以材料為核心探索 : 透過抑制聚集以穩定人類降鈣素
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2024
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人類降鈣素(human calcitonin, hCT)是一種由32個胺基酸所組成的荷爾蒙胜肽。在生物體內扮演調節血鈣水平的重要角色，可抑制破骨細胞的活動，因此常被作為藥物使用來治療骨骼相關疾病，例如: 佩吉特氏病和骨質疏鬆症等。由於它高聚集傾向的特性，導致其藥物效果受到一定限制，我們的研究目標在於開發新的材料用於穩定該胜肽。根據先前實驗室的研究發現，醣類分子對抑制hCT的聚集具有潛在能力，但需要在高濃度下才能發揮聚集抑制的作用。在這項研究中，我們利用史托伯法 (Stöber process )製備溶膠，並引入額外的葡萄糖，希望能有效地串聯溶液中的葡萄糖分子，藉此降低葡萄糖的濃度或提升抑制聚集的效果。在本研究中，我們參考了相關文獻中的製備條件，以獲得穩定的樣品溶液，之後再進行製備條件的優化。當製備出的材料穩定度提升後，我們才開始調整添加的葡萄糖濃度。首先，透過凝膠滲透色譜(Gel permeation chromatography, GPC)判斷材料的交聯程度，確保其交聯程度相似後，再以傅立葉轉換紅外光譜(Fourier-transform infrared spectroscopy, FTIR)確認葡萄糖是否交聯於材料中。接著，我們利用X射線光電子能譜(X-ray photoelectron spectroscopy, XPS)分析材料之元素比例 ，推算出材料中交聯的葡萄糖量。在硫磺素T(Thioflavin T, ThT)動力學實驗中，我們觀察到材料通過減少共同培育下的hCT纖維數量方式達到抑制hCT聚集的效果，該結果也透過穿透式電子顯微鏡(Transmission electron microscopy, TEM)得到驗證。
Human calcitonin (hCT) is a hormonal peptide composed of 32 amino acids. It plays a crucial role in regulating blood calcium levels in the body by inhibiting the activity of osteoclasts. Therefore, it is often used as a medication to treat bone-related diseases, such as Paget's disease and osteoporosis. Due to its high tendency to aggregate, the efficacy of hCT as a drug is somewhat limited. Our research aims to develop new materials to stabilize this peptide. Previous studies in our laboratory have found that carbohydrate molecules have the potential to inhibit the aggregation of hCT, but they require high concentrations to be effective. In this study, we use the Stöber process to prepare sols and introduce additional glucose, hoping to effectively link the glucosemolecules in the solution, thereby reducing the glucose concentration or enhancing the aggregation inhibition effect. In this study, we referred to the preparation conditions in the relevant literature to obtain a stable sample solution, and then optimized the preparation conditions later. Once the stability of the prepared material was improved, we started to adjust the concentration of added glucose. First, we determined the degree of crosslinking of the materials using gel permeation chromatography (GPC) to ensure similar crosslinking levels. We then used Fourier-transform infrared spectroscopy (FTIR) to confirm whether glucose was crosslinked within the materials. Next, we utilized X-ray photoelectron spectroscopy (XPS) to analyze the elemental composition of the materials and estimate the amount of glucose crosslinked within them. In thioflavin T (ThT) kinetic experiments, we observed that the materials inhibited hCT aggregation by reducing the number of hCT fibers when co-incubated. This result was further verified by transmission electron microscopy (TEM).
Human calcitonin (hCT) is a hormonal peptide composed of 32 amino acids. It plays a crucial role in regulating blood calcium levels in the body by inhibiting the activity of osteoclasts. Therefore, it is often used as a medication to treat bone-related diseases, such as Paget's disease and osteoporosis. Due to its high tendency to aggregate, the efficacy of hCT as a drug is somewhat limited. Our research aims to develop new materials to stabilize this peptide. Previous studies in our laboratory have found that carbohydrate molecules have the potential to inhibit the aggregation of hCT, but they require high concentrations to be effective. In this study, we use the Stöber process to prepare sols and introduce additional glucose, hoping to effectively link the glucosemolecules in the solution, thereby reducing the glucose concentration or enhancing the aggregation inhibition effect. In this study, we referred to the preparation conditions in the relevant literature to obtain a stable sample solution, and then optimized the preparation conditions later. Once the stability of the prepared material was improved, we started to adjust the concentration of added glucose. First, we determined the degree of crosslinking of the materials using gel permeation chromatography (GPC) to ensure similar crosslinking levels. We then used Fourier-transform infrared spectroscopy (FTIR) to confirm whether glucose was crosslinked within the materials. Next, we utilized X-ray photoelectron spectroscopy (XPS) to analyze the elemental composition of the materials and estimate the amount of glucose crosslinked within them. In thioflavin T (ThT) kinetic experiments, we observed that the materials inhibited hCT aggregation by reducing the number of hCT fibers when co-incubated. This result was further verified by transmission electron microscopy (TEM).
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人類降鈣素, 類澱粉蛋白聚集, 史托伯法, human calcitonin, amyloid aggregation, Stöber process