研究生: |
Hailemichael Tegenu Gebrie Hailemichael Tegenu Gebrie |
---|---|
論文名稱: |
開發二硒雙親媒性共聚物的多功能藥物遞送系統用於靶向癌症治療 Multi-responsive drug delivery systems based on di-selenium-containing amphiphilic copolymer for targeting cancer therapy |
指導教授: |
蔡協致
Hsieh-Chih Tsai |
口試委員: |
高震宇
Chen-Yu Kao 陳奕君 Yi-Chun Chen 陳玉暄 Yu-Shuan Chen 何明樺 Ho Ming-Hua 蔡協致 Hsieh-Chih Tsai |
學位類別: |
博士 Doctor |
系所名稱: |
應用科技學院 - 應用科技研究所 Graduate Institute of Applied Science and Technology |
論文出版年: | 2023 |
畢業學年度: | 111 |
語文別: | 英文 |
論文頁數: | 186 |
中文關鍵詞: | 氧化還原反應 、二硒化物鍵 、核心交聯 、生物素 、酸鹼敏感型 |
外文關鍵詞: | Redox- responsive, Diselenide bond, Core cross-linked, Biotin, pH-sensitive |
相關次數: | 點閱:322 下載:0 |
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癌症為一種細胞不受控制且持續增殖的疾病,而癌細胞的病變是因健康細胞中的遺傳物質受到傷害所引發的現象。目前在癌症治療中,手術及化學療法為最常使用的治療方式,但在化學療法中,藥物釋放的調節為目前需要改善的問題之一。為了解決這項問題,人們正在努力設計和製造環境刺激影響的奈米材料,而此舉能將藥物長時間的釋放於目標器官進而達到治療效果。在本論文的第一部分中,研究了由甲氧基聚乙二醇嵌段聚己內酯共對二氧雜環己酮 (Bi(PPCD)-Se2) 構建的熱/氧化還原敏感可注射水凝膠的開發。在1H-NMR、13C-NMR 和 FTIR 分析中,證實了 Bi(PPCD)-Se2 的形成,並且研究了 Bi(PPCD)-Se2 水膠的流變特徵、溶膠-凝膠轉變機制和體外降解特性,也在流變數據中表現水膠具有明顯的黏度 (45 Pa.s)。水膠包覆DOX顯示出 1.3% 的載藥量和 93% 的藥品包覆率 ,DOX@Bi(PPCD)-Se2 水膠在 37°C 和pH 7.4 及10mM GSH 和 5mM H2O2 的環境刺激下,在經過 22 天後釋放了 81.6% 和 85.4%的藥物總量 。在MTT 實驗中,水膠的釋放對於HeLa細胞及HaCaT細胞皆沒有毒性的產生。相比之下,在最大濃度為 10 µg/mL 時,DOX/水膠活性在 HeLa 細胞中產生了 31.3 ± 2.2% 的細胞活化效果。 由於其出色的氧化還原誘導降解性所生成的 Bi(PPCD)-Se2水膠,可用作潛在的藥物遞送生物材料。在本論文的第二個部分中,為了減輕高分子微胞的熱力不穩定性並可提高藥品承載率,我們通過開環聚合和點擊反應,製備了含有 DOX 綴合的 pH 敏感聚合物Methoxy Poly (ethylene oxide)-b-Poly (Aspartate-Hydrazide) Bi(mPEG-P[Asp-(Hyd-DOX)])-Se2 CCMs。 在1H-NMR、13C-NMR、拉曼和 FTIR 分析證實了合成材料的形成,並透過 DLS 測量了微胞顆粒大小及zeta 電位鑑定。 根據DLS非交聯微胞 (NCM) 顯示出比 CCM (~72.63 nm) 更大的顆粒尺寸 (~96.43 nm)。在FE-SEM中研究及觀察了高分子微胞的形態並使用 HeLa 細胞以及斑馬魚胚胎測試合成材料的細胞毒性。結果顯示與 DOX/SN-38@NCM不同,DOX/SN-38@CCMs 在體外顯示出受控制的釋放現象。在最大藥物濃度 (10 µg/mL) 的作用下,HeLa 細胞的抑制率比HaCaT(14.25%) 高 50.90%。使用螢光顯微鏡觀察了 SN-38/DOX 負載高分子微胞的細胞攝取和細胞間藥物釋放的現象。 此外利用 3D 腫瘤球體,研究了腫瘤生長侵襲活動。 總結來說,結合上述的這些優勢,合成具pH 敏感的二硒化物的 CCMs 對於 DOX/SN-38 向癌細胞的調節遞送更具有吸引力。我們將含有二硒化物的Biotin -PEG-SeSe-PBLA製備成含生物素介導的高分子微胞,此微胞具氧化還原敏感性且幾乎完全釋放所包覆之 DOX 特性。Biotin-PEG-SeSe-PBLA 會產生均勻分散的高分子微胞,其流體動力學直徑 (Dh) 為 81.54 nm。 高分子微胞中DOX藥物包覆率(EE) 和藥物於載體之攜帶率( DLC)分別為 74.325% 和 5.983wt%。 在與 0.1% H2O2 和 10 mM GSH 培養 8 小時後,未攜帶藥物載體也具氧化還原反應性。 此外DOX@Biotin-PEG-SeSe-PBLA微胞在模擬腫瘤氧化還原環境中表現出最佳的 DOX 釋放,在 0.1% H2O2 和 10 mM GSH 中 72 小時分別為 ~74% 和 ~ 89%。 在最大藥物濃度 (5 µg/mL) 下,HeLa 細胞的抑制率比 HaCaT (11%) 高 76%。二硒化物連接的Biotin-PEG-SeSe-PBLA 產生生理穩定、循環、癌症特異性和智能高分子微胞,需要進行進一步研究。總體來說,氧化還原敏感的含硒高分子微胞對於調節腫瘤細胞中化學藥物的釋放至關重要。
Cancer is a disease that is defined by unchecked cell proliferation and the absence of cell death, and it is brought on by harm and/or alterations to the genetic material of healthy cells. Surgery, radiation, and chemotherapy are the most often utilized types of cancer therapies. One of the main difficulties with chemotherapy is the controlled release of medicines. To solve this problem, efforts are being made to design and create stimuli-responsive nanomaterials, which will allow for the prolonged controlled release of medications at the site of action. In this dissertation, the first section examined, the development of thermo/redox-sensitive injectable hydrogels constructed from methoxy poly (ethylene glycol)-block poly(-caprolactone-co-p-dioxanone) (Bi(PPCD)-Se2). 