公开(公告)号：US08916540B2

公开(公告)日：2014-12-23

申请号：US12354717

申请日：2009-01-15

Applicant: Chi-Huey Wong ,  Ting-Jen Cheng ,  Che Alex Ma ,  Wei-Chieh Cheng

Inventor： Chi-Huey Wong ,  Ting-Jen Cheng ,  Che Alex Ma ,  Wei-Chieh Cheng

IPC: C07K14/195 ,  C07K14/26 ,  C07K14/315 ,  C07K14/25 ,  C07K14/235 ,  C07K14/32 ,  C07K14/31 ,  C07K14/21 ,  C07K14/33 ,  C07K14/255 ,  C07K14/22 ,  C07K14/265 ,  G01N33/573 ,  A61K31/19 ,  A61K31/715 ,  A61K31/427 ,  A61K31/365

CPC classification number: A61K31/702 ,  A61K31/19 ,  A61K31/365 ,  A61K31/427 ,  A61K31/715 ,  C07K14/195 ,  C07K14/21 ,  C07K14/22 ,  C07K14/235 ,  C07K14/25 ,  C07K14/255 ,  C07K14/26 ,  C07K14/265 ,  C07K14/31 ,  C07K14/315 ,  C07K14/3156 ,  C07K14/32 ,  C07K14/33 ,  G01N33/573 ,  G01N2333/91091 ,  Y02A50/473 ,  Y02A50/475 ,  Y02A50/481

Abstract: Moenomycin inhibits bacterial growth by clocking the transglycosylase activity of class A penicillin-binding proteins (PBPs), which are key enzymes in bacterial cell wall synthesis. The binding affinities of moenomycin A with various truncated PBPs were compared showing that the transmembrane domain is important for moenomycin binding. Full-length class-A PBPs from 16 bacterial species were produced, and their binding activities showed a correlation with the antimicrobial activity of moenomycin against Enterococcus faecalis and Staphylococcus aureus. Moreover, a fluorescence anisotropy-based high-throughput assay was developed and used successfully for identification of transglycosylase inhibitors.

Abstract translation: 霉菌素通过计时细菌细胞壁合成中的关键酶A类青霉素结合蛋白（PBP）的转糖苷酶活性抑制细菌生长。 比较了霉酚霉素A与各种截短的PBP的结合亲和力，结果表明跨膜结构域对于霉酚霉素结合是重要的。 产生了16种细菌种类的全长class-A PBPs，其结合活性与霉酚霉素对粪肠球菌和金黄色葡萄球菌的抗菌活性呈显着相关性。 此外，开发了基于荧光各向异性的高通量测定，并成功地用于鉴定转糖基酶抑制剂。

公开(公告)号：US20150342973A1

公开(公告)日：2015-12-03

申请号：US14578453

申请日：2014-12-21

Applicant: Chi-Huey WONG ,  Ting-Jen CHENG ,  Che Alex MA ,  Wei-Chieh CHENG

Inventor： Chi-Huey WONG ,  Ting-Jen CHENG ,  Che Alex MA ,  Wei-Chieh CHENG

IPC: A61K31/702

CPC classification number: A61K31/702 ,  A61K31/19 ,  A61K31/365 ,  A61K31/427 ,  A61K31/715 ,  C07K14/195 ,  C07K14/21 ,  C07K14/22 ,  C07K14/235 ,  C07K14/25 ,  C07K14/255 ,  C07K14/26 ,  C07K14/265 ,  C07K14/31 ,  C07K14/315 ,  C07K14/3156 ,  C07K14/32 ,  C07K14/33 ,  G01N33/573 ,  G01N2333/91091 ,  Y02A50/473 ,  Y02A50/475 ,  Y02A50/481

Abstract: Moenomycin inhibits bacterial growth by clocking the transglycosylase activity of class A penicillin-binding proteins (PBPs), which are key enzymes in bacterial cell wall synthesis. The binding affinities of moenomycin A with various truncated PBPs were compared showing that the transmembrane domain is important for moenomycin binding. Full-length class-A PBPs from 16 bacterial species were produced, and their binding activities showed a correlation with the antimicrobial activity of moenomycin against Enterococcus faecalis and Staphylococcus aureus. Moreover, a fluorescence anisotropy-based high-throughput assay was developed and used successfully for identification of transglycosylase inhibitors.

