公开(公告)号：WO2012159187A3

公开(公告)日：2013-07-18

申请号：PCT/BR2012000162

申请日：2012-05-28

申请人： UNIV RIO DE JANEIRO ,  UNIV LISBOA ,  INST DE MEDICINA MOLECULAR ,  DE ALMEIDA FABIO CENEVIVA LACERDA ,  DA ROCHA MARTINS IVO CRISTIANO ,  DA POIAN ANDREA THOMPSON ,  DOS SANTOS NUNO FERNANDO DUARTE CORDEIRO CORREIA

发明人： DE ALMEIDA FABIO CENEVIVA LACERDA ,  DA ROCHA MARTINS IVO CRISTIANO ,  DA POIAN ANDREA THOMPSON ,  DOS SANTOS NUNO FERNANDO DUARTE CORDEIRO CORREIA

IPC分类号： C07K14/18 ,  A61K38/04 ,  A61P31/14 ,  C07K7/06 ,  C07K7/08 ,  C07K14/005

CPC分类号： C07K14/005 ,  C12N2770/24111 ,  C12N2770/24122

摘要： Dengue virus (DENV) is a Flavivirus that causes the major human arbovirosis, for which no specific treatment exists. The C protein, the viral capsid protein, is a symmetric homodimeric a-helical protein that interacts with intracellular lipid droplets (LDs) during viral replication, possibly via a alpha2-alpha2' non-polar central patch that has been hypothesized to interact with lipids. The C protein binding sites to LD were identified, revealing a new function for a conserved segment in the N- terminal disordered region, and indicating that conformational selection is involved in recognition. The obtained results showed that C protein positively-charged N-terminal region prompts the interaction with negatively- charged LDs, after which a conformational rearrangement enables the access of the central hydrophobic patch to LD surface. Altogether, the results allowed the design of a peptide with inhibitory activity of C protein-LD binding, paving the way for new drug development approaches against dengue. The ability of this peptide to bind to LDs in a similar fashion to that of the C protein was demonstrated employing different techniques, such as, but not limited to, nuclear magnetic resonance, zeta potential analysis and atomic force microscopy. Moreover, the de novo designed peptides were tested regarding their ability to inhibit DENV C protein binding to LDs, having been shown that it effectively inhibits the DENV C interaction with LDs, a finding of direct applicability for dengue and related Flavivirus originated pathologies.

摘要翻译： 登革热病毒（DENV）是一种黄病毒病毒，可导致主要的人体无症状，无特殊治疗。 病毒衣壳蛋白C蛋白是病毒复制过程中与细胞内脂质液滴（LD）相互作用的对称同二聚体α-螺旋蛋白，可能通过已经假设与脂质相互作用的α2-α2'非极性中心贴片 。 鉴定了C蛋白与LD的结合位点，揭示了N-末端无序区域中保守区段的新功能，并表明构象选择涉及识别。 得到的结果表明，C蛋白带正电的N末端区域促使与带负电荷的LDs相互作用，之后，构象重排使得中心疏水性贴片能够进入LD表面。 总之，结果允许设计具有C蛋白-LD结合的抑制活性的肽，为针对登革热的新药开发方法铺平了道路。 使用不同的技术（例如但不限于核磁共振，ζ电位分析和原子力显微镜）证明了该肽以与C蛋白类似的方式与LD结合的能力。 此外，测试了从头设计的肽对于抑制DENV C蛋白与LD结合的能力，已经显示其有效抑制与LD的DENV C相互作用，这是对登革热和相关的黄病毒起源病理学的直接适用性的发现。

公开(公告)号：WO2012159187A2

公开(公告)日：2012-11-29

申请号：PCT/BR2012/000162

申请日：2012-05-28

申请人： UNIVERSIDADE FEDERAL DO RIO DE JANEIRO - UFRJ ,  UNIVERSIDADE DE LISBOA ,  INSTITUTO DE MEDICINA MOLECULAR ,  DE ALMEIDA, Fabio, Ceneviva, Lacerda ,  DA ROCHA MARTINS, Ivo, Cristiano ,  DA POIAN, Andrea, Thompson ,  DOS SANTOS, Nuno, Fernando, Duarte, Cordeiro, Correia

发明人： DE ALMEIDA, Fabio, Ceneviva, Lacerda ,  DA ROCHA MARTINS, Ivo, Cristiano ,  DA POIAN, Andrea, Thompson ,  DOS SANTOS, Nuno, Fernando, Duarte, Cordeiro, Correia

IPC分类号： C07K14/18

CPC分类号： C07K14/005 ,  C12N2770/24111 ,  C12N2770/24122

摘要： Dengue virus (DENV) is a Flavivirus that causes the major human arbovirosis, for which no specific treatment exists. The C protein, the viral capsid protein, is a symmetric homodimeric a-helical protein that interacts with intracellular lipid droplets (LDs) during viral replication, possibly via a α2-α2' non-polar central patch that has been hypothesized to interact with lipids. The C protein binding sites to LD were identified, revealing a new function for a conserved segment in the N- terminal disordered region, and indicating that conformational selection is involved in recognition. The obtained results showed that C protein positively-charged N-terminal region prompts the interaction with negatively- charged LDs, after which a conformational rearrangement enables the access of the central hydrophobic patch to LD surface. Altogether, the results allowed the design of a peptide with inhibitory activity of C protein-LD binding, paving the way for new drug development approaches against dengue. The ability of this peptide to bind to LDs in a similar fashion to that of the C protein was demonstrated employing different techniques, such as, but not limited to, nuclear magnetic resonance, zeta potential analysis and atomic force microscopy. Moreover, the de novo designed peptides were tested regarding their ability to inhibit DENV C protein binding to LDs, having been shown that it effectively inhibits the DENV C interaction with LDs, a finding of direct applicability for dengue and related Flavivirus originated pathologies.

摘要翻译： 登革热病毒（DENV）是一种黄病毒病毒，可导致主要的人体无症状，无特殊治疗。 C蛋白是病毒衣壳蛋白，是一种对称的同二聚体α-螺旋蛋白，可在病毒复制过程中与细胞内脂滴（LD）相互作用，可能通过已经假设与脂质相互作用的a2-a2'非极性中心斑块 。 鉴定了C蛋白与LD的结合位点，揭示了N-末端无序区域中保守区段的新功能，并表明构象选择涉及识别。 得到的结果表明，C蛋白带正电的N末端区域促使与带负电荷的LDs相互作用，之后，构象重排使得中心疏水性贴片能够进入LD表面。 总之，结果允许设计具有C蛋白-LD结合的抑制活性的肽，为针对登革热的新药开发方法铺平了道路。 使用不同的技术（例如但不限于核磁共振，ζ电位分析和原子力显微镜）证明了该肽以与C蛋白类似的方式与LD结合的能力。 此外，测试了从头设计的肽对于抑制DENV C蛋白与LD结合的能力，已经显示其有效抑制与LD的DENV C相互作用，这是对登革热和相关的黄病毒起源病理学的直接适用性的发现。