Design of the nanointerface for detection, recovery, and regeneration of abnormal proteins

用于异常蛋白质检测、回收和再生的纳米界面设计

基本信息

批准号：
14380412
负责人：
FUJIMOTO Keiji
金额：
$ 7.1万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

Polymeric nanoparicles have been paid to attention as a drug carrier. It is required that carriers are inert and non-toxic and are not accumulated or are degradable in the body. Therefore we selected poly-L-lysine (PLL) that is degradable polypeptide for a backbone of a nanoparticle. Because PLL exhibits structural changes among random, helix, and β-sheet by pH and temperature, we expect the stimuli-sensitive release of drugs from nanoparticles.PLL was allowed to react with poly (ethylene glycol) (PEG) to improve the dispersion-stability and to prolong the lifetime in blood (mPEG-PLL polymer). Then, mPEG-PLL polymer was coupled with phosphatidic acid (PA) that acts as a hydrophobic site (mPEG-PLL-PA nanoparticle). The obtained polymers spontaneously aggregated into a nanoparticulate shape by the hydrophobic association among PA moieties. Whilst mPEG-PLL polymers were assembled to nanoparticles upon the formation of β-sheet structure in high pHs and high temperatures (mPEG-PLL nanoparticle). Both nanoparticles were 10-80nm in size and possessed the positive change. FITC-labeled nanoparticles were internalized into A431 cells. Adriamycin (ADR) that is an anticancer drug was incorporated into mPEG-PLL-PA nanoparticle more than mPEG-PLL nanoparticle because of PA's great hydrophobicity. However, drug release showed an opposite tendency. In pH4, release rate of ADR from mPEG-PLL nanoparticles was faster than that from mPEG-PLL-PA nanoparticles. Thereby survival rate of A431 cells decreased when they were treated with ADR-loaded mPEG-PLL nanoparticles.We applied β-sheet association of the mPEG-PLL polymer to inhibition of protein aggregation. PLL tends to aggregate through β-sheet association by raising pH and temperature. This aggregation could be inhibited by adding the mPEG-PLL polymer. This suggests that the mPEG-PLL polymer is capable of inhibiting aggregation of abnormal proteins that cause conformational diseases.

聚合物纳米粒子已被付费作为毒品携带者出席。要求携带者是惰性和无毒的，并且不累积或在体内降解。因此，我们选择了用于纳米颗粒骨架的可降解多肽的聚多肽（PLL）。由于PLL通过pH和温度表现出随机，螺旋和β-折叠之间的结构变化，因此我们期望药物对纳米颗粒的刺激敏感性释放。PLL被允许与聚（乙二醇）（PEG）反应，以改善分散稳定性，并在血液中延长血液（MPEG-PLL Polymer）。然后，将MPEG-PLL聚合物与充当疏水位点（MPEG-PLL-PA纳米粒子）的磷脂酸（PA）偶联。获得的聚合物自发地通过Pa部分中的疏水结合而自发地汇总成纳米的形状。在高pHS和高温（MPEG-PLL纳米粒子）中形成β-折叠结构后，将MPEG-PLL聚合物组装到纳米颗粒上。这两个纳米颗粒的大小均为10-80nm，并假定正变化。 FITC标记的纳米颗粒被内化为A431细胞。由于PA的疏水性很大，因此是一种抗癌药物的Adriamycin（ADR）比MPEG-PLL纳米颗粒多地掺入MPEG-PLL-PA纳米粒子中。但是，药物释放表现出相反的趋势。在PH4中，MPEG-PLL纳米颗粒的ADR释放速率比MPEG-PLL-PA纳米颗粒的释放速度快。因此，当用ADR负载的MPEG-PLL纳米颗粒处理A431细胞的存活率降低。我们将MPEG-PLL聚合物的β-折叠关联用于抑制蛋白质聚集。 PLL倾向于通过升高pH和温度来通过β-折叠的关联来聚集。可以通过添加MPEG-PLL聚合物来抑制该聚集。这表明MPEG-PLL聚合物能够抑制引起构象疾病的异常蛋白的聚集。