Alzheimer’s disease is a neurological disorder marked by the accumulation of amyloid beta(Aβ)aggregates,resulting from mutations in the amyloid precursor protein.The enzymeβ-secretase,also known asβ-site amyloid p...Alzheimer’s disease is a neurological disorder marked by the accumulation of amyloid beta(Aβ)aggregates,resulting from mutations in the amyloid precursor protein.The enzymeβ-secretase,also known asβ-site amyloid precursor protein cleaving enzyme 1(BACE1),plays a crucial role in generating Aβpeptides.With no targeted therapy available for Alzheimer’s disease,inhibiting BACE1 aspartic protease has emerged as a primary treatment target.Since 1999,compounds demonstrating potential binding to the BACE1 receptor have advanced to human trials.Structural optimization of synthetically derived compounds,coupled with computational approaches,has offered valuable insights for developing highly selective leads with drug-like properties.This review highlights pivotal studies on the design and development of BACE1 inhibitors as anti-Alzheimer’s disease agents.It summarizes computational methods employed in facilitating drug discovery for potential BACE1 inhibitors and provides an update on their clinical status,indicating future directions for novel BACE1 inhibitors.The promising clinical results of Elenbecestat(E-2609)catalyze the development of effective,selective BACE1 inhibitors in the future.展开更多
目的构建BACE1基因干扰质粒,并研究其在neuro-2a细胞中的表达,为以其为靶点的基因治疗提供稳定转染的质粒。方法选择人、小鼠和大鼠的BACE1基因共有序列为干扰靶点,设计3组连接有GFP的干扰质粒,将构建好的质粒转染neuro-2a细胞,通过Real...目的构建BACE1基因干扰质粒,并研究其在neuro-2a细胞中的表达,为以其为靶点的基因治疗提供稳定转染的质粒。方法选择人、小鼠和大鼠的BACE1基因共有序列为干扰靶点,设计3组连接有GFP的干扰质粒,将构建好的质粒转染neuro-2a细胞,通过Real time RT-PCR及Western-blotting的方法分别在RNA及蛋白质水平上检测BACE1的表达,以分析其干扰的效率。结果经酶切及测序证实,插入的DNA片段序列与设计序列完全一致。与空质粒对照组相比,3个干扰靶点对BACE1基因的表达均有不同程度的抑制作用。其中pYr-1.1-siBACE1在mRNA水平及蛋白质水平干扰效果均最好。结论成功构建了BACE1基因的干扰质粒pYr-1.1-siBACE1,并能有效抑制neuro-2a细胞内源性BACE1基因表达,为靶向BACE1基因的治疗提供了有力的工具。展开更多
目的探讨Aβ3-10重复片段质粒免疫接种3月龄APPswe/PSEN1双转基因(AD)鼠脑内BACE1的影响。方法应用Aβ3-10重复片段质粒免疫3月龄APPswe/PSEN1双转基因鼠,同等剂量的PBS免疫对照组及C57BL/6J组小鼠,各组小鼠均在免疫后2、4、6次后通过RT...目的探讨Aβ3-10重复片段质粒免疫接种3月龄APPswe/PSEN1双转基因(AD)鼠脑内BACE1的影响。方法应用Aβ3-10重复片段质粒免疫3月龄APPswe/PSEN1双转基因鼠,同等剂量的PBS免疫对照组及C57BL/6J组小鼠,各组小鼠均在免疫后2、4、6次后通过RT-PCR法检测BACE1 m RNA水平,Western blotting法检测BACE1/GFAP/NF-κB蛋白水平,免疫组化法观察Aβ1-42脑内分布情况。水迷宫检测其行为学改变。结果在免疫2次后BACE1 m RNA表达水平C57BL/6J组<Aβ3-10组<对照组(F=4.649,P=0.021);免疫4次Aβ3-10组<C57BL/6J组<对照组(F=115.683,P=0.001);免疫6次C57BL/6J组<Aβ3-10组<对照组(F=86.600,P<0.001)。BACE1的蛋白表达水平在免疫2、4、6次后C57BL/6J组<Aβ3-10组<对照组(F=432.843,P<0.001;F=57.673,P<0.001;F=26.550,P=0.001),NF-κB的蛋白表达水平在免疫2次后C57BL/6J组<Aβ3-10组<对照组(F=109.127,P<0.001);免疫4,6次后Aβ3-10组<C57BL/6J组<对照组(F=30.301,P<0.001;F=129.967,P<0.001)。GFAP的蛋白表达水平在免疫2、4、6次后C57BL/6J组<Aβ3-10组<对照组(F=27.782,P=0.001;F=26.132,P=0.001;F=26.450,P=0.001);Aβ1-42在脑内的分布C57BL/6J组<Aβ3-10组<对照组(皮质:F=5.395,P=0.021;F=47.135,P=0.000;F=25.306,P=0.000,海马:F=11.023,P=0.002;F=14.936,P=0.001;F=50.132,P=0.000)。总潜伏期C57BL/6J组<Aβ3-10组<对照组(F=8.938,P=0.016;F=5.745,P=0.04;F=7.073,P=0.017)。结论 Aβ3-10重复片段质粒可以影响脑内BACE1的表达,影响Aβ产生,改善空间记忆能力。展开更多