1H-NMR, 13C-NMR, and FTIR analysis confirmed the formation of Bi(PPCD)-Se2. The rheological features, sol-gel transition mechanisms, and in vitro degradation characteristics of Bi(PPCD)-Se2 hydrogels were studied. The rheological data show that the hydrogel has a noticeable viscosity. The hydrogel showed 1.3% of Drug loading content and 93% encapsulation efficiency of Doxorubicin. The DOX@Bi(PPCD)-Se2 hydrogel released 81.6% and 85.4% of its payload after 22 days at 37°C and pH 7.4 with 10mM GSH and 5mM H2O2 stimuli. In HeLa and HaCaT cells, the MTT study revealed no toxicity. In contrast, the DOX/hydrogel activity produced 31.3 ± 2.2% of cell viability in HeLa cells at the maximal concentrations of 10 µg/mL. Due to its excellent redox induced degradability, the generated Bi(PPCD)-Se2 hydrogel may be used as a potential drug delivery biomaterial with a focus on local drug distribution in a sustained way. In our second project in order to alleviate thermodynamic instability of Polymeric micelles and enhancing drug control releasing capacity, we were prepared diselenide containing DOX-conjugated pH-sensitive polymeric prodrug Methoxy Poly (ethylene oxide)-b-Poly(Aspartate-Hydrazide) Bi(mPEG-P[Asp-(Hyd-DOX)])-Se2 CCMs via ring-opening polymerization and click reaction. 1H-NMR, 13C-NMR, Raman, and FTIR analysis confirmed the formation of the synthesized material while the micelle size and zeta potential were measured by DLS. According to DLS, the non-cross-linked micelle (NCMs) displayed a larger (~96.43 nm) particle size than the CCMs (~72.63 nm). Unlike the DOX/SN-38@NCMs, the DOX/SN-38@CCMs displayed a controlled release outline in vitro. At the maximal drug concentration (10 µg/mL), HeLa cells were inhibited 50.90% more than HaCaT (14.25%). Using fluorescence imaging, the cellular uptake and intercellular drug discharge of SN-38/DOX-loaded PMs were studied. Additionally, utilizing 3D tumor spheroids, the tumor growth inhibition activity was investigated. Overall, the combination of all these attractive features makes the synthesized pH-sensitive diselenide containing CCMs attractive for regulated delivery of DOX/SN-38 to cancer cells. We intensified our efforts to produce biotin-mediated PMs from Biotin-PEG-SeSe-PBLA that contain diselenide and have improved redox triggered and nearly complete DOX-releasing property. The self-assembly biotin-PEG-SeSe-PBLA would result in uniformly dispersed PMs with a hydrodynamic diameter (Dh) of 81.54 nm. Significant DOX EE and DLC, 74.325% and 5.983wt%, respectively, were observed in the PMs. The blank PMs micelles showed improved redox reactivity following an 8 h incubation with 0.1% H2O2 and 10 mM GSH. Additionally, DOX@Biotin-PEG-SeSe-PBLA micelles demonstrated optimal DOX release in mimicked tumor redox environment, at ~74% and ~ 89%, in 0.1% H2O2 and 10 mM GSH 72 h, respectively. At the maximal drug concentration (5 µg/mL), HeLa cells were inhibited 76% more than HaCaT (11%). In conclusion, the diselenide-linked Biotin-PEG-SeSe-PBLA produced physiologically stable, long-circulating, cancer-specific, and intelligent PMs that warrant additional in vivo research. Overall, redox sensitive Selenium-containing PMs are essential for the regulated discharge of chemo drug in tumor cells.
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