公开(公告)号：US20100121107A1

公开(公告)日：2010-05-13

申请号：US12506982

申请日：2009-07-21

Applicant: Chi-Huey Wong ,  Che Alex Ma ,  Ting-Jen Rachel Cheng ,  Wei-Chieh Cheng

Inventor： Chi-Huey Wong ,  Che Alex Ma ,  Ting-Jen Rachel Cheng ,  Wei-Chieh Cheng

IPC: C12Q1/48 ,  C07C233/75

CPC classification number: C07C235/64 ,  C07C235/84 ,  C07K2299/00 ,  C12Q1/18 ,  C12Q1/48 ,  G01N33/573 ,  G01N2333/91102 ,  G01N2500/00 ,  G01N2500/04 ,  G01N2500/20

Abstract: The crystal structure at 2.16 Å resolution of the full-length bacterial bifunctional transglycosylase penicillin-binding protein 1b (PBP1b) from Escherichia coli, in complex with its inhibitor moenomycin, is provided. The atomic coordinates of the complex as well as the moenomycin binding site are provided. Three dimensional structures of amino acid residues involved in moenomycin binding and transglycosylation activity are identified. Binding site for peptidoglycan synthesis inhibitors comprising inhibitor-binding site comprises amino acid residues from at least one of transglycosylase (TG), UvrB domain 2 homolog (UB2H) and transmembrane (TM) domains of PBP1b are identified at an atomic level of resolution. Methods for rational drug design based on the atomic coordinates are provided. Methods for screening for antibiotics based on anisotropic binding assay and transglycosylase inhibitor assays are provided. Novel antibiotics based on the screening assays of the invention are disclosed.

Abstract translation: 提供了与其抑制剂新霉素复合的大肠杆菌全长细菌双功能转糖苷酶青霉素结合蛋白1b（PBP1b）的分辨率为2.16的晶体结构。 提供复合物的原子坐标以及霉酚霉素结合位点。 鉴定了涉及霉酚霉素结合和转糖基化活性的氨基酸残基的三维结构。 包含抑制剂结合位点的肽聚糖合成抑制剂的结合位点包含以分辨率的原子级别鉴定的来自转糖苷酶（TG），UvrB结构域2同源物（UB2H）和跨膜（TM）结构域中的至少一种的氨基酸残基。 提供了基于原子坐标的合理药物设计方法。 提供了基于各向异性结合测定法和转糖苷酶抑制剂测定法筛选抗生素的方法。 公开了基于本发明筛选试验的新型抗生素。

公开(公告)号：US20090203641A1

公开(公告)日：2009-08-13

申请号：US12354717

申请日：2009-01-15

Applicant: Chi-Huey Wong ,  Ting-Jen Cheng ,  Che Alex Ma ,  Wei-Chieh Cheng

Inventor： Chi-Huey Wong ,  Ting-Jen Cheng ,  Che Alex Ma ,  Wei-Chieh Cheng

IPC: A61K31/715 ,  C12Q1/48 ,  G01N33/53 ,  C12P21/00 ,  A61K31/365 ,  A61K31/427 ,  A61K31/19

CPC classification number: A61K31/702 ,  A61K31/19 ,  A61K31/365 ,  A61K31/427 ,  A61K31/715 ,  C07K14/195 ,  C07K14/21 ,  C07K14/22 ,  C07K14/235 ,  C07K14/25 ,  C07K14/255 ,  C07K14/26 ,  C07K14/265 ,  C07K14/31 ,  C07K14/315 ,  C07K14/3156 ,  C07K14/32 ,  C07K14/33 ,  G01N33/573 ,  G01N2333/91091 ,  Y02A50/473 ,  Y02A50/475 ,  Y02A50/481

Abstract: Moenomycin inhibits bacterial growth by clocking the transglycosylase activity of class A penicillin-binding proteins (PBPs), which are key enzymes in bacterial cell wall synthesis. The binding affinities of moenomycin A with various truncated PBPs were compared showing that the transmembrane domain is important for moenomycin binding. Full-length class-A PBPs from 16 bacterial species were produced, and their binding activities showed a correlation with the antimicrobial activity of moenomycin against Enterococcus faecalis and Staphylococcus aureus. Moreover, a fluorescence anisotropy-based high-throughput assay was developed and used successfully for identification of transglycosylase inhibitors.

Abstract translation: 霉菌素通过计时细菌细胞壁合成中的关键酶A类青霉素结合蛋白（PBP）的转糖苷酶活性抑制细菌生长。 比较了霉酚霉素A与各种截短的PBP的结合亲和力，结果表明跨膜结构域对于霉酚霉素结合是重要的。 产生了16种细菌种类的全长class-A PBPs，其结合活性与霉酚霉素对粪肠球菌和金黄色葡萄球菌的抗菌活性呈显着相关性。 此外，开发了基于荧光各向异性的高通量测定，并成功地用于鉴定转糖基酶抑制剂。