- 學位論文
開發三階段肺纖維化藥物篩選模型，篩選HDAC6、HDAC8和HDAC6/8抑制劑用於治療TGF-β1誘導的特發性肺纖維化
Establishment of a three-stage platform to screen for newly synthesized HDAC6, HDAC8, and HDAC6/8 inhibitors against TGF-β1-induced idiopathic pulmonary fibrosis
本文將於2028/07/27開放下載。若您希望在開放下載時收到通知，可將文章加入收藏。
摘要
不明原因性特發性肺纖維化 (Idiopathic Pulmonary Fibrosis) 直至目前無法被完全治癒，且病情的進展難以控制，容易造成肺功能持續的惡化，並導致高致死率。「泛組蛋白去乙醯化酶抑制劑」 (Pan HDAC Inhibitor)，例如Suberoylanilide Hydroxamic acid (SAHA)，雖然已經顯示出減緩肺纖維化進程的潛力，但由於是會抑制所有的組蛋白去乙醯化酶，存在靶向外的效應，進而造成許多像是肝、皮膚毒性副作用。因此，具有選擇性的「組蛋白去乙醯化酶抑制劑 (HDAC Inhibitor)」對於減少副作用是有益的，且能較精準地延緩甚至治癒疾病。為了實現這一目標，我們與台大藥學系余兆武老師合作，設計並合成了24種新的HDAC6、HDAC8或雙重HDAC6/8抑制劑，並建立了一個三階段篩選平台，以快速篩選出能夠有效緩解TGF-β1誘導肺纖維化的HDAC抑制劑。這個三階段篩選平台包括小鼠NIH-3T3胚胎纖維母細胞預篩檢作為第一階段、人類肺纖維母細胞（HPF）篩檢作為第二階段。第三階段，則是使用撲類惡注射劑 (Bleomycin) 引發肺纖維化小鼠模型評估領導藥物的預防肺纖維化效果。在第一階段中，從24個合成HDAC抑制劑中，篩選後共得到五個命中藥物 (Hit Compounds)，然後在第二階段中，同時使用人類肺纖維母細胞作為篩選模型並且對五個命中藥物進行Caco-2通透性 (Permeability) 和肝微粒體穩定性 (Liver Microsomal stability) 測試，最終得到了兩個領導化合物 (Lead Compound)。進一步在第三階段中，通過肺纖維化小鼠模型確效兩個領導化合物的抗纖維化作用。這兩個領導化合物能有效抑制TGF-β1誘導的肺纖維細胞增殖和纖維化相關蛋白表現，進而減緩肺纖維化的進程。這一創新的三階段篩選平台將加速發現和降低開發緩解TGF-β1誘導的肺纖維化HDAC抑制劑的成本。此外，選擇性HDAC抑制劑的應用將有助於開發更有效且無毒副作用的藥物，這將有望改善特發性肺纖維化患者的生活質量。
並列摘要
Idiopathic pulmonary fibrosis of unknown etiology cannot be completely cured, and the progression of the disease is difficult to control, leading to a high mortality rate and continuous deterioration of lung function. Although histone deacetylase inhibitors (HDAC inhibitors) such as suberoylanilide hydroxamic acid (SAHA), a pan HDAC inhibitor, have shown potential in slowing the progression of pulmonary fibrosis, they have non-specific effects on all histone deacetylase enzymes, resulting in many side effects such as liver and kidney toxicity. Therefore, selective HDAC inhibitors can be beneficial in reducing side effects and more precisely delaying or even curing pulmonary fibrosis. To achieve this goal, we collaborated with Dr. Chao-Wu Yu at the National Taiwan University School of Pharmacy to design and synthesize twenty-four new HDAC6, HDAC8, or dual HDAC6/8 inhibitors and established a three-stage screening platform to quickly identify HDAC inhibitors that can effectively alleviate TGF-β1-induced pulmonary fibrosis. This three-stage screening platform was employed to evaluate potential drugs for their preventive effect on pulmonary fibrosis. In the first stage, NIH-3T3 mouse embryonic fibroblasts served as the pre-screening model, leading to the identification of five hit compounds out of 24 synthesized HDAC inhibitors. Subsequently, the second stage utilized human pulmonary fibroblasts (HPF) as the screening model, and two leading compounds were selected based on permeability and liver microsomal stability tests. Lastly, the anti-fibrotic effects of the two leading compounds were confirmed in the mouse model of induced pulmonary fibrosis through bleomycin injection in the third stage. These two leading compounds can effectively inhibit TGF-β1-induced pulmonary fibroblast proliferation and fibrosis-related protein expression, thereby slowing the progression of pulmonary fibrosis. This innovative three-stage screening platform will accelerate the discovery and reduce the development costs of HDAC inhibitors for alleviating TGF-β1-induced pulmonary fibrosis. In addition, the application of selective HDAC inhibitors will help develop more effective and less toxic drugs, which is expected to improve the quality of life of patients with idiopathic pulmonary fibrosis.
參考文獻
1. Alvarez, D., et al., IPF lung fibroblasts have a senescent phenotype. Am J Physiol Lung Cell Mol Physiol, 2017. 313(6): p. L1164-L1173.
2. Quinn, C., A. Wisse, and S.T. Manns, Clinical course and management of idiopathic pulmonary fibrosis. Multidiscip Respir Med, 2019. 14: p. 35.
3. Martinez, F.J., et al., Idiopathic pulmonary fibrosis. Nat Rev Dis Primers, 2017. 3: p. 17074.
4. van Manen, M.J., et al., Optimizing quality of life in patients with idiopathic pulmonary fibrosis. Ther Adv Respir Dis, 2017. 11(3): p. 157-169.
5. Kreuter, M., et al., The clinical course of idiopathic pulmonary fibrosis and its association to quality of life over time: longitudinal data from the INSIGHTS-IPF registry. Respir Res, 2019. 20(1): p. 59.