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The Application of REDOR NMR to Understand the Conformation of Epothilone B

The structural information of small therapeutic compounds complexed in biological matrices is important for drug developments. However, structural studies on ligands bound to such a large and dynamic system as microtubules are still challenging. This article reports an application of the solid-state...

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Autores principales: Lee, Jae-Ho, Kim, Moon-Su, Lee, Hyo Won, Lee, Ihl-Young C., Kim, Hyun Kyoung, Kim, Nam Doo, Lee, SangGap, Seo, Hwajeong, Paik, Younkee
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5535963/
https://www.ncbi.nlm.nih.gov/pubmed/28698492
http://dx.doi.org/10.3390/ijms18071472
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author Lee, Jae-Ho
Kim, Moon-Su
Lee, Hyo Won
Lee, Ihl-Young C.
Kim, Hyun Kyoung
Kim, Nam Doo
Lee, SangGap
Seo, Hwajeong
Paik, Younkee
author_facet Lee, Jae-Ho
Kim, Moon-Su
Lee, Hyo Won
Lee, Ihl-Young C.
Kim, Hyun Kyoung
Kim, Nam Doo
Lee, SangGap
Seo, Hwajeong
Paik, Younkee
author_sort Lee, Jae-Ho
collection PubMed
description The structural information of small therapeutic compounds complexed in biological matrices is important for drug developments. However, structural studies on ligands bound to such a large and dynamic system as microtubules are still challenging. This article reports an application of the solid-state NMR technique to investigating the bioactive conformation of epothilone B, a microtubule stabilizing agent, whose analog ixabepilone was approved by the U.S. Food and Drug Administration (FDA) as an anticancer drug. First, an analog of epothilone B was designed and successfully synthesized with deuterium and fluorine labels while keeping the high potency of the drug; Second, a lyophilization protocol was developed to enhance the low sensitivity of solid-state NMR; Third, molecular dynamics information of microtubule-bound epothilone B was revealed by high-resolution NMR spectra in comparison to the non-bound epothilone B; Last, information for the macrolide conformation of microtubule-bound epothilone B was obtained from rotational-echo double-resonance (REDOR) NMR data, suggesting the X-ray crystal structure of the ligand in the P450epoK complex as a possible candidate for the conformation. Our results are important as the first demonstration of using REDOR for studying epothilones.
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spelling pubmed-55359632017-08-04 The Application of REDOR NMR to Understand the Conformation of Epothilone B Lee, Jae-Ho Kim, Moon-Su Lee, Hyo Won Lee, Ihl-Young C. Kim, Hyun Kyoung Kim, Nam Doo Lee, SangGap Seo, Hwajeong Paik, Younkee Int J Mol Sci Article The structural information of small therapeutic compounds complexed in biological matrices is important for drug developments. However, structural studies on ligands bound to such a large and dynamic system as microtubules are still challenging. This article reports an application of the solid-state NMR technique to investigating the bioactive conformation of epothilone B, a microtubule stabilizing agent, whose analog ixabepilone was approved by the U.S. Food and Drug Administration (FDA) as an anticancer drug. First, an analog of epothilone B was designed and successfully synthesized with deuterium and fluorine labels while keeping the high potency of the drug; Second, a lyophilization protocol was developed to enhance the low sensitivity of solid-state NMR; Third, molecular dynamics information of microtubule-bound epothilone B was revealed by high-resolution NMR spectra in comparison to the non-bound epothilone B; Last, information for the macrolide conformation of microtubule-bound epothilone B was obtained from rotational-echo double-resonance (REDOR) NMR data, suggesting the X-ray crystal structure of the ligand in the P450epoK complex as a possible candidate for the conformation. Our results are important as the first demonstration of using REDOR for studying epothilones. MDPI 2017-07-09 /pmc/articles/PMC5535963/ /pubmed/28698492 http://dx.doi.org/10.3390/ijms18071472 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lee, Jae-Ho
Kim, Moon-Su
Lee, Hyo Won
Lee, Ihl-Young C.
Kim, Hyun Kyoung
Kim, Nam Doo
Lee, SangGap
Seo, Hwajeong
Paik, Younkee
The Application of REDOR NMR to Understand the Conformation of Epothilone B
title The Application of REDOR NMR to Understand the Conformation of Epothilone B
title_full The Application of REDOR NMR to Understand the Conformation of Epothilone B
title_fullStr The Application of REDOR NMR to Understand the Conformation of Epothilone B
title_full_unstemmed The Application of REDOR NMR to Understand the Conformation of Epothilone B
title_short The Application of REDOR NMR to Understand the Conformation of Epothilone B
title_sort application of redor nmr to understand the conformation of epothilone b
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5535963/
https://www.ncbi.nlm.nih.gov/pubmed/28698492
http://dx.doi.org/10.3390/ijms18071472
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