文摘Alzheimer’s disease is a neurological disorder marked by the accumulation of amyloid beta(Aβ)aggregates,resulting from mutations in the amyloid precursor protein.The enzymeβ-secretase,also known asβ-site amyloid precursor protein cleaving enzyme 1(BACE1),plays a crucial role in generating Aβpeptides.With no targeted therapy available for Alzheimer’s disease,inhibiting BACE1 aspartic protease has emerged as a primary treatment target.Since 1999,compounds demonstrating potential binding to the BACE1 receptor have advanced to human trials.Structural optimization of synthetically derived compounds,coupled with computational approaches,has offered valuable insights for developing highly selective leads with drug-like properties.This review highlights pivotal studies on the design and development of BACE1 inhibitors as anti-Alzheimer’s disease agents.It summarizes computational methods employed in facilitating drug discovery for potential BACE1 inhibitors and provides an update on their clinical status,indicating future directions for novel BACE1 inhibitors.The promising clinical results of Elenbecestat(E-2609)catalyze the development of effective,selective BACE1 inhibitors in the future.
文摘目的构建BACE1基因干扰质粒,并研究其在neuro-2a细胞中的表达,为以其为靶点的基因治疗提供稳定转染的质粒。方法选择人、小鼠和大鼠的BACE1基因共有序列为干扰靶点,设计3组连接有GFP的干扰质粒,将构建好的质粒转染neuro-2a细胞,通过Real time RT-PCR及Western-blotting的方法分别在RNA及蛋白质水平上检测BACE1的表达,以分析其干扰的效率。结果经酶切及测序证实,插入的DNA片段序列与设计序列完全一致。与空质粒对照组相比,3个干扰靶点对BACE1基因的表达均有不同程度的抑制作用。其中pYr-1.1-siBACE1在mRNA水平及蛋白质水平干扰效果均最好。结论成功构建了BACE1基因的干扰质粒pYr-1.1-siBACE1,并能有效抑制neuro-2a细胞内源性BACE1基因表达,为靶向BACE1基因的治疗提供了有力的工具。
文摘目的探讨Aβ3-10重复片段质粒免疫接种3月龄APPswe/PSEN1双转基因(AD)鼠脑内BACE1的影响。方法应用Aβ3-10重复片段质粒免疫3月龄APPswe/PSEN1双转基因鼠,同等剂量的PBS免疫对照组及C57BL/6J组小鼠,各组小鼠均在免疫后2、4、6次后通过RT-PCR法检测BACE1 m RNA水平,Western blotting法检测BACE1/GFAP/NF-κB蛋白水平,免疫组化法观察Aβ1-42脑内分布情况。水迷宫检测其行为学改变。结果在免疫2次后BACE1 m RNA表达水平C57BL/6J组<Aβ3-10组<对照组(F=4.649,P=0.021);免疫4次Aβ3-10组<C57BL/6J组<对照组(F=115.683,P=0.001);免疫6次C57BL/6J组<Aβ3-10组<对照组(F=86.600,P<0.001)。BACE1的蛋白表达水平在免疫2、4、6次后C57BL/6J组<Aβ3-10组<对照组(F=432.843,P<0.001;F=57.673,P<0.001;F=26.550,P=0.001),NF-κB的蛋白表达水平在免疫2次后C57BL/6J组<Aβ3-10组<对照组(F=109.127,P<0.001);免疫4,6次后Aβ3-10组<C57BL/6J组<对照组(F=30.301,P<0.001;F=129.967,P<0.001)。GFAP的蛋白表达水平在免疫2、4、6次后C57BL/6J组<Aβ3-10组<对照组(F=27.782,P=0.001;F=26.132,P=0.001;F=26.450,P=0.001);Aβ1-42在脑内的分布C57BL/6J组<Aβ3-10组<对照组(皮质:F=5.395,P=0.021;F=47.135,P=0.000;F=25.306,P=0.000,海马:F=11.023,P=0.002;F=14.936,P=0.001;F=50.132,P=0.000)。总潜伏期C57BL/6J组<Aβ3-10组<对照组(F=8.938,P=0.016;F=5.745,P=0.04;F=7.073,P=0.017)。结论 Aβ3-10重复片段质粒可以影响脑内BACE1的表达,影响Aβ产生,改善空间记忆能力。
