds for nonsense suppression activities on the cell-based model and homozygous Dravet mice model as lead compounds. Specific Aim 2: pharmacoefficacy of hit compounds in Dravet mice.(1)Testing the lead compounds on the Dravet mice for their therapeutic efficacy in seizure control, hyperactivity control, and autism control (2)Verifying safety and toxicity of lead compounds抑制無譯密碼基因轉殖鼠新藥開發non-sense suppressionDravet syndromenew drug development